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Forest ServiceRocky Mountain Research Station and RNGR
R Kasten Dumroese1 Peng Zhang2 Jeremiah R Pinto11US Forest Service and 2Northeast Forestry University Harbin China
Combining Nitrogen and Rhizobia to Improve Nursery Growth of Nitrogen-fixing Plants
8th Western Native Plants Conference 14 November 2019 - Olympia WA
United States Department of Agriculture
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen and Plant Growth
bull Nitrogen usually the most critical element for plant growth
bull Present in soil in organic and inorganic forms
bull Ammonium and nitrate predominant inorganic forms
bull Organic forms (amino acids) can be primary N source
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen and Plant Growth
bull N form can affect plant growth
bull Much work with conifers at least with inorganic forms
bull Organic forms with northern latitude conifers and others
bull Sometimes better growth sometimes not
bull Few studies with organic forms and broadleaves and forbs
bull Organics having lower leaching potential
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen Runoff
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen Plant Growth amp Rhizobia
bull Rhizobia bacteria colonize some plant roots produce nodules and convert atmospheric N into ammonia in exchange for plant carbohydrates
bull Harsh site implications
bull Conventional wisdom applications of high N suppress nodule development Low N can improve nodulation
bull Data with Acacia koa indicates good nodule development despite high N applications
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Dumroese et al (2009)
Height
Dumroese et al (2011a)
Nodule number
Nodule biomass
Previous studies framed the questions
Potential for Using Biochar in Container Media Used to Grow Native Plants
Converting forest biomass to bioenergycreates biochar waste which might be used to help grow native plants for restoration
Nursery costs may be reduced plant quality improved and carbon sequestered below ground at minimal cost Variety of species
Dumroese et al (2011b 2018)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Biochar
bull Quality characteristics vary by feed stock and process
bull Conversion of organic matter in the absence of oxygenndash Pyrolysis ndash 300 to 700 oC less energy more biochar no ash
ndash Gasification ndash 700 oC more energy less biochar some ash
bull Gasification may have improved cation exchange capacity
bull Nearly all plant response research done with pyrolysis
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash Two StudiesN Sourcebull Black locust and lupinebull 5 nitrogen treatments
ndash Organic vs inorganic
bull 2 and 1 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
Biochar Sourcebull Black locustbull 3 biochar treatments
ndash Pyrolysis vs gasification
bull 2 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash 15N Theory
bull Most N is 14Nbull ldquoTagrdquo fertilizer by adding minute 15Nbull Determine 15N14N for plant only using fertilizer bull If plant has active rhizobia that add 14N to the system from the
atmosphere ratio will be lowerbull Expressed as δ 15N bull Higher δ 15N means less N supplied by rhizobia
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
NH4
NO3
NH4N
O3
L-ar
ginine
Cont
rol
Sho
ot b
iom
ass
(g)
00
01
02
03
04
05
ab ab
aa
b
United States Department of Agriculture
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen and Plant Growth
bull Nitrogen usually the most critical element for plant growth
bull Present in soil in organic and inorganic forms
bull Ammonium and nitrate predominant inorganic forms
bull Organic forms (amino acids) can be primary N source
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen and Plant Growth
bull N form can affect plant growth
bull Much work with conifers at least with inorganic forms
bull Organic forms with northern latitude conifers and others
bull Sometimes better growth sometimes not
bull Few studies with organic forms and broadleaves and forbs
bull Organics having lower leaching potential
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen Runoff
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen Plant Growth amp Rhizobia
bull Rhizobia bacteria colonize some plant roots produce nodules and convert atmospheric N into ammonia in exchange for plant carbohydrates
bull Harsh site implications
bull Conventional wisdom applications of high N suppress nodule development Low N can improve nodulation
bull Data with Acacia koa indicates good nodule development despite high N applications
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Dumroese et al (2009)
Height
Dumroese et al (2011a)
Nodule number
Nodule biomass
Previous studies framed the questions
Potential for Using Biochar in Container Media Used to Grow Native Plants
Converting forest biomass to bioenergycreates biochar waste which might be used to help grow native plants for restoration
Nursery costs may be reduced plant quality improved and carbon sequestered below ground at minimal cost Variety of species
Dumroese et al (2011b 2018)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Biochar
bull Quality characteristics vary by feed stock and process
bull Conversion of organic matter in the absence of oxygenndash Pyrolysis ndash 300 to 700 oC less energy more biochar no ash
ndash Gasification ndash 700 oC more energy less biochar some ash
bull Gasification may have improved cation exchange capacity
bull Nearly all plant response research done with pyrolysis
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash Two StudiesN Sourcebull Black locust and lupinebull 5 nitrogen treatments
ndash Organic vs inorganic
bull 2 and 1 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
Biochar Sourcebull Black locustbull 3 biochar treatments
ndash Pyrolysis vs gasification
bull 2 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash 15N Theory
bull Most N is 14Nbull ldquoTagrdquo fertilizer by adding minute 15Nbull Determine 15N14N for plant only using fertilizer bull If plant has active rhizobia that add 14N to the system from the
atmosphere ratio will be lowerbull Expressed as δ 15N bull Higher δ 15N means less N supplied by rhizobia
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
NH4
NO3
NH4N
O3
L-ar
ginine
Cont
rol
Sho
ot b
iom
ass
(g)
00
01
02
03
04
05
ab ab
aa
b
United States Department of Agriculture
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen and Plant Growth
bull N form can affect plant growth
bull Much work with conifers at least with inorganic forms
bull Organic forms with northern latitude conifers and others
bull Sometimes better growth sometimes not
bull Few studies with organic forms and broadleaves and forbs
bull Organics having lower leaching potential
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen Runoff
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen Plant Growth amp Rhizobia
bull Rhizobia bacteria colonize some plant roots produce nodules and convert atmospheric N into ammonia in exchange for plant carbohydrates
bull Harsh site implications
bull Conventional wisdom applications of high N suppress nodule development Low N can improve nodulation
bull Data with Acacia koa indicates good nodule development despite high N applications
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Dumroese et al (2009)
Height
Dumroese et al (2011a)
Nodule number
Nodule biomass
Previous studies framed the questions
Potential for Using Biochar in Container Media Used to Grow Native Plants
Converting forest biomass to bioenergycreates biochar waste which might be used to help grow native plants for restoration
Nursery costs may be reduced plant quality improved and carbon sequestered below ground at minimal cost Variety of species
Dumroese et al (2011b 2018)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Biochar
bull Quality characteristics vary by feed stock and process
bull Conversion of organic matter in the absence of oxygenndash Pyrolysis ndash 300 to 700 oC less energy more biochar no ash
ndash Gasification ndash 700 oC more energy less biochar some ash
bull Gasification may have improved cation exchange capacity
bull Nearly all plant response research done with pyrolysis
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash Two StudiesN Sourcebull Black locust and lupinebull 5 nitrogen treatments
ndash Organic vs inorganic
bull 2 and 1 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
Biochar Sourcebull Black locustbull 3 biochar treatments
ndash Pyrolysis vs gasification
bull 2 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash 15N Theory
bull Most N is 14Nbull ldquoTagrdquo fertilizer by adding minute 15Nbull Determine 15N14N for plant only using fertilizer bull If plant has active rhizobia that add 14N to the system from the
atmosphere ratio will be lowerbull Expressed as δ 15N bull Higher δ 15N means less N supplied by rhizobia
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
NH4
NO3
NH4N
O3
L-ar
ginine
Cont
rol
Sho
ot b
iom
ass
(g)
00
01
02
03
04
05
ab ab
aa
b
United States Department of Agriculture
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen Runoff
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen Plant Growth amp Rhizobia
bull Rhizobia bacteria colonize some plant roots produce nodules and convert atmospheric N into ammonia in exchange for plant carbohydrates
bull Harsh site implications
bull Conventional wisdom applications of high N suppress nodule development Low N can improve nodulation
bull Data with Acacia koa indicates good nodule development despite high N applications
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Dumroese et al (2009)
Height
Dumroese et al (2011a)
Nodule number
Nodule biomass
Previous studies framed the questions
Potential for Using Biochar in Container Media Used to Grow Native Plants
Converting forest biomass to bioenergycreates biochar waste which might be used to help grow native plants for restoration
Nursery costs may be reduced plant quality improved and carbon sequestered below ground at minimal cost Variety of species
Dumroese et al (2011b 2018)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Biochar
bull Quality characteristics vary by feed stock and process
bull Conversion of organic matter in the absence of oxygenndash Pyrolysis ndash 300 to 700 oC less energy more biochar no ash
ndash Gasification ndash 700 oC more energy less biochar some ash
bull Gasification may have improved cation exchange capacity
bull Nearly all plant response research done with pyrolysis
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash Two StudiesN Sourcebull Black locust and lupinebull 5 nitrogen treatments
ndash Organic vs inorganic
bull 2 and 1 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
Biochar Sourcebull Black locustbull 3 biochar treatments
ndash Pyrolysis vs gasification
bull 2 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash 15N Theory
bull Most N is 14Nbull ldquoTagrdquo fertilizer by adding minute 15Nbull Determine 15N14N for plant only using fertilizer bull If plant has active rhizobia that add 14N to the system from the
atmosphere ratio will be lowerbull Expressed as δ 15N bull Higher δ 15N means less N supplied by rhizobia
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
NH4
NO3
NH4N
O3
L-ar
ginine
Cont
rol
Sho
ot b
iom
ass
(g)
00
01
02
03
04
05
ab ab
aa
b
United States Department of Agriculture
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Nitrogen Plant Growth amp Rhizobia
bull Rhizobia bacteria colonize some plant roots produce nodules and convert atmospheric N into ammonia in exchange for plant carbohydrates
bull Harsh site implications
bull Conventional wisdom applications of high N suppress nodule development Low N can improve nodulation
bull Data with Acacia koa indicates good nodule development despite high N applications
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Dumroese et al (2009)
Height
Dumroese et al (2011a)
Nodule number
Nodule biomass
Previous studies framed the questions
Potential for Using Biochar in Container Media Used to Grow Native Plants
Converting forest biomass to bioenergycreates biochar waste which might be used to help grow native plants for restoration
Nursery costs may be reduced plant quality improved and carbon sequestered below ground at minimal cost Variety of species
Dumroese et al (2011b 2018)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Biochar
bull Quality characteristics vary by feed stock and process
bull Conversion of organic matter in the absence of oxygenndash Pyrolysis ndash 300 to 700 oC less energy more biochar no ash
ndash Gasification ndash 700 oC more energy less biochar some ash
bull Gasification may have improved cation exchange capacity
bull Nearly all plant response research done with pyrolysis
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash Two StudiesN Sourcebull Black locust and lupinebull 5 nitrogen treatments
ndash Organic vs inorganic
bull 2 and 1 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
Biochar Sourcebull Black locustbull 3 biochar treatments
ndash Pyrolysis vs gasification
bull 2 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash 15N Theory
bull Most N is 14Nbull ldquoTagrdquo fertilizer by adding minute 15Nbull Determine 15N14N for plant only using fertilizer bull If plant has active rhizobia that add 14N to the system from the
atmosphere ratio will be lowerbull Expressed as δ 15N bull Higher δ 15N means less N supplied by rhizobia
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
NH4
NO3
NH4N
O3
L-ar
ginine
Cont
rol
Sho
ot b
iom
ass
(g)
00
01
02
03
04
05
ab ab
aa
b
United States Department of Agriculture
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Dumroese et al (2009)
Height
Dumroese et al (2011a)
Nodule number
Nodule biomass
Previous studies framed the questions
Potential for Using Biochar in Container Media Used to Grow Native Plants
Converting forest biomass to bioenergycreates biochar waste which might be used to help grow native plants for restoration
Nursery costs may be reduced plant quality improved and carbon sequestered below ground at minimal cost Variety of species
Dumroese et al (2011b 2018)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Biochar
bull Quality characteristics vary by feed stock and process
bull Conversion of organic matter in the absence of oxygenndash Pyrolysis ndash 300 to 700 oC less energy more biochar no ash
ndash Gasification ndash 700 oC more energy less biochar some ash
bull Gasification may have improved cation exchange capacity
bull Nearly all plant response research done with pyrolysis
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash Two StudiesN Sourcebull Black locust and lupinebull 5 nitrogen treatments
ndash Organic vs inorganic
bull 2 and 1 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
Biochar Sourcebull Black locustbull 3 biochar treatments
ndash Pyrolysis vs gasification
bull 2 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash 15N Theory
bull Most N is 14Nbull ldquoTagrdquo fertilizer by adding minute 15Nbull Determine 15N14N for plant only using fertilizer bull If plant has active rhizobia that add 14N to the system from the
atmosphere ratio will be lowerbull Expressed as δ 15N bull Higher δ 15N means less N supplied by rhizobia
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
NH4
NO3
NH4N
O3
L-ar
ginine
Cont
rol
Sho
ot b
iom
ass
(g)
00
01
02
03
04
05
ab ab
aa
b
United States Department of Agriculture
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Potential for Using Biochar in Container Media Used to Grow Native Plants
Converting forest biomass to bioenergycreates biochar waste which might be used to help grow native plants for restoration
Nursery costs may be reduced plant quality improved and carbon sequestered below ground at minimal cost Variety of species
Dumroese et al (2011b 2018)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Biochar
bull Quality characteristics vary by feed stock and process
bull Conversion of organic matter in the absence of oxygenndash Pyrolysis ndash 300 to 700 oC less energy more biochar no ash
ndash Gasification ndash 700 oC more energy less biochar some ash
bull Gasification may have improved cation exchange capacity
bull Nearly all plant response research done with pyrolysis
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash Two StudiesN Sourcebull Black locust and lupinebull 5 nitrogen treatments
ndash Organic vs inorganic
bull 2 and 1 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
Biochar Sourcebull Black locustbull 3 biochar treatments
ndash Pyrolysis vs gasification
bull 2 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash 15N Theory
bull Most N is 14Nbull ldquoTagrdquo fertilizer by adding minute 15Nbull Determine 15N14N for plant only using fertilizer bull If plant has active rhizobia that add 14N to the system from the
atmosphere ratio will be lowerbull Expressed as δ 15N bull Higher δ 15N means less N supplied by rhizobia
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
NH4
NO3
NH4N
O3
L-ar
ginine
Cont
rol
Sho
ot b
iom
ass
(g)
00
01
02
03
04
05
ab ab
aa
b
United States Department of Agriculture
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Introduction ndash Biochar
bull Quality characteristics vary by feed stock and process
bull Conversion of organic matter in the absence of oxygenndash Pyrolysis ndash 300 to 700 oC less energy more biochar no ash
ndash Gasification ndash 700 oC more energy less biochar some ash
bull Gasification may have improved cation exchange capacity
bull Nearly all plant response research done with pyrolysis
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash Two StudiesN Sourcebull Black locust and lupinebull 5 nitrogen treatments
ndash Organic vs inorganic
bull 2 and 1 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
Biochar Sourcebull Black locustbull 3 biochar treatments
ndash Pyrolysis vs gasification
bull 2 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash 15N Theory
bull Most N is 14Nbull ldquoTagrdquo fertilizer by adding minute 15Nbull Determine 15N14N for plant only using fertilizer bull If plant has active rhizobia that add 14N to the system from the
atmosphere ratio will be lowerbull Expressed as δ 15N bull Higher δ 15N means less N supplied by rhizobia
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
NH4
NO3
NH4N
O3
L-ar
ginine
Cont
rol
Sho
ot b
iom
ass
(g)
00
01
02
03
04
05
ab ab
aa
b
United States Department of Agriculture
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash Two StudiesN Sourcebull Black locust and lupinebull 5 nitrogen treatments
ndash Organic vs inorganic
bull 2 and 1 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
Biochar Sourcebull Black locustbull 3 biochar treatments
ndash Pyrolysis vs gasification
bull 2 mg N per weekbull 75 container capacitybull Morphological assessmentbull 15N assessmentbull Commercial inoculants
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash 15N Theory
bull Most N is 14Nbull ldquoTagrdquo fertilizer by adding minute 15Nbull Determine 15N14N for plant only using fertilizer bull If plant has active rhizobia that add 14N to the system from the
atmosphere ratio will be lowerbull Expressed as δ 15N bull Higher δ 15N means less N supplied by rhizobia
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
NH4
NO3
NH4N
O3
L-ar
ginine
Cont
rol
Sho
ot b
iom
ass
(g)
00
01
02
03
04
05
ab ab
aa
b
United States Department of Agriculture
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Methods ndash 15N Theory
bull Most N is 14Nbull ldquoTagrdquo fertilizer by adding minute 15Nbull Determine 15N14N for plant only using fertilizer bull If plant has active rhizobia that add 14N to the system from the
atmosphere ratio will be lowerbull Expressed as δ 15N bull Higher δ 15N means less N supplied by rhizobia
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
NH4
NO3
NH4N
O3
L-ar
ginine
Cont
rol
Sho
ot b
iom
ass
(g)
00
01
02
03
04
05
ab ab
aa
b
United States Department of Agriculture
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
NH4
NO3
NH4N
O3
L-ar
ginine
Cont
rol
Sho
ot b
iom
ass
(g)
00
01
02
03
04
05
ab ab
aa
b
United States Department of Agriculture
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
NH4
NO3
NH4N
O3
L-ar
ginine
Cont
rol
Sho
ot b
iom
ass
(g)
00
01
02
03
04
05
ab ab
aa
b
United States Department of Agriculture
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
NH4NO3 ndash Without inoculation NH4NO3 ndash With inoculation
United States Department of Agriculture
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form and inoculation significant
bull N fertilization increased morphological traits
bull Inoculation improved growth
Leaf width Shoot Root
Per
cent
age
Incr
ease
0
100
200
300
400
500
600
Results ndash Lupine
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Control ndash No inoculation no N Control ndash Inoculation no N
United States Department of Agriculture
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Box Plots
Top 25 of values
Bottom 25 of values
50 of values
Median
Mean
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Interpretation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Lupine ndash N Form
N C
onte
nt (m
g pe
r tis
sue
type
)
0
5
10
15
20
0
50
100
150
200
15
N x
100
0
0
150
300
450
600
750
900
0
5
10
15
Shoot Root Shoot Root
A B
C D
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Variables
bull Heightbull RCDbull Leaf biomassbull Stem biomassbull Shoot biomassbull Root biomassbull Nodule biomassbull SR
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Non-inoc Inoculated
Hei
ght (
cm)
0
10
20
30
40
50No N fertilizerN fertilizer
Results ndash Black Locust
United States Department of Agriculture
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Ammonium sulfate
Left inoculation + N Right ammonium sulfate
Controls
Left inoculation Right no inoculation
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Shoo
t bio
mas
s (g
)
0
1
2
3
4
5
6
a aa a
b
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
NH4
NO3
NH4N
O3L-
argin
ineCo
ntro
l
Nod
ule
biom
ass
(g)
00
01
02
03
04
a
abb
b
c
Results ndash Black Locust
United States Department of Agriculture
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
bull N form inoculation and interaction significant
ndash Exceptions Shoot-to-root ratio unaffected RCD and nodule biomass unaffected by interaction
bull Inoculation + N fertilization yielded biggest seedlings
bull N fertilization improved growth no differences among N forms
ndash Exception Nodule biomass affected by N form
bull Inoculation improved growth
Results ndash Black Locust
United States Department of Agriculture
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
N fertilizer (far left is the no N fertilizer control)
Inoculated + N fertilizer (far left is the no N fertilizer control)
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash N Form
Arginine(organic)
NH4NO3(inorganic)
Inoculated
Control
150
200
250
300
N C
onte
nt (m
g pe
r tis
sue
type
)
0
2
4
6
8
10
12
14
15
N x
100
0
600
750
900
1050
1200
1350
0
2
4
6
8
10
12
Leaf Stem Root Leaf Stem Root
A B
C D
Zhang et al (2019)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Hei
ghtc
m
0
20
30
40
50a
b b
cc
c
Roo
t-col
lar d
iam
eter
mm
0
3
4
5
aab bcbcd
cd d
Leav
es b
iom
ass
g
00
5
10
15
20
25ab
c
dd d
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-Ino
cula
tion
Gas
ifica
tion-
Inoc
ulat
ion
asifi
catio
n-N
on-I
nocu
latio
n
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-I
nocu
latio
n
Stem
bio
mas
sg
00
2
4
6
8
10a a
bb
bb
Col 2 Col 4 Col 2 Col 4 Col 2 Col 4 R
oot b
iom
ass
g
00
1
2
3
4
5
6
7a
b b
bcbc
c
Nod
ule
biom
ass
g
00
1
2
3
4
5a
bb
cc
c
Pyro
lysis
-Ino
cula
tion
Pyro
lysis
-Non
-inoc
ulat
ion
Gas
ifica
tion-
Inoc
ulat
ion
Gas
ifica
tion-
Non
-inoc
ulat
ion
Cco
ntro
l-Ino
cula
tion
Con
trol-N
on-in
ocul
atio
n
Tota
l pla
nt b
iom
ass
g
0
1
2
3
4ab
c
dd d
Inoculated
Control
HeightRCDBiomass
NoduleRootShootLeafTotal
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
Pyr
olys
isg
N c
onte
ntg
0
20
40
60
80
100 InoculationNon-inoculation
Leaf Stem Root
Gas
ific
atio
ng
0
20
40
60
80
100
Con
trol
0
20
40
60
80
100
Pyro
lysi
s
0
100
200
300
400
500
600
Gas
ific
atio
nδ
15N
times10
00
0
100
200
300
400
500
600
Con
trol
0
100
200
300
400
500
600
Leaf Stem Root
Pyrolysis
No biochar
Gasification
Inoculated
Control
Sun et al (in prep)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Results ndash Black Locust ndash Biochar
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Conclusions
bull N fertilization without inoculation improved growth N form did not matter
bull Inoculation without N fertilization improved growth
bull For black locust N fertilization + inoculation improved growth more than either factor alone
bull N-fixing Rhizobium colonize and fix N well despite nursery conditions of robust N fertilization
bull Biochar from gasification can further stimulate N-fixation
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
United States Department of Agriculture
Inoculation ndash DIY
Wilkinson and Janos (2014)
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
Thank you
kasdumroeseusdagov
The views expressed were strictly those of the author and do not necessarily represent the positions or policy of the US
Department of Agriculture
References (Note these are included in the conference handout)
Dumroese RK Jacobs DF Davis AS 2009 Inoculating Acacia koa with Bradyrhizobiumand applying fertilizer in the nursery effects on nodule formation and seedling growth HortScience 44 443ndash446 httpswwwfsusdagovtreesearchpubs32844
Dumroese RK Heiskanen J Englund K Tervahauta A 2011 Pelleted biochar chemical and physical properties show potential use as a substrate in container nurseries Biomass amp Bioenergy 35 2018ndash2027httpswwwfsusdagovtreesearchpubs37521
Dumroese RK Davis AS Jacobs DF 2011 Nursery response of Acacia koa seedlings to container size irrigation method and fertilization rate Journal of Plant Nutrition 34 877ndash887 httpsfsusdagovtreesearchpubs37495
Dumroese RK Pinto JR Heiskanen J Tervahauta A McBurney KG Page-Dumroese DS Englund K 2018 Biochar can be a suitable replacement for sphagnum peat in nursery production of Pinus ponderosa seedlings Forests 9 article 232 21 phttpswwwfsusdagovtreesearchpubs56147
Sun Q Dumroese RK Liu Y In prep Interaction of biochar type and rhizobia increases the growth and biological nitrogen fixation of Robinia pseudoacacia seedlings
Wilkinson KM Janos DP 2014 Chapter 13 Beneficial microorganisms In Wilkinson KM Landis TD Haase DL Daley BF Dumroese RK editors Tropical Nursery Manual A Guide to Starting and Operating a Nursery for Native amp Traditional Plants Washington DC USDA Forest Service Agriculture Handbook 732 256-262httpsrngrnetpublicationstropical-nursery-manual
Zhang P Dumroese RK Pinto JR 2019 Organic or inorganic nitrogen and rhizobia inoculation provide synergistic growth response of a leguminous forb and tree Frontiers in Plant Science ndash Plant Nutrition 10 article 1308 12 phttpswwwfsusdagovtreesearchpubs59084
United States Department of Agriculture
Forest ServiceRocky Mountain Research Station and RNGR
