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Magnaporthe oryzae avirulence gene AvrPib and application thereof

A non-toxic gene and rice blast fungus technology, applied in the field of genetic engineering, can solve the problem of unstable resistance of disease-resistant varieties

Active Publication Date: 2015-02-18
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the diversity and variability of the blast fungus population, the resistance of resistant varieties is unstable, so that the problem of susceptibility of resistant varieties has not been resolved.

Method used

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  • Magnaporthe oryzae avirulence gene AvrPib and application thereof
  • Magnaporthe oryzae avirulence gene AvrPib and application thereof
  • Magnaporthe oryzae avirulence gene AvrPib and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Avirulent Gene of Magnaporthe grisea AvrPib Bitcloning of

[0037] (1) AvrPib Construction of locus genetic map

[0038] In order to discover and identify the avirulence gene of Magnaporthe grisea AvrPib , using the blast fungus strain CHL357 (mating type MAT1-1 ; for containing Pib The rice cultivar IRBLb-B exhibited toxicity; Hua et al. 2012, Theor Appl Genet 125: 1047-1055) and CHL42 (mating type MAT1-2 ; showed no toxicity to IRBLb-B; Zhai et al. 2014, Plos One 9: e98067) 83 offspring ascospore strains obtained from crossing were inoculated into rice variety IRBLb-B (Kobayashi et al. 2007, JARQ 41: 31-37) , the results showed that there were 41 avirulent (non-pathogenic) strains and 42 virulent (pathogenic) strains in this mapping group, and the segregation ratio was 1:1. It was inferred that the avirulence of CHL42 to rice variety IRBLb-B was controlled by a pair of dominant genes.

[0039] In order to quickly determine the chromosomal locatio...

Embodiment 2

[0073] Example 2 AvrPib The gene structure, protein sequence and mutants of

[0074] (1) AvrPib Inference of gene structure: using the step-by-step method for AvrPib The DNA sequence of the gene was determined, and the coding region of the gene was predicted by software such as DNAStar. Wherein, SEQ ID NO: 1 and SEQ ID NO: 2 are respectively AvrPib Genomic DNA and cDNA sequences. AvrPib Genomic DNA was 2618 bp in length, and its full-length cDNA was 637 bp, containing an open reading frame of 225 bp, and the 5' and 3' untranslated regions were 322 bp and 90 bp, respectively. By comparing genomic DNA and cDNA, it was found that the open reading frame of the gene has only one exon and no intron ( image 3 a).

[0075] (2) AvrPib Presumption of protein sequence: using software such as DNAStar to AvrPib The protein sequence encoded by the gene was predicted, and the result is shown in SEQ ID NO:3. AvrPib It encodes a protein polypeptide consisting of 75 amino acid resid...

Embodiment 3

[0096] Example 3 AvrPib Application of Specific Molecular Markers in Population Monitoring of Magnaporthe grisea

[0097] (1) AvrPib Design of specific molecular markers: obtained by using Example 2 AvrPib Genomic DNA sequence (SEQ ID NO: 1) pair AvrPib Genomic sequences on both sides of the site were compared and analyzed. The results showed that the genome sequences on both sides of it had high specificity and great variability, especially the transposon in its 5'upstream region ( Pot2 , Pot3 ) insertion will result in loss of its function. Therefore, 2 pairs of primers were designed in its coding region and its two sides ( AvrPib F1 / R1; AvrPib F2 / R2), as its specific molecular marker ( Figure 5 a).

[0098] The primer sequences are as follows:

[0099] AvrPib F1: GGACAAGGGAGGCAAATCTAAC

[0100] AvrPib R1: ATGCCGACAATGCGAGGTAT

[0101] AvrPib F2: TGGAGAAGACTTTGATGC

[0102] AvrPib R2: GAACGCATAATGGCAACTA

[0103] (2) AvrPib Application of specific mol...

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Abstract

The invention relates to the technical field of gene engineering, and particularly discloses magnaporthe oryzae avirulence gene AvrPib and application thereof. The avirulence gene comprises a DNA (deoxyribonucleic acid) sequence, with a promoter, as shown in SEQIDNO:1 or a cDNA (complementary deoxyribonucleic acid) sequence as shown in SEQIDNO:2. Molecular targets of a novel pesticide can be designed on the basis of the structure of the gene and application test results thereof; by covalently introducing the gene and relevant anti-disease genes into host plants, such as rice, new disease-resistant varieties can be bred; specific molecular markers produced according to the gene sequence can be applied to monitoring field magnaporthe grisea groups; monitoring results obtained can be used for guiding the reasonable layout of the disease-resistant varieties; the gene has a wide application value, and has great significance in rice production and breeding research.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a non-toxic gene of rice blast fungus AvrPib and its application. Background technique [0002] Rice ( Oryza sativa L. ) is an important food crop and an important industrial raw material for the survival of billions of people in the world, and it is also the host of various diseases and insect pests. by pathogenic fungi Magnapothe oryzae The rice blast caused by rice blast is one of the most devastating diseases in rice production in the world, causing 10-30% rice yield loss every year. Moreover, the fungus can also infect more than 50 kinds of gramineous plants such as wheat and cereals. [0003] 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. However, due to the diversity and variabil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/37C12N15/31C12N15/82A01H5/00C12Q1/68C12Q1/04C12R1/645
CPCC07K14/37C12N15/8205C12N15/8282C12Q1/6895C12Q2600/156
Inventor 潘庆华张树林何丽云杨先锋王玲
Owner SOUTH CHINA AGRI UNIV
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