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Trichoderma sp. capable of inhibiting growth of citrus black spot bacteria and application thereof

A Trichoderma, black spot technology, applied in the direction of application, plant growth regulator, microbial-based methods, etc., can solve problems such as endangering human health, polluting the environment, and pesticide residues

Active Publication Date: 2020-04-10
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the production process, traditional pesticide control often pollutes the environment, and there are pesticide residues in citrus fruits, which endanger human health

Method used

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  • Trichoderma sp. capable of inhibiting growth of citrus black spot bacteria and application thereof
  • Trichoderma sp. capable of inhibiting growth of citrus black spot bacteria and application thereof
  • Trichoderma sp. capable of inhibiting growth of citrus black spot bacteria and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1 strain separation

[0022] Soil samples were collected from all over the country and separated by the selective medium dilution plate method. The process is as follows: Weigh 10g of soil samples, put them into a 250mL triangular flask filled with small steel balls and 90mL of sterile water, and place them on a shaker at 120r min -1 Shake for 10 minutes to fully disperse the soil sample to form a 10-fold dilution; then draw 1 mL of the 10-fold dilution, add it to a centrifuge tube of 9 mL of sterile water, and mix well; then take 1 mL from it and add it to 9 mL of sterile water In a centrifuge tube of water, mix thoroughly to prepare a 1000-fold dilution. Pour the melted PDA culture medium into a petri dish, 15mL per dish; after the culture medium is cooled to form a flat plate, draw 0.1mL of the soil sample diluted 1000 times and add it to the PDA medium plate, and then use sterile coating Spread the diluted solution evenly with a device, and repeat 3 time...

Embodiment 2

[0027] Embodiment 2 strain identification

[0028] The colony morphology of PDA medium and the characteristics of conidiophore branches and spores were classified.

[0029] On PDA medium, at 25°C, 12h / 12h light and dark culture strains for more than 7 days, stick a small piece of transparent tape on the edge of the colony, then remove the tape and put it in glycerol lactic acid on the glass slide, add 1 Drop glycerol lactic acid, add a cover glass for inspection (lactic acid 500mL, glycerol 1000mL, distilled water 500mL).

[0030] Strain A94 grew rapidly on PDA medium, with an average diameter of more than 1.5 cm per day. The initial hyphae are white, and the aerial hyphae are well developed. With the massive production of spores, the colonies gradually turned dark green ( figure 1 ). Conidiophores erect or curved, smooth. The branches are irregular, the branches are nearly right-angled, and the bottle stems are solitary, opposite, and whorled. The bottle stem is obvious...

Embodiment 3

[0031] Example 3 Molecular identification

[0032] (1) DNA extraction

[0033] 1) After culturing the A94 strain on a PDA plate at 22°C for 7 days, scrape the mycelium on the plate with a toothpick, and put it into a sterilized 1.5mL centrifuge tube containing 300μL extraction buffer;

[0034] The formula of the extraction buffer is: 1M KCl, 100mM Tris-HCl, 10mM EDTA, pH=8.0;

[0035] 2) Grind the picked hyphae with an electric grinder, then add 300 μL of extraction buffer, and shake vigorously for 2 minutes;

[0036] 3) Centrifuge at 10000rpm for 10min;

[0037] 4) Aspirate the supernatant, transfer to another new centrifuge tube, and discard the precipitate;

[0038] 5) Add an equal volume of isopropanol (analytical grade) to the supernatant, gently invert and mix several times, then centrifuge at 12000rpm for 10min to precipitate nucleic acid;

[0039]6) Gently pour off the supernatant, and invert the centrifuge tube containing the precipitate on absorbent paper to drai...

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Abstract

The invention discloses trichoderma sp. capable of inhibiting growth of citrus black spot bacteria and an application thereof. The trichoderma sp. is screened and separated and is named as Trichodermasp, a strain number of the trichoderma asperellum is A94, and a preservation number is CCTCC NO: M2019869. The strain A94 can efficiently secrete ligninase, has a very good degradation effect on plant lignin, has an efficient parasitic effect on citrus black spot bacteria, and also can resist salt. The trichoderma sp. has inhibiting and parasitic decomposition effects on pathogenic bacteria, andalso has a strong decomposition effect on plant residues, so that the infection source of the citrus black spot disease is greatly reduced, and the prevention and treatment effect is greatly improved.

Description

technical field [0001] The invention relates to the technical field of plant pathogen prevention and control, in particular to a Trichoderma aculeatus capable of inhibiting the growth of citrus black spot fungus and an application thereof. Background technique [0002] Citrus is an important fruit. There are many pathogenic bacteria that harm citrus, which poses a huge threat to the safe production of citrus. Among them, citrus black spot fungus is an important pathogen on citrus, causing citrus black spot disease. Citrus black spot fungus can infect citrus leaves, shoots, branches and fruits. Susceptibility to new shoots causes tree vigor to weaken and new shoots to die; leaf susceptibility affects photosynthesis; fruit susceptibility causes dense black spots to form on the fruit surface, seriously affecting the appearance quality, greatly reducing the commercial value of the fruit, causing serious damage Economic losses. Citrus black spot fungus can also carry out sapro...

Claims

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Application Information

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IPC IPC(8): C12N1/14A01N63/38A01P3/00A01G13/00C12R1/885
CPCA01G13/00C12N1/145C12R2001/885
Inventor 王洪凯李红叶林福呈王秀青
Owner ZHEJIANG UNIV
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