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Rice blast resistance gene Pi1 functional specificity molecular marker and application thereof

A resistance gene and molecular marker technology, applied in the field of agricultural biology, can solve problems such as being unsuitable for identification of germplasm resources or screening of resistance resources, unsuitable for large-scale resistance identification and MAS work, etc. volume and low cost

Inactive Publication Date: 2016-09-21
BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

However, the markers used are located in the resistance gene Pi1 There is a certain physical distance between the flanks and the target gene. They can only be used for polymorphism analysis between specific parents, and are not suitable for identification of germplasm resources or screening of resistance resources.
Although Pan Qinghua et al. (2012) developed Pi1 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 MAS work

Method used

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  • Rice blast resistance gene Pi1 functional specificity molecular marker and application thereof
  • Rice blast resistance gene Pi1 functional specificity molecular marker and application thereof
  • Rice blast resistance gene Pi1 functional specificity molecular marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Rice blast resistance gene Pi1 Gene-specific molecular markers and their primer design and detection

[0033] (1) Analysis of Pi1 insertion / deletion (insertion-deletion, InDel) site:

[0034] Downloaded from public database Pi1 and Pik-m , Pik , Pik-p , Pik-s , Pi7 with Pik-h Partial genome sequences of rice donor varieties, sequenced varieties Nipponbare (Nipponbare) and 9311 have no corresponding genome sequences, sequence alignment and screening for the Pi1 site Pi1 A specific insertion / deletion (InDel) difference site that is specific and can be distinguished from other rice blast allelic resistance genes at this site. Rice blast resistance genetic variety IRBL1-CL with gene-specific InDel ( Pi1 ) and the resistant variety IRBLk-Ka ( Pik ), resistant variety IRBLkp-K60 ( Pik-p ), the resistant variety IRBLkm-Ts ( Pik-m ), resistant variety IRBLkh-K3 ( Pik-h ), the resistant variety IRBLks-S ( Pik-s ) and the resistant variety IRBL...

Embodiment 2

[0063] Embodiment 2: resistance gene Pi1 Gene-specific molecular markers in identifying Pik Application of different rice blast resistance genes in gene cluster regions.

[0064] According to the polymorphic PCR product bands, the target gene can be Pi1 with other Pik The resistance genes on the gene cluster were distinguished. Such as figure 1 As shown, depending on the stripe position, the Pi1 Distinguished from other resistance genes identified at this locus, 220 bp indicates that it contains Pi1 209 bp indicates other resistance genes that contain this site, and other bands indicate functional genes that do not contain this site. It can be seen that the test results are consistent with the design analysis, indicating that the resistance gene Pi1 Gene-specific molecular markers can be identified in Pi1 gene and Pik Gene family alleles and other aspects have been applied.

Embodiment 3

[0065] Embodiment 3: resistance gene Pi1 Detection application of gene-specific molecular markers in other rice varieties

[0066] Select 12 representative rice varieties, in order: Zhenshan 97B, Co39, Fuhui 838, Gufeng B, Jinnante 43B, Nante, Anfeng B, Teqing 2, Baoxie 123B, Nanjing 11 , Zhuzhen B, Bantam Nantes.

[0067] Genomic DNA was extracted respectively, and using this as a template, PCR amplification, enzyme digestion and electrophoresis detection were performed according to the method of Example 1. According to the size of the digested product (band), the resistance gene can be Pi1 Distinguished from other rice blast resistance genes. Such as figure 2 As shown, the resistance spectrum phenotype is Pi1 Type individuals (carrying rice blast resistance gene Pi1 The presence of Pi1-InDel gene-specific molecular markers can be detected in the disease-resistant variety IRBL1-CL (samples in lane 1), while the molecular markers cannot be detected in individuals with o...

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Abstract

The invention provides a rice blast resistance gene Pi1 functional specificity molecular marker and application thereof. The molecular marker in a specificity banding pattern with a rice blast resistance gene Pi1 is amplified from a rice genome DNA through primer pairs of SEQ ID NO.1 and SEQ ID NO.2. The rice blast resistance gene Pi1 functional specificity molecular marker has very important application value, and by means of the marker, the utilization efficiency of the gene in germplasm resource screening, marker-assisted selection breeding, gene pyramiding breeding and transgenosis breeding can be improved.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, in particular to a rice blast resistance gene Pi1 Functional molecular marker Pi1-InDel and its method and application. Background technique [0002] by Magnaporthe grisea ( Magnapothe oryzae ) caused by rice blast is the most important disease that endangers the stable yield of rice, and it causes serious food losses in 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); therefore, mining and rational utilization are widely used. Spectrum resistance genes are an important way to obtain long-lastin...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q1/686C12Q2600/13C12Q2600/156C12Q2565/125
Inventor 田大刚陈松彪胡昌泉陈在杰陈子强林艳王锋
Owner BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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