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SNP Molecular Marker and Application for Detecting Rice BPH3 Gene Resistance to Brown Planthopper

A molecular marker and rice technology, applied in the field of molecular biology, can solve problems such as human harm, environmental pollution, and small differences

Active Publication Date: 2021-07-06
HUAZHI RICE BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main types of markers used in literature reports are SSR and InDel markers, which have the disadvantages of low polymorphism rate and small differences in breeding
The EB or polyacrylamide used in the detection process is easy to pollute the environment and cause harm to the human body

Method used

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  • SNP Molecular Marker and Application for Detecting Rice BPH3 Gene Resistance to Brown Planthopper
  • SNP Molecular Marker and Application for Detecting Rice BPH3 Gene Resistance to Brown Planthopper
  • SNP Molecular Marker and Application for Detecting Rice BPH3 Gene Resistance to Brown Planthopper

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The development of the SNP molecular marker of embodiment 1 rice Bph3 gene

[0042] According to the location information of the gene, the brown planthopper resistance gene Bph3 is located in the region of rice chromosome 4 6939046-6969039 (MSU7.0). Take the gene interval as the center and expand 50kb to both sides to extract the SNP site, and select according to the PIC value and whether there are other SNP sites in the 50bp around the SNP site. At the same time, according to the comparison between the cloned Bph3 gene sequence and the Nipponbare sequence, select the SNP site, and use BatchPrimer3 to design primers for it. For these SNP markers, KASP reaction verification was performed on the Bph3 gene donor materials RH, IR56 and other rice varieties without Bph3, and the SNP marker K_040503 with good co-segregation and amplification effect with the resistant donor material was selected. For the above SNP marker development flow chart, see figure 1 .

[0043] The pri...

Embodiment 2

[0060] The application of the SNP molecular marker of embodiment 2 rice Bph3 gene

[0061] For the molecular marker design of the present invention, the KASP reaction can be used for high-throughput detection of Bph3 gene resistance to brown planthopper in rice materials, and the primer combination designed in Example 1 is used. The SNP marker K_040503 was verified by natural population using 95 materials. The 95 accessions included susceptible control materials, common hybrid rice and core rice breeding materials. Marked in natural group typing results such as figure 2 As shown, in addition to Xieyou 716, which detected the heterozygous Bph3 genotype, other susceptible control materials, common hybrid rice and core rice breeding materials were detected as homozygous bph3 genotype without BPH resistance. It can be seen that the SNP marker K_040503 detection of the Bph3 gene locus is a highly specific resistance locus, which can be conveniently and efficiently used to identi...

Embodiment 3

[0062] Example 3 Genetic location of SNP markers and phenotypic verification of segregated populations

[0063] 1. Marker genetic position verification

[0064] Utilize the RIL population of 79 Bph3 strains and 10 SNP markers with polymorphisms in the parents and the Bph3-specific marker K_040503 developed in Example 1 of the present invention for genetic position verification, and the marker K_040503 of the present invention is mapped to rice Chromosome 4 at 0.8cM (see image 3 ).

[0065] 2. Marker phenotype verification

[0066] Using the marker K_040503 to genotype the F2 population of 1327 (Bph3 donor) and C594, compared with the genotype data of the SSR marker, the population material and the SSR marker data came from Longping Hi-Tech, as shown in Table 4, the consistency of the two markers was relatively high. Well, the feasibility and accuracy of the SNP molecular marker detection method developed in the present invention has been verified again, and it can be used ...

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Abstract

The invention provides a SNP molecular marker and application for detecting rice brown planthopper resistance gene Bph3. The invention discloses a SNP marker K_040503 with good co-separation and amplification effect with the rice brown planthopper resistance gene Bph3. The marker detects base 6903106 of rice chromosome 4, and the polymorphism is C / G. The sequence of the marker K_040503 primer developed based on KASP technology is shown in SEQ ID No.1‑3. The SNP molecular marker of the present invention detects the Bph3 gene site as a highly specific site, and can be conveniently and efficiently used to identify whether a rice variety contains the Bph3 gene. The application method of the SNP molecular marker provided by the invention is accurate, reliable, and easy to operate, and is suitable for identification of Bph3 gene and assisted selection breeding.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a SNP molecular marker of rice brown planthopper resistance gene Bph3 and a method for detecting the molecular marker. Background technique [0002] Rice is an important food crop in my country. Brown planthopper is a monophagous pest of rice. It sucks phloem juice through the stylet and damages rice plants, resulting in slow growth, delayed tillering, and increased hollow grains. The brown planthopper is also the carrier of some rice viruses such as grass stunt virus and leaf dwarf virus, which seriously affects the production and safety of rice. At present, the control of BPH mainly relies on chemical control, which not only increases the production cost and the insecticide resistance of pests, but also pollutes the environment. Therefore, the use of host resistance to breed resistant varieties of BPH is considered to be an effective way to control its damage. [00...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 彭佩江南郑秀婷梁毅贺治洲
Owner HUAZHI RICE BIO TECH CO LTD