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Bursaphelenchus xylophilus effector protein BxSCD1 and application thereof

A technology of effector protein, pine xylophilus, applied in the field of molecular biology

Active Publication Date: 2021-07-23
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a migratory plant parasitic nematode, pine xylophilus secretes effectors to inhibit PAMP-triggered basic defense responses and functional analysis of cysteine ​​during infection.

Method used

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  • Bursaphelenchus xylophilus effector protein BxSCD1 and application thereof
  • Bursaphelenchus xylophilus effector protein BxSCD1 and application thereof
  • Bursaphelenchus xylophilus effector protein BxSCD1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] 1. Transient expression of pine wood nematode candidate effector BxSCD1 in Nicotiana benthamiana

[0028] 1. Carrier construction

[0029] Using the eDNA of B. xylophilus as a template, specific primers were designed, and the BxSCD1 gene was amplified by polymerase chain reaction (PCR) using the Phanta Max Super-Fidelity DNA Polymerase (Vazyme, Nanjing, China) kit. The PCR amplification system is: 2*Phanta Max Super-Fidelity DNA Polymerase 25μL, 2×Phanta Max Buffer 1μL, dNTP Mix 1μL, forward and reverse primers 2μL, cDNA template 1-5μL, distilled water (Distillation-distillation H 2 O,ddH 2 O) Make up to 50 μL. The PCR reaction conditions were: 98°C for 5min, 98°C for 30s, 55°C for 30s, 72°C for 1min / kb, after 33 cycles, 72°C for 10min. Afterwards, the homologous recombination kit Clone Express II One Step Cloning Kit (Vazyme, Nanjing, China) was used to connect its complete coding region to the viral expression vector PVX (pGR107-3*HA). After the ligation system is...

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Abstract

The invention discloses a bursaphelenchus xylophilus effector protein BxSCD1 and application thereof, and belongs to the technical field of molecular biology. The amino acid sequence of the bursaphelenchus xylophilus effector protein BxSCD1 is as shown in SEQ ID No. 1, and the nucleotide sequence for expressing the protein is as shown in SEQ ID No. 2. The BxSCD1 is mainly located to cytoplasm, cysteine on the 117 site and cysteine on the 153 site take an important effect on structural stability of the BxSCD1, and the BxSCD1 can inhibit cytonecrosis and basic defense response caused by a bursaphelenchus xylophilus primordial associated molecular pattern (PAMP) BxCDP1 in nicotiana benthamiana and host pine trees. According to the invention, the interaction between the bursaphelenchus xylophilus and the host pine trees is discussed from inhibition of the plant immunoreaction by the effector secreted by the bursaphelenchus xylophilus, and the bursaphelenchus xylophilus effector protein BxSCD1 has the important significance for revealing the pathogenesis of the bursaphelenchus xylophilus, early detecting the occurrence of the bursaphelenchus xylophilus disease and the like.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to the effector protein BxSCD1 of pine wood nematode and its application. Background technique [0002] During the interaction between the pathogen and the host, when the pathogen infects the host and successfully invades the plant, it will secrete effectors to inhibit the immune response of the plant, enhance the disease susceptibility of the host plant, and promote the plundering of host nutrients by the pathogen , such as in the feeding process of plant pathogenic nematodes, will secrete salivary proteins to plant cells to interfere with the host's defense. Most of the resistance genes in plants can encode a class of proteins containing NBS-LRR domains, which function in cells and cause second-level immune responses. NBS-LRR proteins can specifically and directly recognize effectors, similar to receptors and ligands. Pathogen effectors can also modify the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C12N15/12C12N15/84C12N1/21A01H6/82A01H5/00A01H7/00C12R1/01
CPCC07K14/4354C12N15/8285Y02A40/146
Inventor 吴小芹温彤玥叶建仁李煜
Owner NANJING FORESTRY UNIV
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