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Rice Blast Resistance Gene pi36 Co-dominant Molecular Marker and Its Application

A rice blast resistance gene and molecular marker technology, applied in biochemical equipment and methods, DNA/RNA fragments, microbial determination/inspection, etc., can solve the problems of cumbersome resistance identification, many manpower and costs, and inaccurate results , achieve the effect of shortening the breeding cycle, low cost and simple operation

Active Publication Date: 2022-03-22
YUAN LONGPING HIGH TECH AGRI CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In conventional disease resistance breeding, the identification of resistance is cumbersome, requires a lot of manpower and cost, and the identification is easily affected by external conditions, resulting in inaccurate results, so the breeding efficiency is low

Method used

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  • Rice Blast Resistance Gene pi36 Co-dominant Molecular Marker and Its Application
  • Rice Blast Resistance Gene pi36 Co-dominant Molecular Marker and Its Application
  • Rice Blast Resistance Gene pi36 Co-dominant Molecular Marker and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Development of Pi36 Gene Molecular Marker

[0032] The rice blast resistance gene Pi36 is located on the short arm of rice chromosome 8. Through sequencing the genome region of Pi36 of rice varieties known to be resistant to or susceptible to blast disease such as Kasalath, 9311, and Nipponbare, and comparing and analyzing the NCBI database online, two specific genes were found. Sexual SNP loci, wherein SNP1 is located at base 2886707 on chromosome 8 of the Nipponbare sequence, the resistant allele of Pi36 is G, and the susceptible allele is A (such as figure 1 Shown in A); SNP2 is located at chromosome 2888118 of No. 8 Nipponbare sequence, the resistant allele of Pi36 is G, and the susceptible allele is A (such as figure 1 shown in B). Two pairs of dominant markers were developed for these two SNP loci, which were used to identify disease-resistant alleles and susceptible alleles respectively. The combination of two pairs of dominant markers can realize the ...

Embodiment 2

[0033] Example 2 Design of detection primers for co-dominant molecular markers of Pi36 gene

[0034] Detection primers were designed for the two Pi36 gene-specific molecular markers developed in Example 1, and the principle of primer design was as follows.

[0035] First, design specific primers for amplifying the disease-resistant allele, and use the complementary base C of the disease-resistant allele G base at the SNP1 site as the 3' end of the reverse primer to design the primer Pi36-RR, and at the 3' end At the third base, the C base is changed to an A base to introduce a mismatch, and then a matching forward primer Pi36-RF is designed upstream of it, and the primers Pi36-RR and Pi36-RF (SEQ IDNO: 3-4 ) can only specifically amplify the Pi36 disease-resistant allele and produce a 345bp band, while the Pi36 susceptible allele has no band when amplified;

[0036] Then, design specific primers for amplifying the susceptible allele, design the forward primer Pi36-SF with the...

Embodiment 3

[0040] Example 3 Establishment of rice blast resistance gene Pi36-specific molecular marker detection method

[0041] According to the detection primers of the two molecular markers of Pi36 designed in Example 2, the reaction program and reaction system of PCR were designed, and through continuous optimization, the following reaction program and system were determined:

[0042] PCR reaction system (10μL): 1ul 10×PCR reaction buffer, 0.8ul 10mM dNTP, 0.15μL for all 4 primers (10μM), 0.1ul Taq DNA polymerase (2.5U / ul), 2ul DNA template, ddH 2 O supplemented to 10ul.

[0043] The PCR reaction conditions were: pre-denaturation at 95°C for 5 minutes; denaturation at 95°C for 30 seconds, annealing at 56°C for 30 seconds, extension at 72°C for 45 seconds, a total of 32 cycles; extension at 72°C for 8 minutes.

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Abstract

The present invention provides co-dominant molecular markers of rice blast resistance gene Pi36, including marker SNP1 and / or marker SNP2, wherein the polymorphic site contained in marker SNP1 is located at base 2886707 corresponding to rice Nipponbare No. 8 chromosome. The base is G or A; the polymorphic site contained in the marker SNP2 is located at the base corresponding to rice Nipponbare No. 8 chromosome 2888118, and the base is G or A. Specific amplification primers were designed for the molecular markers, and the Pi36 genotype was detected by PCR amplification. The molecular marker of the present invention is co-dominant, and can effectively distinguish three different genotypes. The molecular marker can be used for breeding rice blast-resistant rice, shortens the breeding cycle of rice blast-resistant rice, accelerates the breeding speed, and reduces the breeding cost. Simple operation, low cost, short cycle and so on.

Description

technical field [0001] The invention belongs to the fields of molecular biology and plant molecular breeding, and in particular relates to a co-dominant molecular marker of rice blast resistance gene Pi36 and its application. Background technique [0002] Rice, as the main food crop in my country, is threatened by pests and diseases all the year round, among which rice blast is one of the most serious diseases. In various rice producing areas in China, rice blast occurs every year. When the disease and insects are prevalent, the yield can be reduced by 10% to 20%, and in severe areas it can reach 40% to 50%, or even no harvest. Extensive use of rice blast-resistant agents can alleviate the harm of rice blast, but it also greatly increases the cost of rice production and pollutes the environment. Practice has shown that breeding rice blast-resistant varieties is the most economical and effective measure to control rice blast. However, due to the large number of rice blast ph...

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 杨远柱邓钊刘兰兰王凯秦鹏符辰建严天泽周延彪
Owner YUAN LONGPING HIGH TECH AGRI CO LTD
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