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Phytophthora cinnamomi effector protein Avh57 and application thereof

A technology of effector protein and Phytophthora, applied in the fields of application, angiosperm/flowering plants, biochemical equipment and methods, etc., can solve problems such as different structure of different effector proteins, achieve the goal of reducing plant mortality and significant application value Effect

Active Publication Date: 2021-07-16
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is recorded in the prior art that the Phytophthora camphora effector protein Avh87 gene can induce plant cell death induced by the apoptotic precursor protein Bax, but only one effector protein is far from enough to study the infection mechanism of Phytophthora camphora pathogenic process. Not enough, and the structure of different effector proteins is also completely different

Method used

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  • Phytophthora cinnamomi effector protein Avh57 and application thereof
  • Phytophthora cinnamomi effector protein Avh57 and application thereof
  • Phytophthora cinnamomi effector protein Avh57 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Acquisition of the gene encoding effector protein Avh57 of Phytophthora camphora

[0049] The system analysis was carried out in the software MEGA7, using the method of Neighbor-Joining. Such as figure 1 As shown, this figure provides the evolutionary relationship and degree of association between P.cin_Avh57 and Phytophthora in the figure, and provides a reference for the research of P.cin_Avh57 gene.

[0050] Then, the Phytophthora camphora strain was selected as the test material, and the Phytophthora camphora genome sequence was analyzed according to the reported effector protein gene information, and the effector protein genes in the entire genome of Phytophthora camphora were obtained. Then design primers Avh57-F and Avh57-R according to the obtained gene fragments to amplify and screen the obtained target gene fragments.

[0051] 1. Using CTAB-SDS method to extract high-quality Phytophthora camphora genomic DNA

[0052] Cultivate Phytophthora camphor...

Embodiment 2

[0066] Example 2 Analysis of the expression pattern of Avh57 gene during the infection of apple by Phytophthora camphora

[0067] 1. Phytophthora camphora strains infecting apples

[0068] The mycelium blocks stored in the refrigerator at 4°C were placed on V8 solid medium for activation, and cultured at 25°C for 24-36h. Place a piece of sterilized filter paper of appropriate size on the bottom of the sterilized Petri dish (diameter 9cm) and moisten it with sterilized water. Sterilize the sterile operating table. Wipe the surface of the apples with alcohol cotton to sterilize them. Use the outer flame of the alcohol lamp to sterilize the edge of the cutter, and cut about 1cm on the four sides of the apple 2cubes, remove the pulp. Pick out the colony and put it on the sterilized filter paper and stuff it into the incision on the surface of the apple. Only three sides are stuffed, and the other side is used as a control of blank solid medium. Plug the incision with absorben...

Embodiment 3

[0082] Example 3 Transient expression of Avh57 gene in tobacco

[0083] 1. Construction of PVX recombinant expression vector

[0084] (1) The target gene fragment Avh57 was amplified by SmaⅠ restriction PCR, and the inserted fragment was recovered, with a size of about 567 bp.

[0085] Reaction system: ddH 2 O (33 μL), SmaI (2 μL), plasmid (10 μL), 10×cut Buffer (5 μL). 37°C, 30min.

[0086] (2) Ligate with pGR107 vector cut with the same restriction enzymes, and transform Escherichia coli DH5α.

[0087] (3) The transformed DH5α was screened for Kan resistance, the obtained colonies were shaken overnight at 37°C, and the plasmid was extracted.

[0088] (4) The recombinant plasmid was digested and identified with restriction endonuclease SmaI. The recombinant plasmids that were initially identified correctly after enzyme digestion were sent to GenScript Biotechnology Co., Ltd. for sequencing. The sequence shows that the recombinant plasmid inserted into the DNA fragment o...

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Abstract

The invention discloses an effector protein Avh57 from phytophthora cinnamomi as well as a coding gene and an application thereof. The protein has an amino acid sequence shown in SEQ ID NO: 1 in a sequence table. Experiments prove that the protein controlled and expressed by the gene can participate in invasion process of phytophthora cinnamomi into nicotiana benthamiana, and the gene has the function of inhibiting plant cell death induced by proapoptotic protein Bax. The phytophthora cinnamomi effector protein of the present invention has important significance on enriching molecular mechanism data information about interaction of plant pathogenic oomycetes and hosts, and establishing a comprehensive prevention and control technical strategy against plant pathogenic oomycetes diseases.

Description

technical field [0001] The invention belongs to the technical field of Phytophthora camphorae effectors, and in particular relates to Phytophthora camphorae effector protein Avh57 and its application. Background technique [0002] Phytophthora cinnamomi (Phytophthora cinnamomi) is a globally distributed and highly invasive soil-borne pathogen. It can be saprophytic, parasitic or obligate in different forms in soil and plant tissues, through surface and groundwater flow, and plant roots. Spread by contact, animal movement, and human activities (road building, logging, mining, etc.), it has an extremely broad host range. [0003] Effector proteins are a class of exocrine protein molecules secreted by pathogens that can change the cell structure and metabolic pathways of host plants, thereby promoting successful infection of host plants or triggering host defense responses. Many plant pathogens secrete effector proteins, such as those found in bacteria, fungi, oomycetes, and n...

Claims

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Application Information

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IPC IPC(8): C07K14/37C12N15/31C12N15/84A01H5/00A01H6/82
CPCC07K14/37C12N15/8205C12N15/8282
Inventor 戴婷婷李亚星陈贞鹏焦彬彬
Owner NANJING FORESTRY UNIV
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