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CAPS molecular marker based on tomato yellow leaf curl virus disease resistance gene Ty-3

A technology of tomato yellowing curved leaves and molecular markers, applied in the field of agricultural biotechnology engineering, can solve problems such as false positives

Active Publication Date: 2016-06-01
QINGDAO AGRI UNIV
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  • Summary
  • 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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  • CAPS molecular marker based on tomato yellow leaf curl virus disease resistance gene Ty-3
  • CAPS molecular marker based on tomato yellow leaf curl virus disease resistance gene Ty-3
  • CAPS molecular marker based on tomato yellow leaf curl virus disease resistance gene Ty-3

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Development and verification of the CAPS marker based on the Ty-3 gene

[0033] 1. Test materials

[0034] The disease-resistant inbred line A45 and the susceptible inbred line A39 used in this experiment were detected by the known molecular marker P6-25, and A45 was a resistant homozygous (Ty-3 / Ty-3), while A39 was a susceptible homozygous (ty-3 / ty-3). A45 is the female parent, and A39 is the male parent to prepare a hybrid combination to obtain F1, and then self-cross to obtain F2 segregation population.

[0035] 2. Vaccination identification

[0036] 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 whiteflies, kill the poisonous whiteflies after two weeks, and then transplant the seedlings into the solar greenhouse or field, and observe Condition results.

[0037] 3...

Embodiment 2

[0052] Example 2 Molecular marker-assisted selection of Ty-3 gene using CAPS marker primers

[0053] Utilize the Ty-3-CAPS marker based on Ty-3 developed in Example 1 for the BC that the research group is going to transfer the Ty-3 gene 1 48 strains of generation population materials (the susceptible parent P32 with high soluble solid content is the female parent, the inbred line P57 containing the Ty-3 gene is the male parent, and its F 1 Backcross generation with P32 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-CAPS was heterozygous positive (332bp, 221bp and 112bp specific bands were present at the same time) and 21 individual plants were numbered 4, 5, 8, 11, 12, 13, 14, 15, 17, 23, 25, 27, 29, 30, 32, 33, 34, 35, 36, 41 and 48 individual plants; the genotype detected by marker Ty-3-CAPS was Negative (with 221bp and 112bp specific bands at the same time) 27 indivi...

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Abstract

The invention relates to a CAPS molecular marker based on tomato yellow leaf curl virus disease resistance gene Ty-3. The nucleotide sequence of the molecular marker is disclosed as SEQ ID No.1. The invention also relates to a primer pair for amplifying the CAPS marker based on tomato yellow leaf curl virus disease resistance gene Ty-3. The sequences of the primer pair are as follows: the sequence of the forward primer is 5'TTGCCACATTAAGCAGAACG-3'; and the sequence of the reverse primer is 5'-TGACGGTCATTGAATGTGCT-3'. The marker is a codominant marker based on PCR (polymerase chain reaction) technique, and can obviously lower the cost and save the labor as compared with the RFLP, AFLP, and other markers. Besides, the marker is completely based on the disease-resistant gene Ty-3, is coseparated from the Ty-3 gene, and thus, does not have the problem of false positive when being used for detection. When being used for seedling stage selection, the marker can reduce the workload, and can avoid the difficulty in accurately screening out the individual plant with the disease-resistant gene due to the insufficient inoculation, thereby accelerating the breeding progress.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology engineering, and particularly relates to a CAPS 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 tomato produc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 王辉王富李文丽董盼盼
Owner QINGDAO AGRI UNIV
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