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Radopholus similis associated bacteria with potential of genetic transformation

A technology for banana nematodes and associated bacteria, applied in the direction of bacteria, nematicides, biocides, etc., can solve the problem of no biocontrol effect of banana nematode associated bacteria, and achieve strong genetic transformation potential, good application prospects, and stable associated relationships Effect

Active Publication Date: 2018-05-11
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no research report on the associated bacteria of banana nematodes and their biocontrol effects

Method used

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  • Radopholus similis associated bacteria with potential of genetic transformation
  • Radopholus similis associated bacteria with potential of genetic transformation
  • Radopholus similis associated bacteria with potential of genetic transformation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Acquisition of Pseudomonas fluorescens pf36, an associated bacterium of the banana borer nematode

[0032] 1. Isolation and purification of the bacteria associated with the banana borer nematode and the endophytic bacteria of the carrot callus

[0033] (1) Isolation of bacteria associated with the banana borer nematode

[0034] In order to ensure that the isolated bacteria have a stable companionship with the nematode spp., 10 populations of stencils from different host plants (Table 1) were subcultured on the carrot callus for more than 3 times. The subcultured banana borer nematodes (mixed nematodes) were collected and placed in 1.5 ml sterilized centrifuge tubes (about 1000). Centrifuge at 4500rpm for 45s, discard the supernatant, wash the nematodes repeatedly 3 times, then grind the nematodes collected at the bottom of the tube thoroughly with a sterilized glass rod, add 1ml of sterile water and shake well, absorb 50μl of the grinding solution and spread ...

Embodiment 2

[0071] Example 2 Application of Pseudomonas fluorescens associated with banana nematode in the control of banana nematode

[0072] 1. Introduction of protease pase4 gene into bananas to wear the associated bacteria - Pseudomonas fluorescens

[0073] (1) Conversion method

[0074] The protease pase4 gene was introduced into the target bacteria Pseudomonas fluorescens pf36 through vector transformation and conjugation transposition respectively. The nucleotide sequence of the protease pase4 gene is shown in SEQ ID NO.1.

[0075] ① Vector transformation method

[0076] Using the recombinant plasmid pase4-pEASY connected with the protease pase4 gene preserved in the laboratory as a template, use the following primers to amplify the protease pase4 gene.

[0077] Upstream primer M1:

[0078]

[0079] The lower horizontal line is the EcoR I restriction site, and the wavy line is the ribosome binding site;

[0080] Downstream primer M2:

[0081]

[0082] The lower horizont...

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Abstract

The invention discloses radopholus similis associated bacteria with the potential of genetic transformation. The associated bacteria are Pseudomonas fluorescens (pf36), and are preserved in GuangdongMicrobial Culture Collection Center on November 8, 2017, and the preservation number is GDMCC No:60278. One dominant associated bacterium is separated and screened from multiple populations of radopholus similis, and the dominant associated bacterium is the Pseudomonas fluorescens (pf36) which has the very stable associated relationship with the radopholus similis, a genetically modified target bacterium is provided for mediated expression of nematicidal protease genes, and due to the fact that the associated relationship of the target bacterium and the radopholus similis is improved to express the nematicidal protease genes, the target bacterium can be used for biological control over the radopholus similis, and the associated bacteria are safe to crops. Accordingly, a new idea and methodare provided for control over the plant nematode, that is to say, biological control over nematode is achieved by means of nematode associated bacterium mediated nematicidal protease genes.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to a bacterium associated with the nematode nematode punctatus with genetic modification potential. Background technique [0002] Plant nematode disease is a plant disease caused by the attack and parasitism of plant parasitic nematodes. Injured plants can affect normal growth and development due to invading nematodes absorbing nutrients in the body; secretions during nematode metabolism can also stimulate the cells and tissues of host plants, resulting in plant deformities, and control by natural enemies. Reduce the production and quality of agricultural products. Among them, Radopholus similis, also known as banana root rot, is the most dangerous obligate parasitic nematode in bananas, flower planting and many commercial crops. Its host range is very wide. There are more than 350 kinds of host plants reported. . In addition to serious harm to bananas, peppers and co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/21A01N63/00A01P5/00C12R1/39
CPCA01N63/00C12N9/50C12N1/205C12R2001/39
Inventor 谢辉王东伟陈德强丁善文其他发明人请求不公开姓名
Owner SOUTH CHINA AGRI UNIV
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