Molecular marker for identifying resistance traits of Phyricularia oryzae, identification method and application

A rice blast and molecular marker technology, applied in the field of molecular biology, can solve the problems of cumbersome operation, age-related influence of phenotype identification, dependence on detection of resistant alleles, etc., so as to improve detection efficiency and simple identification operation method. , low cost effect

Inactive Publication Date: 2018-05-11
LIAONING PROVINCIAL SALINE ALKALI LAND UTILIZATION & RES INST
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the above technical problems, the present invention provides a molecular marker, identification method and application for identification of rice blast resistance traits, which overcomes the defects that phenotypic identification is greatly affected by age and complicated operation, and also overcomes the current Pib Detection of resistant and sensitive alleles of genes relies on sequencing or the defects of multiple PCR detection

Method used

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  • Molecular marker for identifying resistance traits of Phyricularia oryzae, identification method and application
  • Molecular marker for identifying resistance traits of Phyricularia oryzae, identification method and application
  • Molecular marker for identifying resistance traits of Phyricularia oryzae, identification method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Using the above-mentioned molecular marker Pib-TPAP to identify the rice blast fungus resistance traits of the tested rice materials, specifically includes the following steps:

[0033] The test materials are the rice variety Nipponbare and the rice variety Wuyunjing 27, which are the control materials in the Pib gene cloning experiment, in which Nipponbare carries the rice blast allele type Pib, and Wuyunjing 27 carries the rice blast resistance allele Type Pib.

[0034] S1, Genomic DNA was extracted from two materials, Nipponbare and Wuyunjing 27, respectively, and the genomic DNA was extracted using the Quanjin Plant DNA Extraction Kit (operate according to the kit instructions).

[0035] S2, use the genomic DNA obtained in S1 as a template, and use Pib-TPAP-O-F, Pib-TPAP-O-R, Pib-TPAP-I-F, and Pib-TPAP-I-R as primers for PCR amplification.

[0036] The PCR amplification reaction system is 20 μL: including DNA 1 μL (1ng / μL); Pib-TPAP-O-F 0.5 μL, Pib-TPAP-O-R 0.5 μL, P...

Embodiment 2

[0043] According to the research of Liu Yang et al. (Liu Yang. Molecular marker-assisted selection and application of rice blast resistance PIB gene [D]. Sichuan Agricultural University, 2007.), the Pib resistance allele has significant resistance to blast fungus physiological race ZB13. sex. Example 2 The DH population produced by hybridization of resistant-sensitivity allelic varieties was used as the test material, and the accuracy and practicability of the invention were verified by inoculating ZB13 blast fungus physiological race and comparing the genotype and phenotype.

[0044] The DH population consists of 80 stable lines obtained from rice blast-susceptible allele type Nipponbare (male parent) hybrid rice blast-resistant allele type rice variety Wuyunjing No. 27 (female parent). The Pib allele types of 80 stable lines were identified by marking Pib-TPAP, and compared with the two-year inoculation phenotype in 2013-2014. The results are shown in Table 2. The results in...

Embodiment 3

[0048] Example 3 Using the molecular marker Pib-TPAP to detect the genotype of the offspring of the RIL population

[0049] According to the research of Liu Yang et al. (Liu Yang. Molecular marker-assisted selection and application of rice blast resistance PIB gene [D]. Sichuan Agricultural University, 2007), the Pib resistance allele has significant resistance to the rice blast fungus physiological race ZB13 . Example 3 The RIL population produced by hybridization of varieties with resistant-sensitivity alleles was used as the test material, and the accuracy and practicability of the invention were verified by inoculating ZB13 rice blast fungus physiological race and comparing the genotype and phenotype.

[0050] The RIL population is the susceptible allele type Koshihikari (male parent) crossed with the disease-resistant allele type Wuyunjing 27 (female parent), 160 stable lines obtained by single-seed transmission method, through the marker Pib -TPAP identified the Pib all...

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Abstract

The invention belongs to the technical field of molecular biology and discloses a molecular marker for identifying resistance traits of Phyricularia oryzae, an identification method and an application. The molecular marker is Pib-TPAP and contains Pib-TPAP-O-F with a nucleotide sequence represented as SEQ ID NO.1, Pib-TPAP-O-R with a nucleotide sequence represented as SEQ ID NO.2, Pib-TPAP-I-F with a nucleotide sequence represented as SEQ ID NO.3 and Pib-TPAP-I-R with a nucleotide sequence represented as SEQ ID NO.4. The molecular marker can be used for directly identifying two allelotypes ofMagnaporthe oryzae resistance of a Pib gene, on the basis of a PCR (polymerase chain reaction) technology, the two disease-resistant allelotypes can be accurately identified by one-time PCR without sequencing or enzyme digestion, so that detection efficiency of a genetic locus is greatly improved, and accuracy rate reaches 100%.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and specifically relates to a molecular marker for identifying rice blast resistance traits, an identification method and an application. Background technique [0002] Rice blast caused by Magnaporthe oryzae is one of the important diseases of rice, which can cause a large reduction in rice yield, and in severe cases cause grain failure, seriously affecting the food safety production in my country. Historical practice has proved that the selection of blast-resistant rice varieties is the most effective solution to reduce the damage of rice blast. Since the identification of rice blast resistance is cumbersome and greatly affected by age, molecular marker-assisted selection of rice blast-resistant varieties can greatly improve the selection efficiency without being affected by the external natural environment. best means. [0003] At present, 24 rice blast resistance genes have been su...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/13C12Q2600/156C12Q2600/172C12Q2531/113
Inventor 毛艇张丽丽刘妍赵一洲李鑫张战李旭倪善军付雪娇黄河于小彭荆华宋双李振宇刘福才
Owner LIAONING PROVINCIAL SALINE ALKALI LAND UTILIZATION & RES INST
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