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

Pending Publication Date: 2020-05-08
SUZHOU MIDI BIOTECH CO LTD +1
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AI Technical Summary

Problems solved by technology

[0004] Sf9 cells are a clonal culture derived from Spodoptera frugiperda (FallArmyworm) IPLB-Sf21-AE cells used in suspension cultures for rapid expansion of cell stocks, propagation of baculovirus and production of recombinant proteins , but in vitro cell culture will produce virus mutations, which may lead to a decrease in the virulence of the produced virus
[0005] Although the acquisition of polyhedrosis virus is still through wild discovery and collection, this method is low in efficiency, high in cost, expensive, time-consuming, and cannot meet the needs of modern environmental pest outbreaks and prevention and control.

Method used

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  • Domestication method and application of nuclear polyhedrosis virus adapted to spodoptera frugiperda
  • Domestication method and application of nuclear polyhedrosis virus adapted to spodoptera frugiperda
  • Domestication method and application of nuclear polyhedrosis virus adapted to spodoptera frugiperda

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preparation example Construction

[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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Abstract

The invention discloses a domestication method and application of a nuclear polyhedrosis virus adapted to spodoptera frugiperda. The domestication method includes the following steps: performing UV irradiation on an autographa californica nuclear polyhedrosis virus, applying the treated virus to artificial feed of the spodoptera frugiperda to obtain virus-containing feed, feeding hungry spodopterafrugiperda larvae with the virus-containing feed, collecting poisonous insects or dead insects, performing crushing, performing filtration, and performing centrifugal collection to obtain a free-state autographa californica nuclear polyhedrosis virus; and performing iteration culture and amplification alternately on the free-state autographa californica nuclear polyhedrosis virus by using isolated cells and insects of the spodoptera frugiperda to obtain a domesticated autographa californica nuclear polyhedrosis virus, and improving viral titer of the domesticated virus to the spodoptera frugiperda by an insect cultivating method to obtain the nuclear polyhedrosis virus adapted to the spodoptera frugiperda. The autographa californica nuclear polyhedrosis virus provided by the invention hasthe characteristics of strong pertinence, long control acting time, no side effects, and difficulty in producing drug resistance by pests, and the virus domestication efficiency is fast and efficient.

Description

technical field [0001] The invention relates to a domestication and preparation method of a nuclear polyhedrosis virus, in particular to a method for domestication and industrial production of a nuclear polyhedrosis virus suitable for Spodoptera frugiperda and its application, and belongs to the technical field of biological pesticides. Background technique [0002] Spodoptera frugiperda (J.E.Smith), also known as fall armyworm, belongs to the family Noctuidae of the order Lepidoptera. Native to tropical and subtropical regions of America, it is an important pest on local corn. With the increasing frequency of international trade activities, fall armyworm has invaded 44 African countries south of the Sahara and Asian countries such as India, Bangladesh, Sri Lanka, and Myanmar. It began to invade my country in 2019, causing serious harm to my country's agricultural production. . [0003] The Food and Agriculture Organization of the United Nations and the World Health Organiz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A01N63/40A01P7/04C12R1/93
CPCC12N7/00A01N63/00C12N2710/14151C12N2710/14171C12N2710/14131
Inventor 滕小锘孙祥明袁秀丽金亮
Owner SUZHOU MIDI BIOTECH CO LTD
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