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 pro

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

Example Embodiment

[0048] Example 1 Efferfer protein AVH57 encoding gene

[0049] System analysis is performed in software MEGA7, using neighbor-joining methods. Such as figure 1 As shown, this figure provides the evolutionary relationship between P.CIN_AVH57 and Phytophthia in the figure, and provides a reference for the study of the P.CIN_AVH57 gene.

[0050] Then, the sequential strain is selected as the participating material, and the genome sequence in the mildew genome is analyzed according to the reported effector protein gene information, and the effector protein gene in the mildew pyrofy gain is obtained. Then, the target gene fragment obtained by the primer AVH57-F, AVH57-R, amplified screener is then designed according to the obtained gene fragment.

[0051] 1. Extract high-quality Etuta genome DNA with CTAB-SDS method

[0052] Steppermushfami plant in solid V8 medium plate culture (formula: 170ml V8 vegetable juice plus 1.6G calcium carbonate, mix, 2000 rpm centrifuge 5 min taken, add 15...

Example Embodiment

[0066] Example 2AVH57 gene expression mode analysis in the process of manfrontimusca infection

[0067] 1. Ethanfurus infected apple

[0068] The bacterial block preserved at 4 ° C refrigerators was activated on the V8 solid medium, 25 ° C culture 24-36 h. The sterilization petri dish (9 cm diameter) placed a suitable size sterilized filter paper, and wets with sterilization water. Sterilization of sterile operators. Wipe the surface with alcohol cotton, sterilize. Use a sterile sterilizer to sterilize the knife with a lightweight lamp, cut about 1cm in the apple 2The small square, remove the flesh. Pick the colonies out place in the filter paper of the sterilized bacteria to the cutting of the surface of the apple, only three sides, as well as a control of blank solid medium. Use a degreasing cotton plug in the incision, sprinkle the sterilized tap water. Put it into the incubation in a constant temperature incubator with a tray. After infection of 24h, 48h, 72h, 96h, collected b...

Example Embodiment

[0082] Example 3AVH57 gene expression transiently in tobacco

[0083] 1.PVX recombinant expression vector construction

[0084] (1) Sma I-excreted PCR amplification target gene fragment AVH57, recovered insertion piece, with a size of about 567 bp.

[0085] Reaction system: DDH 2 O (33 μL), Sma I (2 μL), plasmid (10 μL), 10 x Cut Buffer (5 μL). 37 ° C, 30 min.

[0086] (2) Connect the PGR107 vector of the same enzyme to transform E. coli DH5α.

[0087] (3) The transformed DH5α was screened by Kan resistance, and the plasmid was extracted after the colonies were extracted after 37 ° C shaker overnight.

[0088] (4) The recombinant plasmid was subjected to the recombinant plasmid using restriction endonuclease SMAI. The correct recombinant plasmid was sequencing in the correct recombinant plasmid to send the enzyme. The recombinant plasmid of the DNA fragment of the SEQ ID NO: 2 was inserted between the enzyme digestion site SmaI of the PGR107 vector was named PGR107 / AVH57 (plasmi...

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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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