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Entomopathogenic nematode symbiotic bacteria strains and applications for inhibiting various plant fungal diseases

A technology for entomopathogenic nematodes and plant fungal diseases, applied in the fields of plant growth regulators, applications, microorganism-based methods, etc., can solve food safety problems, environmental problems, problems affecting human health and foreign exchange earnings from agricultural exports, and achieve sustainable effects. The effect of long period, low requirements on culture conditions, environmental protection and non-toxicity

Active Publication Date: 2022-04-29
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the widespread use of chemical pesticides has brought a series of environmental and food safety issues; it has seriously affected human health and the export of agricultural products

Method used

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  • Entomopathogenic nematode symbiotic bacteria strains and applications for inhibiting various plant fungal diseases
  • Entomopathogenic nematode symbiotic bacteria strains and applications for inhibiting various plant fungal diseases
  • Entomopathogenic nematode symbiotic bacteria strains and applications for inhibiting various plant fungal diseases

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] The screening of embodiment 1 bacterial strain

[0058] The entomopathogenic nematode symbiotic bacteria strain NN6 of this embodiment belongs to Xenorhabdus bovineii, and the preservation number is: CCTCC NO: M 2020097, which was deposited in the China Center for Type Culture Collection on April 29, 2020.

[0059] The Burger's pathogenic bacteria (Xenorhabdus bovineii) NN6 of the present embodiment obtains as follows:

[0060] Infect the 5th instar larvae of Mellonella mellonella with entomopathogenic nematodes. After the larvae die, soak them in 75% alcohol by volume for 30s, wipe the surface of the larvae with 0.5% NaClO solution by mass percentage, and dry the larvae with sterile filter paper Cut off the 5th or 6th pair of gastropods of Mellonella mellonella with sterilized scissors, use a pipette gun to suck up the hemolymph exuded from the wound and drop it on the NBTA+SP plate medium, use inoculation Carry out four-section lines on the ring, place it in a bioche...

Embodiment 2

[0063] Embodiment 2 Preparation of entomopathogenic nematode symbiotic bacteria fermentation broth

[0064] The preparation method of the fermentation broth of the entomopathogenic nematode symbiotic bacteria of the present embodiment is carried out according to the following steps:

[0065] 1) Use a 1mL tip to dip the bacterial suspension from the tube of entomopathogenic nematode symbiotic NN6 glycerol bacteria stored at -80°C, streak on the NBTA+SP plate medium, and culture at 28°C for 24 hours to obtain a single colony;

[0066] 2) Pick blue-green primary colonies and inoculate them into LB+SP liquid medium, and shake and culture at 28°C for 24 hours to obtain bacterial liquid;

[0067] 3) Inoculate the bacterial solution cultivated in 2) with a 5% inoculation amount in the YS shake flask fermentation medium for 24 hours with shaking to obtain a seed solution;

[0068] 4) The seed liquid obtained in 3) was inoculated in the same medium with a 5% inoculum amount and contin...

Embodiment 3

[0069] Example 3 Preparation of bacterial cell suspension and sterile filtrate of entomopathogenic nematode symbiotic bacteria

[0070] The separation of the components of the fermentation broth of the entomopathogenic nematode symbiotic bacteria NN6 described in this embodiment is carried out according to the following steps:

[0071] 1) Centrifuge the fresh fermented liquid obtained in Example 2 at 10,000 rpm for 10 min to obtain bacterial cell sediment and supernatant;

[0072] 2) The bacterial cell precipitate obtained by centrifugation is diluted and mixed with sterile water to obtain a bacterial cell suspension;

[0073] 3) The supernatant obtained by centrifugation is sterilized by filtration with a filter membrane with a pore size of 0.22 μm;

[0074] 4) Check the number of supernatant colonies in the filtered supernatant with the plate coating method again, and it is less than 10 2 When CFU / mL, the obtained supernatant was considered as sterile filtrate.

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Abstract

The invention discloses an entomopathogenic nematode symbiotic bacterial strain and its application for inhibiting various plant fungal diseases. The bacterial strain is a highly virulent strain NN6 of Pathogenobacter burgdorferi, and the preservation number is: CCTCC NO: M 2020097, April 29, 2020 Preserved in China Center for Type Culture Collection. The pathogenic bacteria burgdorferi NN6 of the present invention has stronger ability to inhibit the growth of Fusarium oxysporum watermelon-specific type and cucumber-specific type, Fusarium graminearum, Magnaporthe oryzae, Botrytis cinerea, Phytophthora capsici mycelium growth, and with With the increase of volume concentration, the antibacterial effect is remarkable. The bacterial strain also has the effect of inhibiting the germination of Fusarium spores, and the inhibitory effect is remarkable. The bacterium has the characteristics of low requirements for culture conditions, long duration, environmental protection and non-toxicity, etc. It is of great significance for protecting the ecological environment and improving the added value of agricultural products. In addition, it has the advantage of a wider antibacterial spectrum, which makes it very good. Market development and application prospects.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an entomopathogenic nematode symbiotic bacterial strain, in particular to an entomopathogenic nematode symbiotic bacterial strain for inhibiting various plant fungal diseases and its application. The present invention uses a biological control method to control the growth of plant pathogenic fungi, It is suitable for the production of green and pollution-free agricultural products in the modern ecological agricultural economy. Background technique [0002] Fusarium wilt of watermelon, commonly known as "dead seedling disease", is a disease of the vascular system caused by Fusarium oxysporum infection. It is a very serious disease in watermelon production at home and abroad, and has become the main obstacle restricting the production of watermelon in my country. Fusarium oxysporum can survive for 5-6 years, even more than 10 years without the host. In bacteria-carrying soil, the rate o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/20A01P3/00C12R1/01
CPCC12N1/20A01N63/20C12R2001/01C12N1/205
Inventor 张克云张潘杰包浩然柳李旺窦振国王浩肖振华肖宇辉柳王亮林建徐良熊阳杰高国富阿德勒·迪曼
Owner NANJING AGRICULTURAL UNIVERSITY
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