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Pi15 resistance gene of rice blast, and application

A rice blast resistance gene technology, applied in the field of cloning of rice blast resistance gene Pi15, can solve the problems of cloning rice blast resistance gene and achieve the effect of shortening the breeding time

Inactive Publication Date: 2007-10-10
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Before the application of the present invention was filed, there was no report of cloning rice blast resistance gene on rice chromosome 1

Method used

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  • Pi15 resistance gene of rice blast, and application
  • Pi15 resistance gene of rice blast, and application
  • Pi15 resistance gene of rice blast, and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Genetic Analysis and Preliminary Mapping of Rice Blast Resistance Gene Pi15

[0045] The present invention is for 3 Fs derived from the hybrid combination of japonica rice disease-resistant variety GA25 and 3 indica rice disease-susceptible varieties AS20-1, IR36 and Q61. 2 Populations were inoculated with strains CHL0416 and CHL0670 that showed distinct non-affinity / affinity responses to both parental varieties, and the results showed that the three F 2 The segregation ratios of resistant plants and susceptible plants in the population were all consistent with 3R:1S, so it was inferred that the resistance of GA25 to the above three inoculated strains was controlled by a pair of dominant genes. to these 3 F's 2 The linkage analysis of the resistance genes corresponding to the population showed that they were controlled by the same dominant gene, so the three F 2 From the population, a large mapping population consisting of 76 resistant individuals and 504 s...

Embodiment 2

[0047] Example 2: Construction of an electronic contig covering the Pi15 site

[0048] Since RAPD marks BAPi15 782 and BAPi15 486 Closely linked to the Pi15 locus, therefore, after RAPD markers achieved chromosomal landing, a contig covering the Pi15 locus was constructed using the genome sequence of the reference variety Nipponbare as a reference, which was composed of BAC clone AP005879, PAC clone AP005593, AP005811 and AP005700 and other 4 clones (Figure 2).

Embodiment 3

[0049] Example 3: Fine Mapping of Rice Blast Resistance Gene Pi15

[0050] In order to further fine-tune the Pi15 locus, we developed PCR-based markers in the gene region of interest based on the constructed contigs. Using the online software tools ORF Finder and BLASTP to search the sequence of this region, 9 ORFs with LRR or NBS structures were obtained. Then, these 9 ORFs were used as CRG, and 9 pairs of specific primers were designed on their respective flanking sequences, and PCR amplification was carried out respectively. No bands could be amplified initially with rTaq DNA polymerase. After exploration, we adopted the cocktail method, combining Ex Taq and Probest Taq DNA polymerases to better solve the problem of mismatch between primers and template DNA, so that all CRGs were amplified. The results showed that: 6 CRG markers such as CRG1, CRG2, CRG3, CRG4, CRG5 and CRG6 showed good specificity (only the target band appeared), while the other 3 CRG markers were discarded...

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PUM

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Abstract

This invention discloses rice Magnaporthe grisea resistance gene Pi15 nucleotide sequence; amino acid polypeptide sequence coded by rice Magnaporthe grisea resistance gene Pi15, and its application. Magnaporthe grisea resistance gene Pi15 is a member of resistance gene family nonTIR-NBS-LRR, and is a constitutive expression gene. Magnaporthe grisea resistance gene Pi15 can be used to breed disease-resistant rice or plants through genetic transformation, or used as a molecular marker in plant breeding.

Description

Technical field [0001] The invention relates to the field of genetic engineering, and specifically to the cloning of a rice blast resistance gene Pi15 and its application. Background technique [0002] During the growth process, plants are often attacked by a variety of pathogens, and plants adopt a variety of defense strategies to protect themselves and avoid being invaded by them. One of the most important defense mechanisms in plants is the ability to recognize the presence of obligate pathogenic microorganisms and activate their own defense response system. Plant recognition of pathogenic bacteria is mediated by disease resistance genes. Therefore, the analysis and research on the structure of disease resistance gene products is the basis for understanding the plant disease resistance mechanism, and also has important guiding significance for the prevention and control of plant diseases. [0003] To date, more than 40 disease resistance genes have been isolated from 3 ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82
Inventor 潘庆华林菲王玲
Owner SOUTH CHINA AGRI UNIV
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