IMPROVING RESISTANCE TO FUSARIUM WILT RACE THREE OF TOMATO Jessica Chitwood-Brown Tomato Breeders Roundtable Meeting 2018

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1 IMPROVING RESISTANCE TO FUSARIUM WILT RACE THREE OF TOMATO Jessica 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

2 Outline Fusarium wilt of tomato Available resistance and its challenges Improving available resistance Novel resistance Improved Fol3 resistant cultivars

3 Tomatoes in Florida Photo credit: J. Chitwood-Brown

4 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 Photo credit: Three races Photo credit: J. Chitwood-Brown

5 Genetic resistance I Race 1 I-2 Race 2 I-3 Race 3 S. pimpinellifolium S. pimpinellifolium S. pennellii

6 Genetic resistance I Race 1 I-2 Race 2 I-3 Race 3 S. pimpinellifolium S. pimpinellifolium S. pennellii

7 Genetic resistance I Race 1 I-2 Race 2 I-3 Race 3 S. pimpinellifolium S. pimpinellifolium S. pennellii Focusing on race 3 (Fol3) resistance

8 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

9 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

10 Genetic resistance I-3 cultivars available since the 1990 s Associated with negative traits Increased sensitivity to bacterial spot Reduced fruit size 1999

11 Genetic resistance I-3 cultivars available since the 1990 s Associated with negative traits Increased sensitivity to bacterial spot Reduced fruit size

12 Genetic resistance I-3 cultivars available since the 1990 s Associated with negative traits Increased sensitivity to bacterial spot Reduced fruit size years of breeding later

13 Genetic resistance I-3 cultivars available since the 1990 s Associated with negative traits Increased sensitivity to bacterial spot Reduced fruit size years of breeding later Linkage drag or pleiotropy?

14 Approach Eliminate linkage drag with I-3 Identify and utilize novel resistance

15 Approach Eliminate linkage drag with I-3 Identify and utilize novel resistance

16 Linkage drag Linkage drag or pleiotropy? Pleiotropy I-3 itself Linkage drag linked genes from wild species

17 Linkage drag Linkage drag or pleiotropy? Pleiotropy I-3 itself Linkage drag linked genes from wild species

18 Eliminate linkage drag I-3

19 Eliminate linkage drag

20 Eliminate linkage drag

21 Eliminate linkage drag

22 Eliminate linkage drag

23 Eliminate linkage drag

24 Eliminate linkage drag

25 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 Fla Fla Introgression Lg/Lg Lg/Min Min/Min -/- Min/- Min/Min Photo credit: J. Chitwood-Brown

26 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 Fla Fla Introgression Lg/Lg Lg/Min Min/Min -/- Min/- Min/Min Photo credit: J. Chitwood-Brown

27 Eliminate linkage drag Genotype Fla Fla Introgression Bacterial Average Spot Rating Fruit Size (g) Lg/Lg 5.8 A 132 A Lg/Min 5.6 AB 149 AB Min/Min 5.2 BC 154 BC -/ Min/ Min/Min

28 Eliminate linkage drag Genotype Fla Fla Introgression Bacterial Average Spot Rating Fruit Size (g) Lg/Lg Lg/Min Min/Min / Min/- 4.9 NS 110 Min/Min

29 Eliminate linkage drag Genotype Fla Fla Introgression Bacterial Average Spot Rating Fruit Size (g) Lg/Lg Lg/Min Min/Min / Min/- 4.9 NS 110 Min/Min Linkage with negative effects likely broken.

30 Approach Eliminate linkage drag with I-3 Identify and utilize novel resistance

31 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. Li Photo credit: J. Chitwood-Brown

32 Novel Fol3 resistance Molecular markers used to select away from known resistance genes Spring resistance from 26 accessions advanced to BC5F1 Choosing families for mapping Clear phenotypes Segregation ratios Phylogenetic relationships Photo credit: J. Chitwood-Brown Photo credit: J. Chitwood-Brown

33 Novel Fol3 resistance Resistance from Healthy Infected Ratio LA :1 LA :1 LA :1 LA :1 LA :1 LA :1 LA :1

34 Novel Fol3 resistance Phylogenetic relationships Different accessions belong to different clusters Accessions from different clusters chosen Credit: T.G. Lee

35 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

36 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-Brown Photo credit: J. Chitwood-Brown

37 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

38 QUESTIONS?

39 Recombinant inbred lines Tomato Chromosome 7 Marker: 7g5862 7g5951 7g6096 I-3CAPS 7g501 7g728 7g916 7g923 7g6362 7g6363 SLG-1 7g957 Physical position: Fla a Fla. 7217D 51 b Recombinant Lines I-3 Fla c bb6 Control / Haplotype Pct. Disease Severity R14 R24 R15 R08 R09 R07 R11 R17 R12 R18 PP 62 a LL 43 b PP 41 LL 37 PP 51 a LL 36 b PP 52 a LL 37 b PP 61 a LL 43 b PP 55 a LL 48 b PP 57 LL 55 PP 65 LL 62 PP 57 LL 58 PP 44 LL 43

40 R14 R24 R15 R08 R09 R07 R11 R17 R12 R18 Recombinant inbred lines PP 62 a LL 43 b PP 41 LL 37 PP 51 a LL 36 b PP 52 a LL 37 b PP 61 a LL 43 b PP 55 a LL 48 b PP 57 LL 55 PP 65 LL 62 PP 57 LL 58 PP 44 LL 43 I-3

41 Horsfall-Barratt Scale Score % Diseased Tissue 1 0% 2 0-3% 3 3-6% % % % % % % % % %

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