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A mutant strain of Trichoderma suitable for compounding with hymexazol to control melon wilt, bacterial agent and its application

A technology of mutant strains and oxamidil, which can be used in applications, fungicides, biocides, etc., and can solve problems such as weak

Active Publication Date: 2021-04-27
HUNAN AGRI BIOTECH RES CENT +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, some existing Trichoderma strains are generally not very resistant to hymexazol. The higher the EC50 value, the more resistant to hymexazol, but the strains with good tolerance are not necessarily good, because generally good tolerance Poor sporulation ability, but strains with excellent sporulation ability and antagonism to hymexazol are needed in the process of biocontrol

Method used

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  • A mutant strain of Trichoderma suitable for compounding with hymexazol to control melon wilt, bacterial agent and its application
  • A mutant strain of Trichoderma suitable for compounding with hymexazol to control melon wilt, bacterial agent and its application
  • A mutant strain of Trichoderma suitable for compounding with hymexazol to control melon wilt, bacterial agent and its application

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Experimental program
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Effect test

Embodiment 1

[0040] Embodiment 1: creation of Trichoderma T45 mutant

[0041] The wild-type Trichoderma T45 was isolated from a highly antagonistic Trichoderma isolated from the watermelon rhizosphere soil in Gaoqiao Town, Changsha City for 6 years. Using EMS mutagenesis technology to create T45 mutants: Trichoderma T45 was cultured in PDA medium for 7 days and then diluted to 10 6 -10 7 cfu / ml spore concentration; take 100uL of EMS solution (pH 7.0, concentration 0.4M) and 50uL of the above spore suspension and mix them well in a sterilized 1.5mL EP tube, and shake for 4.5h in the dark, at a temperature of 28°C. After the treatment, add 50uL terminator to terminate the mutagenesis reaction; spread the treated spore suspension on a PDA plate containing 200ug / ml hymexazol active ingredient, place it at 28°C for 2-3 days, and record the colony growth respectively situation, select a single colony with strong growth potential, and subculture for 5 times.

[0042] A total of 35 mutant Trich...

Embodiment 2

[0043] Embodiment 2: Screening and identification of Trichoderma asperellum M45a

[0044] Using the plate confrontation method, inoculate Trichoderma (mutant bacteria and natural isolates) and Fusarium oxysporum (FON, FOM and FOC) on PDA medium plates respectively, and place the plates in a 28°C incubator for 72 hours of activation. Then use a hole puncher to take 0.5cm to face-to-face culture on the PDA plate. The diameter of the plate is 90mm, and the inoculation distance of the two bacteria is 50mm. Each of them is repeated 3 times, cultivated at 28°C for 5 days, and then measures the radius of the colony. Trichoderma is effective against Fusarium oxysporum The relative biocontrol activity of bacteria is expressed by the following formula:

[0045] Inhibition rate (%)Ir=(R-Ri) / R×100,

[0046] R: growth radius of the control Fusarium,

[0047] Ri: The growth radius of Fusarium in confrontation culture of Trichoderma.

[0048] Antagonistic ability is an important factor to...

Embodiment 3

[0052] Embodiment 3: Hymexazol affects the sporulation of Trichoderma

[0053] Dilute an equal amount of Trichoderma using a hemocytometer (the total amount of initial spores is 10 6 cfu) were respectively placed on 50ml PD medium with hymexazol content concentrations of 50, 100, 200, 400, 800 and 1000ug / ml, with wild bacteria T45 as the control, repeated three times, and cultured at 28°C and 280rmp / min On the 7th day, the sporulation number of Trichoderma was measured by blood counting method, and the total sporulation amount of each treatment was calculated to determine the effect of hymexazol on sporulation ability of different mutants.

[0054] The result shows (table 2), compares the processing that does not add hymexazol, and 1000ug / ml hymexazol makes the sporulation yield of Trichoderma T45 and M45a decline respectively by 80 and 45 times of magnitude, but difference is not significant, while other There is a significant difference in the decline of the sporulation of ...

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Abstract

The invention discloses a Trichoderma asperellum mutant strain (Trichoderma asperellum) M45a suitable for compounding with hymexazol to prevent and control melon wilt, and the preservation number is CCTCC NO: M2019513. Compared with other types of Trichoderma, the hymexazol-resistant ability of the strain is significantly improved, which can enrich the biological control strain resources of watermelon wilt and has broad application prospects. The present invention also finds that the synergistic control strategy of hymexazol and Trichoderma M45a has a synergistic effect, and the dosage of hymexazol is significantly reduced compared with the dosage of other compound bacterial agents in the prior art, which can realize the reduction of chemical The pollution targets of fungicides to the environment indicate that the bacteria and their fungicides have good development and application prospects in related fields such as biopesticides and microbial preparations. The invention has important theoretical and practical significance for solving the continuous cropping obstacles of watermelon and other melons.

Description

technical field [0001] The invention belongs to the technical field of microorganisms and prevention and control of diseases and insect pests, and in particular relates to a mutant strain of Trichoderma suitable for compounding with hymexazol to prevent and control melon wilt, an inoculation agent and applications thereof. Background technique [0002] Trichoderma (Trichoderma) belongs to the half-knowledge phylum Hyphospora, which is generally recognized as a beneficial biological and antifungal fungus in the world, and widely exists in soil, rotten wood and vegetable substrates. Trichoderma has strong survivability and wide adaptability. It has antagonistic effects on various plant pathogens. It can prevent and control soil-borne diseases and some leaf diseases of various plants, enhance plant stress resistance and repair the environment polluted by agrochemicals. increasingly important role in sustainable agriculture. At present, more than 60 countries on five continents...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14A01N63/38A01N43/80A01P3/00C12R1/885
CPCA01N43/80C12N1/14C12N1/145C12R2001/885
Inventor 张屹梁志怀魏林朱菲莹肖姬玲何英唐炎英
Owner HUNAN AGRI BIOTECH RES CENT