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Entomopathogenic nematode symbiotic bacteria and application thereof

A nematophile and bacillus technology, which is applied to entomopathogenic nematode symbiotic bacteria and its application fields, and can solve problems such as unclear insecticidal spectrum and the like

Inactive Publication Date: 2010-11-24
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

All published or published literatures indicate that the symbiotic bacteria of entomopathogenic nematodes or the insecticidal proteins secreted by symbiotic bacteria have oral insecticidal activity against insects, but the insecticidal spectrum is still unclear, especially the supernatants of such bacteria have not been found The metabolites are active against insects or mite eggs

Method used

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  • Entomopathogenic nematode symbiotic bacteria and application thereof
  • Entomopathogenic nematode symbiotic bacteria and application thereof
  • Entomopathogenic nematode symbiotic bacteria and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Embodiment 1, the acquisition of Xenobacterium nematophila HB408 bacterial strain

[0022] From 1997 to 2006, the present inventors isolated a plurality of entomopathogenic nematode strains from soil samples collected from different regions in Hebei Province by trapping the wax moth. Isolate the symbiotic bacteria of the nascent type according to the following method: infect the 5th instar larvae of Mellonella mellonella with entomopathogenic nematodes, after the death of the worm body, soak with 75% (volume percent concentration) alcohol for 10min, rinse 3 times with sterile water, and use Blot the moisture on the worm body with sterile filter paper. Use sterilized tweezers to clamp the head and tail of the worm body, then cut off a gastropod with sterilized scissors, and drop the outflowing body fluid directly into the NBTA identification medium (nutrient agar 45g, tribasic tetrazolium chloride azole 0.04g, bromothymol blue 0.025g, water 1000mL, pH 7.2-7.4), after ma...

Embodiment 2

[0027] Embodiment 2, the oral insecticidal activity of pathogenic bacteria nematophila HB408 bacterial strain and the determination of insecticidal spectrum

[0028] Streak the Xenobacterium nematophila HB408 strain on the NBTA medium (nutrient agar 45g, tribasic tetrazolium chloride 0.04g, bromothymol blue 0.025g, water 1000mL, pH 7.2-7.4) plate, Cultivate at 28°C-30°C for 36h-48h, pick a single primary colony and insert it into 20mL beef broth medium (beef peptone 10g, beef extract 3g, sodium chloride 5g, water 1000mL, pH 7.2-7.4, high pressure at 121°C Sterilize for 30 minutes), place at 28°C-30°C, 200rpm / min constant temperature shaking culture overnight, to obtain seed bacterial liquid. Then the seed bacteria liquid was transferred to 200mL beef broth culture medium according to the inoculation amount of 1% (volume ratio 1:100), and the shaking culture was continued for 48h, and the HB408 fermented bacteria liquid obtained was directly used for the determination of insect...

Embodiment 3

[0037] Example 3, Insecticidal activity of the supernatant and thalline in the fermentation broth of Xenobacterium nematophila HB408

[0038] Prepare the Xenobacterium nematophila HB408 bacterial liquid according to the method in Example 2, then centrifuge part of the HB408 bacterial liquid at 10000r / min and 4°C for 15min, draw the supernatant and filter it through a 0.45 μm filter membrane to obtain the supernatant of the HB408 bacterial liquid The clear liquid is ready for use. After centrifugation, the resulting precipitate was washed twice with an appropriate amount of PBS buffer solution (0.02mol / L, pH 7.4) at 10000r / min and 4°C for 15min, and the resulting bacteria were washed with PBS buffer solution (0.02mol / L, pH 7.4 ) was resuspended to the same volume of bacterial cell suspension as the original bacterial solution of HB408 for later use.

[0039] Leaves of cabbage (feeding Plutella xylostella larvae) or corn leaves (feeding migratory locust) were respectively in th...

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Abstract

The invention discloses entomopathogenic nematode symbiotic bacteria and an application thereof. The entomopathogenic nematode symbiotic bacteria are xenorhabdus nematophila CGMCC No.3004. The fermentation liquor of the bacterial strain, the supernate of the fermentation liquor, and the ethyl acetate extracts of the thallus or of the supernate of the fermentation liquor of thallus has insecticidal action or ovicidal action on multiple agricultural pests.

Description

technical field [0001] The invention relates to an entomopathogenic nematode symbiotic bacterium and its application, in particular to an entomopathogenic nematode symbiotic bacterium and its application in the production of insecticide and / or ovicide. Background technique [0002] Entomopathogenic nematode symbionts are a class of Gram-negative bacteria present in the intestinal tract of entomopathogenic nematodes, belonging to the family Enterobacteriaceae (Hominick WM. Wallingford: CABI Publishing UK, 2002.266-264), including the genus Pathogenus ( Xenorhabdus) and Photorhabdus, which are symbiotic with nematodes of Steinernema and Heterorhabditis, respectively (Chalabaev S., Turlin E., Bay S., et al.Applied and Environmental Microbiology, 2008, 74: 1717-1725; Forst S., Dowds B., Boemare N., et al. Annual review of microbiogy, 1997, 51: 47-72.). [0003] In nature, such bacteria exist in the intestines of nematodes at the third instar infection stage. As the nematodes in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P7/04A01P7/02C12R1/01
Inventor 王勤英孔繁芳宋萍南宫自艳曹克强
Owner HEBEI AGRICULTURAL UNIV.
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