Embed Size (px)
Citation preview
1
Screening, identification and characteristics
of a quinclorac degrading strain Arthrobacter
sp. MC-10
CHEN De-xin Ph.D.
Qingzhou Tobacco Research Institute of CNTC
Email:[email protected]
Tel:(86)532 66715507 Fax:(86)532 88701032
15 Oct 2014 Québec Canada
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
Occurrence of tobacco quinclorac injury
Control methods of tobacco quinclorac injury
Research on quinclorac degrading bacteria
2
Screening, identification and characteristics of a quinclorac
degrading strain Arthrobacter sp. MC-10
Main Content
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
3
1 Occurrence of tobacco quinclorac injury
1.1 Quinclorac Profile
Formula: C10H5CI2NO2;
Name: 3,7-dichloroquinoline-8- carboxylic aeid
Hormone type herbicides, mainly used in paddy
Thermal stability, light stability, stable in acidic medium
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
4
1 Occurrence of tobacco quinclorac injury
1.2 Long vinca, serious impact
Half-life is 35.6 d
No significant effect on the growth of
tobacco after 269 d
Injury symptoms: Leaves curl back,
become narrow and leathery; In severe
cases, loss of economic value
Residues
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
7
2 Control methods
Physical control: Activated carbon adsorption
Chemical control: Quicklime neutralize
Pharmacy management: Ultra-sensitive protein, Brassinolide
Cultivation measures: Crop rotation, deep overburden
Control methods can not prevent injury effectively
How to control tobacco quinclorac injury more simple and
rapidly
Degrading bacteria filter
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
3 Research of quinclorac degrading bacteria
8
3.1 Screening of degrading bacteria MC-10
100mL 50mg/L
quinclorac
inorganic salt
culture
Add in
28℃, 150
r/minculti
vatd 7d
Adapter
50mg/L
quinclorac
inorganic
salt culture
5% inoculum 5g soil
samples
Separation of
pure strains
enrichment culture four
times
Expand
trainning in
LB culture
Preliminary get 12
degrading bacteria
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
3 Research of quinclorac degrading bacteria
9
3.2 Determination of degradation
High performance
liquid chromatography
(HPLC)
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
3 Research of quinclorac degrading bacteria
10
3.2 Determination of degradation
Strain
No.
Quinclorac
concentration
after 7 days
(mg/L）
Degradation
rate Strain No.
Quinclorac
concentration
after 7 days
(mg/L）
Degradatio
n rate
MC-01 15.15 23.7% MC-07 15.81 20.4%
MC-02 16.51 16.9% MC-08 5.19 73.5%
MC-03 17.03 14.3% MC-09 17.85 10.2%
MC-04 17.99 9.5% MC-10 1.63 91.3%
MC-05 6.07 69.1% MC-11 8.51 56.9%
MC-06 8.25 58.2% MC-12 11.35 42.7%
CK 19.89 -
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
11
3 Research of quinclorac degrading bacteria 3.3 HPLC chromatograms of MC-10
MC - 10
degradation
quinclorac
Control quinclorac
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
3 Research of quinclorac degrading bacteria
12
3.4 Identification of MC-10
Colony morphology of MC-10
Morphology
Electron microscopy
Molecular Biology
Arth
robacte
r sp.
TEM images of
MC-10
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
13
MC-10 (KJ619485)
Arthrobacter mysorens (JDXVZ4J3015)
Arthrobacter arilaitensis (JDWR1MZG014)
Arthrobacter bergerei (JDXE6GBR014)
Arthrobacter gangotriensis (JDXMGYCS015)
Arthrobacter woluwensis (JDX71G7Y014)
Arthrobacter globiformis (JDXTH1W9014)
Arthrobacter stackebrandtii (JDX9A2VP014)
Arthrobacter phenanthrenivorans (JDXE5NUD015)
Arthrobacter chlorophenolicus (JDXGT54S014)
Arthrobacter aurescens (JDXB2BMZ014)
Arthrobacter nicotinovorans (JDXX0VJU015)
Arthrobacter crystallopoietes (JDXJ4KJ2015)
Arthrobacter luteolus (JDXUVES7014)
Arthrobacter castelli (JDXFJWV3015)
Arthrobacter cumminsii (JDXKFYRN014)
52
99
98
100
97
66
67
79
52
0.005
MC-10 (KJ619485)
Phylogenetic tree of Strain MC-10
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
3 Research of quinclorac degrading bacteria
3.5 Characterization of MC-10
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
15
Optimum degradation
temperature：30℃
Optimum degradation
pH：7
Optimum inoculum：5%
Optimum
degradation
conditions
3 Research of quinclorac degrading bacteria
3.5 Characterization of MC-10 2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
3 Research of quinclorac degrading bacteria
Reduce the toxic effects on root, promote root
development
Alleviate the effects of quinclorac injury, alleviate
leaf curl
16
3.6 Alleviate the effects of quinclorac injury
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
17
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
18
A:Normal tobacco plants; B:Toxic soil and MC-10;
C: Toxic soil
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
1. Field Application of MC-10
2. Industrialized breeding
2 Problem
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
20
2014
_AP
35_C
henD
exin
Con
gres
s201
4 -
Doc
umen
t not
pee
r-re
view
ed b
y C
OR
ES
TA
