Metarhizium anisopliae MAYX130921 and application thereof

A technology of scarab metarhizium anisopliae and black beetle, applied in the field of microorganisms, to achieve the effects of rapid growth, strong pathogenicity, and high spore germination rate

Inactive Publication Date: 2015-11-11
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to provide a kind of Metarhizium anisopliae MAYX130921 and its application, aiming at overcoming the deficiency that the existing Metarhizium anisopliae can not parasitize the black beetle, prevent and treat the black beetle by means of biotechnology, avoid or reduce the Resistance problems caused by irrational use of chemical pesticides

Method used

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  • Metarhizium anisopliae MAYX130921 and application thereof
  • Metarhizium anisopliae MAYX130921 and application thereof
  • Metarhizium anisopliae MAYX130921 and application thereof

Examples

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] In Eshan County, Yuxi City, Yunnan Province, the diseased Diablo spp. infected by an entomogenic fungus was collected.

[0014] Gill beetle larvae.

[0015] Sabouraud medium (SDAY): 1% peptone + 1% yeast powder + 4% glucose + 1.5-2% agar powder + 1000ml water.

[0016] 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.

[0017] Culture conditions: Culture in a 28°C light (12L:12D) incubator. After the colonies are formed, transfer to the SDAY slant of the test tube, cultivate for 3 to 5 days, and transfer to a 4°C refrigerator for storage.

[0018] 1.1.2 Isolation and purification of pathogenic bacteria

[0019] Isolation: Isolate the pathogenic bacteria from the dead body of the sick...

Embodiment 2

[0028] Embodiment two, the biological characteristics of the purified bacterial strain of Metarhizium anisopliae

[0029] 2.1 Materials and methods

[0030] 2.1.1 Tested strains

[0031] Select a plate with vigorous growth and uniform growth after purification as the test strain. The hyphae were inoculated on SDAY again, and cultivated in a constant temperature light (12L:12D) incubator at 28°C.

[0032] 2.1.2 Determination of colony growth rate and sporulation

[0033] Take a plate of Metarhizium anisopliae cultured in advance and punch holes with an 8mm puncher, inoculate it on a new medium, do three repetitions, and place it in a constant temperature light (12D:12L) incubator at 28°C for cultivation , measure its diameter regularly every day and record it until the colony is overgrown with the culture medium. Use a hole puncher with a diameter of 8mm to take the bacterium cake at the same position of the culture medium, add 1% Tween-80 and 20ml sterile water, wash the s...

Embodiment 3

[0044] Example 3, Indoor Toxicity Determination of Metarhizium anisopliae MAYX130921 to Scaraba larvae

[0045] 3.1. Materials and methods

[0046] 3.1.1 Source of tested insects

[0047] The 2nd instar larvae of the black beetle were reared with potato cubes in an incubator with a temperature of (25±1)°C, a humidity of (70±5)%, and a photoperiod of 0L:24D in our laboratory. The second-instar larvae of the black beetle, which are disease-free and of the same size, were selected as test insects.

[0048] 3.1.2 Preparation of spore suspension

[0049] Get the well-growing Purified Scarab anisopliae Metarhizium anisopliae, 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 6 、10 5 、10 4 The spore suspensions with five...

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Abstract

The invention provides metarhizium anisopliae MAYX130921 and an application thereof. The metarhizium anisopliae MAYX130921 is preserved on Dec 4th, 2014 in China General Microbiological Culture Collection Center with the preservation number of CGMCC No.10139. The metarhizium anisopliae MAYX130921 is a bacterial strain initially separated from holotrichia parallela larvas, is easy to culture and can grow rapidly, sporulation quantity is high, and spore germination rate is high; meanwhile, conidiospores of the metarhizium anisopliae MAYX130921 have strong pathogenicity on the holotrichia parallela larvas, are environment-friendly and hardly produce drug resistance and can be widely applied to prevention and control of holotrichia parallela.

Description

Technical field: [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a Metarhizium anisopliae MAYX130921 and an application thereof. Background technique: [0002] The dark gill beetle Holotrichiaparallela Motschulsky is an important agricultural underground pest. Its adults (scarabs) feed on the underground leaves of harmful crops. The larvae live in the soil for a long time and feed on the underground rhizomes and tubers (stems) of plants, causing the invasion of microorganisms such as bacteria in the soil. Dyeing causes underground rhizomes to rot. The control methods for the larvae of the black beetle were once dominated by chemical control. Although the chemical control reduced the damage of grubs to a certain extent, it could not cure the damage of grubs. Moreover, the improper use of chemical pesticides aggravated the pollution of soil and water systems. , pesticide residues in agricultural products also exceeded the st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A01P7/04C12R1/645
Inventor 陈斌杜广祖张立敏肖关丽和淑琪郑亚强昝庆安桂富荣李正跃
Owner YUNNAN AGRICULTURAL UNIVERSITY
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