| 摘要 |
The present disclosure is related to a mass production method of Bacillus thuringiensis Cry spores and proteins, as well as an inexpensive and unconventional liquid culture media prepared from agro-industrial byproducts for Lepidoptera control in crucifers. The liquid culture medium comprises a mixture of soy flour and tomato puree in combination with mineral salts to prepare an initial inoculum (or fermentation) medium and a fermentation medium for mass production of spores and Cry proteins. The invention refers to a culture medium which can be used as initial inoculum, said inoculum comprising 2.0 to 10.0 g of mashed tomato, 0.1 to 1 g of soy flour, 0.1 to 1 g of sucrose, 0.01 to 1.0 g of calcium carbonate, 0.1 to 0.5 g of monobasic potassium phosphate, 0.001 to 0.01 g manganese sulfate, 0.0 to 0.001 g of magnesium sulfate, 0.001 to 0.01 g of ferrous sulfate and 0.001 to 0.01 g of calcium chloride per 100 ml of each medium. The invention also relates to the same culture medium, whic h can be used for mass production and consists of 20 to 100 g of mashed tomato, 1 to 10 g of soy flour, 1 to 10 g of sucrose, 0.5 to 10 g of calcium carbonate, 1 to 10 g monobasic potassium phosphate, 0.01 to 0.1 g manganese sulfate, 0.0 to 0.001 g of magnesium sulfate, 0.01 to 0.1 g of ferrous sulfate and 0.01 to 0.1 g of calcium chloride per 1000 ml of medium. The invention further relates to a production method which comprises inoculating the mixture of Bacillus thuringiensis strains in the selected culture medium to prepare an initial fermentation inoculum or primary fermentation under the following conditions; 20° C -35° C temperature, 100 rpm in 500 rpm of agitation and an initial pH of 5 -9, as well as the use of this initial inoculum or primary fermentation for the fermentation of mass production of spores and Cry proteins to level reactor, under the conditions of 0.1vvm 2 vvm of aeration, 20° C -35° C temperature, 100 rpm to 500 rpm stirring speed and a pH of 5 to 9. The Conc entrations of each component in the culture media of the present invention are more effective in the production of spores and highly toxic Cry proteins with a longer effect for pest control, thereby ensuring a control of adults, nymphs, larvae and eggs of insect pests, especially of the Lepidoptera group and on cruciferous crops. |
