InDel molecular marker based on TYLCV (tomato yellow leaf curl virus) resistance gene Ty-3

A tomato yellowing curved leaf and molecular marker technology, which is applied in the field of agricultural biotechnology engineering, can solve the problems of false positives and achieve the effects of strong specificity, shortened breeding years, and high stability

Active Publication Date: 2016-06-08
QINGDAO AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is a certain genetic distance between these markers and disease-resistant genes, so a certain proportion of false positives will occur during screening.

Method used

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  • InDel molecular marker based on TYLCV (tomato yellow leaf curl virus) resistance gene Ty-3
  • InDel molecular marker based on TYLCV (tomato yellow leaf curl virus) resistance gene Ty-3
  • InDel molecular marker based on TYLCV (tomato yellow leaf curl virus) resistance gene Ty-3

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Development and verification of InDel markers based on Ty-3 gene

[0031] 1. Test materials

[0032] The disease-resistant inbred line P1 and the susceptible inbred line P2 used in this experiment were confirmed by the detection of the known molecular marker P6-25. Fit (ty-3 / ty-3). P1 is the female parent, and P2 is the male parent to prepare a hybrid combination, after obtaining F1, and then selfing to obtain F2 segregation population.

[0033] 2. Vaccination identification

[0034] The resistance identification adopts the whitefly inoculation identification method, specifically referring to the method of Yang Xiaohui (2012). At the stage of 2-3 true leaves of the seedlings, place them in a growth room with poisonous whitefly, kill the poisonous whitefly after two weeks, and then transplant the seedlings into the solar greenhouse or field, and observe Condition results.

[0035] 3. Development and verification of InDel molecular markers

[0036] A. Deve...

Embodiment 2

[0050] Example 2 Molecular marker-assisted selection of Ty-3 gene using InDel marker primers

[0051] Utilize the InDel marker primer (Ty-3-InDel) based on Ty-3 developed in Example 1 to the BC that the research group prepares to transduce the Ty-3 gene 1 48 strains of generation population materials (the susceptible parent P31 with high soluble solid content is the female parent, the inbred line P55 containing the Ty-3 gene is the male parent, and its F 1 Generation and P31 were backcrossed to obtain BC 1 ) for marker-assisted selection. the result shows( Figure 4 ): Among the 48 individual plants, the genotype detected by the marker Ty-3-InDel was heterozygous positive (there were 510bp and 260bp specific bands at the same time) and 19 individual plants were numbered 1, 2, 3, 5, No. 9, 13, 14, 16, 18, 21, 29, 30, 33, 37, 39, 40, 42, 43 and No. 47 individual plants; the genotype detected by the marker Ty-3-InDel was negative (only 260bp Specific bands) 29 individual plan...

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Abstract

The invention relates to an InDel molecular marker based on a TYLCV (tomato yellow leaf curl virus) resistance gene Ty-3. The nucleotide sequence of the molecular marker is represented as SEQ ID No.1. The invention further relates to a primer pair for amplifying the InDel molecular marker based on the TYLCV resistance gene Ty-3. Sequences of the primer pair are as follows: a forward primer sequence: 5'-TGCAGGAACAGAATGATAGAAAA-3', and a reverse primer sequence: 5'-GCTCAGCATCACCTGAGACA-3'. The molecular marker is totally based on the TYLCV resistance gene Ty-3. According to the developed InDel molecular marker based on the TYLCV resistance gene Ty-3, the marker is totally based on the resistance gene Ty-3, is completely linked with the resistance gene Ty-3 and is a codominant molecular marker. When the marker is applied to molecular marker-assisted selection of the TYLCV resistance gene Ty-3, the accuracy of selection can be greatly improved, the breeding cycle can be shortened, and accordingly, the breeding process is accelerated.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology engineering, and particularly relates to an InDel molecular marker based on tomato yellow leaf curl virus disease resistance gene Ty-3, primers for amplifying the molecular marker, a detection method using the molecular marker and its application. Background technique [0002] Tomato (Solanum lycopersicum L.) belongs to Solanaceae (Solanaceae) and is an annual herbaceous plant of Solanaceae. Tomato originates from the western region of South America and is one of the important vegetable crops widely cultivated in the world, and is deeply loved by consumers. With the expansion of tomato cultivation area and the diversification of cultivation methods, the problem of tomato pests and diseases has become increasingly serious. In recent years, tomato yellow leaf curl virus disease has commonly occurred in tomato cultivation in protected areas around the world, causing huge economic losses to t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
CPCC12Q1/6895C12Q2600/156
Inventor 王富王辉李文丽董盼盼
Owner QINGDAO AGRI UNIV
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