Pea downy mildew resistance gene fragment or gene marker and application

A technology of gene markers and gene fragments, applied in the field of plant biotechnology science, can solve problems such as economic loss and affecting the growth and development of pea plants, and achieve the effects of shortening the breeding period, improving breeding efficiency, and shortening the excavation cycle

Inactive Publication Date: 2015-04-29
CHANGSHU ZHILIN FRUITS & VEGETABLES PROFESSIONAL COOP
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Problems solved by technology

Downy mildew seriously affects the normal growth and development p

Method used

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  • Pea downy mildew resistance gene fragment or gene marker and application
  • Pea downy mildew resistance gene fragment or gene marker and application
  • Pea downy mildew resistance gene fragment or gene marker and application

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Embodiment Construction

[0015] The identification of disease-resistant genes plays an important role in the theoretical research of crop disease-resistant genetics and the selection of disease-resistant varieties. This method can quickly identify downy mildew resistance genes in pea. The specific implementation process is as follows:

[0016] 1. DNA extraction: Take an appropriate amount of young leaves of pea, dry them with silica gel, grind them into powder, and extract the total genomic DNA by CTAB method. The extracted DNA was detected by an ultraviolet spectrophotometer, and samples with an OD260 / OD280 value of 1.8-2.0 were selected and stored at -20°C for future use.

[0017] 2. Amplification and cloning of homologous fragments of disease resistance genes: according to the amino acid sequences GG(L / I / V / M / S)GKTT and G(L / N ) PL (A / T / S) (L / V), designed a pair of degenerate primers: NBS15'-GKTT GGIGGIWSIGGIAARACIACG-3' and NBS25'-GCIGCIIARIGGRTTICC-3'. Take 50 ng of pea genomic DNA as a template...

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Abstract

The invention relates to a pea downy mildew resistance gene fragment or gene marker and an application. Specifically, one pea downy mildew resistance gene fragment or gene marker is rapidly separated by using an anti-disease gene homologous sequence method, and the sequencing shows that the nucleotide length of the gene fragment or the gene marker is 528bp. By adopting the pea downy mildew resistance gene fragment or the gene marker, the pea downy mildew resistance gene discovery period is effectively shortened, and rapid identification on a downy mildew gene is facilitated; corresponding co-separation functional markers (SNP, SCAR and the like) can be developed through the identified downy mildew gene, and rapid auxiliary molecular marker selection on the downy mildew gene can be also implemented with high accuracy; with the combination of other anti-disease gene molecular markers, multi-resistance breeding materials can be innovated, the breeding years are reduced, and the breeding efficiency is improved; and basis is made for expressing the pea downy mildew resistance gene molecule mechanism.

Description

technical field [0001] The invention utilizes the homologous sequence method of the disease-resistant gene to clone pea downy mildew-resistant gene fragments or convert them into molecular markers, etc., and mainly relates to conventional hybrid breeding of pea, molecular marker-assisted breeding, positioning and molecular markers of downy mildew-resistant genes. Mapping, cloning of downy mildew resistance gene and analysis of regulation mechanism of disease resistance gene belong to the field of plant biotechnology science. Background technique [0002] Downy mildew is one of the most important diseases of pea production. Mainly damage the leaves. In the early stage of the disease, brown lesions appear on the leaves, and the spore layer of the mold cluster is born on the back or surface of the leaves, white to lavender. There are relatively more spore layers on the back of leaves. Tender slightly suffer more. The lavender mold layer on the back of the leaves can cover th...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 钱孝英钱春桃
Owner CHANGSHU ZHILIN FRUITS & VEGETABLES PROFESSIONAL COOP
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