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Wild vitis quinquangularis anti-disease gene and application thereof

A disease-resistant gene, hair grape technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problem that the quality is not as good as European grape varieties

Active Publication Date: 2017-04-26
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

my country has abundant wild grape resources, some of which are highly resistant to disease, such as Shang-24 (S-24), but its quality is not as good as that of European grape varieties

Method used

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  • Wild vitis quinquangularis anti-disease gene and application thereof
  • Wild vitis quinquangularis anti-disease gene and application thereof
  • Wild vitis quinquangularis anti-disease gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: VqWRKY52 response to hormones and tissue-specific and spatio-temporal specific expression in Arabidopsis

[0037] In previous studies, the applicant found that the expression of VqWRKY52 was induced by powdery mildew of grapevine. In order to determine its response to SA and JA, the expression of VqWRKY52 was analyzed under the treatment of these two hormones. hours, VqWRKY52 could be strongly induced by SA, but not by MeJA ( figure 1 A).

[0038] In order to further verify this result, the 2107bp promoter region of VqWRKY52 was cloned and fused in front of the GUS reporter gene to form a Pro VqWRKY52 : GUS reporter structure, transiently transformed tobacco to analyze promoter activity in response to hormones, using Pro 35S :GUS as a control. The results showed that the VqWRKY52 promoter could be induced by SA but not by JA ( figure 1 B), which is the same as figure 1 The results of A are consistent.

[0039] To look at tissue-specific and spatio-tempo...

Embodiment 2

[0040] Example 2: Response of VqWRKY52 Transgenic Arabidopsis T3 Generation Seedlings to Powdery Mildew

[0041] To further determine the role of VqWRKY52 in plant defense, three T3 transgenic lines (#28, #30, #33), Arabidopsis wild-type (WT) and pad4 mutant were inoculated against Arabidopsis powdery mildew, see For its response to pathogenic bacteria, the inventor will inoculate powdery mildew with 3-4 week-old adult seedlings, observe different strains of active oxygen, dead cell accumulation and pathogenic bacteria growth, count the conidia quantity of single colony of powdery mildew, quantitative The relative expression of disease resistance-related genes was analyzed. The results showed that three transgenic strains (#28, #30, #33) had mild disease status after powdery mildew inoculation compared with wild control (WT), and the disease status of pad4 mutant was the most serious ( image 3 A). Three transgenic lines (#28, #30, #33) showed strong powdery mildew-induced c...

Embodiment 3

[0045] Example 3: Response of VqWRKY52 Transgenic Arabidopsis T3 Seedlings to Pseudomonas syringae

[0046] To determine the role of VqWRKY52 in plant resistance to Pseudomonas syringae, three T3 generation transgenic lines (#28, #30, #33) and Arabidopsis wild-type (WT) were inoculated with Pseudomonas syringae, see For its response to pathogenic bacteria, 3-4 week-old adult seedlings were inoculated with pathogenic bacteria, and different strains of reactive oxygen species, accumulation of dead cells and growth of pathogenic bacteria were observed, and the number of pathogenic bacteria in leaves 3 days after inoculation was counted. Relative expression of disease-related genes ( Figure 6, 7). The results showed that the three transgenic lines (#28, #30, #33) were milder than the wild control (WT) after being inoculated with pathogenic bacteria ( Figure 6 A). Simultaneously, count the amount of growth of bacteria in different genotypes 3 days after inoculation, and find t...

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Abstract

The invention discloses a wild vitis. quinquangularis anti-disease gene and application thereof. The full length of a complete open reading frame sequence is 1095bp, and 364 amino acids are encoded. Moreover, a pCAMBIA2300-35S-VqWRKY52 over-expression vector is constructed, a model plant, namely, arabidopsis thaliana is imported by an inflorescence infection method, and over-expression is performed in the arabidopsis thaliana, so that the resistance of the arabidopsis thaliana to powdery mildew and pseudomonas syringae parasitic in living organisms is improved remarkably, and the resistance to saprophytic botrytis cinerea is lowered. A wild vitis. quinquangularis-24 anti-disease gene VqWRKY52 participates in an SA-mediated anti-disease signal path, can enhance the anaphylactic reaction, can be used for enhancing the resistance of plants to the fungus powdery mildew and the pseudomonas syringae parasitic in the living organisms, and plays a role in regulating and controlling the anaphylactic reactions of the plants through regulation and control of the expression of the VqWRKY52 gene, so that the reactions of the plants to different pathogenic bacteria are regulated and controlled.

Description

technical field [0001] The invention relates to the technical field of plant disease-resistant gene identification and genetic engineering, in particular to a wild grape disease-resistant gene and its application. Background technique [0002] Unlike animals, plants have mobile defense cells and an acquired immune system, which can only rely on the signals emitted by each plant cell and by the site of infection. During the long-term evolution process with plants, plant pathogenic bacteria have evolved a variety of life strategies to help them infect plants, and formed a set of their own unique immune system, in which transcription factors play a very important regulatory role . [0003] WRKY transcription factors are one of the more important types of transcription factors, which play a positive or negative regulatory role in the entire plant immune system network. WRKY transcription factor is a unique transcriptional regulatory factor found in plants. The N-terminus of it...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8281C12N15/8282
Inventor 王西平王现行涂明星
Owner NORTHWEST A & F UNIV
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