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LbKN1 gene derived from Lubao No.1 and application of LbKN1 gene

A gene and genetic modification technology, applied in the field of molecular biology and plant protection, can solve the problems of production and application in trouble, loss of production and application value, and demise.

Active Publication Date: 2020-08-18
INST OF PLANT PROTECTION SHANDONG ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, after a long period of artificial cultivation and preservation, the multinucleated spores of the "Lubao No. 1" strain increased, and the pathogenicity of the fungal agent to dodder was severely reduced, and even lost its production and application value.
The severe degradation of the strain put the production and application of "Lubao No. 1" in a difficult situation, almost on the verge of extinction

Method used

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  • LbKN1 gene derived from Lubao No.1 and application of LbKN1 gene
  • LbKN1 gene derived from Lubao No.1 and application of LbKN1 gene
  • LbKN1 gene derived from Lubao No.1 and application of LbKN1 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Cloning of genes related to pathogenicity

[0020] 1.1 Acquisition of positive mutants

[0021] The "Lubao No. 1" strain can be obtained through commercial purchase, such as the China Center for Microorganism Culture Collection (CMCC), and the platform number is Bio-17420. After the above-mentioned bacterial strains were activated, they were cultured on a 9cm PDA culture plate. After 10 days, the conidia on the surface of the colony were washed with 6 mL / dish of sterile water. After filtration, the concentration of spores in the filtrate was observed under a microscope through a hemocytometer, and diluted to The concentration of spores is 1×10 5 pc / mL, spare.

[0022] A single colony of Agrobacterium AGL-1 containing the freshly activated binary vector pBHt2 was picked, inoculated in 2 mL of LB liquid medium (kanamycin 50 μg / mL), and cultured at 220 rpm and 28 °C for 1 day. Take 250 μL of the shaken Agrobacterium solution and place it in 50 mL of MM medium ...

Embodiment 3

[0037] Example 3 Phenotypic functions of mutant strains

[0038] With the wild-type bacterial strain of Lubao No. 1 and the obtained in embodiment 1 wxya The deletion mutant strain was activated on PDA medium, and after 6 days of culture at 28 °C, the edge of the colony was picked and cultured on a new PDA medium. After 7 days of culture, it was used to complete the analysis of the mutant's biological properties, sporulation and pathogenicity Determination.

[0039] Refer to Example 1.2 to obtain the relevant method of highly pathogenic mutants to verify the control effect of the strain. After 10 days of inoculation, the remaining dodders in the same area after the control of the mutant and the wild type were picked, weighed, and compared with those of the clean water control group. Control effect (%) = (weight of dodder in control group - weight of dodder in inoculation group) / weight of dodder in control group. The inoculation groups were mutant and wild type, respective...

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Abstract

The invention provides an LbKN1 gene derived from Lubao No.1. An amino acid sequence coded by the LbKN1 gene is shown as SEQ ID NO: 1. The LbKN1 gene can be used for modifying a colletotrichum acutatum dodder specialized type so as to prevent and treat dodder, and the modification is deletion, knockdown or knockout. The LbKN1 gene provided by the invention can be used for regulating and controlling the pathogenicity of colletotrichum acutum dodder seed specialized type (Colletotrichum acutum f. Sp. Cutanea) on the dodder seeds, so that the pathogenicity of the colletotrichum acutum dodder seedspecialized type (Colletotrichum acutum f. Cutanea) on the dodder seeds, specifically, the spore production capacity of the colletotrichum acutatum dodder specialized strain without the LbKN1 gene isremarkably higher than that of a wild strain. The LbKN1 gene participates in a biocontrol bacterium pathogenicity related process, and can be applied to the field of gene engineering for improving the pathogenicity of Lubao No.1 and creating an efficient biocontrol bacterium agent.

Description

technical field [0001] The invention belongs to the field of molecular biology and plant protection, and specifically relates to a wxya Gene and application thereof, specifically related to a special type of dodder derived from anthracnose oxysporum ( Colletotrichum acutatum f. sp. cuscuta ) of "Lubao One" wxya Genes and their applications. Background technique [0002] According to statistics from the National Agricultural Technology Extension Service Center, in 2017, the area of ​​weeds in my country reached 1.38 billion mu, and the direct economic loss caused by weed damage was close to 100 billion yuan. In 2016, the sales of herbicides in my country was 1.972 billion US dollars, accounting for 46% of the total sales of pesticides in my country, ranking first among all kinds of pesticides. Herbicides have become one of the important factors that seriously threaten my country's food and environmental safety. Therefore, the research and development and demand for en...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N1/21A01N63/20A01P13/02C12R1/01
CPCC07K14/195A01N63/20
Inventor 李健李美高兴祥房锋
Owner INST OF PLANT PROTECTION SHANDONG ACAD OF AGRI SCI
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