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Loofah brown spot-resistant gene segments or gene markers and application

A gene fragment and gene marker technology, applied in the field of plant biotechnology science, can solve problems such as economic loss, affecting the growth and development process of loofah plants, and achieve the effects of shortening breeding years, improving breeding efficiency, and improving identification efficiency.

Inactive Publication Date: 2015-04-29
南农大(常熟)新农村发展研究院有限公司
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Problems solved by technology

Brown spot disease has seriously affected the normal growth and development process of loofah plants, bringing great economic losses to production

Method used

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  • Loofah brown spot-resistant gene segments or gene markers and application
  • Loofah brown spot-resistant gene segments or gene markers and application
  • Loofah brown spot-resistant gene segments or gene markers and application

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Embodiment Construction

[0017] 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 the brown spot resistance gene of loofah. The specific implementation process is as follows:

[0018] 1. DNA extraction: Take an appropriate amount of young leaves of loofah, 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.

[0019] 2. Amplification and cloning of the homologous fragment of the disease resistance gene: according to the amino acid sequence GG(L / I / V / M / S)GKTT and G(L / N) of the conserved region of the known NBS-LRR type disease resistance gene ) PL (A / T / S) (L / V), designed a pair of degenerate primers: NBS15'-GKTT GGIGGIWSIGGIAARA...

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Abstract

The invention mainly relates to loofah brown spot-resistant gene segments or gene markers and an application. Primarily, 3 loofah brown spot-resistant gene segments or gene markers are quickly separated by a resistance gene homologous sequence method; and according to sequencing, the nucleotide length of the gene segments or gene markers is 498bp, 501bp and 497bp respectively. According to the invention, the discovery cycle of the loofah brown spot-resistant gene is effectively shortened, and rapid identification of the brown spot gene is facilitated; corresponding co-segregation functional markers (SNP, SCAR and the like) are developed by use of the identified brown spot gene, and the brown spot gene also can be quickly applied to the marker-assisted selection of the brown spot-resistant gene while the accuracy is high; in combination with other molecular markers on disease-resistant gene, multiple resistant breeding materials can be created, the breeding period is shortened, and the breeding efficiency is improved; and a foundation is laid for the elaboration of the molecular mechanism of the loofah brown spot-resistant gene.

Description

technical field [0001] The invention utilizes the homologous sequence method of disease-resistant genes to clone loofah anti-brown spot disease gene fragments or convert them into molecular markers, etc., and mainly relates to conventional hybrid breeding of loofahs, molecular marker-assisted breeding, positioning and molecular markers of loofah-resistant brown spot genes. Mapping, cloning of anti-brown spot gene and analysis of regulation mechanism of disease-resistant gene belong to the field of plant biotechnology science. Background technique [0002] Brown spot is one of the most important diseases in loofah production. Mainly damages leaves. Lesions are brown to taupe, round or long to irregular, with a diameter of 0.5-13 mm. The edge of the lesion is obvious or not obvious, sometimes there is a chlorotic to yellow halo, mildew is rare. At sunrise in the morning or sunset in the evening, silver-gray luster can be seen on the lesions, which is caused by the reflectio...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 钱孝英钱春桃
Owner 南农大(常熟)新农村发展研究院有限公司