Resistance gene Pi36 of rice blast and its use

A rice blast and resistance gene technology is applied in the field of isolation, cloning and application of the rice blast resistance gene Pi36, and the effect of shortening the breeding time is achieved.

Inactive Publication Date: 2009-11-04
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 that the rice blast resistance gene on the 8th chromosome of rice was cloned

Method used

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  • Resistance gene Pi36 of rice blast and its use
  • Resistance gene Pi36 of rice blast and its use
  • Resistance gene Pi36 of rice blast and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Genetic Analysis and Preliminary Mapping of Rice Blast Resistance Gene Pi36

[0039] Such as figure 1 , the present invention clones the resistance gene Pi36 by using the map-based cloning method. First, in order to discover and identify new resistance genes to rice blast, F from two hybrid combinations of indica rice resistant variety Q61 and japonica rice susceptible variety Aichixu and Lijiangxintuanheigu (LTH) were studied. 2 Group (F 2 -1 and F 2 -2), inoculate the bacterial strain CHL39 showing a clear non-affinity / affinity response to the parental species. The results show that the 2 F 2 The segregation ratio of resistant plants and susceptible plants in the population was 3:1. It was inferred that the resistance of Q61 to the inoculated strain CHL39 was controlled by a pair of dominant genes.

[0040] In order to quickly determine the chromosomal location of the main resistance gene, an F population consisting of 40 resistant and 580 susceptibl...

Embodiment 2

[0041] Example 2: Fine Mapping and Electron Physical Mapping of New Rice Blast Resistance Gene Pi36

[0042] In order to precisely determine the position of the Pi36 locus, we used the reference sequences in the public database to develop 5 polymorphic markers, 1 STS marker R36STS2 and 4 candidate resistance gene (candidate resistance gene, CRG) markers, CRG1 , CRG2, CRG3 and CRG4, and 580 strains of highly susceptible F 2 Individuals were subjected to linkage analysis. The results showed that the Pi36 locus was finely localized in a region of 0.6 cM between CRG2-RM5647 and completely co-segregated with the markers CRG3 and CRG4. In order to construct the physical map of this locus, we used the bacterial artificial chromosome (BAC) and P1-mediated artificial chromosome (P1-derived artificial chromosome, PAC) clones of the reference species Nipponbare, and through bioinformatics analysis (bioinformatics analysis, BIA), and an electron physical map of this site was constructed...

Embodiment 3

[0043] Example 3: Predictive Annotation and Sequence Analysis of the New Rice Blast Resistance Gene Pi36 Candidate Gene

[0044]In order to determine the candidate gene of Pi36, we use the reference sequence of Nipponbare, through three gene prediction software RiceGAAS (http: / / ricegaas.dna.affrc.go.jp), Gramene (http: / / 143.48.220.116 / resources / ) and Softberry's FGENESH (http: / / www.softberry.com) carried out gene prediction and annotation analysis on the target gene region, and initially identified the candidate resistance genes of Pi36 as Pi36-1, Pi36-2 and Pi36-3, among which Pi36-3 includes Pi36-1 and Pi36-2. Therefore, the function of Pi36-3 was confirmed by the following gene function complementation experiment.

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Abstract

The invention discloses a nucleotide sequence of a new rice resistance gene Pi36 and its coded amino acid polypeptide sequence and application. This gene belongs to the nonTIR-NBS-LRR resistance gene family and is a constitutively expressed gene. The invention also relates to the use of the gene to transform rice or other plants to breed disease-resistant varieties and the application of the molecular marker produced according to the gene sequence in breeding.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the isolation and application of a rice blast resistance gene Pi36. Background technique [0002] Plants are often attacked by various pathogens during the growth process, and plants adopt various defense strategies to protect themselves and avoid being attacked by them. One of the most important defense mechanisms in plants is the ability to recognize the presence of obligate pathogenic microorganisms and initiate 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-resistant gene products is the basis for understanding the mechanism of plant disease resistance, and also has important guiding significance for the prevention and control of plant diseases. [0003] So far, more than 40 disease resistance genes have been isolated from 3 mono...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63
Inventor 潘庆华王玲林菲刘新琼
Owner SOUTH CHINA AGRI UNIV
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