Domestication method and application of nuclear polyhedrosis virus adapted to spodoptera frugiperda
A nuclear polyhedron and Spodoptera frugiperda technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, applications, etc., can solve the problems of inability to adapt to pest outbreaks and control, virus virulence decline, virus mutation, etc. problems, achieve human, animal and other biological safety, accelerate virus differentiation, and highly targeted effects
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[0025] In view of the technical problems existing in the prior art, the inventor of this case was able to propose the technical solution of the present invention after long-term research and a large amount of practice, which is mainly suitable for indoor domestication and preparation of nuclear polyhedrosis virus of Spodoptera frugiperda, The virus prepared by the method is the Autographa californica nuclear polyhedrosis virus cultivated through in vitro culture of sf9 insect cells (spodoptera frugiperda cells) and insect bodies alternately acclimated. The method specifically includes the steps of virus domestication and preparation, expanded production of animal cell reactors, cell collection and crushing for producing Autographa californica nuclear polyhedrosis virus, preparation of dosage forms, and the like.
[0026] The technical solution, its implementation process and principle will be further explained as follows.
[0027] One aspect of the embodiments of the present i...
Embodiment 1
[0061] Variation treatment of nuclear polyhedrosis virus by ultraviolet irradiation intensity:
[0062] The name of the biopesticide virus applied to the infestation of Spodoptera frugiperda is Autographa californica nuclear polyhedrosis virus, and the treatment includes the following steps:
[0063] (1) Use the stock solution of Autographa californica nuclear polyhedrosis virus, dilute it with water 15 times (equivalent to 66 times of the normal use concentration) and treat it with ultraviolet irradiation, the intensity of the ultraviolet irradiation is 50~70μW / cm 2 , the irradiation time is 1-2 hours;
[0064] (2) Preparation of Spodoptera frugiperda larvae: artificially raise a large number of Spodoptera frugiperda larvae, climate incubator settings: temperature 26±0.5°C, humidity 60±5%, light: dark=14:10, select 3-4 instars Spodoptera frugiperda larvae were starved for 12 to 24 hours;
[0065] (3) First use the virus treatment feed obtained in step (1), and feed the norm...
Embodiment 2
[0089] Insect isolated cells and in vivo culture 1:1 cross virus domestication method:
[0090] The name of the biopesticide virus that is applied to the control of African Spodoptera frugiperda in the present embodiment is Autographa californica nuclear polyhedrosis virus, and the domestication of this virus and the preparation method of biopesticide include the following steps:
[0091] (1) Infect for the first time, use the original powder of Autographa californica nuclear polyhedrosis virus, carry out ultraviolet irradiation with ultraviolet rays, and the irradiation intensity is 60 μ W / cm 2 , the irradiation time is 1 hour, then dilute 20 times with water, drop it on the feed and feed the larvae of Spodoptera frugiperda. Larvae will have an obvious poisoning reaction, appetite and vitality will be significantly reduced), every 50 to 60 heads in one tube (50ml centrifuge tube), add 5-10ml of cell culture medium to each tube, filter after grinding, centrifuge, and take the ...
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