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Rice blast resistance gene Pi5 function specificity molecular marker and application thereof

A rice blast resistance gene technology, applied in the field of rice blast resistance gene Pi5 function-specific molecular markers, can solve the problems of unclear distribution and restrictions on the wide application of resistance genes, and achieve clear and simple band patterns and high product quality. Good separation effect and good stability

Active Publication Date: 2016-11-23
苏州顶德生物科技有限公司
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AI Technical Summary

Problems solved by technology

However, the distribution of Pi5 in rice germplasm resources and varieties in my country is still unclear, which restricts the wide application of this resistance gene in breeding.

Method used

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  • Rice blast resistance gene Pi5 function specificity molecular marker and application thereof
  • Rice blast resistance gene Pi5 function specificity molecular marker and application thereof

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Effect test

Embodiment 1

[0022] Embodiment 1: Rice blast resistance gene Pi5 gene-specific molecular marker and primer design and detection thereof

[0023] (1) Analysis of Pi5 insertion / deletion (insertion-deletion, InDel) site:

[0024] Download the published Pi5 genome sequence from the public database (Genebank accession numbers are: EU869185, EU869186), design primers for the Pi5 site, and screen Pi5-specific alleles that can be distinguished from other rice blast alleles at this site Insertion / deletion (InDel) differential sites for resistance genes. The multiple sequence alignment results of the rice blast resistance gene Pi5 with gene-specific InDel and other varieties containing non-Pi5 functional genes are as follows, wherein the missing two bases are the identified insertion / deletion of the Pi5 gene (InDel ) difference sites.

[0025] IRBL5-M: TATATTTTTTCTGTCCCTAAAATAGTTTTTTTT--CTATTACTGTCATTCATGTGACTCATTTTGTTTTTTAG

[0026] Zegu: TATATTTTTTCTGTCCCTAAAATAGTTTTTTTT--CTATTACTGTCATTCATGTGAC...

Embodiment 2

[0066] Example 2: Application of resistance gene Pi5 gene-specific molecular marker in identifying Pi5 gene.

[0067] According to the polymorphic PCR product bands, the target gene Pi5 can be distinguished. figure 1 It is the amplification of 12 representative varieties using the developed marker JJ801. According to the identification basis of the marker, IRBL5-M(Pi5), Zhonghua 16, Yueyezhan, Huajienuo, Minghui 63, Guichao Both No. 2 and Lianjing No. 7 are positive. figure 2 The results showed that the Pi5 gene-specific molecular marker can distinguish both alleles that do not contain this site and alleles that contain this site. That is to say, for all rice varieties carrying Pi5, the electrophoresis detection band appears at 93bp, while other alleles at this site appear at 95bp, and those not including this site appear as no band. It can be seen that the accuracy of the test results has been improved compared with the previous markers, indicating that the resistance gene...

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Abstract

The invention discloses a rice blast resistance gene Pi5 function specificity molecular marker named as Pi5InDel and application of the rice blast resistance gene Pi5 function specificity molecular marker. The nucleotide sequence of the rice blast resistance gene Pi5 function specificity molecular marker is shown in the sequence SEQ ID NO.1. The rice blast resistance gene Pi5 function specificity molecular marker is high in specificity, is low in cost and high in throughput in practical application, and can be widely applied groups with different genetic backgrounds to screen rice germplasm containing the rice blast resistance gene Pi. By means of the molecular marker, the utilization efficiency of the molecular marker in molecular marker auxiliary selective breeding, gene pyramiding breeding and transgene breeding can be improved.

Description

technical field [0001] The invention relates to the field of agricultural biotechnology, in particular to the field of rice blast technology, in particular to a functional specific molecular marker of rice blast resistance gene Pi5 and its application. Background technique [0002] Rice is one of the most important food crops in the world, and the economic losses caused by rice blast amount to billions of dollars every year (Zheng Zhao et al., 2009). Almost all the main rice producing areas in my country are harmed by rice blast, with an annual incidence area of ​​3.3-5.7 million hectares and a loss of hundreds of millions of kilograms of rice. From the perspective of food security and rice genetics and breeding, breeding disease-resistant varieties is one of the most cost-effective methods to control rice blast. However, due to the rapid pathogenic variation of rice blast physiological races, the resistance of single resistant varieties will gradually lose (Yin Desuo et al...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 杨立明罗玉明纪剑辉方继朝刘永锋
Owner 苏州顶德生物科技有限公司
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