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Isaria farinose (Holmskiold)Fries IFMZ130719 and application thereof

A technology of IFMZ130719 and Isyria clavula, applied in the field of microorganisms, to achieve the effects of large spore production, simple cultivation, and less resistance to antibiotics

Inactive Publication Date: 2016-01-20
云南省烟草公司大理州公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a kind of IFMZ130719 and its application, aiming at overcoming the deficiency that the existing I. farina can not be found to be able to parasitize the larvae of silk beetle (Maladerasp.), and to prevent and control the silk beetle by means of biotechnology. Scarlet beetle (Maladera sp.) larvae, to avoid or reduce the resistance problem caused by unreasonable use of chemical pesticides

Method used

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  • Isaria farinose (Holmskiold)Fries IFMZ130719 and application thereof
  • Isaria farinose (Holmskiold)Fries IFMZ130719 and application thereof
  • Isaria farinose (Holmskiold)Fries IFMZ130719 and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Embodiment one, the isolation of pathogenic bacteria, identification

[0011] 1.1 Materials and methods

[0012] 1.1.1 Materials

[0013] Diseased silk beetle larvae infected by an entomogenic fungus were collected in Mengzi City, Yunnan Province.

[0014] PDA medium (potato glucose medium): 200 g of peeled potatoes, 20 g of glucose, 15-20 g of agar, and 1000 mL of distilled water.

[0015] Sterile operating conditions: All utensils and utensils are sterilized in a high-temperature autoclave (121°C, 30min), and inoculation and other operations are performed in an ultra-clean workbench.

[0016] Culture conditions: Culture in a 25°C light (12L: 12D) incubator. After the colonies are formed, transfer to the test tube PDA slope, culture for 2 to 3 days, and transfer to a 4°C refrigerator for storage.

[0017] 1.1.2 Isolation and purification of pathogenic bacteria

[0018] Isolation: Isolate the pathogenic bacteria from the dead body of the sick silk beetle larvae. Th...

Embodiment 2

[0027] Embodiment two, biological characteristics of the purified bacterial strain of I. farinae

[0028] 2.1 Materials and methods

[0029] 2.1.1 Tested strains

[0030] Select a plate with vigorous growth and uniform growth after purification as the test strain. The hyphae were inoculated on PDA again, and cultured in a constant temperature light (12L:12D) incubator at 25°C.

[0031] 2.1.2 Determination of colony growth rate and sporulation

[0032] Take a plate of I. farinae that has been cultivated in advance, punch holes with an 8mm puncher, inoculate it on another fresh medium, do three repetitions, and place it in a constant temperature light (12D:12L) incubator at 25°C Cultured in medium, the diameter was regularly measured and recorded every day until the colony was overgrown with the culture medium. Use a hole puncher with a diameter of 8mm to take the fungus cake at the same position of the culture medium, add 1% Tween-80 and 20mL sterile water, wash the spores ...

Embodiment 3

[0044] Embodiment three, the indoor toxicity determination of the purified strain of I. farinae to the silk beetle

[0045] 3.1. Materials and methods

[0046] 3.1.1 Source of tested insects

[0047] Tobacco plants were planted in pots indoors, and the silk beetles were raised. The healthy and active third-instar larvae of the same size were selected as test insects. After the third-instar larvae were inoculated, they were placed in a petri dish filled with potato slices (R =12cm), were raised under the conditions of 25±1°C and a photoperiod of 0L:24D.

[0048] 3.1.2 Preparation of spore suspension

[0049] Get the purified silty Isyriatium clavium that grows well and wash the conidia with sterile water and filter to configure 10 8 Spores / ml conidia suspension, use a sterile capillary pipette to take a drop of the filtrate onto a hemocytometer, count the spores under a microscope and record the data, and dilute to 10 with sterile water containing 0.5% Tween 80 8 、10 7 、10...

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Abstract

The invention provides Isaria farinose (Holmskiold)Fries IFMZ130719 and application thereof. The Isaria farinose (Holmskiold)Fries IFMZ130719 is preserved in China General Microbiological Culture Collection Center on December 4, 2014, with an accession number of CGMCC No. 10137. The Isaria farinose (Holmskiold)Fries IFMZ130719 is a strain separated from a larva body of Maladera sp. on tobacco for the first time, is simple to culture, grows fast and has a great sporulation quantity and a high spore germination rate. The conidiospore of the Isaria farinose (Holmskiold)Fries IFMZ130719 has powerful pathogenicity on Maladera sp. harm to tobacco, is environment-friendly and pollution-free, hardly incurs drug resistance and can be widely applied to prevention and treatment of larva of Maladera sp.

Description

Technical field: [0001] The invention belongs to the technical field of microorganisms, and in particular relates to Isariafarinose (Holmskiold) Fries] IFMZ130719 and its application. Background technique: [0002] The silk beetle Maladerasp. belongs to the family Coleoptera and is an important leaf-eating pest on tobacco in Yunnan Province. It is widely distributed in Yunnan Province, and the tobacco planted in mountainous areas is seriously harmful. Flue-cured tobacco grows from transplanting seedlings to clusters For a long time and a long period of prosperity, they are all harmed by the silk beetle. The leaves are bitten into holes and notches, and fields with a large number of insect populations in the peak period occur. The leaves of the tobacco plants are eaten up, leaving only the veins and forming a broom shape. loss, quality degradation. Because the insect feeds on the tobacco rhizosphere with larvae and survives, adults can eat tobacco leaves, and because the lar...

Claims

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Application Information

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IPC IPC(8): C12N1/14A01N63/04A01P7/04C12R1/645
Inventor 王新中徐发华陈斌韩非杜广祖肖关丽和淑琪郑亚强王德勋
Owner 云南省烟草公司大理州公司
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