CN110951697A

CN110951697A - Prinsepia utilis nuclear polyhedrosis virus and production method thereof

Info

Publication number: CN110951697A
Application number: CN201911184544.7A
Authority: CN
Inventors: 王金昌; 靳亮; 占智高
Original assignee: INSTITUTE OF MICROBIOLOGY JIANGXI ACADEMY OF SCIENCES
Current assignee: Suzhou Womei Biology Co ltd
Priority date: 2019-11-27
Filing date: 2019-11-27
Publication date: 2020-04-03
Anticipated expiration: 2039-11-27
Also published as: CN110951697B

Abstract

The invention provides a flat moth Nuclear Polyhedrosis Virus, which is a flat moth Nuclear Polyhedrosis Virus Nanchang strain (Thosea sinensis Nuclear Polyhedrosis Virus-Nanchang, TsNPV-NC), the Virus is preserved in a China center for type culture collection in 2019, 10 and 25 days, and the preservation number is CCTCC NO: v201975. The TsNPV-NC virus provided by the invention has stronger toxicity, has good development and application prospects, and has application values in preparation of insecticides and prevention and control of Plutella xylostella. The invention also provides a method for producing TsNPV-NC by using the flat spiny moth larvae fed with the artificial feed.

Description

Prinsepia utilis nuclear polyhedrosis virus and production method thereof

Technical Field

The invention belongs to the technical field of viruses, and particularly relates to a flat spiny moth nucleopolyhedrosis virus and a production method thereof.

Background

The flat leaf moth (Thosea sinensis Walker) belongs to the family of Choristidae in Lepidoptera, and is also called black-spot leaf moth and hot pepper. The flat spiny moth is widely distributed (23 provinces in China and some countries in southeast Asia are distributed), and has edible impurities (the hosts are 71 kinds in 37 families), and the main hosts comprise tea-oil trees, tea trees, walnuts, persimmons, jujubes, apples, pears, Chinese tallow trees, sweetgum, Chinese ash, poplar, boxwood, willow, sweet osmanthus, chinaberry, cinnamomum camphora, paulownia, tung tree, sycamore, camptotheca acuminata, gingkoes, mulberries, Chinese chestnuts and other forest trees and fruit trees.

The flat spiny moth occurs for 2-3 generations in south Chang in one year, and the larval instar is as follows: 2 days for the first instar, 3-4 days for the second instar, 5-6 days for the third instar, 7-8 days for the fourth instar, 6-9 days for the fifth instar, 5-10 days for the sixth instar, and the period of the larva is about 28-40 days, and the damage time is as long as half a year from 5 months to 10 months. The flat spiny moth reaches 43 plants of 30 families on the host of Nanchang, is one of the main leaf-eating pests in Jiangxi province, leaves of the young larva can eat into a net shape, leaf veins are remained, the young larva can eat into notches and holes after 4 years, and only leaf stalks and main veins are remained. After 6 years, the food intake is greatly increased, and the whole leaves are completely eaten, so that the fruit yield is reduced. The loss of the blade is 10 to 15 percent under normal conditions, and can reach more than 30 percent when the loss is serious; aggregation and spawning of the Ophiopogon laticifera and mutual attraction among larvae lead to high-density aggregation of the Ophiopogon laticifera larvae on a certain host, and when the Ophiopogon laticifera eats leaves of a smooth whole tree, the tree dies.

Except that the larvae of the Sporisria punctata harm trees, the larvae touch the skin of people to cause dermatitis, the skin of the pricked parts has severe burning pain, serious people have severe pain, a few of the Sporisria punctata can be accompanied with symptoms such as headache, dizziness, nausea, vomit, palpitation, discomfort of the whole body, swelling and pain of local lymph nodes and the like, and the allergens of the Sporisria punctata can cause allergic asthma. Therefore, the method has economic value in production and has important significance in guaranteeing the physical and psychological health of human beings.

Chua Xiuyu et al found Spodoptera frugiperda nuclear Polyhedrosis Virus (TsNPV) for the first time in Beijing at 7 months 1964, and subsequently in Jiangxi, Hebei, Anhui, Hunan, Shandong. The virus can be spread in the Plectranthus niveus population through horizontal diffusion and vertical transmission, the natural infection rate is generally 43%, and can reach more than 70-87.2% at high speed, even 100%, and the virus is an important biological factor for controlling the Plectranthus niveus population quantity. Toxicity determination, host specificity test, toxicity test and field test are carried out on TsNPV by a biocontrol room of the institute of plant protection of agricultural sciences of Anhui province, and through the indoor and field tests, the virus has strong toxicity and host specificity, has high control effect on the Ophiophaga litura (the Achiophaga litura larvae in different instars are sprayed by three concentrations of 6 hundred million PIB/ml, 0.6 hundred million PIB/ml and 0.06 hundred piB/ml for 15 days, the death rate of 2 instars is 100 percent, 100 percent and 66.7 percent, the death rate of 3 instars is 100 percent, 100 percent and 96.2 percent, the death rate of 4 instars is 100 percent, the death rate of 5 instars is 100 percent, 96.8 percent and 85.9 percent, the death rate of 6 instars is 33.8 percent, 18.9 percent and 42.8 percent), has certain infectivity on the Ophiophaga moth and the Ophiophaga, and is nontoxic to silkworm and white rats. The method for preventing and controlling the flat moth by spraying does not need expensive equipment, is simple and easy to implement, has low cost and is convenient to popularize and apply. It can be seen that TsNPV has strong toxicity to Ophiophaga littoralis, and TsNPV has no infectivity, pathogenicity and other toxicity to human and young spinal animals (biocontrol chamber of institute of plant protection of agronomy academy of agricultural sciences, Anhui, etc.. Ophiophaga litura nuclear polyhedrosis virus research [ J ] microbiological report, 1979(5): 14-16), and can form viruses in pest populations by horizontal diffusion and vertical transmission.

In recent years, the flat spiny moth is abused to cause disasters in various districts, roads, orchards and tea gardens of Nanchang, auxiliary means such as manually catching the flat spiny moth larvae, removing worm cocoons, trapping and killing adults by lamps and the like can be adopted for preventing and treating the flat spiny moth, and chemical pesticides are mainly used at present. However, the use of a large amount of chemical pesticides not only causes the pesticide residue in forest areas to exceed the standard, but also damages the natural enemies of pests (mantis, lygus lucorum, prodigiosus too clitocybe kummenseng, prodiginis bilineata, ichthyophthirius multifiliis, trichogramma, and the like) while killing the pests, breaks down the ecological balance, easily causes the drug resistance of the pests, and is not beneficial to the sustainable control of the pests. The best approach is to adopt biological control, wherein spraying TsNPV virus preparation is the most effective approach.

Disclosure of Invention

The invention aims to provide a prodigiosin nucleopolyhedrosis virus with stronger toxicity and a production method thereof.

In order to achieve the above object, the present invention provides a sphingas mori Nuclear Polyhedrosis Virus (TsNPV), which is a sphingas mori Nuclear Polyhedrosis Virus strain (TsNPV-NC) deposited in the chinese typical culture collection at 10-25 months in 2019 with a collection number of CCTCC NO: v201975.

The invention also provides application of the prinsepia utilis nuclear polyhedrosis virus in preventing and treating the prinsepia utilis.

The invention also provides a pesticide containing the flat moth nucleopolyhedrosis virus.

The invention also provides a production method of the above-mentioned flat moth nucleopolyhedrosis virus, including,

(1) collecting the flat spiny moth and feeding the flat spiny moth by using artificial feed, wherein the artificial feed comprises 80-100 g of cooked soybean meal and 80-10 g of cooked wheat flour0g, 10-20 g of yeast powder, 50-70 g of cane sugar, 1-1.5 g of glycine, 10-15 g of agar, 0.5-1.5 g of Vc and composite V_B0.5-1.5 g of ethylparaben, 0.3-0.8 g of 10% sodium hydroxide and 600-1000 mL of tap water;

(2) feeding the larvae of the Sporisria insignis with an artificial feed containing the Sporisria insignis nuclear polyhedrosis virus, and starting feeding a nontoxic artificial feed after feeding for 24-36 hours until all the Sporisria insignis die.

(3) Collecting the platypodium platypomum cadavers to obtain the platypodium platypomum nuclear polyhedrosis virus.

Preferably, in the step (1), the larvae of the Plutella xylostella are raised in an intelligent illumination incubator, the temperature is controlled to be 28 +/-1 ℃, the relative humidity is 80-90%, and the light cycle L: d ═ 16: 8, the adult stage is carried out in a greenhouse, the temperature is 20-32 ℃, the average temperature is 25 ℃, the relative humidity is 80-90%, and natural illumination is carried out.

Preferably, in the step (2), dead bodies of the larvae of the tonsillitis are taken, soaked in sterilized 0.02mol/L PBS (pH 7.2) for 10min, vigorously shaken or vortexed for 1min, filtered by 3 layers of gauze, the filtrate is subjected to differential centrifugation for several times, firstly centrifuged at 800r/min for 5min, the supernatant is taken, then centrifuged at 4000 r/min for 5min, the supernatant is discarded, and sterile water is suspended until relatively pure polyhedrosis are obtained, so that the tonsillitis nucleopolyhedrosis virus is obtained.

Preferably, in step (2), the concentration of the Spodoptera frugiperda nucleopolyhedrovirus is 1 × 10⁵And PIB/mL, wherein the larva age of the flat moth is 5-6.

The concentration of the Prinsepia utilis nuclear polyhedrosis virus provided by the invention is 1 multiplied by 10⁷PIB/mL、1×10⁶PIB/mL、 1×10⁵PIB/mL、1×10⁴PIB/mL、1×10³The larvae of the Plutella xylostella die within 2d after the PIB/mL infects the larvae of the Plutella xylostella, the death rate of the larvae of the Plutella xylostella is correspondingly improved along with the increase of the infection dose, and the death rate is 1 multiplied by 10⁷PIB/mL、 1×10⁶PIB/mL、1×10⁵And after 3 concentrations of PIB/mL infect the larvae of the plutella xylostella within 8-9 days, the death rate of the larvae of the plutella xylostella reaches 100%. 1X 10⁷PIB/mL、1×10⁶PIB/mL、1×10⁵The half-to-death time of 3 concentrations of PIB/mL is 4dRight, 1X 10⁴PIB/mL、1×10³PIB/mL、1×10²PIB/mL、1×10¹Half-to-dead times of 4 concentrations of PIB/mL were 6d, 7d, 8d and 8d, respectively; therefore, the TsNPV virus provided by the invention has stronger toxicity than the TsNPV separated from plant protection research institute biocontrol rooms and the like of the agricultural academy of sciences of Anhui province, and has good development and application prospects.

The flat moth Nuclear Polyhedrosis Virus is prepared from flat moth Nuclear Polyhedrosis Virus Nanchang strain (Thoseasinensis Nuclear Polyhedrosis Virus-Nanchang, TsNPV-NC), the preservation date is 2019, 10 and 25 days, and the preservation number is CCTCC NO: v201975, classified and named as Spodoptera litura Nuclear Polyhedrosis Virus Nanchang strain TsNPV-NC (Thosea sinensis Nuclear Polyhedrosis Virus Nanchang), and the name of the depositary unit is: china center for type culture Collection, the collection center addresses are: wuhan university in Wuhan, China.

Drawings

FIG. 1 is a diagram of a typical "liquefied" condition after death of a virus infected larva as in example 1;

FIG. 2 is a mirror image of the TsNPV polygon in example 1;

FIG. 3 is a drawing of a 200kV cryo-transmission electron microscope of TsNPV sections;

Detailed Description

In order to explain the technical solutions, structural features, and technical effects achieved by the present invention in detail, the following detailed description and the accompanying drawings are given.

In the following examples, TsNPV virus was isolated from cadavers of naturally dead larvae of Buxus Davidiana in the park of the Exi lake wetland in Nanchang, Jiangxi; the larvae of the Sphaeria indica to be tested are healthy larvae of Sphaeria indica grown in Buxus Davidiana in wetland parks of Nanchang, Jiangxi; purchasing a Xitianlong mini storage box as a pest cultivation vessel; the artificial feed adopts leaf branches of Buxus sinica (washed with sterile water for 3 times, air dried, and irradiated with ultraviolet rays for 15min) before feeding.

Example 1 TsNPV Virus isolation, multiplication, extraction and identification

TsNPV virus stock solution (separated from Aixi lake wetland male city in Nanchang, Jiangxi)Dead insect bodies of euonymus japonicus flat moth larvae naturally died) diluted to 1 × 10 with sterile water⁶And (3) spraying the PIB/mL solution onto branches with leaves of the Buxus sinica Diels by using a plastic sprinkling can, drying in the shade, putting into a Happy dragon mini storage box, and accessing collected 4-6 th instar Bombycis Mori larvae. 28 ℃, light week 9: 15 breeding (light breeding for 9h and dark breeding for 15h) until all the larvae of the Plutella xylostella die. Observing every day, feeding for 48h, replacing with fresh Buxus Dayesii branches with leaves, taking out in time, and storing in a refrigerator at 4 deg.C.

Then, dead insect bodies of the larvae of the flat moth are taken, soaked for 10min by sterilized 0.02mol/L PBS (pH7.2), vigorously shaken or vortexed for 1min, filtered by 3 layers of gauze, and subjected to differential centrifugation for several times (firstly, centrifugation is carried out at 800r/min for 5min, supernatant is taken out, centrifugation is carried out at 4000 r/min for 5min, the supernatant is discarded, and sterile water is suspended) until purer polyhedra are obtained. Under an optical microscope, counting and photographing are carried out by using a blood counting chamber, and the virus liquid is stored in a refrigerator at 4 ℃.

Quickly fixing the purified virus polyhedra with special glutaraldehyde for an electric mirror, rinsing with 0.1mol/L PBS, fixing with 1% osmium tetroxide, and rinsing with 0.1mol/L PBS; using 30% -100% acetone to dehydrate in a gradient way; embedding and polymerizing by using epoxy resin; and positioning the semi-thin slices and slicing by using an ultrathin slicer. After uranium acetate and catalpol lead solution are subjected to double staining, the prepared section copper net is placed under a 200KV freezing transmission electron microscope FEI Tecnai G20, the accelerating voltage is 200V, and an electron microscope picture is taken.

The results are shown in figure 1, figure 2 and figure 3, the pleiothis obliquus larvae are infected with virus, the appetite is reduced, the actions are slow, the diseased larvae turn yellow in the early stage and finally turn yellow brown, grey brown or black brown, after the larvae die, the bodies turn soft, the tissues in the bodies are liquefied, the epidermis is loose and easy to break, milky white to brown pus juice flows out, the odor is not generated, and the typical 'liquefied' disease is shown in figure 1. The liquefied blood cells and fat tissues of the larvae of the flat moth are tabletted, so that the nuclear polyhedrosis virus proliferated in cell nuclei can be observed, and the TsNPV amplification success of the flat moth nuclear polyhedrosis virus can be observed.

The larvae of the Plutella xylostella are infected with TsNPV virus for 8d and all the larvae die, the collected larvae are soaked in PBS, TsNPV is extracted by differential centrifugation after violent oscillation, virus polyhedra with strong refractivity are visible by microscopic examination of an optical microscope, the microscopic examination result of the TsNPV is shown in figure 2, and the virus polyhedra with strong refractivity are shown in the figure.

The virus is identified by an ultrathin section electron microscope in an electron microscope room of the institute of Wuhan virus of Chinese academy of sciences, and the polyhedrosis virus is an irregular polyhedron from pentagon to nearly circular. The virions are rod-latent and the single particles are embedded in the protein, see FIG. 3. From the disease state of the infected larvae, the morphology and size of polyhedra and virions, the virus was identified as a nuclear polyhedrosis virus in the family Baculoviridae as fourth reported by the International Committee for viral Classification.

Example 2 bioassay of TsNPV Virus

TsNPV virus was diluted to 1X 10 gradient with sterile water⁷PIB/mL，1×10⁶PIB/mL，1×10⁵PIB/mL，1×10⁴PIB/mL，1×10³PIB/mL，1×10²PIB/mL，1×10¹6 concentrations of PIB/mL. And (4) respectively soaking branches with leaves of the Buxus sinica Maxim with each concentration of TsNPV virus liquid, airing, and carrying out feeding experiments. Each group of the obtained mixture is divided into a group, a certain amount of larvae of Plutella xylostella are placed in each group, the groups are respectively placed in a sterilized container, the container is tightly tied and sealed by using wet gauze, sterile water is coated for comparison, and the steps are repeated for 2 times. After 24h of infection, fresh clean leaves were added to continue feeding. And observing and recording according to the test requirements. Mortality was calculated, median time to death was calculated for each different virus concentration, and the resulting data were analyzed for variance using SPSS software.

The results are shown in Table 1, from which it can be seen that 1X 10⁷PIB/mL、1×10⁶PIB/mL、1×10⁵PIB/mL、 1×10⁴PIB/mL、1×10³The larvae of the Plutella xylostella die within 2d after the PIB/mL infects the larvae of the Plutella xylostella, the death rate of the larvae of the Plutella xylostella is correspondingly improved along with the increase of the infection dose, and the death rate is 1 multiplied by 10⁷PIB/mL、1×10⁶PIB/mL、 1×10⁵And after 3 concentrations of PIB/mL infect the larvae of the plutella xylostella within 8-9 days, the death rate of the larvae of the plutella xylostella reaches 100%. 1X 10⁷PIB/mL、1×10⁶PIB/mL、1×10⁵Half-to-dead times of 3 concentrations of PIB/mL were allAbout 4d, 1 × 10⁴PIB/mL、1×10³PIB/mL、1×10²PIB/mL、1×10¹Half-to-dead times of 4 concentrations of PIB/mL were 6d, 7d, 8d and 8d, respectively; therefore, the separated TsNPV virus has stronger toxicity than the TsNPV separated by a plant protection research institute of agricultural academy of sciences of Anhui province, and has good development and application prospects.

TABLE 1 mean corrected mortality of TsNPV lethal to Trichoplusia ni larvae infection

The SPSS software is used for variance analysis, the test result of the effect between subjects is shown in table 2, and the sig value of the virus concentration, the death time and the virus concentration-death time is 0.0000 to less than 0.01, which shows that the difference of the lethal effect of different concentrations of TsNPV on the larvae of the Plutella xylostella is obvious.

TABLE 2 examination of the Effect between subjects

Example 3 establishment of method for producing TsNPV virus by feeding Spodoptera frugiperda with artificial feed

3.1 feeding Flat thorn moth with artificial feed

Preparing artificial feed: cooked soybean powder 90g, cooked wheat flour 90g, yeast powder 15g, cane sugar 60g, glycine 1.25g, agar 12g, Vc and composite V_B1g of each, 0.5g of ethylparaben, 37.5mL of 10% potassium hydroxide and 800mL of tap water. Dissolving agar, adding ethylparaben and potassium hydroxide, stirring, adding semen glycines powder, semen Tritici Aestivi powder and yeast powder, stirring, cooling to about 60 deg.C, adding vitamins, stirring, and packaging.

Obtaining a flat moth source: collecting aged larvae of Oplophora platyphylla from field, feeding into pupation with host plant with leaf branches, feeding into adult with honey water until mating and oviposition, collecting ovum blocks, sterilizing with formaldehyde, and collecting hatched larvae as insect source. Feeding conditions are as follows: feeding the larvae in an intelligent illumination incubator, controlling the temperature to be 28 +/-1 ℃, controlling the relative humidity to be 80-90%, and controlling the light cycle L: d ═ 16: 8, the adult stage is carried out in a greenhouse, the temperature is 20-32 ℃, the average temperature is 25 ℃, the relative humidity is 80-90%, and natural illumination is carried out.

Feeding larvae newly hatched from the plutella xylostella to pupate by using artificial feed for the plutella xylostella and host plant branches with leaves respectively, feeding the adults by using honey water after the pupae eclosion becomes adults, mating and laying eggs, calculating indexes of 6-instar larvae length, weight, larva development history, pupal weight, pupal period, eclosion rate, adult weight, egg laying amount and the like of the plutella xylostella fed by using the artificial feed for the plutella xylostella and the host plant branches with leaves respectively, evaluating the nutritional status of the feed, and determining the influence of the artificial feed on the growth and development of the plutella xylostella. Feeding conditions are as follows: feeding the larvae in an intelligent illumination incubator, controlling the temperature to be 28 +/-1 ℃, controlling the relative humidity to be 80-90%, and controlling the light cycle L: d ═ 16: 8, the adult stage is carried out in a greenhouse, the temperature is 20-32 ℃, the average temperature is 25 ℃, the relative humidity is 80-90%, and natural illumination is carried out.

The results are shown in Table 3, the average body length of the six-instar larvae is 21mm, the average weight of the six-instar larvae is 1.1g, the development period is 28-35 d, the average pupa weight is 0.55g, the pupal period is 9-15 d, the eclosion rate is 98%, and the average egg laying amount of each female moth is 116 grains.

TABLE 3 growth and development results of Onychid moth fed with artificial feed and host plant

3.1 determination of optimal conditions for TsNPV proliferation

TsNPV virus was diluted to 1X 10 gradient with sterile water⁷PIB/mL，1×10⁶PIB/mL，1×10⁵PIB/mL，1×10⁴PIB/mL，1×10³PIB/mL，1×10²6 concentrations of PIB/mL, ready for use. Respectively selectTaking 600 heads of 4, 5 and 6 instar flat moth larvae, dividing the feed into 1800 small blocks (approximately 1 head of larva for 1 d), inoculating 10 microliter of diluted virus into each block of feed, feeding 100 heads of flat moth larvae of one instar at each virus concentration, feeding nontoxic feed from the 2 nd, and feeding at 27 ℃ until all flat moth larvae die. During the period, attention is paid to observation every day, and the larvae of the flat moth which is died of illness are taken out in time and stored in a refrigerator at 4 ℃. And after all larvae of the flat spiny moth die, collecting dead bodies respectively, and calculating the half-lethal time of each virus concentration.

Taking dead insect bodies of the flat moth larvae, soaking the dead insect bodies in sterilized 0.02mol/L PBS (pH 7.2) for 10min, strongly shaking or whirling for 1min, filtering by using 3 layers of gauze, and carrying out differential centrifugation on the filtrate for several times (firstly centrifuging at 800r/min for 5min, taking supernatant, then centrifuging at 4000 r/min for 5min, discarding supernatant, and suspending in sterile water) until pure polyhedra are obtained. Under an optical microscope, counting by using a blood counting chamber, and calculating the single-head toxicity yield of the flat moth larvae in different instars.

The results are shown in Table 4, using 1X 10⁵TsNPV produced by feeding TsNPV venom of 5 th instar or 6 th instar to PIB/mL can meet the production requirement.

TABLE 4 Spodoptera frugiperda TsNPV proliferation table

The larva of the flat moth is fed with artificial feed (the natural feed is limited by the growing season of the feed plant), so that conditions for factory, mechanization and automation of virus production can be created. The artificial feed for feeding insects has the following advantages: (1) the feed is not limited by seasons and can be fed all the year round; (2) the feed is not limited by hosts and land and can be fed in large quantities; (3) can reduce or avoid the interference of factors such as microbial infection, pesticide and the like; (4) experimental insects with regular and consistent physiological standards are easily obtained; (5) can control the weaving of feed to research the feeding and development of insects.

The concentration of the Prinsepia utilis nuclear polyhedrosis virus provided by the invention is 1 multiplied by 10⁷PIB/mL、1×10⁶PIB/mL、 1×10⁵PIB/mL、1×10⁴PIB/mL、1×10³The larvae of the Plutella xylostella die within 2d after the PIB/mL infects the larvae of the Plutella xylostella, the death rate of the larvae of the Plutella xylostella is correspondingly improved along with the increase of the infection dose, and the death rate is 1 multiplied by 10⁷PIB/mL、 1×10⁶PIB/mL、1×10⁵And after 3 concentrations of PIB/mL infect the larvae of the plutella xylostella within 8-9 days, the death rate of the larvae of the plutella xylostella reaches 100%. 1X 10⁷PIB/mL、1×10⁶PIB/mL、1×10⁵The half-to-death time of 3 concentrations of PIB/mL is about 4d and 1 × 10⁴PIB/mL、1×10³PIB/mL、1×10²PIB/mL、1×10¹Half-to-dead times of 4 concentrations of PIB/mL were 6d, 7d, 8d and 8d, respectively; therefore, the TsNPV virus provided by the invention has stronger toxicity than the TsNPV separated from plant protection research institute biocontrol rooms and the like of the agricultural academy of sciences of Anhui province, and has good development and application prospects.

The above disclosure is only a preferred embodiment of the present invention, and should not be taken as limiting the scope of the invention, so that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.

Claims

1. A Spodoptera frugiperda nuclear polyhedrosis virus, comprising: the Virus is a Nanchang strain of a plutella xylostella Nuclear Polyhedrosis Virus (Thosea sinensis Nuclear Polyhedrosis Virus-Nanchang, TsNPV-NC), is preserved in the China center for type culture collection in 2019 and 10 and 25 months, and has the preservation number of CCTCC NO: v201975.

2. The use of the Autographa californica nuclear polyhedrosis virus of claim 1 for controlling Autographa californica.

3. An insecticide, characterized by: comprising the Autographa californica nuclear polyhedrosis virus of claim 1.

4. The method for producing a Autographa californica nuclear polyhedrosis virus according to claim 1, wherein: comprises the steps of (a) preparing a mixture of a plurality of raw materials,

(1) collecting Oplophora platyphylla, feeding with artificial feed, and making into artificial feed bagComprises 80-100 g of cooked soybean powder, 80-100 g of cooked wheat flour, 10-20 g of yeast powder, 50-70 g of cane sugar, 1-1.5 g of glycine, 10-15 g of agar, 0.5-1.5 g of Vc and compound V_B0.5-1.5 g of ethylparaben, 0.3-0.8 g of 10% sodium hydroxide and 600-1000 mL of tap water;

(2) feeding the larvae of the Ophiophaga ni with an artificial feed containing the Ophiophaga ni nuclear polyhedrosis virus, and after feeding for 24-36 hours, starting feeding the nontoxic artificial feed until all the Ophiophaga ni dies.

5. The method for producing a Spodoptera frugiperda nuclear polyhedrosis virus according to claim 4, comprising: in the step (1), feeding the larvae of the Plutella xylostella in an intelligent illumination incubator, controlling the temperature to be 28 +/-1 ℃, the relative humidity to be 80-90%, and the light cycle L: d ═ 16: 8, the adult stage is carried out in a greenhouse, the temperature is 20-32 ℃, the average temperature is 25 ℃, the relative humidity is 80-90%, and natural illumination is carried out.

6. The method for producing a Spodoptera frugiperda nuclear polyhedrosis virus according to claim 4, comprising: in the step (2), dead insect bodies of the larvae of the plutella xylostella are taken, soaked in sterilized 0.02mol/L PBS (pH 7.2) for 10min, strongly oscillated or vortexed for 1min, filtered by 3 layers of gauze, the filtrate is subjected to differential centrifugation for several times, centrifuged at 800r/min for 5min, the supernatant is taken, centrifuged at 4000 r/min for 5min, the supernatant is discarded, and sterile water is suspended until relatively pure polyhedrosis obtained, so that the plutella xylostella nuclear polyhedrosis virus is obtained.

7. The method for producing a Spodoptera frugiperda nuclear polyhedrosis virus according to claim 4, comprising: in the step (2), the concentration of the flat moth nucleopolyhedrosis virus is 1 x 10⁵And PIB/mL, wherein the larva age of the flat moth is 5-6.