Purpureocillium lilacinum with high pathogenicity to oilseed rape sclerotinia sclerotiorum and application thereof

A technology for lilac sclerotiorum and rape sclerotinia, which is applied in the field of microorganisms to achieve the effects of easy cultivation, easy preparation and fast reproduction

Inactive Publication Date: 2017-07-25
河南省农业科学院园艺研究所 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current research on S. sclerotiorum biocontrol bacteria is mostly in the laboratory stage, and there is no strain that can effectively control S. sclerotiorum in agricultu...

Method used

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  • Purpureocillium lilacinum with high pathogenicity to oilseed rape sclerotinia sclerotiorum and application thereof
  • Purpureocillium lilacinum with high pathogenicity to oilseed rape sclerotinia sclerotiorum and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0017] Example 1: Acquisition of P. lilacensis strain pt361

[0018] The original strain of P. lilacinus pl36-1 Mr. Wang Mingzu isolated it from root-knot nematode in Hubei Province in 1991 (refer to the "Preliminary Report on the Research of Root-knot Nematode Egg-parasitic Fungus" published in "Journal of Phytopathology" in 1991), and it is preserved in the Phytopathological Nematode Research of Huazhong Agricultural University room. pt361 is a mutant strain obtained by genetic transformation of T-DNA insertion mediated by Agrobacterium tumefaciens.

[0019] For the method of genetic stability of transformants, refer to the method of Zeng Daxing. Roughly as follows: Inoculate the transformed strain of P. lilacinus on a PDA plate without glufosinate and carboxyl, culture at 28°C in the dark for 3-5 days, then transfer to a refrigerator at 4°C for 2 days, and then 28°C, constant temperature and dark culture for 3-5 days, use a puncher to pick out the mycelium at the edge of...

Embodiment 2

[0020] Example 2: Antagonistic effect of the metabolites of P. lilacinosa strain pt361 on Sclerotinia sclerotiorum (PDA plate)

[0021] Preparation of the fermentation broth of P. lilacinium: Into a 250mL Erlenmeyer flask containing 150mL of PDB medium, insert a 0.5cm diameter Pt361 bacterial block of P. Take the fermentation broth, centrifuge at 10000r / min for 15min, take the supernatant and filter it with a bacterial filter. The resulting liquid is the stock solution (100%). Dilute 10%, 30% and 50% with sterile water as needed.

[0022] PDA plate preparation: Cool the melted PDA medium to about 30°C, take 2mL of the above-mentioned bacteria solution of different concentrations and add them to 8mL of cooled PDA medium, mix evenly, pour the plate, and let it stand for 1-2h.

[0023] Inoculation: use a hole puncher with a diameter of 0.5 cm to punch holes on the edge of the mycelium of Sclerotinia sclerotiorum that has grown for 3 days on the glass petri dish, and use an inoc...

Embodiment 3

[0025] Example 3: Antagonistic effect of metabolites of P. lilacinus strain pt361 on Sclerotinia sclerotiorum (rape isolated leaves)

[0026] Preparation of the fermentation broth of P. lilacinium: Into a 250mL Erlenmeyer flask containing 150mL of PDB medium, insert a 0.5cm diameter Pt361 bacterial block of P. Take the fermentation broth, centrifuge at 10000r / min for 15min, take the supernatant and filter it with a bacterial filter. The resulting liquid is the stock solution (100%). Dilute to 10%, 30% and 50% with sterile water as needed.

[0027] Inoculation: Select healthy leaves with the same leaf age and size, wash them with sterile water, and set aside. Soak rapeseed leaves in the above-mentioned fermented liquid of different concentrations for about 20 minutes. Inoculate 2 sclerotinia bacterium blocks with a diameter of 0.5 cm in the middle of both sides of the blade, and cultivate them with moisture at 25° C. for 3 days. The incidence was observed, the incidence was...

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Abstract

The invention relates to purpureocillium lilacinum with high pathogenicity to oilseed rape sclerotinia sclerotiorum and application thereof. The strain is purpureocillium lilacinum pt361, and can be developed into a fungi-proofing preparation to be used for biological control of sclerotinia rot of colza. Compared with an existing chemical agent for controlling oilseed rape sclerotinia sclerotiorum, the strain has the advantages of being efficient, low in toxicity, low in residue amount, free of pollution and capable of reducing the possibility of drug resistance. Besides high pathogenicity to oilseed rape sclerotinia sclerotiorum, the strain is also characterized in high culture speed, large sporulation quantity, high spore germination rate, easy preparation and great market application prospects.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a Porphyra lilacinum ( Purpureocillium lilacinum ) and its applications. Background technique [0002] Sclerotinia sclerotiorum is caused by Sclerotinia sclerotiorum ( Sclerotinia sclerotiorum (Lib.) de Bary) is a worldwide disease that mainly occurs in relatively cool and humid areas where rapeseed grows. In the UK, Sclerotinia can cause up to 50% yield loss in rapeseed; in Germany, rapeseed stalks can be as high as 70% in severe years. In my country’s winter rape production areas, rape sclerotinia is a frequent disease, ranking first among the three major diseases of rape, the incidence rate is generally 10-30%, and it can reach more than 80% in severe years, and the yield is reduced by 11-73%. Reduced by 1 to 5%. The major rape production areas in the middle and lower reaches of the Yangtze River and the southeast coast, including Jiangsu, Anhui, Shanghai, Zhe...

Claims

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Application Information

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IPC IPC(8): C12N1/14A01N63/04A01P3/00C12R1/645
CPCA01N63/30C12N1/145C12R2001/645
Inventor 史宣杰杨凡肖炎农马凯赵秀山孙文喜李艳蔡毓新
Owner 河南省农业科学院园艺研究所
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