Unlock instant, AI-driven research and patent intelligence for your innovation.

Rice blast resistance gene Pik-s functional specific molecular marker, as well as method and application thereof

A technology of resistance gene and molecular marker, applied in the field of agricultural biology

Inactive Publication Date: 2012-09-05
SOUTH CHINA AGRI UNIV
View PDF0 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] During the fine mapping of Pik-s, the applicant found that in the genomic region containing the Pik-s locus, the Pik-s donor variety Shin 2 (Wang et al.2009.Characterization of rice blast resistance genes in the Pik cluster and fine mapping of the Pik-p locus. Phytopathology 99: 900-905) has a similar genome structure to the donor varieties of Pik-m, Pik and Pik-p, that is, there is a large insertion between the reference sequence Nipponbare / missing

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Rice blast resistance gene Pik-s functional specific molecular marker, as well as method and application thereof
  • Rice blast resistance gene Pik-s functional specific molecular marker, as well as method and application thereof
  • Rice blast resistance gene Pik-s functional specific molecular marker, as well as method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1: Alignment of Pik-s allele coding region sequence and identification of SNP

[0063]Amplified by conventional PCR (Yuan et al.2011. The Pik-p resistance to Magnaporthe oryzae in rice is mediated by a pair of closely linked CC-NB S-LRR genes. Theor Appl Genet 122:1017-1028; Zhai et al. al.2011. The isolation and characterization of Pik, a rice blast resistance gene which emerged after rice domestication. New Phytologist 189: 321-334), sequencing (Shanghai Yingjun Biotechnology Co., Ltd., Guangzhou Branch), from 5 antibiotics Diseased rice varieties [Shin 2 (Pik-s donor variety), Tsuyuake (Pik-m donor variety), Kusabue (Pik donor variety), K60 (Pik-p donor variety); Wang et al.2009.Characterization of rice blast resistance genes in the Pik cluster and fine mapping of the Pik-p locus.Phytopathology 99:900-905], K3 (Pik-h donor species; Xu et al.2008.Efficient authentic fine mapping of the rice blast resistance gene Pik-h in the Pik cluster, using new Pik-h-di...

Embodiment 2

[0064] Example 2: Pik-s functionally specific molecular markers and their primer design and detection

[0065] According to dCAPS (Derived Cleaved Amplified Polymorphic Sequences; Neff et al. 2002. Web-based primer design for single nucleotide polymorphism analysis. Trends in Genetics 18: 613-615) and CAPS (Cleaved Amplified Polymorphic Sequences; procedure for mapping Arabidopsis mutations using co-dominant ecotype-specific PCR-based markers.Plant J 4:403-410) marker design principle, according to the dCAPS marker principle, design the function with mismatched bases at the Piks-1SNP site Specific upstream primer Piks-1FNP-F, and functionally specific downstream primer Piks-1FNP-R is designed at 100-200 bp downstream of it, as shown in SEQ ID NO.1 and SEQ ID NO.2 respectively; according to the principle of CAPS labeling , design functionally specific upstream and downstream primers Piks-2FNP-F and Piks-2FNP-R within the range of 100bp upstream and downstream of the Piks-2SNP s...

Embodiment 3

[0086] Example 3: Application of the resistance gene Pik-s function-specific molecular marker in the identification of different rice blast resistance genes

[0087]Nine rice lines containing different rice blast resistance genes were collected, namely: Shin 2 (Pik-s donor variety; Wang et al.2009.Characterization of rice blast resistance genes in the Pik cluster and fine mapping of the Pik -p locus. Phytopathology 99:900-905); K3 (Pik-h donor species; Xu et al. 2008. Efficient authentic fine mapping of the rice blast resistance gene Pik-h in the Pik cluster, using new Pik-h -differentiating isolates. Molecular Breeding 22:289-299); IRBLkh-K3, IRBLk-Ka, IRBLkm-TS, IRBLkp-K60, IRBL7-M, IRBL 1-CL, IRBLks-S [Kobayashi et al.2007.Development of new Sets of international standard differential varieties for blast resistance in rice (Oryza sativa L.). JARQ 41:31-37] Genomic DNA, and using this as a template, using Piks-1FNP, Piks-2FNP and Piks-2FNP described in Example 2 PCR amplifi...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a rice blast resistance gene Pik-s functional specific molecular marker, as well as a method and application thereof. The molecular marker consists of Piks-1FNP, Piks-2FNP and Piks-3FNP, which are nucleotide sequences amplified from SEQ ID NO.1 and SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4, and SEQ ID NO.5 and SEQ ID NO.6 of rice total DNA (ribonucleic acid) respectively by primers. Three single base differences different from the Pik-s functional specificity of an affected allele and other rice blast genes can be obtained by analysis with a method of comparing Pik-s allele sequences of a plurality of rice blast resistance genes with the Piks-1 and Piks-2 sequences, and the primers can be obtained through design, and the rice DNA is amplified to respectively obtain the Pik-s functional specific molecular markers Piks-1FNP, Piks-2FNP and Piks-3FNP. The rice blast resistance gene Pik-s functional specific molecular marker can be used for screening and identifying a functional resistance gene in massive rice germplasm resources, and can be applied to molecular marker assisted selective breeding, gene pyramiding breeding and transgenic breeding.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to a rice blast resistance gene Pik-s function-specific molecular marker PiksFNP and its method and application. Background technique [0002] Rice is one of the most important food crops in the world, and more than half of the population uses rice as a staple food. Rice blast caused by Magnapothe oryzae is one of the most serious diseases to rice production, causing serious grain loss every year. From the point of view of environmental protection and agricultural sustainable development, the breeding and utilization of disease-resistant varieties is the safest and most effective way to control rice blast. Traditional rice resistance breeding relies on the identification of resistance phenotypes, which not only requires breeders to have rich experience in inoculation and investigation, but is also easily affected by environmental and human factors. The identifi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 潘庆华陈彩霞翟纯林菲王玲
Owner SOUTH CHINA AGRI UNIV