Molecular marker method for identifying tomato yellow leaf curl virus resistance gene Ty-1

A technology of tomato yellowing curved leaves and molecular markers, applied in the field of agricultural biology, can solve the problems of low linkage degree of marker genes, many operation procedures, and the need for enzyme digestion, etc., to achieve simple electrophoretic band patterns, improve accuracy, and speed up The effect of the breeding process

Active Publication Date: 2015-06-24
上海孙桥现代农业联合发展有限公司 +1
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Problems solved by technology

It aims to solve the technical problems in the prior art that there are many marking procedures for the resistance gene Ty-1 of tomato yellow leaf curl virus, enzyme digestion is required, and the degree of linkage with the marker gene is not high

Method used

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  • Molecular marker method for identifying tomato yellow leaf curl virus resistance gene Ty-1

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Embodiment 1

[0021] Example 1, Screening of Molecular Marker Primers

[0022] The BSA method was used to construct the disease-resistant gene pool and the disease-susceptible gene pool for the screening of molecular marker primers.

[0023] Zamir et al. located the yellow leaf curl virus resistance gene Ty-1 on chromosome 6, and the RAPD marker and CAPS marker linked to this gene have been screened out. According to the positioning result, tomato genome sequence and SSR genetic map, the present invention newly designs a series of marker primers. Then, polymorphism screening was carried out for the designed series of marker primers against disease-resistant materials and non-disease-resistant materials. As a result of the screening, only one pair of primers can amplify the polymorphism, which can be used as molecular marker primers. The molecular marker primer can amplify a 690bp fragment in the disease-resistant material containing the Ty-1 site; it can amplify a 409bp fragment in the no...

Embodiment 2

[0026] Example 2, identification of yellow leaf curl virus resistance gene using screened molecular marker primers

[0027] 1. Extraction of tomato genomic DNA by CTAB method

[0028] The materials to be identified are tomato materials of the following varieties: Shenfen 8, Shenfen 918, Puhong 9, Puhong 10, Shenfen V-1, Dongnong 721, Xianzheng Hongzhu.

[0029] Collect the new leaves of the tomato material of the above-mentioned varieties, and extract DNA with the CTAB method. The specific extraction method is as follows:

[0030] (1) Take 1-50g of fresh plant material and grind it into powder in liquid nitrogen. (2) Transfer the frozen powder into a pre-cooled centrifuge tube, immediately add an equal volume (w / v) of 2×CTAB extraction buffer, and incubate at 65°C for 10-20 minutes, shaking from time to time. (3) Add an equal volume of chloroform / isoamyl alcohol, gently invert the centrifuge tube to mix, and centrifuge at 12000 r / min for 10-20 minutes at room temperature. (...

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Abstract

The invention discloses a molecular marker method for identifying a tomato yellow leaf curl virus resistance gene Ty-1. The method comprises the following steps: extracting a tomato gene group DNA; performing PCR amplification on the extracted tomato gene group DNA by using a molecular marker primer and performing electrophoretic analysis on the obtained PCR amplification product, representing that the yellow leaf curl virus resistance gene Ty-1 in a plant is a disease-resistant plant if a stripe with the length of 690bp is generated in an electrophoretogram, and representing that the plant is a plant system containing ty-1 gene site and is not a disease-resistant plant if the stripe with the length of 409bp is generated in the electrophoretogram. The electrophoretogram in the marker method is simple in stripe form, easy to identify, can be directly applied to marker assistant selection and can improve the identification accuracy; and moreover, the time is saving, and the material resources and the labor force are reduced.

Description

technical field [0001] The invention relates to the technical field of agricultural biology, and specifically relates to a molecular marker method for identifying the resistance gene Ty-1 of tomato yellow leaf curl virus. Background technique [0002] Tomato yellow leaf curl virus is Chinese tomato yellow leaf curl virus (Tomato yellow leaf curl virus, TYLCV), which belongs to Geminiviridae. Compared with other common viral diseases on vegetables (such as mosaic leaf, fern leaves, stripe spot and other viral diseases), tomato yellow leaf curl virus disease has the characteristics of sudden outbreak, rapid expansion, strong harm and difficult treatment. It is a devastating plant disease in tomato production at present, and it is a disease that seriously threatens tomato production all over the world. Breeding disease-resistant varieties has become the main technical means to control the disease. [0003] Marker-assisted selection is an important part of breeding, speeding u...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 刘士辉张辉朱为民姚永康马新海金静孙洪助阎君刘娜于力杨学东
Owner 上海孙桥现代农业联合发展有限公司
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