fine mapping of the tomato yellow leaf curl virus resistance …tgc.ifas.ufl.edu/presentations/11...
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Fine Mapping of the Tomato Yellow Leaf
Curl Virus Resistance Gene Ty-2 on
Chromosome 11 of Tomato
Xiaohui Yang, Jay Scott, and Sam Hutton
Gulf Coast Research & Education Center
University of Florida
43rd Tomato Breeders Roundtable Meeting
October 9 -11, 2011
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Overview
Tomato yellow leaf curl virus disease (TYLCV)
• The first monopartite begomovirus of the family
Geminiviradae.
• First reported in Israel in 1939.
• Serious losses to tomato production in tropical and
subtropical regions of the world.
• Vector: Sweet potato whitefly, Bemisia tabaci (Genn.).
• Disease symptoms: Leaf curling, yellowing and plant
stunting.
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Species Chromosome Reference
Ty-1 S. chilense 6 Zamir et al., 1994
Ty-2 S. habrochaites 11 Hanson et al., 2000
Ty-3 S. chilense 6 Ji et al., 2007
Ty-4 S. chilense 3 Ji et al., 2009
Ty-5 S. peruvianum 4 Anbinder et al., 2009
Overview
• TYLCV Resistance Genes:
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Ty-2 gene introgressions
Chl.11
(Hanson et al., 2000)
19 cM
89.0 T0302
92.0 TG26
84.0 TG36
103.0 TG393
84.0 TG36
(Ji et al., 2009)
6.5 cM
82.5 C2_At1g07960
89.0 T0302
92.0 TG26
103.0 TG393
82.3 C2_At2g28250
(Ji et al., 2007b)
8.0 cM
89.0 T0302
92.0 TG26
84.0 TG36
103.0 TG393
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Bipartite begomovirus disease severity for
combinations of genes Ty3 and Ty2
(from Mejia et al., 2010 TGC report) Guatemala
F1 families and Parents
Ty-2 /
Ty-3 / Ty-2 /
Ty-3 - Ty-2 -
Ty-3 / Ty-2 +
Ty-3 - Ty-2 -
Ty-3 +
Ty-2 -
Ty-3 -
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Ty-3 resistance Ty-2 resistance Horizon
(susceptible)
•TYLCV inoculation (TYLCV-Is )
Florida
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Research Objectives
• Further delimit the Ty-2 introgression to a
shorter interval in the target region.
• Evaluate efficacy of Ty-2 in combination with
Ty-3 and Ty-4.
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Materials and Methods
• Population derived from Heinz H9205:
• F2 plants heterozygous for Ty-2 introgression
selected and F3 seed used for TYLCV evaluation.
• Ty-2 resistant control: homozygous for Ty-2.
• ‘Horizon’ = susceptible control.
Plant material
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• Seedlings exposed to viruliferous whiteflies for
TYLCV for two weeks (Griffiths and Scott, 2001)
• Whiteflies killed prior to transplanting to pots or
the field
• Disease severity rated ~60 days after inoculation
Materials and Methods
Inoculation
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Materials and Methods
Markers
87.0 cM cLEN-11-F24
89.0 cM T0302
84.0 cM TG36
85.0 cM T0386
82.3 cM C2At2g28250
cL1
cL2
C2At3g52090
M1
M2
M3
(Tomato-EXPEN 2000)
82.5 cM C2At1g07960
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F(LOD3) markers
Genetic distance
(cM)
Physical distance
(Mb)
TG36 84.0 SL2.40ch11 51.490
TG105 90.0 SL2.40ch11 52.068
1 cM = ~100kb
~500kb
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• Fall 2009: Screened ~4,000 F2 plants for recombination
~30 recombinants
Ty-2 Fine Mapping
Recombinant screen
Genotype
Flanking Molecular markers
No. of recombinants
C2At1g07960 (82.5 cM)
T0302 (89 cM)
Ⅰ / - 8
Ⅱ - / 5
Ⅲ / + 11
Ⅳ + / 6
+: homozygous for S. habrochaites alleles; -: homozygous for S. lycopersicum alleles; /: heterozygous alleles.
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TYLCV Evaluation
Spring 2010
• Grew progeny of recombinants (categories I and II)
• Inoculated seedlings
• Greenhouse evaluation of virus symptoms
Fall 2010
• Grew out progeny (categories III and IV)
• Screened each plant with markers and selected RILs
• Inoculated RILs
• Greenhouse evaluation of virus symptoms
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Introgression patterns for resistant RILs are shown in green.
Introgression patterns for susceptible RILs are shown in red.
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Ty-2 fine mapping with additional
population
• Spring 2011: Screened ~7,000 F2 plants for recombination
~127 total recombinants (C2At1g07960 and T0302)
~26 informative recombinants (C2At1g07960 and M1)
• Fall 2011:
• Marker screens on progeny of informative recombinants
• Selected RILs and Inoculated
• Disease severity evaluations
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Introgression patterns for homozygous resistant RILs are shown in green.
Introgression patterns for homozygous susceptible RILs are shown in red.
Some part of heterozygous introgression are shown in yellow.
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Markers Recombinant
C2At1g07960 +
TG36 /
cL2 /
C2At3g52090 /
M1 /
No. of Progeny
17 53 21
+ + +
+ / -
Confusing genotype?
No. of Progeny
17 53 21
+ + +
+
+
+
+ / -
No. of Progeny
17 53 21
+ + +
+ /
+ /
+ /
+ / -
No. of Progeny
17 53 21
+ + +
+ / /
+ / /
+ / /
+ / -
+: homozygous for S. habrochaites alleles; -: homozygous for S. lycopersicum alleles; /: heterozygous alleles.
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Summary
• The Ty-2 gene mapped to
<3.0 cM region (<300 Kb).
• Severe recombination
suppression between
C2_At1g07960 and
C2_At3g52090.
<3.0 cM
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Future Work
• Efforts underway to sequence
resistant material for the Ty-2
region.
• Candidate gene identification.
Line Ty-2 Ty-3 Ty-4
1 - - -
2 + - -
3 - + -
4 - - +
5 / - -
6 - / -
7 - - /
8 - + +
9 - + /
10 - / +
11 - / /
12 / - /
13 / / -
14 / / /
• Test resistance of Ty-2, Ty-3, Ty-4
gene combinations.
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