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Preparation method of compound bacterium fermentation product for preventing and treating plant phytophthora, compound bacterium fermentation product and application thereof

A technology for fermentation products and compound bacteria, applied in the directions of botanical equipment and methods, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of single strain source, low control effect, single disease prevention mechanism, etc. The effect of the control effect

Active Publication Date: 2020-08-07
TOBACCO RES INST CHIN AGRI SCI ACAD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The single source of bacterial strains, poor control effect, and backward preparation processing are the main bottlenecks restricting the development of microbial pesticides
At present, the strains selected for microbial pesticides are mostly single strains or a combination of a few strains. The disease prevention mechanism is single, and the control effect is low and unstable. The use of strains from different sources and mechanisms of action has become an important development direction of microbial pesticides.
In terms of microbial fermentation, most of the strains are fermented individually or compounded. The co-cultivation technology that has emerged in recent years has not been reported in the field of biocontrol.

Method used

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  • Preparation method of compound bacterium fermentation product for preventing and treating plant phytophthora, compound bacterium fermentation product and application thereof
  • Preparation method of compound bacterium fermentation product for preventing and treating plant phytophthora, compound bacterium fermentation product and application thereof
  • Preparation method of compound bacterium fermentation product for preventing and treating plant phytophthora, compound bacterium fermentation product and application thereof

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preparation example Construction

[0032] The invention provides a preparation method of a compound bacterial fermentation product for preventing and treating Phytophthora phytophthora, comprising the following steps: (1) carrying out spore culture of Trichoderma marinum, inoculating the obtained spores into a shaking medium for shaking culture, and obtaining Trichoderma marine shock culture; (2) Bacillus subtilis is carried out seed culture, and the obtained seed culture is inoculated into the shock culture medium for shock culture, and the shock culture of Bacillus subtilis is obtained, and the shock culture of Bacillus subtilis is mixed with The shaking culture of Trichoderma marinum obtained in step (1) is mixed and then co-fermented to obtain a co-fermented product, and the co-fermented product is centrifugally filtered to obtain a composite bacterial fermentation product.

[0033] In the present invention, the marine Trichoderma preferably includes Trichoderma aculeatus or Trichoderma reesei, the Trichoder...

Embodiment 1

[0044] 1. Screening culture of marine Trichoderma

[0045] (1) Trichoderma maritima was statically cultured on potato dextrose agar (PDA) medium at 28°C for 7 days.

[0046] (2) Take the small pieces of mycelia grown from the above culture in 200 mL of nutrient broth (NB) medium, shake and culture at 28° C. and 180 rpm for 15 days to obtain Trichoderma marinum fermentation broth, and set up 3 replicates.

[0047] 2. Marine Trichoderma Activity Assay

[0048] Preparation of marine Trichoderma metabolite solution

[0049] After 15 days, use gauze to separate the culture solution and the bacteria in the Trichoderma marinum fermentation broth, extract the culture solution with 1.5 times the volume of ethyl acetate and concentrate it to dryness, and use a 1:1 mixture of methanol and dichloromethane to extract the culture solution. 100 mL was soaked for 3 days and then filtered. After the filtrate was concentrated, the aqueous phase was extracted with 1.5 times the volume of ethyl...

Embodiment 2

[0059] Construction of an interactive culture system between Trichoderma aculeatus HG1 and Bacillus subtilis Tbp55

[0060] (1) Inoculate HG1 on NA (NB medium plus 20g / L agar) medium, culture at 28°C for 3 days, wash out the spores with sterile water, and make the spore concentration reach 10 5 CFU / mL, then take 1mL into 50mL NB liquid medium, shake and culture at 28°C and 180rpm for 48h, and obtain the shaking culture with mycelium growing;

[0061] (2) Inoculate Tbp55 on NA medium and culture for 24h, then inoculate into 50mL NB medium, culture at 28°C and 180rpm for 24h, and adjust the bacterial concentration to 10 5 CFU / mL, 100 μL was added to the 50 mL shaking culture grown for 48 hours in step (1), and co-fermented at 28° C. and 180 rpm for 6 days to obtain a co-fermentation product of HG1 and Tbp55.

[0062] (3) The co-fermentation product of HG1 and Tbp55 obtained in step (2) was filtered twice with a water membrane to obtain a complex bacterial fermentation product, ...

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Abstract

The invention belongs to the technical field of biological prevention and treatment, and particularly relates to a preparation method of a compound bacterium fermentation product for preventing and treating plant phytophthora, the compound bacterium fermentation product and application thereof. The preparation method comprises the following steps: (1), performing spore culturing on marine trichoderma spp., and inoculating the obtained spores into an oscillation culture medium for oscillation culture to obtain a marine Trichoderma spp. oscillation culture; (2), performing seed culturing on bacillus subtilis, inoculating the obtained seed culture into an oscillation culture medium for oscillation culture, mixing the bacillus subtilis oscillation culture with the marine trichoderma spp. oscillation culture obtained in the step (1), carrying out co-fermentation to obtain a co-fermentation product, and centrifuging and filtering the co-fermentation product to obtain a compound bacterium fermentation product. The compound bacterium fermentation product prepared by the invention remarkably reduces the growth of plant phytophthora, and has a better prevention and treatment effect on plantfungal diseases compared with the prevention and treatment effect of a single strain.

Description

technical field [0001] The invention belongs to the technical field of biological control, and in particular relates to a preparation method of a complex bacterial fermentation product for preventing and treating plant phytophthora, a complex bacterial fermentation product and an application thereof. Background technique [0002] Phytophthora oomycetes are important plant pathogens, infecting almost all dicotyledonous plants and some grasses, and according to statistics, cause global economic losses of nearly 50 billion US dollars every year. Due to the wide range of hosts and multiple transmission routes, and the fact that pathogenic bacteria can form oospores and other dormant bodies to survive in the soil for a long time, so far, the prevention and control of Phytophthora is still a worldwide problem. At present, the production of prevention and control agents is mainly metalaxyl, which specifically inhibits RNA polymerase Ⅰ. The drug has a long service life and a relativ...

Claims

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Application Information

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IPC IPC(8): C12P1/02C12P1/04A01N63/22A01N63/38A01P3/00C12R1/885C12R1/125
CPCC12P1/02C12P1/04A01N63/22A01N63/38C12R2001/125C12N1/205C12R2001/885C12N1/145
Inventor 赵栋霖张成省李义强袁源邹平张鹏王晓强绪扩
Owner TOBACCO RES INST CHIN AGRI SCI ACAD
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