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PARMS marker based on SNP mutation of coding region of rice blast resistance gene Pid2 and application

A rice blast and resistance gene technology, applied in the field of PARMS marker and application, can solve the problems of inappropriate large-scale genotype analysis, use of toxic substances, low degree of automation, etc. Easy to grasp effect

Pending Publication Date: 2021-04-30
GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Aiming at the shortcomings in the prior art that the detection of markers for preventing and controlling rice blast is time-consuming, cumbersome to operate, low in efficiency, low in automation, uses toxic substances, and is not suitable for large-scale genotype analysis, the present invention provides a method based on rice blast resistance. The PARMS marker and application of the SNP mutation in the coding region of the sex gene Pid2. The present invention uses the first-generation sequencing to detect the difference of Pid2 in the highly resistant rice blast material Y127 and the susceptible variety Nipponbare, and finds that an A→G mutation occurs at the 554bp site in the coding region of the gene. Introduce the difference between these two bases into the designed forward primer, and add two different fluorescent marker universal primers on this basis, and develop the fluorescent functional molecular marker of rice blast resistance gene Pid2 based on PARMS technology, which is convenient for Pid2 Application of Genes in Molecular Breeding of Rice Resistance

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  • PARMS marker based on SNP mutation of coding region of rice blast resistance gene Pid2 and application
  • PARMS marker based on SNP mutation of coding region of rice blast resistance gene Pid2 and application

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Effect test

Embodiment 1

[0036] The PARMS marker and application based on the SNP mutation in the rice blast resistance gene Pid2 coding region include the following steps:

[0037] 1) Identification of blast resistance of rice materials: Using Nipponbare as a control, 226 rice materials were artificially inoculated at the seedling stage to identify the resistance to blast disease, and the material Y127 resistant to blast disease races ZA9 and ZA13 was screened out;

[0038] 2) Analysis of Pid2 gene structure: The DNA of fresh leaves of Y127 and Nipponbare was extracted by CTAB method, amplified in segments, purified and then sent to the biological company for sequencing. The software Vector NTI 11 was used to analyze the difference of Pid2 gene in the two rice materials. A→G mutation occurred at the 555bp site in the coding region of the gene (see attached figure 1 );

[0039] 3) Design and synthesis of Pid2 gene functional markers: According to the G→A mutation at position 555 of the Pid2 coding r...

Embodiment 2

[0060] The PARMS marker and application based on the SNP mutation in the rice blast resistance gene Pid2 coding region include the following steps:

[0061] 1) Identification of blast resistance of rice materials: using Nipponbare as a control, 200 rice materials were artificially inoculated at the seedling stage to identify the resistance to blast disease, and the material Y127 resistant to blast disease races ZA9 and ZA13 was screened out;

[0062] 2) Analysis of Pid2 gene structure: The DNA of fresh leaves of Y127 and Nipponbare was extracted by CTAB method, amplified in segments, purified and then sent to the biological company for sequencing. The software Vector NTI 11 was used to analyze the difference of Pid2 gene in the two rice materials. An A→G mutation occurred at the 555bp site in the coding region of the gene;

[0063] 3) Design and synthesis of Pid2 gene functional markers: According to the G→A mutation at position 555 of the Pid2 coding region, a PARMS molecular...

Embodiment 3

[0076] The PARMS marker and application based on the SNP mutation in the rice blast resistance gene Pid2 coding region include the following steps:

[0077] 1) Identification of blast resistance of rice materials: Using Nipponbare as a control, 250 rice materials were artificially inoculated at the seedling stage to identify the resistance to blast disease, and the material Y127 resistant to blast disease races ZA9 and ZA13 was screened out;

[0078] 2) Analysis of Pid2 gene structure: The DNA of fresh leaves of Y127 and Nipponbare was extracted by CTAB method, amplified in segments, purified and then sent to the biological company for sequencing. The software Vector NTI 11 was used to analyze the difference of Pid2 gene in the two rice materials. A→G mutation occurs at the 555bp site in the coding region of the gene;

[0079] 3) Design and synthesis of Pid2 gene functional markers: According to the G→A mutation at position 555 of the Pid2 coding region, a PARMS molecular mark...

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Abstract

The invention discloses a PARMS marker based on SNP mutation of a coding region of a rice blast resistance gene Pid2 and an application. According to the invention, the difference of Pid2 in a high rice blast resistance material Y127 and an infected variety Nipponbare is detected by using first-generation sequencing, A->G mutation occurs at a 554bp site of the coding region of the gene, the difference of the two bases is introduced into a designed forward primer, and on the basis, two different fluorescence labeling universal primers are added, so that a fluorescence functional molecular marker of the rice blast resistance gene Pid2 based on the PARMS technology is developed, and application of the Pid2 gene to rice resistance molecular breeding is facilitated.

Description

【Technical field】 [0001] The invention belongs to the technical field of genetic engineering, and relates to a PARMS marker based on the SNP mutation of the rice blast resistance gene Pid2 coding region and its application. 【Background technique】 [0002] Rice blast is a devastating fungal disease of rice caused by Magnaporthe oryzae, which seriously affects the yield and quality of rice planting areas in the world, thus threatening global food security. Rice is one of the main food crops that human beings depend on for survival. It is Staple food for more than half of the world's population. Rice blast is one of the most devastating fungal diseases in rice production in the world. So far, rice blast has been reported in more than 80 countries, among which Asia and Africa are the most serious. It is estimated that enough rice is lost to rice blast to feed 60 million people each year. [0003] As a main and traditional method of pest control, chemical control will not only ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2600/172C12Q2531/113C12Q2535/101C12Q2563/107C12Q2545/113
Inventor 阎勇陈彩虹粟学俊杨行海韦宇李冬秀梁曼玲
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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