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Application of suppressing or mutating tfiiaγ gene in crop disease resistance

A gene and gene expression technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problem that the basic transcription factors of the host are not clear about the function

Inactive Publication Date: 2020-01-17
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But whether this process requires other proteins to participate, or how the host's basic transcription factors play a role in this process is not clear

Method used

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  • Application of suppressing or mutating tfiiaγ gene in crop disease resistance
  • Application of suppressing or mutating tfiiaγ gene in crop disease resistance
  • Application of suppressing or mutating tfiiaγ gene in crop disease resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Sequence and structural analysis of the TFIIAγ gene

[0035] 1. Prediction of TFIIAγ gene structure

[0036] The applicant used a DNA sequence (2944bp-3345bp) of a TALE gene pthXo1 (GenBank registration number: ACD58243) of bacterial blight PXO99 as a bait protein to screen the cDNA library of rice variety Zhonghua 11 (Yuan et al., 2010), and screened A cDNA sequence was searched in the nucleotide database GenBank (http: / / www.ncbi.nlm.nih.gov) with the BLAST method (Altschul et al. 1997), and it was found that the cDNA sequence was similar to a gene from the rice variety Nipponbare located on chromosome 5 The sequences above are highly homologous. In the rice genome database TIGR (http: / / rice.plantbiology.msu.edu / ), this rice sequence is annotated as a gene whose number is LOC_Os05g01710, which is the gene OsTFIIAγ5 involved in the present invention.

[0037] 2. Isolation and cloning of OsTFIIAγ5 gene from rice variety Zhonghua 11

[0038] Rice Zhonghua 11...

Embodiment 2

[0049] Example 2: Suppressing the expression of the rice OsTFIIAγ5 gene to verify its function in rice disease resistance

[0050] 1. Construction of expression-suppressing genetic transformation vector

[0051] The present invention uses RNA interference (RNA interference, RNAi) technology (Smith et al., 2000) to verify the function of the gene by inhibiting the expression of the OsTFIIAγ5 gene in rice variety Zhonghua 11. The main mechanism of this technical approach is to connect a partial fragment of the target gene in the form of inverted repeats to a vector capable of expressing double-stranded RNA (double strand RNA, dsRNA), and introduce the vector into plants through genetic transformation. The obtained transformed plants express a large number of dsRNAs homologous to partial fragments of the target gene. These dsRNAs rapidly form short interfering RNA (short interfering RNA, siRNA). These siRNAs are complementary to the transcripts (mRNA) of the target gene, and un...

Embodiment 3

[0073] Example 3: Point mutation of rice OsTFIIAγ5 gene endows it with function in broad-spectrum disease resistance

[0074] 1. Point mutation of rice OsTFIIAγ5 gene endows it with broad-spectrum resistance to bacterial blight

[0075] Rice families IRBB5 and IR24 are near-isogenic lines. IR24 carries the OsTFIIAγ5 gene, and the 39th amino acid of the encoded OsTFIIAγ5 protein is valine (V); IRBB5 carries the mutated OsTFIIAγ5 gene, namely the recessive xa5 gene. The recessive xa5 gene encodes a mutant OsTFIIAγ5 protein (OsTFIIAγ5 V39E or xa5), the 39th amino acid is mutated from valine to glutamic acid (E). At the booting stage, inoculate the Philippine bacterial blight races PXO99, PXO339, PXO61, PXO71, PXO79, PXO86, PXO112, PXO341 and PXO347, the Japanese bacterial blight race T7174, and the Korean bacterial blight races. Species KACC10331, Physiological races of bacterial blight of China Zhe173, FuJ23, KS-1-21 and YN11. The results showed that the resistance of IRBB5 ...

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Abstract

The invention belongs to the field of plant gene engineering, and particularly relates to an application of inhibiting or mutant TFIIA (transcription factor IIA) gamma subunit genes to crop disease resistance, and functional verification in bacterial pathogen interaction of plants and genes with transcription activator-like effectors (TALE). The genes include rice Os TFIIA gamma 5 genes, pepper Ca TFIIA gamma genes h and tomato S1 TFIIA gamma genes. The TFIIA gamma subunit genes are negative regulatory factors in reaction of plants resisting pathogens with the TALE, rice has broad-spectrum resistance to Xanthomonas oryzae pv. oryzae and Xanthomonas oryzae pv. Oryzicola when Os TFIIA gamma 5 gene expression is suppressed or Os TFIIA gamma 5 genes are mutated, yield traits are unaffected by the mutation Os TFIIA gamma 5 genes, the resistance of tomatoes to Xanthomonas campestris pv. Vesicatoria can be improved when S1 TFIIA gamma gene expression is suppressed in the tomatoes, and the resistance of peppers to Xanthomonas vesicatorium Dowson can be improved when Ca TFIIA gamma gene expression is inhibited in the peppers.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering. It specifically relates to the application of inhibiting or mutating TFIIAγ gene in crop disease resistance. The application relates to the application of plant transcription factor IIAγ subunit (transcription factor IIAgamma subunit, TFIIAγ) gene in antibacterial disease resistance. TFIIAγ gene is a negative regulator in disease resistance response. The ability of the plants inhibiting the expression of TFIIAγ gene to carry the transcription activator-like effector (transcription activator-like effector, TALE) was significantly improved. Background technique [0002] Xanthomonas (Xanthomonas) is a gram-negative genus of prokaryotic organisms Pseudomonas family, all bacteria of the genus Xanthomonas are plant pathogenic bacteria, it includes many pathogenic varieties, and is a worldwide pathogenic bacteria , can harm more than 400 types of plants in at least 27 plant genera (...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/46A01H6/82
CPCC07K14/415C12N15/8205C12N15/8216C12N15/8281C12N2810/10C12N2830/34C12N2830/36
Inventor 袁猛王石平
Owner HUAZHONG AGRI UNIV
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