Applications of double-SNP marker in detection of Tomato yellow leaf curl disease-resistant gene

A technology of tomato yellowing curved leaves and disease resistance genes, applied in the field of plant molecular marker preparation, can solve the problems of SNP gene marker reporting without Ty-1 gene, achieve high accuracy and stability, and improve efficiency

Inactive Publication Date: 2014-06-18
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] So far, there is no report

Method used

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  • Applications of double-SNP marker in detection of Tomato yellow leaf curl disease-resistant gene
  • Applications of double-SNP marker in detection of Tomato yellow leaf curl disease-resistant gene
  • Applications of double-SNP marker in detection of Tomato yellow leaf curl disease-resistant gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Obtaining the cDNA of tomato Ty--1 and ty--1 and the gDNA sequence of ty--1 The establishment of double SNP markers

[0042] (1) Refer to the publication number WO2012125025A1 (http: / / www.google.com.hk / patents / WO2012125025A1?cl=en) to find the cDNA sequence of Ty-1 and its susceptible allele ty-1 (see figure 1 ), the reference (Verlaan MG et al.2013) obtained the gDNA sequence of the susceptible gene ty-1 and its position on the whole genome (chromosome 6 30850904-30879542, composed of Solyc06g051170, Solyc06g051180 and Solyc06g051190), and used for the development of SNP markers.

[0043] (2) Through MultAlin online sequence comparison software (http: / / multalin.toulouse.inra.fr / multalin / multalin.html), the anti-susceptible cDNA sequences were compared, and 54 SNP sites were found ( figure 1 shaded bases). Among them, the bases at positions 313 and 314 of Ty-1 and ty-1 are respectively GT and CG (named NL2) see figure 1 a Bordered bases, the bases at 1159, ...

Embodiment 2

[0058] Example 2 The genotyping and field inoculation verification of the test material with NL2-3 double SNP markers

[0059] (1) NL2-3 test for genotyping of test materials

[0060] The tomato materials used for testing genotyping (not for the purpose of genetic breeding) came from commonly used tomato materials collected by the tomato research group of Huazhong Agricultural University. The results of SNP molecular marker identification are shown in Figure 6 (Where Marker is DS5000, lane 1. is the known disease-resistant homozygous material LA1969, 2. is the known susceptible material Moneymaker (according to the article published by Verlaan M G et al. in 2013), 3. is the known heterozygous disease-resistant material commodity Species F1 generation Huafan No. 11, 4. Swimming lanes 3-19 are the test materials). The screening results showed that lanes 4, 9, and 15 were homozygous disease-resistant genotype Ty-1 / Ty-1, and lanes 2, 5, 6, 8, 10, 12, 13, 14, 16, 17, 18, and 19 w...

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Abstract

The invention belongs to the technical field of plant molecular marker technology, and particularly relates to a double-SNP marker of a gene Ty-1 related to resistance to tomato yellow leaf curl virus disease, cDNA sequences of disease-resistant gene Ty-1 and disease-infected gene (namely allele gene) ty-1 and positions thereof on a whole genome can be obtained from the reported WO2012125025 document. Through sequence comparison, the disease-infected and disease-resistant specific primers NL2 and NL3 can be designed respectively by selecting two multielement SNP-locus GT/CG and GTG/ACA, and are mixed to form a NL2-3 double-SNP marker primer. The three types of Ty-1 and ty-1 can be detected by the NL2-3SNP marker through once PCR and gel electrophoresis display.

Description

technical field [0001] This patent belongs to the technical field of plant molecular marker preparation, and specifically relates to the application of a double SNP molecular marker in the detection of tomato yellow leaf curl disease resistance gene Ty-1. The simple and applicable disease resistance gene marker prepared by the invention can be used in vegetables In particular, marker-assisted selection of tomato disease resistance genes. Background technique [0002] Tomato is one of the most important vegetable crops in the world. It has developed into one of the 30 crops with the highest yield in the world and is widely planted in the north and south of my country. However, with the continuous expansion of protected cultivation areas and the continuous extension of cultivation time, many diseases are showing a trend of increasing year by year, and new diseases are constantly appearing, which has caused serious losses to my country's tomato production. [0003] In recent y...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2565/125
Inventor 叶志彪葛乃蓬李汉霞王涛涛张俊红张余洋杨长宪
Owner HUAZHONG AGRI UNIV
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