Molecular marker primer for aided identification of red pepper gray mold resistance and application of molecular marker primer

An auxiliary identification and molecular marker technology, applied in the field of capsicum biology, to achieve the effect of early breeding and major application prospects

Inactive Publication Date: 2016-02-03
崔青青
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A large number of studies on molecular markers closely linked to resistance genes have been reported, such as apples (Xue et al. see the report

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] An inbred line with high resistance to gray mold 'Longjiao 5', (an inbred line with high resistance to gray mold sold by Gansu Academy of Agricultural Sciences), pepper variety EarlyCalwonder (an inbred line with high susceptibility to gray mold ) The above-mentioned two pepper varieties have been widely used by pepper breeders and can be purchased or obtained from the above-mentioned units without restriction. The inbred line 'Longjiao 5' (male parent) with high resistance to gray mold and the inbred line 'Early Calwonder' (female parent) with high susceptibility to gray mold were crossed to obtain the F1 generation. The F1 generation was self-crossed to obtain the F2 generation segregation population (142 F2 generation individuals). F2 generation individual plants were selfed to obtain F3 generation family seeds. According to the resistance segregation characteristics identified by artificial inoculation of each F3 generation family, the corresponding F2 generation in...

Embodiment 2

[0027]The sequence of the specific detection gene fragment closely linked with the pepper gray mold resistance gene was obtained. After recovering and purifying the 2-item AFLP bands from the polypropylene gel, they were ligated with the pEASY-T1 vector. The ligation system: PCRproducts4μl, pMD-Tvoctor1μl , solutionI5μl, total volume10μl. Add the above reagents into a 0.5 centrifuge tube in an ice bath, flick to mix well, centrifuge briefly, and connect overnight (4°C). Take out the centrifuge tube of 50 μl of competent bacterial suspension from the -80°C refrigerator, put it on the ice box to thaw slowly, add about 10 μl of all the ligated products, flick the tube wall to mix well, and place it on the ice-water mixture for 30 minutes; Quickly heat-shock in a water bath at 42°C for 90s; quickly place it on ice for 3-5 minutes, do not shake the centrifuge tube during this period; add 700 μl LB liquid medium without AmP, transfer to 37°C, and culture on a shaker at 150r / min 1-1...

Embodiment 3

[0029] Develop and identify SCAR primers for Botrytis cinerea resistance from specific gene fragment sequences. According to the sequencing results, remove the carrier and linker sequences, use the remaining sequences to combine with restriction enzyme sites to design SCAR primers, and use the newly developed SCAR primers according to the AFLP primer numbering rules. The marker is named CARpi-1-200, where C represents the SCAR marker, the subscript 200 indicates the length of the amplified fragment of the SCAR marker, and the primer sequence is:

[0030] CARpi-1-200(F): 5'-GTCTGCTCCAACTTCCTCT-3';

[0031] CARpi-1-200(R): 5'-CAGTTCAGCATCATCAATCG-3';

[0032] Using the disease-resistant gene pool, the susceptible gene pool, the DNA of the parents, and the DNA of each individual plant that makes up the resistant-susceptible pool, use the newly synthesized CARpi-1-200 to amplify, and only amplify in the susceptible gene pool and the female parent. A specific band with a size of 2...

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PUM

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Abstract

The invention discloses a molecular maker primer for aided identification of red pepper gray mold resistance and an application of the molecular marker primer. A reagent for aided identification of red pepper gray mold resistance and / or aided identification of red pepper gray mold resistance, provided by the invention, is a molecular maker primer pair CARpi-1-200, and is a specific primer pair consisting of nucleotides. By using the primer pair, provided by the invention, for aided identification of red pepper gray mold resistance and / or aided screening of red peppers with gray mold resistance, compared with a field identification result, the disease-resistant coincidence rate reaches 90%. During red pepper seed cultivation, by using the molecular maker disclosed by the invention, the genes for resisting gray molds, in red pepper breeding materials, can be subjected to molecular maker aided selection before the attack of the gray molds in a seedling stage, thereby improving the breeding efficiency and accelerating the breeding progress.

Description

technical field [0001] The invention relates to a marker primer of a gray mold resistance gene molecule of pepper, belongs to the field of pepper biotechnology, and specifically relates to a molecular marker primer of a gray mold resistance gene in a pepper Gansu Longjiao strain and an application thereof, which can be used for identification and screening. Background technique [0002] Capsicum (Capsicum pp.) is one of the most important vegetable crops in the world. It has many varieties, high yield, rich nutrition and wide application. It plays an important role in people's daily life. In China, after decades of development, the chili planting industry has become the second largest vegetable crop in my country. According to statistics from the United Nations Food and Agriculture Organization, the annual sowing area of ​​chili peppers in my country was 750,000 hectares in 2012, and the annual output of fresh peppers and dried peppers 16.31 million tons, the sown area and ou...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 崔青青
Owner 崔青青
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