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Kit for biological control of mikania micrantha by use of mikania micrantha wilt virus and method

The technology of Mikania micrantha and a kit is applied in the field of comprehensive prevention and control of alien invasive pests, which can solve the problems of high cost, environmental pollution, high labor intensity, etc., achieve good social benefits, avoid environmental pollution, and achieve good control effects.

Active Publication Date: 2013-10-23
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, manual removal is labor-intensive, low in efficiency, high in cost, and difficult to eradicate; chemical control is easy to cause environmental pollution, and biological control is the most promising method to control alien invasive species
The Mikania micrantha wilt virus discovered so far can cause symptoms such as wilting of Mikania micrantha leaves, internode dwarfing, top wilting, etc., and severely inhibit the growth of Mikania micrantha.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Isolation and screening of Mikania wilt virus:

[0048] 1) Take the diseased Mikania micrantha plant leaves identified by molecular biology methods to contain Mikania wilt virus, add 2 times the volume (w / v) of pre-cooled 0.5mol / L potassium phosphate buffer (KPB) to the diseased leaves at 25-27°C , pH7.5, containing 0.1% mercaptoethanol by volume fraction, 0.01% Triton X-100 by mass fraction, and 2.5% sucrose by mass fraction), homogenate for 2 min, stand at 4°C for 1 h, centrifuge at 8000 g for 20 min, and take the supernatant solution, add PEG, NaCl and Triton X-100 to make the concentration reach 6%, 0.1mol / L, and 2.5% respectively, stir at 4°C for 6h, centrifuge at 8500g for 5min, take the precipitate, and use 0.01mol / L KPB (pH7.5, containing 0.01% Triton X-100 mass fraction), fully suspended, centrifuged at 7000g for 10min, absorbed supernatant and placed in a centrifuge tube, precipitated and resuspended and centrifuged, repeated 2 times, combined supernatant , th...

Embodiment 2

[0052] Mikania wilt virus biocontrol kit for Mikania micrantha, comprising the following components:

[0053] 1) Mikania wilt virus toxin (stored at -20°C);

[0054] 2) 0.5mol / L potassium phosphate buffer (KPB, pH7.5, containing 0.1% mercaptoethanol by volume, 0.01% Triton X-100 by mass, and 2.5% sucrose by mass);

[0055] 3) Polyethylene glycol (PEG-6000);

[0056] 4) Sodium chloride (NaCl);

[0057] 5) Triton X-100;

[0058] 6) 0.01mol / L KPB (pH7.5, containing 0.01% Triton X-100 by mass);

[0059] 7) Carborundum (400-600 mesh);

[0060] 8) 0.01mol / L phosphate buffer (pH7.0).

Embodiment 3

[0062] A method for biologically controlling Mikania micrantha using the kit of Example 2, comprising the following steps:

[0063] (1) Dilute the toxin of Mikania micrantha wilt virus with phosphate buffer, and inoculate it with conventional juice at 25-27°C. When the broad bean seedlings have 3 to 4 true leaves, the first and second leaves on the top Sprinkle a layer of quartz sand evenly on the true leaves, dip the inoculation solution with a brush and rub it gently on the inoculated leaves, and then rinse off the remaining substances on the leaves with distilled water. Mikania wilt virus propagates on broad beans, and the diseased leaves are collected after 7-10 days;

[0064] (2) Add 30-50mL 0.01mol / L phosphate buffer (pH7.0) per 1g of fresh diseased leaves, grind thoroughly, centrifuge at 3000r / min for 15min, and take the supernatant for later use;

[0065] (3) Add 0.5g of 400-600 mesh corundum per 100mL of supernatant, mix well, and use a spray gun to spray and inocula...

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PUM

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Abstract

The invention discloses a kit for biological control of mikania micrantha by use of mikania micrantha wilt virus and a method, and belongs to the technical field of comprehensive prevention and control of invasive alien pests. According to the invention, a strain of the mikania micrantha wilt virus is obtained by separating and screening, and an expanding and propagation method of the mikania micrantha wilt virus is disclosed. Through use of the kit for control of the mikania micrantha by use of the mikania micrantha wilt virus, diffusion of the mikania micrantha can be effectively controlled through spraying inoculation of the mikania micrantha wilt virus, shortcomings of control of the mikania micrantha by use of physical and chemical methods can be overcome, and environmental pollution can be avoided. The kit for biological control of the mikania micrantha by use of the mikania micrantha wilt virus has the advantages of convenience and quickness, strong specificity, time and labor saving, and good control effect, and can obtain good social benefits, economic benefits and ecological benefits.

Description

technical field [0001] The invention belongs to the technical field of comprehensive prevention and control of alien invasive pests, and in particular relates to a kit and method for biologically controlling Mikania micrantha wilt virus (MMWV). Background technique [0002] Mikania micrantha H.B.K. is a perennial vine belonging to the genus Mikania Willd in the Compositae family. The biological invasion of Mikania micrantha has caused serious harm to the ecological environment in South China, and the control of Mikania micrantha has become a worldwide problem. Control techniques include manual and mechanical removal, chemical control of herbicides, biological control using natural enemies and pathogenic bacteria, etc. Biological control is an effective method to control the invasion of harmful organisms. Among them, manual removal is labor-intensive, low in efficiency, high in cost, and difficult to eradicate; chemical control is easy to cause environmental pollution, and b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N7/02A01N63/00A01P13/00C12R1/94
Inventor 王瑞龙曾任森郑子豪任勇张晖宋圆圆苏贻娟
Owner SOUTH CHINA AGRI UNIV
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