Thosea sinensis nuclear polyhedrosis virus and production method thereof
A nuclear polyhedron and production method technology, applied in the field of viruses, can solve problems such as sustainable control of unfavorable pests, excessive pesticide residues, and damage to pests
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Embodiment 1
[0025] Example 1 TsNPV Virus Isolation, Proliferation, Extraction and Identification
[0026] Dilute the stock solution of TsNPV virus (isolated from the natural dead larvae of Euonymus euphratica in Aixi Lake Wetland Park, Nanchang City, Jiangxi Province) with sterile water to 1×10 6PIB / mL, sprayed with a plastic watering can on the branches of Euonymus euphratica with leaves, dried in the shade, put them into the Xitianlong mini storage box, and inserted the collected 4-6 instar larvae. 28°C, feeding at 9:15 in the photoperiod (9 hours in the light and 15 hours in the dark) until all the larvae of the scorpion moth died. Pay attention to observation every day during this period, after feeding for 48 hours, replace them with fresh branches of Euonymus euphratica with leaves, and take out the dead larvae in time and store them in a refrigerator at 4°C.
[0027] Subsequently, take the dead larvae of the larvae, soak them in sterilized 0.02mol / L pH7.2 PBS for 10 minutes, shake ...
Embodiment 2
[0032] Bioassay test of embodiment 2 TsNPV virus
[0033] The TsNPV virus was serially diluted with sterile water to 1×10 7 PIB / mL, 1×10 6 PIB / mL, 1×10 5 PIB / mL, 1×10 4 PIB / mL, 1×10 3 PIB / mL, 1×10 2 PIB / mL, 1×10 1 6 concentrations of PIB / mL. The branches with leaves of Euonymus euphratica were soaked with each concentration of TsNPV virus solution, and then dried to carry out the feeding experiment. Each concentration was taken as a group, each group was put into a certain number of larvae of the flat moth, placed in sterilized containers, sealed tightly with wet gauze, coated with sterile water as a control, and repeated 2 times. After 24 hours of infection, fresh and clean leaves were added to continue feeding. Observe and record according to the test requirements. Calculate the mortality rate and calculate the lethal time of different virus concentrations, and the data obtained are analyzed by variance using SPSS software.
[0034] The results are shown in Table ...
Embodiment 3
[0040] The establishment of the method for the production of TsNPV virus of embodiment 3 artificial feed rearing flathead moth
[0041] 3.1 Rearing flat gnat moths on artificial diet
[0042] Preparation of artificial feed: cooked soybean powder 90g, cooked wheat flour 90g, yeast powder 15g, sucrose 60g, glycine 1.25g, agar 12g, Vc and compound V B Each 1g, ethylparaben 0.5g, 10% potassium hydroxide 37.5mL, tap water 800mL. After melting the agar, add ethylparaben and potassium hydroxide while it is hot, stir evenly, add soybean flour, wheat flour and yeast powder, keep stirring, add vitamins when the temperature drops to about 60°C, continue stirring evenly, and pack immediately .
[0043] Acquisition of the source of the scorpion moth: collect the mature larvae of the scorpion moth from the wild, feed them with the leafy branches of the host plant until they pupate, wait for them to emerge into adults, feed the adults with honey water until they mate and lay eggs, collect ...
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