Purpureocillium lilacinum Pth11 gene and functional verification method thereof

A kind of lilac violaceum, gene technology, applied in the field of lilac violaceum Pth11 gene and its function verification, can solve the problems of research, lack of gene level function elaboration and the like

Active Publication Date: 2022-07-05
INST OF PLANT PROTECTION SICHUAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no in-depth study on the process of P. lilacensis invading nematode eggs, and there is a lack of functional elucidation at the gene level.

Method used

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  • Purpureocillium lilacinum Pth11 gene and functional verification method thereof
  • Purpureocillium lilacinum Pth11 gene and functional verification method thereof
  • Purpureocillium lilacinum Pth11 gene and functional verification method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0080] In this example, a gene Pth11-rg1, which is homologous to the transmembrane protein coding gene of rice blast, was screened from the transcriptome and genome sequencing data of P. lilacinus, and its cDNA sequence is shown in SEQ ID NO. 1, and its nucleic acid sequence As shown in SEQ ID NO.3.

[0081] The following steps were used to prepare a genetically engineered strain ΔPth11-rg1 knocking out part of Pth11-rg1 and to verify its gene function:

[0082] ①Collect conidia after routine culture of lilac spore, and adjust the concentration of spore solution to 1×10 5 CFU / mL. Inoculate 100 μL of the above spore liquid into 100 mL of TG medium, and shake at 28° C. and 150 rpm for 24-36 hours until tiny hyphal balls appear (the hyphae at this time are young hyphae). The mycelia were collected by filtration with 4 layers of sterile lens paper, rinsed with a small amount of 0.7M NaCl solution to remove excess medium, and transferred to a 50 mL sterilized Erlenmeyer flask.

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Embodiment 2

[0108] In this example, a gene Pth11-rg2, which is homologous to the transmembrane protein encoding gene of rice blast, was screened from the transcriptome and genome sequencing data of P. lilacinus, and its cDNA sequence is shown in SEQ ID NO. 2, and its nucleic acid sequence As shown in SEQ ID NO.4.

[0109] The following steps were used to prepare a genetically engineered strain ΔPth11-rg2 knocking out part of Pth11-rg2 and to verify its gene function:

[0110] ③ Collect conidia after routine culture of lilac spore, and adjust the concentration of spore fluid to 1×10 5 CFU / mL. Inoculate 100 μL of the above spore liquid into 100 mL of TG medium, and shake at 28° C. and 150 rpm for 24-36 hours until tiny hyphal balls appear (the hyphae at this time are young hyphae). The mycelia were collected by filtration with 4 layers of sterile lens paper, rinsed with a small amount of 0.7M NaCl solution to remove excess medium, and transferred to a 50 mL sterilized Erlenmeyer flask.

...

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Abstract

The invention provides two Purpureocillium lilacinum genes Pth11-rg1 and Pth11-rg2 which are homologous with a rice blast transmembrane protein coding gene pth11, the Pth11 gene is subjected to function shielding by utilizing a gene knockout technology to obtain deltaPth11-rg1 and deltaPth11-rg2 gene engineering strains, and the functions of the Pth11-rg1 and Pth11-rg2 genes of the Purpureocillium lilacinum are verified by utilizing the gene engineering strains. Therefore, primary identification of functions on the gene level can be efficiently carried out.

Description

technical field [0001] The present application relates to the field of biological control of diseases and insect pests, and in particular, to the Pth11 gene of Pseudomonas lilac and its function verification method. Background technique [0002] Plant-parasitic nematodes infest various crops including rice, wheat, maize, soybean, vegetables, tobacco, fruit trees, and forest trees, and are estimated to cause approximately US$80 billion in losses worldwide each year (Jones et al 2013). Paecilomyces lilacinus (Purpureocillium lilacinum) Pathogenic nematodes. The bacterium is widely distributed and has strong adaptability, and is an important application and research object for nematode biological control at home and abroad. [0003] At home and abroad, there are commercial products of lilac spore preparations, which are used to control root-knot nematodes and cyst nematodes, and have good effects. However, the process of the invasion of nematode eggs by P. lilace has not bee...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N1/15C12Q1/686C12Q1/06C12Q1/04A01K67/033C12R1/79
CPCC07K14/37C12Q1/686C12Q1/06C12Q1/04A01K67/033C12Q2521/107C12Q2563/107C12Q2545/101Y02A50/30
Inventor 姬红丽谢家廉杨芳徐幸于文娟
Owner INST OF PLANT PROTECTION SICHUAN ACAD OF AGRI SCI
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