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

A rice blast and resistance gene technology, applied in the field of PARMS markers and applications, can solve the problems of inapplicability to large-scale genotype analysis, low degree of automation, use of toxic substances, etc., and achieve reliable preparation of detection results, easy to grasp, error-prone effect

Pending Publication Date: 2021-05-18
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

[0006] Aiming at the disadvantages of marker detection for preventing and controlling rice blast in the prior art, such as time-consuming, cumbersome operation, low efficiency, low degree of automation, use of toxic substances, and unsuitability for large-scale genotype analysis, the present invention provides a method based on rice The PARMS marker and application of the SNP mutation in the blast resistance gene Pita coding region. The present invention uses next-generation sequencing to detect the difference of Pita between the high blast resistance material Y244 and the susceptible variety Nipponbare, and finds that G→T occurs at the 17bp site in the gene coding region mutation

Method used

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

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Embodiment 1

[0035] A PARMS marker and application based on the SNP mutation of the rice blast resistance gene Pita coding region, comprising the following steps:

[0036] 1) Identification of blast resistance of rice materials: using Nipponbare as a control, 226 rice materials were artificially inoculated at the seedling stage for identification of blast resistance, and the material Y224 resistant to blast disease races ZB1 and ZB9 was screened out;

[0037] 2) Analysis of Pita gene structure: The DNA of fresh leaves of Y224 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 differences of Pita genes in the two rice materials, and found A G→T mutation occurred at the 17bp site in the coding region of the gene (see attached figure 1 );

[0038] 3) Design and synthesis of Pita gene functional markers: According to the G→A mutation at position 555 of the Pita...

Embodiment 2

[0059] A PARMS marker and application based on the SNP mutation of the rice blast resistance gene Pita coding region, comprising the following steps:

[0060] 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 Y224 resistant to blast disease races ZB1 and ZB9 was screened out;

[0061] 2) Analysis of Pita gene structure: The DNA of fresh leaves of Y224 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 differences of Pita genes in the two rice materials, and found A G→T mutation occurred at the 17bp site in the coding region of the gene (see attached figure 1 );

[0062] 3) Design and synthesis of Pita gene functional markers: According to the G→A mutation at position 555 of the...

Embodiment 3

[0075] A PARMS marker and application based on the SNP mutation of the rice blast resistance gene Pita coding region, comprising the following steps:

[0076] 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 Y224 resistant to blast disease physiological races ZB1 and ZB9 was screened out;

[0077] 2) Analysis of Pita gene structure: The DNA of fresh leaves of Y224 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 differences of Pita genes in the two rice materials, and found A G→T mutation occurred at the 17bp site in the coding region of the gene (see attached figure 1 );

[0078] 3) Design and synthesis of Pita gene functional markers: According to the G→A mutation at posit...

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Abstract

The invention discloses a PARMS marker based on SNP mutation in a coding region of rice blast resistance gene Pita and an application. The difference of the Pita in a high-resistance rice blast material Y244 and a susceptible variety Nipponbare is detected by utilizing first-generation sequencing, and G-T mutation occurs at a 17bp site of the gene coding region. The difference of two basic groups is introduced into a designed forward primer, and two different fluorescent labeling universal primers are added on the basis, so that the fluorescent functional molecular marker of the rice blast resistance gene Pita based on the PARMS technology is developed, and the application of a Pita gene in 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 Pita 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
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2600/13
Inventor 韦宇粟学俊陈彩虹杨行海阎勇李冬秀梁曼玲
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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