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Phytophthora cinnamomum effector protein SCR93258 and application thereof

A technology of effector protein and Phytophthora, which is applied in the fields of application, angiosperm/flowering plants, and the use of vectors to introduce foreign genetic materials, etc. It can solve the problems of failure to achieve ideal control effects, difficulties in the prevention and control of Phytophthora camphora, and increase the burden on farmers, etc. question

Active Publication Date: 2022-07-01
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to causing huge economic losses in agriculture, forestry and horticulture, the inadvertent introduction of this pathogen has disastrous consequences for natural ecosystems and biodiversity
At home and abroad, spraying pesticides is mainly used to control the prevalence and harm of Phytophthora camphora, but the ideal control effect has not been achieved, which not only increases the burden on farmers, but also causes environmental pollution.
Although the growth of Phytophthora camphora can be inhibited by using environmentally friendly methods such as mulching and composting to allow other microorganisms to proliferate and secrete cellulase in the mulch, the distribution scale of Phytophthora camphora and the ability to survive in soil or asymptomatic plants for several years , so that the control of Phytophthora camphora disease still faces great challenges and difficulties

Method used

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  • Phytophthora cinnamomum effector protein SCR93258 and application thereof
  • Phytophthora cinnamomum effector protein SCR93258 and application thereof
  • Phytophthora cinnamomum effector protein SCR93258 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Acquisition of the gene encoding the effector protein SCR93258 of Phytophthora cinnamomea

[0046] The strain of Phytophthora cinnamomum was selected as the test material, and the genome sequence of Phytophthora cinnamomum was analyzed according to the reported effector protein gene information, and the effector protein gene in the whole genome of Phytophthora cinnamomum was obtained. Then, primers p1 and p2 are designed according to the obtained gene fragments, and the obtained target gene fragments are amplified and screened.

[0047] 1. Using CTAB-SDS method to extract high-quality Phytophthora cinnamomum genomic DNA

[0048] Phytophthora cinnamomum strains were cultured on solid V8 medium plate (recipe: 170ml V8 vegetable juice plus 1.6g calcium carbonate, mixed, centrifuged at 2000rpm for 5min to take the supernatant, add purified water to 1L, add 15g agar powder, autoclave Bacteria pot for 20min standby), after culturing for 3 days in a 25°C biochemical...

Embodiment 2

[0062] Example 2 Analysis of the expression pattern of effector SCR93258 in the process of Phytophthora cinnamomea infecting apples

[0063] 1. Phytophthora cinnamomum strains infect apples

[0064] The mycelial blocks stored in the refrigerator at 4°C were placed on V8 solid medium for activation, and cultured at 25°C for 24-36 hours. A suitable size of sterile filter paper was placed at the bottom of the sterile petri dish (diameter 9 cm) and moistened with sterile water. Sterilize on sterile bench. Wipe the surface of the apple with alcohol cotton to sterilize it. Use the outer flame of an alcohol lamp to sterilize the edge of the cutter, cut the apple into small cubes of about 1cm2 on all four sides, and remove the pulp. Pick out the colonies and put them on sterilized filter paper and plug them into the incision on the surface of the apple. Only three sides are plugged, and the other side is used as a control of blank solid medium. Plug the incision with absorbent cot...

Embodiment 3

[0075] Example 3 Transient expression of effector SCR93258 in tobacco

[0076] 1. Construction of PVX recombinant expression vector

[0077] (1) The target gene fragment SCR93258 was amplified by SmaI digestion and PCR, and the inserted fragment was recovered with a size of about 291 bp.

[0078] Reaction system: ddH 2 O (33 μL), SmaI (2 μL), plasmid (10 μL), 10×cut Buffer (5 μL). 37℃, 30min.

[0079] (2) ligated with pGR107 vector digested by the same enzyme, and transformed into Escherichia coli DH5α.

[0080] (3) The transformed DH5α was screened for Kan resistance, and the colonies were obtained and the plasmids were extracted after shaking at 37°C overnight.

[0081] (4) The recombinant plasmid was identified by restriction endonuclease SmaI. The recombinant plasmids that were initially identified correctly by enzyme digestion (two target bands of about 10098 bp and 291 bp in size were obtained) were sent to GenScript Bio Co., Ltd. for sequencing. The recombinant pl...

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Abstract

The invention discloses an SCR (Selective Catalytic Reduction) effector protein SCR93258 from phytophthora cinnamomum as well as a coding gene and application of the SCR effector protein SCR93258. The amino acid sequence of the protein is SEQ ID No.1, and the nucleotide sequence for coding modified amino acid is SEQ ID No.2. Experiments prove that the protein expressed under the control of the gene can participate in the process that phytophthora infects nicotiana benthamiana, and plant cell death induced by apoptosis precursor protein Bax is caused. The invention has important significance for enriching a molecular mechanism of interaction of plant phytophthora cinnamomum and hosts and establishing a comprehensive prevention and control technology of plant phytophthora cinnamomum diseases.

Description

technical field [0001] The invention belongs to the technical field of Phytophthora cinnamomum effector, and in particular relates to a Phytophthora cinnamomum effector protein SCR93258 and its application. Background technique [0002] Pathogens can secrete an effector protein, which is a class of exocrine protein molecules that alter host plant cell structure and metabolic pathways to promote successful infection of host plants or initiate host defense responses. Many plant pathogens secrete effector proteins, such as those found in bacteria, fungi, oomycetes, and nematodes. There are many types of effectors in plant pathogens, mainly including apoplast effectors and cytoplasmic effectors. Apoplast effectors include elicitors, necrosis and ethylene-inducing peptide 1-like proteins (NLPs), PcF / SCR proteins, Cellulose binding elicitor and lectin-like , CBEL) and enzyme inhibitors. Cytoplasmic effectors include the RxLR and CRN (Crinkler) protein families. [0003] During...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/37C12N15/31C12N15/82C12N5/10C12N1/21A01H5/00A01H6/82
CPCC07K14/37C12N15/8263Y02A50/30
Inventor 戴婷婷周紫薇焦彬彬何海彬夏宏明
Owner NANJING FORESTRY UNIV