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Black spot disease resisting candidate gene sequence of dowing grape as one of China vitis wild species and use

A candidate gene, the technology of hairy grapes, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of low hybridization efficiency, poor disease resistance, and limited application

Inactive Publication Date: 2005-10-19
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

European grapes have poor disease resistance, and American grape varieties and their varieties have strong disease resistance, but the "fox" flavor of the fruit and its dominant genetic characteristics limit its further application; the hybridization efficiency of Muscadine grapes and European grapes is low

Method used

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  • Black spot disease resisting candidate gene sequence of dowing grape as one of China vitis wild species and use
  • Black spot disease resisting candidate gene sequence of dowing grape as one of China vitis wild species and use
  • Black spot disease resisting candidate gene sequence of dowing grape as one of China vitis wild species and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The Chinese wild grape grapes 83-4-96 and Shang-24, which were preserved in the grape germplasm resources garden of Northwest Agriculture and Forestry University, have been identified for their resistance to black pox (He Puchao et al., Chinese Agricultural Sciences, 1991; Wang Y et al, HortScience, 1998; Wang Y et al, Vitis, 1995) and 64 interspecific hybrid progenies of European grape pink rose and longan and 176 self-crossed progenies of disease-resistant hybrids were used as test materials. Genomic DNA was extracted by the improved CTAB method. According to the obtained Design specific primers for anti-black pox candidate gene sequences, carry out PCR amplification, according to the performance of specific DNA fragments in hybrid materials (Fig. 1, swimming lane 5-23; Fig. 28), contrast the anti-black pox field As a result of the identification, a DNA fragment closely linked with the anti-black pox gene of the grapevine and a DNA fragment of the anti-black pox gene w...

Embodiment 2

[0046] The wild species of Vitis vinifera Nanzheng-1, Taishan-12, 83-4-96 preserved in the grape germplasm resource nursery of Northwest Agriculture and Forestry University (He Puchao et al., Chinese Agricultural Sciences, 1991; Wang Y et al, HortScience , 1998; Wang Y et al, Vitis, 1995) were crossed with European grape varieties Seedless White, Royal Autumn, Kyoho, etc. to obtain hybrid plants resistant to black pox. The above-mentioned grape material genomic DNA was extracted by the improved CTAB method, primers were designed according to the DNA fragment closely linked with the anti-black pox gene obtained in Example 1, and the above-mentioned grape material genomic DNA was amplified by PCR. The hybrid materials of the fragments are black pox-resistant hybrid plants (Figure 29), or carriers of the black pox-resistant gene, which can be used as materials and breeding germplasm for further breeding of disease-resistant varieties.

Embodiment 3

[0048] Field natural identification and field inoculation identification were used to identify the resistance to black pox of the grape hybrid materials in Example 2, and to obtain disease-resistant hybrids and susceptible hybrids. According to the primers designed by the DNA fragment closely linked with the black disease resistance gene of the grapevine obtained in Example 2, the above-mentioned grape hybrid material is carried out to PCR amplification, according to the amplification and separation of the DNA fragment in the hybrid (Fig. 1, swimming lane 5 -23; Fig. 28; Fig. 29), combined with the field identification results, using MAPMAKER software to locate and map the anti-black pox gene, and obtain the linkage map of the disease-resistant gene in the grapevine, for further identification of the anti-black pox gene by map cloning Base.

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Abstract

The present invention relates to conventional grape crossbreeding, molecular marker assisted breeding, and gene location and mapping method and technology. By means of resistant geneanalogus (RGAs), the present invention obtains 26 black spot disease resisting candidate gene DNA segments of Chinese V. quinquangularis Read and these segments are cloned and sequenced and have size of 492-550 bp. Black spot disease resisting marker assisted breeding is performed based on these sequences as the probes for detecting black spot disease resisting gene. Black spot disease resisting gene is located and mapped based on the distribution of these segments in the hybrid progeny of Chinese V. quinquangularis Read and European V. Or, specific primer is designed, and the technology of rapid amplification of cDNA ends (RACE) is adopted in cloning the black spot disease resisting gene of Chinese V. quinquangularis Read, so as to improve grape variety in gene engineering technology.

Description

technical field [0001] The invention relates to methods and technologies for conventional hybrid breeding of grape grapes, molecular marker-assisted breeding, gene positioning and mapping, and gene cloning, and in particular to methods and technologies for anti-black pox candidate gene sequences and their applications. Background technique [0002] Grape black pox (Elsinoe ampelina (de bary) shear) is native to Europe and North Africa, and is one of the important fungal diseases that harm European grapes. At present, the prevention and treatment of black pox is mainly based on chemical control. However, chemical control not only increases production costs, causes environmental pollution, but also may cause variation of grape disease physiological races and drug resistance of pathogenic bacteria, and at the same time reduce grapes and other diseases. Yield and quality of processed products (wine, raisins, etc.). A large number of scientific research and production practices ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/65C12N15/82C12Q1/68
Inventor 王跃进王西平
Owner NORTHWEST A & F UNIV
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