Isaria fumosorosea fermentation and spore powder preparation process

Abstract

The invention discloses an isaria fumosorosea fermentation and spore powder preparation process. The method comprises three stages of a liquid fermentation process, a solid fermentation process and adrying and crushing process, compared with the prior art, a low-value solid-liquid culture medium is adopted; a culture medium formula and a fermentation process are optimized; the method has the advantages that the spore production amount is large, the production period is short, the cost is low, the produced spores have certain environmental stress tolerance and the like, meanwhile, the preparedspore powder has higher survival rate and longer storage period by adopting a low-temperature drying process, and the method is particularly suitable for large-scale production of isaria fumosoroseaaerial spores and hypha mixtures.

Description

technical field

[0001] The invention relates to the field of microbial pesticides, in particular to a process for fermenting and preparing spore powder of I. fumigatus. Background technique

[0002] Entomopathogenic fungi, as a class of entomopathogenic microorganisms, play a very important role in the biological control of pests, among which the application effect of fungal pesticide technology is the most significant. Fungal pesticides have the advantages of a wide range of hosts, significant efficacy, environmental safety, and no resistance to pests, so they have broad application prospects.

[0003] Isaria fumosorosea is an important pathogenic fungus of hydatids, which has high pathogenicity to whitefly, aphid, psyllid and other piercing-sucking mouthparts pests. At present, there are many kinds of I. fumigatus biocontrol products in foreign countries, accounting for about 6% of all fungal pesticide products. At present, there is no commercial preparation of I. fumiga...

Examples

Embodiment 1

[0088] Preparation of inoculum by liquid phase fermentation:

[0089] Mix 150 mesh rice flour and deionized water to prepare 40g / L rice milk, add 1% potassium dihydrogen phosphate (molecular formula: KH 2 PO 4 ), 0.2% calcium chloride (molecular formula: CaCl 2 ), 0.1% manganese sulfate (molecular formula: MnSO 4 ). Insert 5% of the initial inoculum into the sterilized liquid medium, set the temperature of the shaker at 28°C, and rotate at 180rpm, cultivate for 3 days, filter the mycelia and residual medium, and adjust the concentration of spores to 10 7 spores / ml to prepare the fermentation inoculum.

[0090] Preparation of spore powder by solid-phase fermentation:

[0091] Take 500g of rice and add an equal amount of deionized water to soak for 10 hours, then drain and sterilize. Prepare 0.1% vitamin B2 solution and 1% arginine solution and sterilize, add 5 parts by mass of vitamin B to the sterilized rice culture medium 2 solution and 5 parts by mass of arginine solu...

Embodiment 2

[0096] Preparation of inoculum by liquid phase fermentation:

[0097] Mix 150 mesh rice flour and deionized water to prepare 40g / L rice milk, add 1.5% potassium dihydrogen phosphate (molecular formula: KH2PO4), 0.1% calcium chloride (molecular formula: CaCl 2 ), 0.1% manganese sulfate (molecular formula: MnSO4). Insert 5% of the initial inoculum into the sterilized liquid medium, set the shaker temperature at 25°C, and cultivate at 150rpm for 3 days, filter the mycelium and medium and adjust the concentration to 10 7 spores / ml to prepare the fermentation inoculum.

[0098] Preparation of spore powder by solid-phase fermentation:

[0099] Take 500g of rice and add an equal amount of deionized water to soak for 10 hours, then drain and sterilize. Prepare 0.1% vitamin B2 solution and 1% arginine solution and sterilize them, add 10 parts by mass of vitamin B to the sterilized rice culture medium respectively 2 solution and 5 parts by mass of arginine solution were evenly mixed...

Embodiment 3

[0103] Preparation of inoculum by liquid phase fermentation:

[0104] Mix 200 mesh rice flour and deionized water to prepare 50g / L rice milk, add 1.2% potassium dihydrogen phosphate (molecular formula: KH2PO4), 0.2% calcium chloride (molecular formula: CaCl 2 ), 0.3% manganese sulfate (molecular formula: MnSO4). Add 5% of the initial inoculum into the sterilized liquid medium, set the shaker temperature at 30° C., and cultivate at 180 rpm for 3 days. Filter mycelium and medium and adjust the concentration to 10 7 spores / ml to prepare the fermentation inoculum.

[0105] Preparation of spore powder by solid-phase fermentation:

[0106] Take 500g of rice and add an equal amount of deionized water to soak for 12 hours, then drain and sterilize. Prepare 0.1% vitamin B2 solution and 1% arginine solution and sterilize, add 5 parts by mass of vitamin B to the sterilized rice culture medium 2 solution and 5 parts by mass of arginine solution were evenly mixed and placed on a steri...

technical field

[0001] The invention relates to the field of microbial pesticides, in particular to a process for fermenting and preparing spore powder of I. fumigatus. Background technique

[0002] Entomopathogenic fungi, as a class of entomopathogenic microorganisms, play a very important role in the biological control of pests, among which the application effect of fungal pesticide technology is the most significant. Fungal pesticides have the advantages of a wide range of hosts, significant efficacy, environmental safety, and no resistance to pests, so they have broad application prospects.

[0003] Isaria fumosorosea is an important pathogenic fungus of hydatids, which has high pathogenicity to whitefly, aphid, psyllid and other piercing-sucking mouthparts pests. At present, there are many kinds of I. fumigatus biocontrol products in foreign countries, accounting for about 6% of all fungal pesticide products. At present, there is no commercial preparation of I. fumiga...

Examples

Embodiment 1

[0088] Preparation of inoculum by liquid phase fermentation:

[0089] Mix 150 mesh rice flour and deionized water to prepare 40g / L rice milk, add 1% potassium dihydrogen phosphate (molecular formula: KH 2 PO 4 ), 0.2% calcium chloride (molecular formula: CaCl 2 ), 0.1% manganese sulfate (molecular formula: MnSO 4 ). Insert 5% of the initial inoculum into the sterilized liquid medium, set the temperature of the shaker at 28°C, and rotate at 180rpm, cultivate for 3 days, filter the mycelia and residual medium, and adjust the concentration of spores to 10 7 spores / ml to prepare the fermentation inoculum.

[0090] Preparation of spore powder by solid-phase fermentation:

[0091] Take 500g of rice and add an equal amount of deionized water to soak for 10 hours, then drain and sterilize. Prepare 0.1% vitamin B2 solution and 1% arginine solution and sterilize, add 5 parts by mass of vitamin B to the sterilized rice culture medium 2 solution and 5 parts by mass of arginine solu...

Embodiment 2

[0096] Preparation of inoculum by liquid phase fermentation:

[0097] Mix 150 mesh rice flour and deionized water to prepare 40g / L rice milk, add 1.5% potassium dihydrogen phosphate (molecular formula: KH2PO4), 0.1% calcium chloride (molecular formula: CaCl 2 ), 0.1% manganese sulfate (molecular formula: MnSO4). Insert 5% of the initial inoculum into the sterilized liquid medium, set the shaker temperature at 25°C, and cultivate at 150rpm for 3 days, filter the mycelium and medium and adjust the concentration to 10 7 spores / ml to prepare the fermentation inoculum.

[0098] Preparation of spore powder by solid-phase fermentation:

[0099] Take 500g of rice and add an equal amount of deionized water to soak for 10 hours, then drain and sterilize. Prepare 0.1% vitamin B2 solution and 1% arginine solution and sterilize them, add 10 parts by mass of vitamin B to the sterilized rice culture medium respectively 2 solution and 5 parts by mass of arginine solution were evenly mixed...

Embodiment 3

[0103] Preparation of inoculum by liquid phase fermentation:

[0104] Mix 200 mesh rice flour and deionized water to prepare 50g / L rice milk, add 1.2% potassium dihydrogen phosphate (molecular formula: KH2PO4), 0.2% calcium chloride (molecular formula: CaCl 2 ), 0.3% manganese sulfate (molecular formula: MnSO4). Add 5% of the initial inoculum into the sterilized liquid medium, set the shaker temperature at 30° C., and cultivate at 180 rpm for 3 days. Filter mycelium and medium and adjust the concentration to 10 7 spores / ml to prepare the fermentation inoculum.

[0105] Preparation of spore powder by solid-phase fermentation:

[0106] Take 500g of rice and add an equal amount of deionized water to soak for 12 hours, then drain and sterilize. Prepare 0.1% vitamin B2 solution and 1% arginine solution and sterilize, add 5 parts by mass of vitamin B to the sterilized rice culture medium 2 solution and 5 parts by mass of arginine solution were evenly mixed and placed on a steri...

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