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Gene marker of major gene Bph14 for resisting brown planthopper in rice and application thereof

A technology of genetic markers and resistance to brown planthoppers, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems affecting selection efficiency and accuracy, and achieve reliable results, cost savings, and experimental repeatability Good results

Inactive Publication Date: 2014-01-15
江西省农业科学院水稻研究所
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Problems solved by technology

[0005] So far, 26 main genes for resistance to BPH have been reported in rice, among which Bph14 has been successfully cloned. At present, in molecular marker-assisted selection breeding, the selection of this gene mainly uses markers linked to it, such as MRG2329, MRG2346, MRG2684, R1925 and R2443, etc., but gene markers have not been reported, which to some extent affects the selection efficiency and accuracy of using Bph14 in the breeding process for resistance to brown planthoppers

Method used

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  • Gene marker of major gene Bph14 for resisting brown planthopper in rice and application thereof
  • Gene marker of major gene Bph14 for resisting brown planthopper in rice and application thereof
  • Gene marker of major gene Bph14 for resisting brown planthopper in rice and application thereof

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Embodiment Construction

[0022] 1. Test materials

[0023]

[0024] 2. DNA extraction

[0025] Get rice leaf at seedling stage, the extraction of genomic DNA is with reference to the CTAB method of Scott et al. (1988), improves as follows:

[0026] 1) Take about 0.1g leaves, put them in a 2.0mL centrifuge tube, add 1 steel ball and 500μL 2×CTAB DNA extract (20g CTAB, 12.1g Tris-Base, 7.44g EDTA-Na, 81.9g NaCl, dissolved in 1L distilled water Middle), use a tissue grinder to crush the leaves, bathe in water at 65°C for 30 minutes, and vibrate and mix several times during the period;

[0027] 2) Take out the centrifuge tube from the water bath, add about 500 μL of chloroform (CHCl 3 ), fully shake and mix, and centrifuge at 10000rpm for 5min;

[0028] 3) Transfer the supernatant to a new 1.5mL centrifuge tube, add isopropanol 0.7 to 1 times the volume of the supernatant, mix the centrifuge tube slightly and place it in a -20°C refrigerator for half an hour, then centrifuge at 10,000rpm 5min;

[...

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Abstract

The invention relates to a gene marker of a major gene Bph14 for resisting brown planthopper in rice and application thereof. The gene marker Bph14-M1 of Bph14 has the following sequences of forward and reverse primers: a forward primer of (5'-3') AGCCACTTGGTGAACTTATT, and a reverse primer of (5'-3') GATTGACGATGAGGAGACTT. The application method includes 1) extracting rice genomic DNA; carrying out PCR amplification on the rice genomic DNA by using the forward and reverse primers; and carrying out electrophoresis detection on amplification products. The invention employs the marker to assist selection and can accurately transform the brown planthopper resistant gene Bph14 in B5 to a brown planthopper susceptible cultivar and greatly improve the breeding efficiency of brown planthopper resistant varieties.

Description

technical field [0001] The invention relates to a gene marker of a rice variety and an application thereof, in particular to a gene marker of the main gene Bph14 for resistance to brown planthopper in rice and an application thereof. Background technique [0002] The brown planthopper is one of the most serious pests that harm rice production. In the past decade, the brown planthopper has broken out year after year in my country, causing an average annual loss of 2.77 million tons of rice production. In 2010, the Ministry of Agriculture’s Weekly Report on the Prevention and Control of Major Crop Diseases and Insects (No. 23) reported that the cumulative occurrence of brown planthoppers in my country in that year was 302 million mu, which seriously threatened the high and stable yield of rice. It is the most economical, effective and environment-friendly way to control the damage of brown planthopper by using the brown planthopper-resistant gene to breed brown planthopper-res...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 沈显华严松黄仁良沈林军熊宏亮朱珊
Owner 江西省农业科学院水稻研究所
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