CN113331139B - Method for improving BmNPV virus resistance of silkworm offspring and application thereof - Google Patents
Method for improving BmNPV virus resistance of silkworm offspring and application thereof Download PDFInfo
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- CN113331139B CN113331139B CN202110558964.8A CN202110558964A CN113331139B CN 113331139 B CN113331139 B CN 113331139B CN 202110558964 A CN202110558964 A CN 202110558964A CN 113331139 B CN113331139 B CN 113331139B
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- 241000255789 Bombyx mori Species 0.000 title claims abstract description 89
- 241000409811 Bombyx mori nucleopolyhedrovirus Species 0.000 title claims abstract description 34
- 238000000034 method Methods 0.000 title claims abstract description 16
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- 230000013011 mating Effects 0.000 claims abstract description 5
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- 239000000645 desinfectant Substances 0.000 abstract description 9
- 230000036039 immunity Effects 0.000 abstract description 6
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- 235000008708 Morus alba Nutrition 0.000 description 1
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
- A01K67/04—Silkworms
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for resisting BmNPV virus by high silkworm offspring and application thereof, and aims to solve the technical problems of ecological deterioration and disinfectant virus resistance caused by chemical disinfectants in the prior art. The method of the invention comprises the following steps: feeding the silkworm chrysalis or the later chrysalis in 18-22 deg.c, mating with moth and spawning; storing the silkworm eggs obtained by spawning as silkworm seeds; and incubating the silkworm seeds to obtain BmNPV virus-resistant silkworm offspring. The method of the invention avoids environmental pollution and virus resistance to chemical disinfectants, can effectively improve the capability of treating the BmNPV virus resistance of the offspring of silkworms, reduce the death rate of the offspring of silkworms, improve the cocoon yield of the offspring of silkworms and improve the immunity of the offspring of silkworms.
Description
Technical Field
The invention relates to the technical field of silkworm breeding, in particular to a method for improving BmNPV virus resistance of silkworm offspring and application thereof.
Background
Silkworm is a lepidoptera silk-secreting insect taking mulberry leaves as food, and the whole life of the silkworm passes through different development stages in terms of egg, larva, pupa and adult morphology and physiological function. Silkworm nuclear polyhedrosis virus (BmNPV) is one of the most sensitive and common pathogens of silkworm species, and the silkworm is suppuratively and quickly killed after infection. In recent years, the number of silkworm eggs eliminated by the overtoxic in China is high every year, so that huge economic losses of the silkworm industry are caused, and the direct economic losses caused by silkworm eggs can reach tens of millions of yuan.
The silkworm nuclear polyhedrosis virus (BmNPV) particle is in a rod shape, belongs to the baculovirus family, the baculovirus genus and the silkworm cell nuclear polyhedrosis virus species, and is a representative species of the mononuclear capsid nuclear polyhedrosis virus. The outer envelope, which contains double-stranded DNA molecules, during infection, the virion is assembled by tissue using the replicated DNA and the synthesized protein, the envelope is obtained from the host cell membrane by budding, after which the viral infection propagates throughout the host and secondary infection can be caused by the tracheal system of the silkworm, resulting in death of the host.
At present, screening of resistant varieties by a breeding method faces many limitations such as long research and development period, poor directionality, difficulty in considering economic characters while improving resistance, and the like; most silkworm farms mainly disinfect silkworm farms and tools through a disinfectant for silkworms, and the pathogen in the environment is purified so as to avoid infection of silkworms. However, prolonged use of disinfectants can lead to the development of viral resistance and contamination of the surrounding environment.
Therefore, there is currently no clear and efficient way to control BmNPV disease.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for improving the resistance of silkworm offspring to BmNPV virus and application thereof, so as to solve the technical problems of ecological deterioration caused by chemical disinfectants and disinfectant virus resistance generation.
In order to solve the technical problems, the invention adopts the following technical scheme:
a method for improving the resistance of silkworm offspring to BmNPV virus is provided, which comprises the following steps:
(1) Feeding the silkworm chrysalis or the later chrysalis in 18-22 deg.c, mating with moth and spawning;
(2) Storing the silkworm eggs obtained by spawning as silkworm seeds;
(3) And incubating the silkworm seeds to obtain BmNPV virus-resistant silkworm offspring.
Preferably, in the step (1), the pupa late stage is an adult stage.
Preferably, in the step (2), the non-diapause eggs in the silkworm eggs are placed to 22-28 ℃ for hatching.
Further, the hastening green condition is: the ambient temperature is 27+/-1 ℃ and the humidity is 80-85%.
Preferably, in the step (2), diapause eggs in the silkworm eggs are refrigerated 36 to 120 hours or several months after oviposition.
The method for improving the resistance of the offspring of the silkworms to BmNPV virus is applied to silkworm breeding.
Compared with the prior art, the invention has the beneficial technical effects that:
1. according to the method, the silkworm in the pupa stage or the adult stage is placed under the low-temperature condition for treatment, so that the capability of treating the BmNPV virus resistance of the offspring of the silkworm can be effectively improved, the death rate of the offspring of the silkworm is reduced, the cocoon yield of the offspring of the silkworm is improved, the immunity of the offspring of the silkworm is improved, and the effect is remarkable.
2. The method is applicable to all silkworm varieties and has universality.
3. The method of the invention does not use chemical disinfectants, and avoids environmental pollution and virus resistance to the chemical disinfectants.
4. The method has low cost, simple and convenient operation and is easy to popularize and apply in the silkworm area.
Drawings
FIG. 1 is a statistical graph of mortality in different temperature treatment groups after 96h of BmNPV virus treatment in the examples;
FIG. 2 is a graph showing the weight statistics of cocoons treated with BmNPV virus in the examples at different temperatures;
FIG. 3 is a statistical graph of the number of nodules formed in the treatment group at different temperatures after 8h of BmNPV virus treatment in the examples;
FIG. 4 is a statistical graph of the migration number of blood cells in the treatment group at different temperatures after BmNPV virus treatment for 8 hours in the example.
Detailed Description
The following examples are given to illustrate the invention in detail, but are not intended to limit the scope of the invention in any way.
The instruments and devices referred to in the following examples are conventional instruments and devices unless otherwise specified; the related reagents and culture mediums are all conventional products on the market unless specified; the test methods are conventional, unless otherwise specified. The silkworm variety is 'large-sized', and the variety has been subjected to silkworm genome sequencing, so that the silkworm variety has basic commonality; the related feeding adopts a Shanghai-constant artificial climate box as a feeding room; the temperature precision is +/-1 ℃ and the humidity precision is +/-5% in the feeding conditions.
Example 1: influence of different temperature treatment on silkworm in pupal stage
(1) Feeding silkworms to five-instar cocoons to form pupas under normal conditions, and then respectively putting the silkworms into a climatic chamber for feeding treatment at 20 ℃ (treatment group 1), 25 ℃ (control group) and 30 ℃ (treatment group 2) until mating of the chemical moths and spawning are carried out;
(2) Respectively placing silkworm eggs at room temperature for preservation, directly placing non-diapause eggs at 25 ℃ for hatching, wherein the silkworm eggs are subjected to hatching under the following conditions: the ambient temperature is 27+/-1 ℃ and the humidity is 80-85%;
(3) Incubating each group of silkworm eggs, collecting ant, feeding under normal feeding conditions, feeding silkworm to five-year first day with 1×10 4 BmNPV virus at units/ml concentration was inoculated, 30 seeds were inoculated in each group, and 3 replicates were set.
(4) Detecting the BmNPV virus resistance of the silkworms: and detecting the influence of BmNPV virus treatment on the mortality rate, the cocoon weight and the immunity of the next generation of silkworms raised at different temperatures.
The mortality calculation is shown below:
mortality (%): the mortality was counted 96 hours after BmNPV virus inoculation, and the number of dead silkworms/the total number of experimental silkworms was multiplied by 100%
The method for calculating the weight of the cocoons comprises the following steps:
the weight of each cocoon was weighed and 10 replicates were performed, and the average value was compared with the normal temperature treatment group.
The immunity detection method comprises the following steps:
From the detection results, the offspring treated with the BmNPV virus at the pupal stage of 20 ℃ (treated group 1) have a mortality rate slightly higher than that of the control group but lower than that of the treated group 2 compared with the offspring treated at the normal temperature of 25 ℃ (control group) and the offspring treated at the normal temperature of 30 ℃ (treated group 2); the weight of each silkworm cocooning (figure 2) is higher than that of the control group and the treatment group 2; the number of nodules and the number of blood cells migrated (fig. 3 and 4) were higher than the control and treatment groups 2, the offspring of treatment group 1 were immune-enhanced.
Example 2: influence of different temperature treatment on silkworms in adult stage
(1) Feeding silkworm to five-instar cocoon moth under normal condition, respectively placing into artificial climatic box for feeding treatment at 20deg.C (treatment group 1), 25deg.C (control group) and 30deg.C (treatment group 2) until mating and spawning,
(2) Respectively placing silkworm eggs at room temperature for preservation, and directly placing non-diapause eggs at 25 ℃ for hatching, wherein the silkworm eggs are subjected to normal hatching conditions: the ambient temperature is 27+/-1 ℃ and the humidity is 80-85%;
(3) Incubating each group of silkworm eggs, collecting ant, feeding under normal feeding conditions, feeding silkworm to five-year first day with 1×10 4 BmNPV virus at units/ml concentration was inoculated, 30 seeds were inoculated in each group, and 3 replicates were set.
(4) Detecting the BmNPV virus resistance of the silkworms: and detecting the influence of BmNPV virus treatment on the mortality rate, the cocoon weight and the immunity of the next generation of silkworms raised at different temperatures. The detection method is the same as in example 1.
From the detection results, the death rate of the offspring treated with BmNPV virus (see figure 1) is obviously reduced after the offspring treated with the adult stage at 20 ℃ are compared with the offspring treated with the adult stage at normal temperature at 25 ℃ (control group) and the offspring treated with the adult stage at 30 ℃ (treatment group 2); the weight of each silkworm cocooning (figure 2) is obviously increased; both the number of nodules and the number of blood cells migrated (fig. 3 and 4) increased significantly, with greatly enhanced immunity in the offspring of treatment group 1.
In conclusion, the resistance of the offspring of the silkworms to BmNPV virus can be enhanced by the feeding treatment at the silkworm pupa stage of 20 ℃, and the cocoon yield is increased; the raising treatment at 20 ℃ in the adult period of the silkworms can greatly enhance the resistance of the offspring of the silkworms to BmNPV virus, obviously reduce the death rate and increase the cocoon yield.
While the present invention has been described in detail with reference to the drawings and the embodiments, those skilled in the art will understand that various specific parameters in the above embodiments may be changed without departing from the spirit of the invention, and a plurality of specific embodiments are common variation ranges of the present invention, and will not be described in detail herein.
Claims (3)
1. A method for increasing resistance of a silkworm offspring to a BmNPV virus, comprising the steps of:
(1) Feeding the silkworm chrysalis or the later chrysalis in 18-22 deg.c, mating with moth and spawning; the later pupa stage is an adult stage;
(2) Storing the silkworm eggs obtained by spawning as silkworm seeds; placing the non-diapause eggs in the silkworm eggs to be subjected to hatching under the conditions of 22-28 ℃ and 80-85% of humidity; diapause eggs in the silkworm eggs are refrigerated after 36-120 hours or months after spawning;
(3) And incubating the silkworm seeds to obtain BmNPV virus-resistant silkworm offspring.
2. The method for improving the resistance of the offspring of silkworms to the BmNPV virus according to claim 1, wherein the hastening conditions are as follows: the ambient temperature was 27.+ -. 1 ℃.
3. The low-temperature treatment of silkworm pupa period or adult period at 18-22 ℃ is applied to improving BmNPV virus resistance of silkworm offspring.
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| CN102433340A (en) * | 2011-11-25 | 2012-05-02 | 西南大学 | Resistance key gene BmPGRP2 of silkworm to nuclear polyhedrosis virus and application thereof |
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| CN102433340A (en) * | 2011-11-25 | 2012-05-02 | 西南大学 | Resistance key gene BmPGRP2 of silkworm to nuclear polyhedrosis virus and application thereof |
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| 黎京才 ; ."桑蚕养殖及病虫害防治技术探究".《南方农业》.2019,第13卷(第30期),第145、151页. * |
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