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Research Article

Gene-Based Markers for the Tomato Yellow Leaf Curl Virus Resistance Gene Ty-3

Plant Breeding and Biotechnology 2016;4(1):79-86.
Published online: February 28, 2016

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*Corresponding author: Byoung-Cheorl Kang, bk54@snu.ac.kr, Tel: +82-2-880-4563, Fax: +82-2-873-2056
• Received: January 12, 2016   • Revised: February 11, 2016   • Accepted: February 11, 2016

Copyright © 2016 The Korean Society of Breeding Science

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Gene-Based Markers for the Tomato Yellow Leaf Curl Virus Resistance Gene Ty-3
Plant Breed. Biotech.. 2016;4(1):79-86.   Published online February 28, 2016
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Gene-Based Markers for the Tomato Yellow Leaf Curl Virus Resistance Gene Ty-3
Image Image Image
Fig. 1 Phenotype of agro-infiltrated plants. Phenotype of susceptible line ‘A39’, resistant line ‘A45’, their F1, and F2 plants 3 weeks after inoculation are shown. Below: Tomato yellow leaf curl virus DNA was amplified using PAPB marker (Deng et al. 1994). S: susceptible, R: resistant.
Fig. 2 Physical locations and sequences of molecular markers developed based on the Ty-3 gene. Genetic structure of Ty-3 gene (A), modified from Verlaan et al. (2013). Genetic sequences of tomato reference genome SL2.50 (Heinz 1706-BG), susceptible line (A39), and resistant line (A45) were aligned. Primer locations of Ty3-HRM1 (B), Ty3-HRM2 (C), and Ty3-SCAR1 (D) are shown as arrows. HRM: high-resolution melting, SCAR: sequence-characterized amplified region.
Fig. 3 Normalized HRM curves of Ty3-HRM1 (A) and Ty3-HRM2 (B) and polymerase chain reaction products of SCAR marker Ty3-SCAR1 (C). M: size marker, S: genotype of susceptible line ‘A39’, R: genotype of resistant line ‘A45’, H: genotype of heterozygous F1 plants, HRM: high-resolution melting, SCAR: sequence-characterized amplified region.
Gene-Based Markers for the Tomato Yellow Leaf Curl Virus Resistance Gene Ty-3

Primer sequences for high-resolution melting (HRM) and the sequence-characterized amplified region (SCAR) analysis.

Marker Sequence (5′-3′) Type Product size (bp)
Ty3-HRM1 TATACGAGGTTCGCCGTCTT
TCTTGATTCTGTCACCATTGAAA
HRM 190
Ty3-HRM2 GAATAATGTCAAACCAAGAACTAGGA
TGTTTTCAGTTTTAAGAGATTTTAGCA
HRM 152
Ty3-SCAR1 GCTCAGCATCACCTGAGACA
TGCAGGAACAGAATGATAGAAAA
SCAR 519 (R)/269 (S)

Segregation for resistance to Tomato yellow leaf curl virus in different populations.

Population Expected ratio of R:S Observed number of plantsz) χ2 P-value

R S
P1 (‘A39’) 0:1 0 15 - -
P2 (‘A45’) 1:0 15 0 - -
F1 1:0 15 0 - -
BC1F1 1:1 41 34 0.66 0.42
F2 3:1 112 38 0.01 0.92

z)R: resistant, S: susceptible.

Co-segregation analysis of Tomato yellow leaf curl virus resistance and genotype of Ty-3.

Population Genotype Phenotypez) Total

R S
BC1F1 R 0 0 0
H 41 1y) 42
S 0 33 33
Total 41 34 75
F2 R 30 0 30
H 82 3y) 85
S 0 35 35
Total 112 38 150

z)R: resistant, S: susceptible.

y)Number of lines did not show the co-segregated results.

Table 1 Primer sequences for high-resolution melting (HRM) and the sequence-characterized amplified region (SCAR) analysis.
Table 2 Segregation for resistance to Tomato yellow leaf curl virus in different populations.

R: resistant, S: susceptible.

Table 3 Co-segregation analysis of Tomato yellow leaf curl virus resistance and genotype of Ty-3.

R: resistant, S: susceptible.

Number of lines did not show the co-segregated results.