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A African Trichoderma harzianum for disease prevention and growth promotion and its application

A technology for strains and diseases, applied in the field of microbial pesticides, can solve the problems of ecological environment pollution of chemical agents, drug resistance of pathogenic bacteria, and sharp decline in biodiversity

Active Publication Date: 2020-03-31
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The long-term large-scale use of chemical agents has caused a series of problems such as ecological environment pollution, sharp decline in biodiversity, and drug resistance of pathogenic bacteria.

Method used

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  • A African Trichoderma harzianum for disease prevention and growth promotion and its application
  • A African Trichoderma harzianum for disease prevention and growth promotion and its application
  • A African Trichoderma harzianum for disease prevention and growth promotion and its application

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Experimental program
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Effect test

Embodiment 1

[0080] Embodiment 1, the isolation and identification of Trichoderma afroharzianum (Trichoderma afroharzianum) TM2-4 in Africa

[0081] 1.1 Strain isolation

[0082] The strain was isolated from soil samples of special habitat in Wulingyuan virgin forest in Hunan, China. Bacterial strains were separated by the dilution plate method, and the specific operation method was as follows: Weigh 10 g of the soil sample and put it into 90 ml of sterile water to vibrate evenly to obtain 10 -1 Concentration of bacterial solution, use a 1ml sterile pipette to draw 1ml 10 -1 Concentration of bacterial solution in a tube of 9ml sterile water, shake well that is 10 -2 Concentration of bacterial solution, the same method, sequentially diluted to 10 -4 . the above 10 -2 、10 -3 and 10 -4 The soil bacteria solution was spread on the PDA plate respectively, dried and placed upside down in an incubator at 25°C for 3-5 days. Pick a single colony that resembles Trichoderma and transfer it to...

Embodiment 2

[0093] Embodiment 2, African Harz Trichoderma (Trichoderma afroharzianum) TM2-4 is to the inhibition or heavy parasitism of testing pathogenic fungus

[0094] Pick Trichoderma afroharzianum (Trichoderma afroharzianum) TM2-4 bacterial cake with a diameter of 7 mm grown on the plate and the bacterial cake of the pathogenic fungus to be tested, and inoculate them symmetrically on the PDA medium at a position 2 cm away from the center, and the control does not inoculate African Trichoderma afroharzianum (Trichoderma afroharzianum) TM2-4 bacteria cake, only inoculate the bacteria cake of the pathogenic fungus tested, each treatment was repeated three times, and cultured at 25°C for 4 days to observe the growth of the pathogenic fungus. The pathogenic fungi tested included: Botrytis cinerea, Monilinia fructicola, Colletotrichum capsici, Fusarium oxysporum f.sp.cucumerinum.

[0095] Under confrontation culture conditions, Trichoderma afroharzianum TM2-4 can inhibit and cover tomato g...

Embodiment 3

[0096] Embodiment 3, Trichoderma afroharzianum (Trichoderma afroharzianum) TM2-4 in vitro control effect on tomato gray mold

[0097] 3.1 Preparation of Trichoderma afroharzianum TM2-4 sterile fermentation filtrate

[0098] Pick the Trichoderma afroharzianum TM2-4 grown on the plate with the inoculation loop and inoculate it into a 300mL Erlenmeyer flask containing 100mL of liquid fermentation medium, and culture it with shaking at 28°C and 180r / min for 2 days to obtain the seed liquid, and press 5 % inoculum size Transfer the seed liquid to a 500mL Erlenmeyer flask containing 100mL liquid fermentation medium, 28°C, 180r / min shaking fermentation culture for 7d, and collect the fermentation broth. The fermented liquid was centrifuged at 12000 rpm for 10 min to take the supernatant, which was sterilized by filtration with a 0.22 μm microporous membrane to obtain a sterile fermented filtrate of Trichoderma afroharzianum TM2-4. The aseptic fermentation filtrate of Trichoderma afr...

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Abstract

The invention discloses a African Trichoderma harzianum for disease prevention and growth promotion and application thereof. The strain number of Trichoderma harzianum is TM2-4, and its registration number in the General Microorganism Center of China Committee for Culture Collection of Microorganisms is CGMCC No.15881. Trichoderma harzianum M2‑4 has inhibitory effects on tomato cinerea, peach brown rot, pepper anthracnose and cucumber wilt on fruit and vegetable crops; Germination and plant growth are promoted. Trichoderma harzianum TM2‑4 has indoleacetic acid (IAA)-producing activity, siderophilic activity, glucanase-producing activity and chitinase-producing activity. Trichoderma harzianum TM2‑4 has a fast propagation speed, can be artificially cultured, and the culture conditions are simple. The highest spore production in the bran corncob nutrient solution solid-state fermentation medium can reach 3.2×10 9 cfu / g solid bacterial agent.

Description

technical field [0001] The invention relates to a disease-preventing and growth-promoting Trichoderma harzianum and its application in the technical field of microbial pesticides. Background technique [0002] The pathogenic bacterium of tomato gray mold is Botrytis cinerea of ​​the subphylum Botrytis. Botrytis cinerea has a wide range of hosts, not only infecting tomatoes, cucumbers, eggplants, peppers, lettuce, cabbage, onions, beans and other vegetables, but also infecting apples, strawberries, citrus, grapes and other fruit trees. [0003] The frequent occurrence of fungal diseases is one of the main factors affecting the development of the protected horticultural fruit and vegetable crop industry. At present, in addition to breeding excellent disease-resistant varieties and strengthening crop cultivation and management, the measures of spraying chemical agents are mainly used for the prevention and control of gray mold diseases of fruits and vegetables. The long-term ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14A01N63/38A01P3/00A01P21/00C12R1/885
CPCA01N63/30C12N1/14C12N1/145C12R2001/885
Inventor 赵娟刘伟成刘霆刘德文张殿朋吴慧玲董丹张涛涛田兆丰
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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