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Purpureocillium lilacinum strain with strong virulence to Botrytis cinerea and application thereof

A technology of Botrytis cinerea and P. lilacinus, applied in the field of microbiology, can solve the problems of effective control of Botrytis cinerea and other problems, and achieve the effects of less drug resistance, fast cultivation speed, and strong pathogenicity

Inactive Publication Date: 2017-07-21
河南省农业科学院园艺研究所 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, at present, the research on the biocontrol bacteria of Botrytis cinerea is mostly in the laboratory stage, and the effective control of Botrytis cinerea has not been achieved in agricultural production. In-depth study of biological potential

Method used

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  • Purpureocillium lilacinum strain with strong virulence to Botrytis cinerea and application thereof
  • Purpureocillium lilacinum strain with strong virulence to Botrytis cinerea and application thereof

Examples

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Effect test

Embodiment 1

[0016] Example 1: Cultivation and domestication of Porphyridium lavender strain pt362

[0017] Porphyridium lavender original strain pl36-1 was isolated from root-knot nematodes in Hubei Province in 1991 by teacher Wang Mingzu (see 1991 published in "Acta Phytopathology" entitled "Preliminary Research Report on Root-knot Nematode Egg Parasitic Fungi" Article), preserved in the Plant Pathology Nematode Research Laboratory of Huazhong Agricultural University. pt362 is a mutant strain obtained by T-DNA insertion genetic transformation mediated by Agrobacterium tumefaciens.

[0018] The method of transformant genetic stability refers to the method of Zeng Daxing. It is roughly as follows: inoculate the transformed strain of Porphyridium lilacinus on a PDA plate that does not contain glufosinate and chlorophyllin. After culturing for 3-5 days in the dark at a constant temperature at 28°C, transfer to a refrigerator at 4°C for 2 days, and then Incubate in the dark at 28°C for 3-5 days....

Embodiment 2

[0019] Example 2: The antagonistic effect of Porphyridium lavender strain pt362 on Botrytis cinerea (PDA plate)

[0020] Preparation of Porphyridium lavender / Botrytis cinerea: Use a punch with a diameter of 0.5 cm to punch the edge of the colony of P. lavender / Botrytis cinerea that has grown full of hyphae on a glass petri dish, and use an inoculation needle The bacterial clumps are inserted into the cooled plate, incubated at 28°C, and set aside.

[0021] PDA plate preparation: Cool the melted PDA medium to about 30°C and pour the plate. Let stand for 1 to 2 hours.

[0022] Inoculation: Use a hole punch with a diameter of 0.5 cm to punch the edge of the hyphae of Botrytis cinerea that has grown for 3 days on a glass petri dish, and use an inoculating needle to insert the bacterial mass into the cooled plate (cross method). Porphyridium lavender was inoculated in the middle of the plate, cultivated at 25°C, observed and counted the inhibitory effect. See the result figure 1 , (A:...

Embodiment 3

[0023] Example 3: Antagonistic effect of metabolites of Porphyridium lavender strain pt362 on Botrytis cinerea

[0024] Preparation of Porphyridium lavender fermentation broth: Put a 0.5 cm diameter Porphyridium lavender into a 250 mL Erlenmeyer flask containing 150 mL of PDB medium, culture it on a shaker at 28°C, 200r / min, and culture for 6 days. Take the fermentation broth, centrifuge at 10000r / min for 15min, take the supernatant and filter it with a bacterial filter. The obtained liquid is the original liquid. Dilute with sterile water to dilute the original solution 8 times.

[0025] Inoculation: Use healthy leaves of the same age and size, wash them with sterile water, and set aside. Soak the tomato leaves in the fermentation broth of different concentrations for about 20 minutes. Inoculate a block of Botrytis cinerea with a diameter of 0.5 cm in the middle of the leaf, and culture it in a humidified condition at 25°C for 3 days. Observe the incidence, count the incidence...

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Abstract

The invention relates to a Purpureocillium lilacinum strain with strong virulence to Botrytis cinerea and application thereof. The strain is Purpureocillium lilacinum pt362 and can be developed into a biocontrol bacterium preparation for biocontrol of Botrytis cinerea. Compared with conventional chemical agents for preventing and treating Botrytis cinerea, the strain provided by the invention has the characteristics of high efficiency, low toxicity, low residues, no pollution and small possibility of incurring drug resistance. The strain has strong virulence to Botrytis cinerea and is fast to culture, great in sporulation quantity, high in the germination rate of spores and easy to prepare; so the strain has good market application prospects.

Description

Technical field [0001] The invention belongs to the technical field of microorganisms, and specifically relates to a Porphyridium lilacinus ( Purpureocillium lilacinum ) Strains and their applications. Background technique [0002] Gray mold is a worldwide important disease. It can infect the flowers, fruits and other green tissues of more than 200 plants, causing plant tissues to rot and lose their usefulness. In recent years, the widespread occurrence of gray mold in China has severely hindered the increase in vegetable production. Among the solanaceous vegetables, tomatoes suffer the most. The prevention and control of tomato gray mold has become a key measure to protect the development of production, which directly determines the yield and economic benefits of tomatoes. Botrytis cinerea mainly overwinters or oversummers on tomatoes as sclerotia in the soil or hyphae and conidia on diseased bodies. The disease is an airborne disease, which mainly relies on the transmission o...

Claims

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Application Information

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IPC IPC(8): C12N1/14A01N63/04A01P3/00C12R1/645
CPCA01N63/30C12N1/145C12R2001/645
Inventor 杨凡史宣杰肖炎农马凯赵秀山李艳蔡毓新王彬高显敏
Owner 河南省农业科学院园艺研究所
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