Streptomyces luteus solid fermentation medium, preparation method and fermentation method thereof, fermentation product, biocontrol product and application

Abstract

The invention provides a Streptomyces flavinus solid fermentation medium, a preparation method and a fermentation method thereof, a fermentation product, a biocontrol product and an application, and relates to the technical field of microbial fermentation. The Streptomyces luteus solid fermentation medium comprises the following components in parts by weight: 40-50 parts of bran, 15-25 parts of rice husk, 25-35 parts of rice bran, 3-7 parts of calcium carbonate and water. The fermentation method of Streptomyces flavinus includes: optionally after seed activation and liquid seed cultivation of Streptomyces luteus strains, fermenting and culturing by using Streptomyces luteus solid fermentation medium to obtain a fermentation product. The present invention alleviates the defects of the existing Streptomyces flavinus with few viable counts and sporulations. The present invention adopts the above-mentioned culture medium for solid fermentation culture, has a short fermentation period and a large number of viable counts, is suitable for industrial production, and obtains a fermentation product It has remarkable effects in preventing and treating bacterial wilt of crops, and improves the quality and yield of crops.

Description

technical field

[0001] The invention relates to the technical field of microbial fermentation, in particular to a Streptomyces luteus solid fermentation medium, a preparation method and a fermentation method thereof, a fermentation product, a biocontrol product and an application. Background technique

[0002] Plant diseases are one of the natural biological disasters that seriously threaten agricultural production. The use of chemical agents not only kills beneficial farmland microorganisms and destroys farmland ecosystems, but also remains in soil, water bodies, agricultural by-products and livestock products, posing a threat to human health. Moreover, many pathogenic bacteria are resistant to drugs, leading to an increase in the frequency and dosage of chemical agents year by year, and the control effect is poor. Biological control has no effect on beneficial microorganisms in the environment, no residue and no pollution, no killing of natural enemies, no drug resistance,...

Examples

preparation example Construction

[0073] According to a second aspect of the present invention, the preparation method of the above-mentioned Streptomyces flavinus solid fermentation medium is provided, comprising the following steps:

[0074] The bran, rice husk, rice bran, calcium carbonate and water are uniformly mixed in a formula amount, and sterilized to obtain a solid fermentation medium of Streptomyces luteus.

[0075] The medium preparation method is simple and suitable for large-scale production.

[0076] Preferably, the sterilization temperature is 110-130°C, and / or the sterilization time is 30-60min, preferably 40min at 121°C.

[0077] Typical but non-limiting examples of sterilization temperatures are 110°C, 115°C, 120°C, 125°C or 130°C.

[0078] Typical but non-limiting sterilization times are, for example, 30 min, 35 min, 40 min, 45 min, 50 min, 55 min or 60 min.

[0079] After being sterilized at a certain temperature, a sterile Streptomyces flavinus solid fermentation medium is obtained.

...

Embodiment 1

[0147] A fermentation method of Streptomyces flavinus, comprising the following steps:

[0148] (a) slant seed activation: Streptomyces flavinus ACCC 40027 was inoculated under aseptic conditions in the soluble starch of 8g / L, the glucose of 12g / L, the yeast powder of 2g / L, the dipotassium hydrogen phosphate of 1g / L , 0.5g / L of magnesium sulfate and 25g / L of agar, pH 7.0, sterilized at 121°C for 25 minutes on a prepared slant medium, cultivated at 28°C, and cultivated for 120h to obtain slant seeds;

[0149] (b) Liquid seed culture: elute the slant seeds obtained in step (a) with sterile water, and use a sterile pipette to inoculate into 8g / L peptone, 12g / L yeast extract powder, 15g / L Glucose and 10g / L calcium carbonate, pH 7.0, sterilized at 121°C for 25min in the liquid medium prepared, the liquid medium is a 1L Erlenmeyer flask with a sample volume of 200mL, an inoculum volume of 15mL / bottle, a culture temperature of 28°C, and a shaking table of 180r / min, culture time 48h...

Embodiment 2

[0153] A fermentation method of Streptomyces flavinus, comprising the following steps:

[0154] (a) slant seed activation: Streptomyces luteus ACCC 40027 was inoculated under aseptic conditions in 12g / L soluble starch, 8g / L glucose, 5g / L yeast powder, 0.5g / L dihydrogen phosphate Potassium, 1g / L of magnesium sulfate and 15g / L of agar, pH 7.5, sterilized at 121°C for 25 minutes on a prepared slant medium, cultivated at 30°C, and cultivated for 96h, to obtain slant seeds;

[0155] (b) Liquid seed culture: the slanted seeds obtained in step (a) were eluted with sterile water, and inoculated with 12g / L peptone, 8g / L yeast extract powder, 25g / L Glucose and 5g / L calcium carbonate, pH 7.5, sterilized at 121°C for 25 minutes in the liquid medium prepared, the liquid medium is a 1L Erlenmeyer flask with a sample volume of 200mL, an inoculum volume of 10mL / bottle, a culture temperature of 30°C, and a shaking table of 180r / min, culture time 30h, obtain liquid seed;

[0156] (c) Solid f...

technical field

[0001] The invention relates to the technical field of microbial fermentation, in particular to a Streptomyces luteus solid fermentation medium, a preparation method and a fermentation method thereof, a fermentation product, a biocontrol product and an application. Background technique

[0002] Plant diseases are one of the natural biological disasters that seriously threaten agricultural production. The use of chemical agents not only kills beneficial farmland microorganisms and destroys farmland ecosystems, but also remains in soil, water bodies, agricultural by-products and livestock products, posing a threat to human health. Moreover, many pathogenic bacteria are resistant to drugs, leading to an increase in the frequency and dosage of chemical agents year by year, and the control effect is poor. Biological control has no effect on beneficial microorganisms in the environment, no residue and no pollution, no killing of natural enemies, no drug resistance,...

Examples

preparation example Construction

[0073] According to a second aspect of the present invention, the preparation method of the above-mentioned Streptomyces flavinus solid fermentation medium is provided, comprising the following steps:

[0074] The bran, rice husk, rice bran, calcium carbonate and water are uniformly mixed in a formula amount, and sterilized to obtain a solid fermentation medium of Streptomyces luteus.

[0075] The medium preparation method is simple and suitable for large-scale production.

[0076] Preferably, the sterilization temperature is 110-130°C, and / or the sterilization time is 30-60min, preferably 40min at 121°C.

[0077] Typical but non-limiting examples of sterilization temperatures are 110°C, 115°C, 120°C, 125°C or 130°C.

[0078] Typical but non-limiting sterilization times are, for example, 30 min, 35 min, 40 min, 45 min, 50 min, 55 min or 60 min.

[0079] After being sterilized at a certain temperature, a sterile Streptomyces flavinus solid fermentation medium is obtained.

...

Embodiment 1

[0147] A fermentation method of Streptomyces flavinus, comprising the following steps:

[0148] (a) slant seed activation: Streptomyces flavinus ACCC 40027 was inoculated under aseptic conditions in the soluble starch of 8g / L, the glucose of 12g / L, the yeast powder of 2g / L, the dipotassium hydrogen phosphate of 1g / L , 0.5g / L of magnesium sulfate and 25g / L of agar, pH 7.0, sterilized at 121°C for 25 minutes on a prepared slant medium, cultivated at 28°C, and cultivated for 120h to obtain slant seeds;

[0149] (b) Liquid seed culture: elute the slant seeds obtained in step (a) with sterile water, and use a sterile pipette to inoculate into 8g / L peptone, 12g / L yeast extract powder, 15g / L Glucose and 10g / L calcium carbonate, pH 7.0, sterilized at 121°C for 25min in the liquid medium prepared, the liquid medium is a 1L Erlenmeyer flask with a sample volume of 200mL, an inoculum volume of 15mL / bottle, a culture temperature of 28°C, and a shaking table of 180r / min, culture time 48h...

Embodiment 2

[0153] A fermentation method of Streptomyces flavinus, comprising the following steps:

[0154] (a) slant seed activation: Streptomyces luteus ACCC 40027 was inoculated under aseptic conditions in 12g / L soluble starch, 8g / L glucose, 5g / L yeast powder, 0.5g / L dihydrogen phosphate Potassium, 1g / L of magnesium sulfate and 15g / L of agar, pH 7.5, sterilized at 121°C for 25 minutes on a prepared slant medium, cultivated at 30°C, and cultivated for 96h, to obtain slant seeds;

[0155] (b) Liquid seed culture: the slanted seeds obtained in step (a) were eluted with sterile water, and inoculated with 12g / L peptone, 8g / L yeast extract powder, 25g / L Glucose and 5g / L calcium carbonate, pH 7.5, sterilized at 121°C for 25 minutes in the liquid medium prepared, the liquid medium is a 1L Erlenmeyer flask with a sample volume of 200mL, an inoculum volume of 10mL / bottle, a culture temperature of 30°C, and a shaking table of 180r / min, culture time 30h, obtain liquid seed;

[0156] (c) Solid f...

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