CN111593008B

CN111593008B - Solid-state fermentation method and application of streptomyces lavendulae NBF715

Info

Publication number: CN111593008B
Application number: CN202010580421.1A
Authority: CN
Inventors: 黄大野; 曹春霞; 姚经武; 杨丹; 张亚妮; 万中义; 郑娇莉; 李飞
Original assignee: Hubei Biopesticide Engineering Research Center
Current assignee: Hubei Biopesticide Engineering Research Center
Priority date: 2020-06-23
Filing date: 2020-06-23
Publication date: 2021-06-29
Anticipated expiration: 2040-06-23
Also published as: CN111593008A

Abstract

The invention belongs to the technical field of crop prevention and control, and particularly discloses a solid-state fermentation method and application of streptomyces lavendulae NBF 715. Compared with the previously reported NBF715 strain solid fermentation method, the method has the advantages of lower cost, simpler and more convenient fermentation method, no infectious microbe infection and complete fermentation, and has wide application prospect in preventing and controlling the seedling damping-off of crop seedbeds by using the main component perlite of the seedling substrate as the fermentation substrate.

Description

Solid-state fermentation method and application of streptomyces lavendulae NBF715

Technical Field

The invention belongs to the technical field of crop prevention and control, and particularly relates to a solid-state fermentation method and application of streptomyces lavendulae NBF 715.

Background

The occurrence of diseases is an important limiting factor for crop production, often resulting in reduced production and even outcrop failure. The use of chemical pesticides is a main means for preventing and controlling crop diseases, and plays a great role in recovering the loss caused by the diseases. Then, due to frequent use of chemical pesticides, pesticide residues and environmental pollution are caused, and the drug resistance of pathogenic bacteria is caused, so that a safer and more effective prevention and control method is urgently needed in production.

The biological prevention and control is safer and is not easy to generate drug resistance, so that the biological prevention and control method becomes an effective means for reducing and replacing chemical pesticides. Streptomyces is an important biocontrol resource, can generate active substances such as various antibiotics, phytohormones and the like, has important effects on improving the disease resistance and stress resistance of plants, and has the potential of developing biological pesticides.

Fungi and streptomycete are used as biocontrol resources, and cannot produce spores under the liquid fermentation condition, so the fermentation methods in the prior production are solid fermentation. The solid fermentation substrate used in the previous fermentation of streptomycete NBF715 is common rice, corn and bran. Although the rice and the corn have higher fermentation level, the cost is higher, the air mixed bacteria are easily polluted, the bran has less gaps, the problem of incomplete fermentation at different positions exists, and the air mixed bacteria are also easily polluted. The problems influence the industrial production and later application and popularization of the streptomyces lavonofuscus NBF 715.

The streptomyces lavonofuscus NBF715 is a bio-control streptomyces which is discovered by the applicant in the earlier stage and has good bacteriostatic effect on various crop diseases, and can be used for controlling tobacco black shank, tomato, eggplant, pepper and ginger bacterial wilt, controlling citrus penicilliosis, controlling tea leaf round spot, and controlling pepper anthracnose and potato late blight.

In order to solve the problems, the applicant provides a solid-state fermentation method of streptomyces lavendulae NBF715, solves the problems of mixed bacteria pollution and incomplete fermentation, and simultaneously finds that a solid preparation prepared by the bacteria has the function of preventing and treating the plant damping-off.

Disclosure of Invention

The invention aims to provide a solid-state fermentation method of streptomyces lavonofuscus NBF715, which is simple, has large spore yield, NO mixed bacteria pollution and suitability for large-scale popularization, and the preservation number of the streptomyces lavonofuscus NBF715 is CCTCC NO: M2017414.

The invention also aims to provide application of the solid-state fermentation method of the streptomyces lavonofuscus NBF715, and the microbial inoculum prepared by the method can be used for preparing granules of the streptomyces lavonofuscus NBF715, and the granules can be used for preventing and treating plant damping-off.

In order to achieve the purpose, the invention adopts the following technical scheme to realize the purpose:

a solid state fermentation process of streptomyces lavendulae NBF715, comprising: mixing the Streptomyces lavendulae NBF715 fermentation liquor with perlite; the preservation number of the streptomyces lavendulae NBF715 is CCTCC NO: M2017414.

In the above method, preferably, the mixing ratio of the streptomyces lavendulae NBF715 fermentation broth to the perlite is as follows: 4mL of: 1-4 g;

in the above method, preferably, the fermentation temperature is 26-30 ℃, and the fermentation time is 12-16 d;

in the above method, preferably, the perlite is tiled, and the tiling height is 5-15 cm;

the particle size of the perlite is as follows: 2mm-7 mm.

The application of the solid-state fermentation method of the streptomyces lavendulae NBF715 comprises the steps of preparing granules of the streptomyces lavendulae NBF715 by using the method; or the preparation method is used for preparing the damping-off preventing granules in the plug seedling process of field crops or the preparation method is used for preparing the culture medium additive.

Compared with the prior art, the invention has the following advantages:

(1) compared with the traditional solid fermentation method for preparing the microbial inoculum, the solid fermentation method for the streptomyces lavonofuscus NBF715 is lower in cost, simpler and more convenient in fermentation method and less prone to contaminating infectious microbes in the air. Is used for preventing and controlling the occurrence of damping-off in the process of plug seedling of crops.

(2) Compared with the current damping-off prevention and control bactericides propamocarb and pyraclostrobin, the damping-off prevention and control bactericide has the same prevention and control effect. Meanwhile, pesticide residue and environmental pollution are avoided.

Drawings

FIG. 1 perlite fermentation Streptomyces lavonofuscus NBF 715.

FIG. 2 vermiculite fermentation Streptomyces lavendulae NBF 715.

FIG. 3 biochar fermentation of Streptomyces lavonoscens NBF 715.

FIG. 4 shows the effect of Streptomyces lavendulae NBF715 granules in preventing and controlling watermelon damping-off.

Detailed Description

In order to better explain the invention, the following further illustrate the main content of the invention in connection with specific examples, but the content of the invention is not limited to the following examples. The technical solutions of the present invention, if not specifically mentioned, are conventional in the art, and the reagents or materials, if not specifically mentioned, are commercially available.

Example 1:

solid-state fermentation method of Streptomyces lavendulae (Streptomyces enissocaisiis) NBF 715:

preparation of Streptomyces lavendulae (Streptomyces enissocaiesilis) NBF715 seed solution: comprises activating slant-stored strain on ISP-2 plate, selecting and inoculating to 100 ml seed culture medium, and culturing on a shaking table at 28 deg.C and 160 rpm for 4 days; obtaining seed liquid; the preservation number of the streptomyces lavendulae NBF715 is CCTCC NO: M2017414.

The formula of the seed culture medium is as follows: 1L of the culture medium comprises 20g of mannitol, 10g of soybean peptone, 0.35g of potassium dihydrogen phosphate, 0.5g of calcium carbonate, 20 mu L of soybean oil and the balance of water.

Liquid fermentation: inoculating 5% of Streptomyces lavendulae (Streptomyces enissocaesilis) NBF715 seed solution into a fermentation culture medium, wherein the temperature is 28 ℃, the stirring speed of the fermentation tank is 500 rpm, the loading amount of the fermentation culture medium is 60% of the tank volume, and fermenting for 5 days.

The formula of the fermentation medium comprises: 3% of mannitol, 3% of soybean peptone, 0.3% of yeast powder, 0.5% of calcium carbonate and 0.5% of sodium chloride; the pH value is 6.5-7.5, and the rest is water.

Sterilizing perlite at high temperature, and uniformly mixing the fermentation liquor and the perlite, wherein the mixing ratio is 4 mL: 1g of the perlite and the perlite are mixed, and the mixture is spread in a plastic box, the height of the perlite is 5cm, the fermentation temperature is 30 ℃, and the fermentation time is 14 d; drying at 100 ℃ for later use; the particle size of the perlite is as follows: 3mm-5 mm.

Sterilized vermiculite powder and biochar (from corn stalks) are used as reference fermentation substrates; the treatment mode and the fermentation condition are the same as those of the perlite.

After fermentation with perlite substrate, the number of spores of Streptomyces enissocaisii (NBF 715) is 6.6 × 10⁹Each spore/g (figure 1), is cultured by separation and streak, has no mixed bacteria, and is fermented uniformly and completely; vermiculite and biochar serving as substrates have poor fermentation effect and cannot effectively fermentFermentation (fig. 2, 3), vermiculite as substrate solid fermentation can not be carried out, and spores can not be obtained (fig. 2); biochar was not able to ferment continuously (fig. 3), producing speckled spores, indicating that only partial fermentation products were produced.

Example 2:

the application of the granules obtained by the solid state fermentation method of the streptomyces lavendulae NBF715 in preventing and treating the watermelon damping-off disease is as follows:

the perlite granules prepared in example 1 were mixed with turf and vermiculite in the proportions turf: vermiculite: the granular formulation is 3:1:0.2(V: V: V) as watermelon culture medium, the proportion medium of common perlite is used as contrast, and the proportion medium of common perlite is sprayed with 1000 times of pyraclostrobin (250g/L missible oil) as chemical agent contrast. Normally culturing watermelon at 25 deg.C after germination, and placing in a greenhouse at a temperature lower than 15 deg.C after growing to two true leaves. And counting the morbidity of the control and treated seedlings after 10 days, and calculating the control effect. Three replicates of 72 seedlings were set up for the experiment.

The morbidity is (number of sown seeds-number of normal seedlings)/number of sown seeds multiplied by 100%

Control effect is (control incidence rate-treatment incidence rate)/control incidence rate x 100%.

Test results show that the seedling culture substrate containing NBF715 granules and the watermelon seedlings with tray treated by pyraclostrobin have the prevention and control effects of 100%.

Test results show that the morbidity of the seedling substrate containing NBF715 granules and watermelon seedlings without damping-off of the plug tray treated by pyraclostrobin is 0, the control effect is 100%, and the control morbidity is 83.33% (fig. 4).

Claims

1. Streptomyces lavonofuscus (A. glaucescens) ((A. glaucescens))Streptomyces enissocaesilis) Application of granules in preparation of preparation for preventing and treating watermelon damping-off, namely streptomyces lavonofuscus (streptomyces lavendulae)Streptomyces enissocaesilis) The preparation method of the granules comprises the following steps: mixing the streptomyces lavendulae fermentation liquor with perlite according to the proportion of 4 mL: 1-4g of perlite, flatly paving the perlite after mixing, wherein the flatly paving height is 5-15cm, and the fermentation temperature after mixingThe fermentation time is 12-16d at the temperature of 26-30 ℃; the particle size of the perlite is as follows: 2mm-7mm, and the preservation number of the streptomyces lavendulae is CCTCC NO: M2017414.