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IMPROVING RESISTANCE TO FUSARIUM WILT RACE
THREE OF TOMATOJessica Chitwood-Brown
Tomato Breeders Roundtable Meeting 2018
J. Lee, S.F Hutton, T. G. Lee, and G.E. Vallad
Gulf Coast Research and Education Center, University of Florida
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Outline
Fusarium wilt of tomato
Available resistance and its challenges
Improving available
resistance
Novel resistance
Improved Fol3 resistant cultivars
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Tomatoes in Florida
Photo credit: J. Chitwood-Brown
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Fusarium wilt in tomato
• Fusarium oxysporum f. sp. lycopersici (Fol)• Soilborne fungus – colonizes vascular tissue
• Forms chlamydospores
• Chlorosis, wilting, stunting, vascular browning, plant death
• Management• Fumigation
• Phase-out of methyl bromide
• Host resistance is most effective
• Three races
Photo credit: http://plantpath.ifas.ufl.edu/u-scout/Tomato/Pages/Fusarium_wilt.html
Photo credit: J. Chitwood-Brown
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Genetic resistance
I
Race 1
S. pimpinellifolium
I-2
Race 2
S. pimpinellifolium
I-3
Race 3
S. pennellii
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Genetic resistance
I
Race 1
S. pimpinellifolium
I-2
Race 2
S. pimpinellifolium
I-3
Race 3
S. pennellii
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Genetic resistance
I
Race 1
S. pimpinellifolium
I-2
Race 2
S. pimpinellifolium
I-3
Race 3
S. pennellii
Focusing on race 3 (Fol3) resistance
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Genetic resistance
• I-3 cultivars available since the 1990’s• Associated with negative traits
• Increased sensitivity to bacterial spot
Photo credit: Li et al. 2018
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Genetic resistance
• I-3 cultivars available since the 1990’s• Associated with negative traits
• Increased sensitivity to bacterial spot
Photo credit: Li et al. 2018
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Genetic resistance
• I-3 cultivars available since the 1990’s• Associated with negative traits
• Increased sensitivity to bacterial spot
• Reduced fruit size
• 1999
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Genetic resistance
• I-3 cultivars available since the 1990’s• Associated with negative traits
• Increased sensitivity to bacterial spot
• Reduced fruit size
• 1999
• 2004
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Genetic resistance
• I-3 cultivars available since the 1990’s• Associated with negative traits
• Increased sensitivity to bacterial spot
• Reduced fruit size
• 1999
• 2004
• 20+ years of breeding later
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Genetic resistance
• I-3 cultivars available since the 1990’s• Associated with negative traits
• Increased sensitivity to bacterial spot
• Reduced fruit size
• 1999
• 2004
• 20+ years of breeding later
• Linkage drag or pleiotropy?
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Approach
Eliminate linkage drag
with I-3
Identify and utilize novel resistance
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Approach
Eliminate linkage drag
with I-3
Identify and utilize novel resistance
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Linkage drag
• Linkage drag or pleiotropy?• Pleiotropy – I-3 itself
• Linkage drag – linked genes from wild species
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Linkage drag
• Linkage drag or pleiotropy?• Pleiotropy – I-3 itself
• Linkage drag – linked genes from wild species
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Eliminate linkage drag
I-3
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Eliminate linkage drag
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Eliminate linkage drag
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Eliminate linkage drag
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Eliminate linkage drag
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Eliminate linkage drag
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Eliminate linkage drag
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Eliminate linkage drag
• Backcrossing this minimal introgression into several elite UF/IFAS breeding lines
• Testing for effect on bacterial spot sensitivity and fruit size
Genotype Introgression
Lg/Lg
Lg/Min
Min/Min
-/-
Min/-
Min/Min
Fla. 7946
Fla. 8059
Photo credit: J. Chitwood-Brown
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Eliminate linkage drag
• Backcrossing this minimal introgression into several elite UF/IFAS breeding lines
• Testing for effect on bacterial spot sensitivity and fruit size
Genotype Introgression
Lg/Lg
Lg/Min
Min/Min
-/-
Min/-
Min/Min
Fla. 7946
Fla. 8059Photo credit: J. Chitwood-Brown
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Eliminate linkage drag
Genotype IntrogressionBacterial
Spot Rating
Average
Fruit Size (g)
Lg/Lg 5.8 132
Lg/Min 5.6 149
Min/Min 5.2 154
-/- 4.8 111
Min/- 4.9 110
Min/Min 4.8 115
Fla. 7946
Fla. 8059
A
AB
BC
A
AB
BC
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Eliminate linkage drag
Genotype IntrogressionBacterial
Spot Rating
Average
Fruit Size (g)
Lg/Lg 5.8 132
Lg/Min 5.6 149
Min/Min 5.2 154
-/- 4.8 111
Min/- 4.9 110
Min/Min 4.8 115
Fla. 7946
Fla. 8059 NS
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Eliminate linkage drag
Genotype IntrogressionBacterial
Spot Rating
Average
Fruit Size (g)
Lg/Lg 5.8 132
Lg/Min 5.6 149
Min/Min 5.2 154
-/- 4.8 111
Min/- 4.9 110
Min/Min 4.8 115
Fla. 7946
Fla. 8059 NS
Linkage with negative effects likely broken.
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Approach
Eliminate linkage drag
with I-3
Identify and utilize novel
resistance
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Novel Fol3 resistance
• S. pennellii demonstrated to have high resistance to fusarium wilt
• 42 S. pennellii accessions from the Tomato Genetic Resource Center (TGRC)• Almost all accessions resistant
• Resistance introgressed into Florida breeding lines via backcrossing• Seedling disease assays, selecting for
resistance
Photo credit: J. Chitwood-Brown
Photo credit: J. Li
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Novel Fol3 resistance
• Molecular markers used to select away from known resistance genes
• Spring 2018 - resistance from 26 accessions advanced to BC5F1
• Choosing families for mapping• Clear phenotypes
• Segregation ratios
• Phylogenetic relationships
Photo credit: J. Chitwood-BrownPhoto credit: J. Chitwood-Brown
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Novel Fol3 resistance
Resistance
fromHealthy Infected Ratio
LA750 45 7 6:1
LA1273 43 12 4:1
LA1302 33 13 3:1
LA1303 42 13 3:1
LA1522 43 13 3:1
LA1809 37 15 3:1
LA1911 44 12 4:1
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Novel Fol3 resistance
• Phylogenetic relationships
• Different accessions belong to different clusters
• Accessions from different clusters chosen
Credit: T.G. Lee
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Novel Fol3 resistance
• Phylogenetic relationships
• Different accessions belong to different clusters
• Accessions from different clusters chosen
Families with resistance from these 4 accessions are being genotyped for mapping and prepared
for characterization studies.
Credit: T.G. Lee
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Impacts
Linkage between I-3 and negative traits likely broken
Continued exploration of novel Fol3 resistance alleles
Combine available and novel genes for more durable resistance
Photo credit: J. Chitwood-BrownPhoto credit: J. Chitwood-Brown
https://www.upi.com/Science_News/2015/08/19/Better-tasting-grocery-store-tomatoes-coming-soon-scientists-say/8801439994501/
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Thank you
• Committee members• Dr. Sam Hutton – advisor• Dr. Tong Geon Lee• Dr. Gary Vallad – plant pathology lab• Dr. Vance Whitaker
• Tomato breeding lab• Rebecca Wente • Dolly, Jose, Judith, Kazuyo, Keri, Nate, Reza, Tim• John and Upinder• Edgar and Jasmine
• Funding• PBGI • USDA Tomato CGC
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QUESTIONS?
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Recombinant inbred lines
Marker:
Physical position: Fla.7228 69 a
Fla. 7217D 51 b
Fla.7169 38 c
PP 44R18
LL 43
PP 57R12
LL 58
PP 65R17
LL 62
PP 57R11
LL 55
PP 55 aR07
LL 48 b
PP 61 aR09
LL 43 b
PP 52 aR08
LL 37 b
PP 51 aR15
LL 36 b
PP 41R24
LL 37
PP 62 aR14
LL 43 b
63.63 63.63 63.63 63.70
Recombinant Lines I-3
bB6
58.62 59.51 60.96 62.77 63.18 63.41 63.59 63.60 63.62
7g916 7g923 7g6362 7g6363 SLG-1 7g957
Tomato Chromosome 7
Control /
Haplotype
Pct. Disease
Severity7g5862 7g5951 7g6096 I-3CAPS 7g501 7g728
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Recombinant inbred lines
Marker:
Physical position: Fla.7228 69 a
Fla. 7217D 51 b
Fla.7169 38 c
PP 44R18
LL 43
PP 57R12
LL 58
PP 65R17
LL 62
PP 57R11
LL 55
PP 55 aR07
LL 48 b
PP 61 aR09
LL 43 b
PP 52 aR08
LL 37 b
PP 51 aR15
LL 36 b
PP 41R24
LL 37
PP 62 aR14
LL 43 b
63.63 63.63 63.63 63.70
Recombinant Lines I-3
bB6
58.62 59.51 60.96 62.77 63.18 63.41 63.59 63.60 63.62
7g916 7g923 7g6362 7g6363 SLG-1 7g957
Tomato Chromosome 7
Control /
Haplotype
Pct. Disease
Severity7g5862 7g5951 7g6096 I-3CAPS 7g501 7g728
I-3
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Horsfall-Barratt ScaleScore % Diseased Tissue
1 0%
2 0-3%
3 3-6%
4 6-12%
5 12-25%
6 25-50%
7 50-75%
8 75-87%
9 87-94%
10 94-97%
11 97-100%
12 100%