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Phytophthora camphora effector protein avh49 and its application

An effector protein, avh49 technology, applied in the fields of application, recombinant DNA technology, angiosperms/flowering plants, etc., can solve the problems of different structure of different effector proteins, achieve the effect of reducing plant mortality and great application value

Active Publication Date: 2022-02-22
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is reported in the prior art that the effector protein Avh87 gene of Phytophthora camphora 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 camphora effector protein avh49 and its application
  • Phytophthora camphora effector protein avh49 and its application
  • Phytophthora camphora effector protein avh49 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Acquisition of the gene encoding effector protein Avh49 of Phytophthora camphora

[0050] The Phytophthora camphora strain was selected as the test material, and the gene sequence of the effector gene Avh49 was obtained according to the whole genome sequence of Phytophthora camphora. Then design primers p1 and p2 according to the obtained gene fragments, and 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 camphora strains on solid V8 medium plate (recipe: 340ml V8 fruit juice plus 3.4g calcium carbonate, mix well with glass rod, filter the liquid with two layers of filter cloth, add nine times the volume of pure water to dilute, take 200ml in 250ml Add 3g of agar powder to the Erlenmeyer flask, and sterilize at 121°C for 20 minutes for later use.) After culturing in a constant temperature incubator at 25°C for 3 days, take 8-12 pi...

Embodiment 2

[0065] Example 2 Homology analysis between different effector proteins

[0066] According to the analysis of the genome information of Phytophthora camphora, the amino acid sequence encoded by Av h49 cloned in this study and Phytophthora camphora (P.cinnamomi), P.cinnamomi var.robiniae and Phytophthora camphora The comparison of other effector molecules of P. cinnamomi var. parvispora showed that the homology between PcinnAvh49 and other effector proteins was not high, such as figure 2 As shown, it can be seen that there is no research reference between effector proteins, and bioinformatics analysis and functional experimental verification of gene proteins are needed to determine the function of each effector protein.

Embodiment 3

[0067] Example 3 Analysis of the expression pattern of Avh49 gene during the infection of apple by Phytophthora camphora

[0068] 1. Phytophthora camphora strains infecting apples

[0069] The mycelium pieces stored in the refrigerator at 4°C were activated on V8 solid medium, and cultured at 25°C for 3 days. Place a piece of sterilized filter paper of appropriate size on the bottom of the sterilized Petri dish and moisten it with sterilized water. Wipe the surface of the apple with an alcohol swab. Use a scalpel to cut small squares about 1 cm2 and 1 cm thick on three sides of the apple to remove the pulp. Pick out the mushroom cake and stuff it into the incision on the surface of the apple, only stuffing two sides, and one side as a blank control of the solid medium. Plug the incision with absorbent cotton and drip with sterile tap water. Pack it into a constant temperature incubator with a tray and cultivate it. After infecting for 24h, 48h, 72h, and 96h respectively, ...

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Abstract

The invention discloses an effector protein Avh49 derived from Phytophthora camphora and its coding gene and application. The protein has the amino acid sequence of SEQ ID NO: 1 in the sequence listing. Experiments have proved that the protein controlled by the gene can participate in the process of Phytophthora camphora infecting Nicotiana benthamiana, and has the function of inducing plant cell death. The invention is of great significance for enriching the molecular mechanism data information of the interaction between the plant pathogenic oomycete and the host, and establishing the comprehensive prevention and control technology strategy of the plant pathogenic oomycete disease.

Description

technical field [0001] The invention belongs to the technical field of Phytophthora camphorae effectors, and in particular relates to Phytophthora camphorae effector protein Avh49 and its application. Background technique [0002] Phytophthora cinnamomi is one of the most destructive plant pathogenic oomycetes known, with a worldwide distribution and a host range of nearly 5000 species. As a kind of semi-biotrophic pathogenic oomycete, Phytophthora camphora acts on biotrophy at the initial stage of infecting the host, and turns into necrotrophy after successful invasion. This means that Phytophthora camphora may manipulate plant cell death through a complex process in the interaction with the host; in the biotrophic stage, the inhibition of host cell death is considered to be an important infection strategy; and in the necrotrophic stage During the transition, plant programmed cell death may be exploited by it to adapt to the pathogenic process and complete the life cycle. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/37C12N15/31C12N15/82A01H5/00A01H6/82
CPCC07K14/37C12N15/8261
Inventor 戴婷婷李亚星吴翠萍周紫薇徐洁莹焦彬彬
Owner NANJING FORESTRY UNIV
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