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Introgression of type IV trichomes and zingiberene into tomato from S. habrochaites,
LA2329: Current statusJohn Snyder, Mohammad Dawood and Ammar Al-Bayati
University of Kentucky
Lexington, KY
snyder@uky.edu
“Figure 1. TYLCV distribution pattern among B. tabaci B and Q biotypes in China. The survey was
carried out in the field season of 2009. The white and red dots represent B. tabaci B and the Q biotype, respectively. The black portion of the dots denotes TYLCV-infected B. tabaci population and its respective infection rate. Inset: Outline map of China showing all the provinces. Surveyed provinces are highlighted in a blue shade.doi:10.1371/journal.pone.0034817.g001”
Pan H, Chu D, Yan W, Su Q, Liu B, et al. (2012) Rapid Spread of Tomato Yellow Leaf Curl Virus in China Is Aided Differentially by Two InvasiveWhiteflies. PLoS ONE 7(4): e34817
Project Objectives
• Transfer the production of high levels of zingiberene and high type IV trichome density from the wild tomato Solanum habrochaites to the cultivated tomato using conventional breeding methods
• Recover good reproductive fitness
• Evaluate resistance to spidermites and other arthropod species in the breeding population
Luckwill, 1943
Trichome diversity in Lycopersicon
Luckwill, 1943
Type IV is present on S. habrochaites
(L. hirsutum) and absent on S.
lycopersicum (L. esculentum)
Type V is present on S.
lycopersicum (L. esculentum)
and absent on S. habrochaites
(L. hirsutum).
The morphology of the type VI gland
differs between the two species.
S. lycopersicum
S. habrochaites
0
200
400
600
800
1000
1200
1400
1600
To
tal N
um
ber
of
Ap
hid
s
Accesssion Number
ES
HS
S
MR
HR
R
Aphid Resistance of S. habrochaites
accessions Beijing, 2002
In previous tests, LA2329 had
intermediate densities of type IV
and type VI trichomes.
LA2329 was resistant to
B. argentifollii
Primary methods
• Crossing scheme is a modified backcross
• 7-epizingiberene measured by gas chromatography (GC)
• Trichome density assessed visually at 50X on abaxial surfaces
min0 2 4 6 8 10
counts
0
20000
40000
60000
80000
100000
120000
FID1 A, (6-23-16\B116.D)
0.3
42
0.4
55
0.4
83
0.5
16
2.6
34
3.1
71
4.1
39
4.8
62
4.9
99
5.2
46
5.3
84
5.7
60
5.8
30
5.9
99
6.5
95
6.8
26
6.9
15
7.2
34
7.3
31
7.5
32
7.6
18
7.7
34
7.8
64
7.9
46
8.1
68
8.2
75
8.4
54
8.6
26
8.7
31
8.8
97
8.9
51
9.0
81
9.8
04
10.5
00
10.9
48
11.2
25
11.4
69
To measure zingiberene, 10 to 20 cm² of leaf tissue is collected and placed in a scintillation vial. Two ml of hexane + internalstandard are added, mixed and then an aliquot is removed for analysisby GC. Leaf area is determined from scanned images of leaf tissue. Zingiberene is expressed as GC area units/ cm².
Internal Std.
Scan of leaflet tissue
~10 % self –fruit set, 10 %
High type IV density
Currently at BC5
Results
LA2329 BC4F2Tomato
IS
IS
ISMT MTZGB
ZGB
• Presence of zingiberene is dominant. High concentration is recessive
• Plants with very low or 0 concentrations of monoterpenes appear to be homozygous for zingiberene synthase
• Transgressive segregation for monoterpenes
Results for BC4F2
• For the BC4F2, the segregation for presence:absence of zingiberenewas 277:115 (2.4:1), X²=3.93
• Concentration of zingiberene ranged from 0 to 1.52 X 10E8 AU/cm² of leaflet tissue
• Mean was 1.0 X 10E7
• Most but not all set fruit in the field
0
10
20
30
40
50
60
70
5.00E+06
1.50E+07
2.50E+07
3.50E+07
4.50E+07
5.50E+07
6.50E+07
7.50E+07
8.50E+07
9.50E+07
1.05E+08
1.15E+08
1.25E+08
1.35E+08
No
. of
See
d/F
ruit
Zin
gib
ere
ne
co
nce
ntr
atio
n (
FID
are
a u
nit
/cm
²)
Genotype
Zingiberene concentration (solid bars, left axis) and seed set (hatched bars, right axis) for 27 selected BC4F2 interspecific genotypes. Of ~550 BC4F2 interspecific hybrids evaluated for type IV trichome density and zingibereneconcentration, the 27 depicted in this graph were those having type IV density and zingiberene concentration approximately equal to or greater than those on the wild resistant donor parent S. habrochaites LA2329. Seeds were obtained from 16 hybrids. Thus frequency of individuals in the BC4F2 having type IV trichome density and zingibereneconcentration similar to those of wild parent, and had the ability to produce seeds was nearly 3%.
O32 fruit - 2017~3.4 x 107 zingiberene/cm2
High IV density~25 seed per fruit
High zingiberene, no IV’s
LINE N Zingiberene (AU/cm²)
H21 29 2.2E+07 A
G85 33 1.7E+07 B
G82 35 1.5E+07 B
H20 34 1.1E+07 C
Zingiberene differences among four BC3F4 families
• Reproductive Fitness
Dwarfing can be extreme
Exerted Stigma!!!BC4F3January 2018
More stigma problems!
Reproductive fitness
• Seeds
• Seeds per fruit
• No. of flowers
• Flowers per truss
• Pollen
• Stigma problems?
• Seed germination
• Pollen stainability is significantly correlated with fruit set (r=0.82).
• Seed germination is a problem.
Some valuable phenotypes?
• Small Stature (~20 inches tall
• Erect, 1 major branch (no staking?)
• Very concentrated set (~50 fruit in two weeks)
• Other phenotypes with value• Very high brix (2x normal)
• High flavor
Columnar Plants
Thanks to:
Belinda Labadie
Mohammad Dawood
Ammar Al-Bayati
Dr. Barbara Liedl
Trichomes on wild and
cultivated eggplant
snyder@uky.edu
A NEW SESQUITERPENE ALCOHOL FROM WILD TOMATO
REPELLED THE TWO-SPOTTED SPIDER MITE, TETRANYCHUS
URTICAE
Mohammad H Dawood: PhD Student
Advise by Dr. John Snyder
Horticulture Department
April 6th 2018
INTRODUCTION• Tomato Solanum lycopersicum is one of the most
economically important world-wide grown vegetable.
• However, tomato is a host for numerous pests that
reduce productivity.
• Tomato breeders have focused more on increasing fruit
quantity and quality and less on enhancing crop
resistance to herbivores.
Ric Bessin,
UKY
• Because of many pathogens and pests, tomato cultivation
in most cases considered a high risk crop.
• Because it needs intensive use of insecticides and
pesticides.
ashmkn.com
Levels of cultivar insect resistance are not enough to
decrease the amount of insecticides applied.
Alternative approach:
decrease the chemical sprays, and
allows for a smaller environmental impact.
Why wild type?
S. habrochaites
Do other compounds present in trichome secretions have
repulsive activities against the two spotted-spider mite
Tetranychus urticae, similar to that of zingiberene?
OBJECTIVES
Ric Bessin,
UKY
MATERIAL AND METHODS
Plant materials
• LA2329 accession Solanum habrochaites
2016-2017
• Florida Materials 2015
Spot test on TLC plate,
UV illumination
Column Chromatography
Silica gel,
MTBE,
Hexane
GC/MS
(GC)
SPIDER MITES (TETRANYCHUS URTICAE
KOCH) AND BIOASSAY
Spider mites
nurtured on bean
Phaseolus vulgaris
L.
Hexane
zingiberene
RESULTS AND DISCUSSION
Florida materials
LA2329 F accessionLA2329 A accession
After Separation
CONCLUSION
• LA2329 (A) which has ZGB-OH had activity against
spider mite behavior more than LA2329 F that included
ZGB only.
• After separation of two major components, the ZGB-OH
affected spider mite behavior more than ZGB.
THANKS
Department of Horticulture
The Role of Trichome Secretions and Densities in Spider Mite Behavior for an Interspecific Population of Tomato
Advisor Dr. John SnyderPresented by
Ammar AL-Bayati, PhD candidate
Overview
Introduction
Hypothesis
Objectives
Materials and Methods
Results
Conclusion
Introduction
There is significant necessity for assay platforms to assess and identify levelsof resistance and insect performance, as demonstrated by recent publicationsreporting this subject.
Our hypothesis: Are behavioral differences of mites associated with thepresence or absence of leaf compounds and trichome densities?
Objectives
1- Observe the differences of mite behavior among a range of hybrids compared to
the positive and negative controls.
2- Identify the interactions between genotypes and leaf surfaces with spider mites
with respect to presence and absence of type IV trichomes.
3- Determine the interaction of zingiberene content on mite performance.
Plant Materials and Methods
An interspecific populationbetween Solanum lycopersicumand a wild relative, Solanumhabrochaites LA2329, used inthis research was maintained inthe greenhouse and field insummer 2017 at the Universityof Kentucky, Lexington, Ky.
13 tomato genotypes (BC3F3 &BC3F4) were selected based onpresence or absence of trichometype IV, and ZGB.
Whole Leaf Bioassay
Parameters Measured
Leaflet position (1-5) and surface infested by mites
Mite webbing score and Feeding damage score (0-3)
Leaflet position of webbing and feeding damage (1-5)
Egg density (Egg No. per cm2 of total leaf area)
ZGB and monoterpene (MTP) content using gas chromatography (GC-FID).
Type IV trichome counting for abaxial and adaxial surfaces using a microgrid.
Data were analyzed by SAS using GLM & CORR procedures.
Results
Mean (n=3) for Trichome Type IV Density per 4.3 mm2 Microgrid
and Phytochemical concentration per cm2
Genotype Generation
Type IV
Density
Lower
Surface
Monoterpene
(AU/cm2 leaf
area)
Zingiberene
(AU/cm2 leaf
area)
LA2329 Wild Parent 45 0.0E+00 4.5E+06
LA2329OH Wild Control 65 0.0E+00 8.6E+06
PI127826 Wild Control 350 0.0E+00 1.2E+07
W126 Tomato 0 2.3E+06 0.0E+00
W129 Tomato Control 0 3.1E+05 0.0E+00
W160 Tomato Parent 0 4.8E+05 0.0E+00
X155 BC3F3 203 6.0E+06 0.0E+00
X166 BC3F3 173 0.0E+00 1.1E+07
X71 BC3F3 208 1.4E+05 5.7E+07
Z116 BC3F3 116 6.6E+06 0.0E+00
Z120 BC3F3 57 1.8E+06 0.0E+00
Z161 BC3F4 0 0.0E+00 2.6E+06
Z58 BC3F4 0 0.0E+00 4.8E+06
Correlations among parameters after Combining Variables by Surface
Results
Spider mite Webbing and Feeding Damage
W129
Z58W
160
X71
W126
Z120Z161
LA2329
LA2329O
H
Z116X155
X166
PI127826
Tomato Genotypes
0.0
0.5
1.0
1.5
2.0
2.5W
eb
bin
g S
co
re,
Day 7
Results
Spider mite Webbing and Feeding Damage
Results
Two-spotted spider mites with eggs
Tetranychus urticae Koch
https://agfax.com/2017/08/04/iowa-corn-soybeans-control-options-for-twospotted-spider-mites/
W129
W160
W126
Z58Z120
Z161
Z116
LA2329
X155
X166
X71
LA2329O
H
PI127826
Tomato Genotypes
0
2
4
6
Day 7
Egg D
en
sity
(N
o/c
m2 l
eaf
are
a),
Results
Susceptible line
Web
Resistant Line
Results
Conclusion
Based on reduced mite success on some of the genotypes, we concluded:
Resistance has been successfully transferred from the wild to the interspecific population.
This bioassay demonstrated behavioral differences of mites associated with the presence or absence of
leaf compounds and trichome densities.
New plant versions that can produce toxic or repellent chemicals on their own can defend themselves
against certain types of arthropods and insects which in turn reduce or eliminate extensive synthetic
pesticide utilization and cost.
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