Method for identifying pathogenicity of fusarium verticillioides indoors

A technology for indoor identification of Fusarium verticillium, applied in the field of indoor identification of pathogenicity of Fusarium pseudoverticillium

Active Publication Date: 2019-07-23
INST OF PLANT PROTECTION HEBEI ACAD OF AGRI & FORESTRY SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no mature method for indoor identification of the ...

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  • Method for identifying pathogenicity of fusarium verticillioides indoors
  • Method for identifying pathogenicity of fusarium verticillioides indoors
  • Method for identifying pathogenicity of fusarium verticillioides indoors

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preparation example Construction

[0039] The present invention does not have special limitation to the composition raw material of described PDA plate culture medium and preparation method, adopts the conventional PDA plate culture medium of this area to get final product; In the specific implementation process of the present invention, described PDA plate culture medium is preferably prepared by comprising the following raw materials Obtain: 200g of potatoes, 20g of glucose, 20g of agar and 1000mL of distilled water.

[0040] The present invention has no special limitation on the inoculation amount of the Fusarium pseudoverticillium; the environment for the cultivation is preferably a dark environment; the time for the cultivation is preferably 3 to 5 days, more preferably 4 days; the temperature for the cultivation is preferably 25 to 30°C, more preferably 28°C.

[0041] After obtaining a single colony of Fusarium pseudoverticillium, the present invention cuts off the bacteria-carrying medium block along the...

Embodiment 1

[0055] Embodiment 1 A kind of indoor identification method of Fusarium pseudoverticillium pathogenicity (inoculation site: corn silk)

[0056] 1. Materials: fresh filaments of variety Zheng 58, Fusarium pseudoverticillium mutant strains (216 strains) to be identified for pathogenicity.

[0057] 2. The method for obtaining the mutant strains of Fusarium pseudorhiza with pathogenicity to be identified

[0058] Cultivation of mutant strains of Fusarium pseudorhiza

[0059] a. Fusarium pseudoverticillium is a strain with pathogenicity to corn ears preserved by the Institute of Plant Protection, Hebei Academy of Agriculture and Forestry Sciences (this strain is the wild-type strain CK).

[0060] b. Establish and optimize the transformation system of the Fusarium pseudomonas mutant with the preserved bacterial strain, the transformation method is as follows:

[0061] ① Transformation of Agrobacterium: Purchase the constructed Agrobacterium tumefaciens AGL-1 (with a bivalent vector...

Embodiment 2

[0087] Example 2 Field inoculation of the weakly pathogenic strain of Fusarium pseudoverticillium screened in Example 1.

[0088] (1) Purpose of the test: back inoculation in the field to screen weak pathogenicity strains among Fusarium verticillium transformants.

[0089] (2) Test material: inbred variety Zheng 58.

[0090](3) Back-inoculation strains: No. 110 weak pathogenicity strain identified in Example 1, No. 38 strong pathogenicity strain (select a bacterial strain randomly from 215 strong pathogenicity strains), wild-type bacterial strain WT (Hebei Province A pathogenic strain to corn ears preserved by the Institute of Plant Protection, Academy of Agriculture and Forestry).

[0091] (4) Tie-back method: use the filament smear method to keep the 1-2cm filament protruding from the bract leaf. Cut off the rest, apply the cultured strains in the petri dish directly on the trimmed filaments, and cover with pollination bags and mesh bags to prevent the invasion of other di...

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Abstract

The invention provides a method for identifying pathogenicity of fusarium verticillioides indoors, and belongs to the technical field of prevention and treatment of corn diseases. The method comprisesthe following steps of (1) culturing the fusarium verticillioides indoors to obtain a fusarium verticillioides single bacterial colony; (2) cutting off a culture medium block having bacteria along the outer edge of the fusarium verticillioides single bacterial colony obtained in the step (1) to obtain bacterium cakes; and (3) taking fresh corn pistils, enabling the taken fresh corn pistils to beattached and put on the bacterium cakes obtained in the step (2) in parallel, performing airtight standing for 5-9d, observing the pathogenesis situation of the corn pistils, and determining the pathogenicity of the fusarium verticillioides. The method disclosed by the invention is operated indoors, the operation course is simple, the work load is small, and human resources can be saved; besides,in the implementation course, temperature and humidity are easy to control and are not liable to influence by external environment, and the experimental result is accurate, stable, reliable and low inerrors. After verification, the identification result is consistent with the identification result of the pathogenicity of land for growing field crops.

Description

technical field [0001] The invention relates to the technical field of corn disease prevention and control, in particular to a method for indoor identification of the pathogenicity of Fusarium pseudoverticillium. Background technique [0002] Maize ear rot is a fungal disease that seriously threatens maize production in various maize producing areas in the world. It has occurred in various parts of our country and is becoming more and more serious. It directly affects the yield and quality of corn, and often causes serious economic losses. [0003] At present, the reported pathogenic fungi of corn ear rot mainly include Fusarium spp., Penicillium spp., Trichoderma spp., Helminthosporium spp. and Aspergillus ( Aspergillus spp.), etc., wherein Fusarium is the main pathogen, and Fusarium verticillioides is the main pathogen that causes corn ear rot. A weakly pathogenic Fusarium pseudoverticillium strain was produced by Agrobacterium-mediated transformation, which has immune p...

Claims

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Application Information

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IPC IPC(8): C12Q1/18
CPCC12Q1/18
Inventor 丁梦军李坡石洁马红霞金戈刘树森张家齐郭宁
Owner INST OF PLANT PROTECTION HEBEI ACAD OF AGRI & FORESTRY SCI
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