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SNP molecular marker for detecting rice brown planthopper resistance gene Bph14 and application thereof

A molecular marker and anti-BPH technology, applied in the field of molecular biology, can solve the problems of environmental pollution, human harm, low polymorphism rate, etc., and achieve the effect of environmental friendliness, easy operation and strong specificity

Active Publication Date: 2018-03-20
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 for detecting rice brown planthopper resistance gene Bph14 and application thereof
  • SNP molecular marker for detecting rice brown planthopper resistance gene Bph14 and application thereof
  • SNP molecular marker for detecting rice brown planthopper resistance gene Bph14 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Development of SNP Molecular Marker of Rice Bph14 Gene

[0041] According to the location information of the gene, the brown planthopper resistance gene Bph14 gene region is located in the region of rice chromosome 3 35693286-35699010 (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 Bph14 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 18 rice varieties containing Bph14 gene donor materials RH, IR56 and other 18 rice varieties without Bph14, and the SNP marker K_030513 with good co-segregation and amplification effect with the resistant donor material was selected. For the above SNP marker development flow char...

Embodiment 2

[0059] Example 2 Application of SNP Molecular Marker of Rice Bph14 Gene

[0060] For the molecular marker design of the present invention, the KASP reaction can be used to detect the Bph14 gene resistance to brown planthopper in rice materials in high throughput, and the primer combination designed in Example 1 is used. SNP marker K_030513 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, except for the heterozygous Bph14 genotype detected in Xieyou 716, other susceptible control materials, common hybrid rice and core rice breeding materials were detected as homozygous Bph14 genotype without BPH resistance. It can be seen that the SNP marker K_030513 detects the Bph14 gene locus as a highly specific resistance locus, which can be conveniently and efficiently used to identify whether the rice var...

Embodiment 3

[0061] Example 3 Genetic location of SNP markers and phenotypic verification of segregated populations

[0062] 1. Marker genetic position verification

[0063] Utilize the RIL population of 79 strains of Bph14 and 11 SNP markers with polymorphisms in the parents and the Bph14-specific marker K_030513 developed in Example 1 of the present invention for genetic position verification, and the marker K_030513 of the present invention is mapped to rice The 79.2cM position of chromosome 3 (see image 3 ).

[0064] 2. Marker phenotype verification

[0065] The genetic phenotype of the marker K_030513 linked to the gene Bph14 in the present invention was verified by using the RIL population of B5 and Mh63. The phenotype identification was entrusted to Wuhan Hetaiqing Biotechnology Co., Ltd., 83.3% of the 12 strains containing Bph14 were above the level of insect resistance, and 83.3% of the 12 strains without Bph14 were susceptible to insects. type (table 4), the phenotype data a...

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Abstract

The invention provides a SNP molecular marker for detecting rice brown planthopper resistance gene Bph14 and application thereof. SNP marker K_030513 having a good rice brown planthopper resistance gene Bph14 co-segregating and amplification effect is disclosed. The marker detects the base at the 35692898 site (MSU7.0) of No. 3 chromosome of rice, the polymorphism is C / G, and the primer sequence of the marker K_030513 developed based on KASP technology is shown as SEQ ID No. 1-3. The SNP molecular marker can be used for detecting a genetic locus with high specificity of the Bph14 gene, and canbe conveniently and efficiently used to identify whether the Bph14 gene is contained in a rice variety. An application method of the SNP molecular marker provided by the present invention is accurate, reliable, and simple in operation, and is applicable to the identification of the Bph14 gene and assisted selective breeding.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a SNP molecular marker of the rice brown planthopper resistance gene Bph14 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. ...

Claims

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Application Information

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Patent Type & Authority Applications(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