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A function-specific molecular marker of rice blast resistance gene pik-p and its application

A resistance gene and molecular marker technology, applied in the field of agricultural biology, can solve the problems of molecular marker-assisted selection breeding that is not suitable for large-scale resistance identification, and achieve the effect of low cost, strong operability and low cost

Active Publication Date: 2021-11-09
BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI +1
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  • Application Information

AI Technical Summary

Problems solved by technology

Although Pan Qinghua et al. (2012) developed Pik-p Function-specific molecular markers, but because the K / N genotype needs to be identified before the SNP marker is used, in fact, there is a certain incompatibility in the genotype, and the identification process requires tedious work such as enzyme digestion identification (Zhai et al. 2010), not suitable for large-scale resistance identification and marker-assisted selection (MAS) work

Method used

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  • A function-specific molecular marker of rice blast resistance gene pik-p and its application
  • A function-specific molecular marker of rice blast resistance gene pik-p and its application
  • A function-specific molecular marker of rice blast resistance gene pik-p and its application

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Experimental program
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Effect test

Embodiment 1

[0035] Embodiment 1: rice blast resistance gene Pik-p Function-specific molecular markers and their primer design and detection

[0036] (1) Pik-p Analysis of insertion-deletion (InDel) sites:

[0037] Download the representative varieties of rice from the public database Pik-p genes and their homologues Pik , Pik-m, Pi1 and Pik-h The genome sequence and the genome sequence of the corresponding regions of the sequenced varieties Nipponbare and 9311, for Pik-p site for sequence comparison, screening Pik-p A specific insertion / deletion (InDel) difference site that can be distinguished from the site and other rice blast allele resistance genes. The genome sequences of the sequenced varieties Nipponbare and 9311 have no corresponding regions, and the rice blast resistance gene variety IRBLkp-K60 with gene-specific InDel ( Pik-p ) and the resistant variety IRBLk-Ka ( Pik ), the resistant variety IRBLkm-Ts ( Pik-m ), resistant variety IRBLkh-K3 ( Pik-h ) and the res...

Embodiment 2

[0061] Embodiment 2: resistance gene Pik-p Gene-specific molecular markers in identifying Pik Application of different rice blast resistance genes in gene cluster regions.

[0062] According to the polymorphic PCR product bands, the target gene can be Pik-p with other Pik The resistance genes on the gene cluster were distinguished. Such as figure 1 As shown, depending on the stripe position, the Pik-p Distinguished from other resistance genes identified at this locus, 359bp indicates that it contains Pik-p Gene, 378bp means that it contains the resistance gene locus, and other bands or no bands mean that it does not contain the functional gene of the locus. It can be seen that the test results are consistent with the design analysis, indicating that the resistance gene Pik-p Gene-specific molecular markers can be identified in Pik-p gene and Pik Gene family alleles and other aspects have been applied.

Embodiment 3

[0063] Embodiment 3: resistance gene Pik-p Detection application of gene-specific molecular markers in other rice varieties

[0064] Table 1 Correspondence figure 2 72 representative varieties in

[0065]

[0066] Genomic DNA was extracted respectively, and using this as a template, PCR amplification and electrophoresis detection were carried out according to the method of Example 1. According to the size of the product (band), the resistance gene can be Pik-p Distinguished from other rice blast resistance genes. Such as figure 2 As shown, the resistance spectrum phenotype is Pik-p Type individuals (carrying rice blast resistance gene Pik-p The specific fragment of Pik-p-InDel gene can be detected in the samples of lane 11, 16 and 62 of the disease-resistant variety IRBLkp-K60, but the molecular marker cannot be detected in individuals with other spectrum resistance phenotypes. It can be seen that the test results are consistent with the design analysis, indicating...

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Abstract

The invention provides a rice blast resistance gene Pik‑p Function-specific molecular markers and their applications, amplified from rice genomic DNA by primer pair SEQ ID NO.1 and SEQ ID NO.2 and rice blast resistance gene Pik‑p Molecular markers with specific band patterns. Rice blast resistance gene provided by the present invention Pik‑p Function-specific molecular markers have important application value. The use of this marker can improve the efficiency of the gene in germplasm resource screening, molecular marker-assisted selection breeding, gene aggregation breeding, and transgenic breeding.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology and relates to a rice blast resistance gene Pik-p Function-specific molecular markers and their applications. Background technique [0002] The rice blast fungus ( Magnapothe oryzae ) caused by rice blast disease has caused serious food loss to almost all major rice producing areas in the world (Qu 1985, Ma et al. 2015). Long-term production practice shows that breeding and utilizing disease-resistant varieties is the safest and most effective way to control rice blast. Moreover, due to the frequent pathogenic variation of rice blast physiological races, the resistance of single resistant varieties will be gradually lost in 3 to 5 years after planting (Yin Desuo et al., 2011); Spectrum resistance genes are an important way to obtain long-lasting, broad-spectrum disease-resistant varieties. The conventional identification method of resistance gene is inoculation identification, but be...

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 田大刚陈松彪王宗华王锋陈在杰陈子强林艳
Owner BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI