CN108477082B - Method for instantly relieving diapause by heat shock of silkworms and delaying incubation by refrigeration - Google Patents
Method for instantly relieving diapause by heat shock of silkworms and delaying incubation by refrigeration Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
- A01K67/04—Silkworms
Abstract
The invention relates to a method for instantly removing diapause and refrigerating delayed hatching by heat shock of silkworms, which comprises the following steps: s1, hatching the silkworm eggs after pickling, and feeding the silkworm eggs to cocoons; s2, collecting male and female moths of the moths which are simultaneously dissolved, mating for 4-5h, then splitting, placing the male moths on silkworm connecting paper, and laying eggs; s3, after spawning, protecting the silkworm egg-containing silkworm; s4, collecting ants and feeding after hatching the first silkworm egg; hatching the silkworm eggs for the second time, placing the silkworm eggs in an intermediate storage for 2 hours after the silkworm eggs are hatched, wrapping the silkworm eggs with a film, placing the silkworm eggs in a cold storage, taking out a preset amount of silkworm eggs every 3 days, placing the taken-out silkworm eggs in the intermediate storage for 2 hours, and placing the silkworm eggs in an environment at 25 ℃ for collecting and feeding the ants; placing the third part of silkworm egg in the intermediate storage for 6h, wrapping with film in a cold storage, taking out a part of silkworm egg every 5 days after 30 days, placing the silkworm egg in the intermediate storage for 24h, hatching, and collecting ant.
Description
Technical Field
The invention belongs to the fields of biotechnology and bioengineering, and relates to a method for immediately removing diapause by heat shock and delaying incubation by refrigeration of silkworms.
background
The silkworm, commonly called silkworm, is a lepidoptera silking insect with high economic value and using mulberry leaves as foodstuff, and belongs to the mulberry silkworm species of the genera Bombycis of the families Bombycidae of the phylum arthropoda. The silkworm passes through 4 morphological and physiological completely different development stages of egg, larva, pupa and adult, and different parts and metabolites of each development stage have wide medicinal value.
Silkworm has an important color physiological phenomenon, namely, the generation number which occurs in one year. The first-pass silkworm variety only goes through one era, the laid eggs enter a long diapause period until the diapause is finished and the development is started in the second year under the condition of proper temperature and humidity; the bivoltine silkworm variety generates two generations in a natural environment within one year, and the breeding generations can be controlled by artificially releasing diapause; the diversified silkworm varieties are more than three generations in one year; the non-diapause diversified variety has no diapause period. The growth period of the larvae of the bivoltine silkworm varieties and the sizes of the silkworm cocoons are between those of a single variety and those of multiple varieties, and the physique, cocoon silk character and the like of the bivoltine silkworm varieties are also suitable for the production requirements of the silkworm industry, so that the bivoltine diapause eggs are commonly used in the production of the silkworm industry, and the bivoltine variety group with excellent production characters and rich varieties is formed.
In recent years, with the growth of agricultural jianianhua, popular science bases, agricultural science and technology parks and the like all over the country, mulberry silkworm culture is increasingly applied to relevant theme venues (for example, beijing agricultural jianianhua, Guangxi Yulin agricultural jianianhua, Jiangsu Yanghe agricultural jianianhua and the like). In order to fully display the silkworm culture in China, a construction unit needs to purchase specific silkworm seed resources from various breeding organizations for display in addition to purchasing common silkworm seeds on the market. As silkworms are living animals, the time of one generation is only 1 or more months, and most agricultural Jianianhua all carry out year-round orchard establishment operation at present, so that specific germplasm resources are required to be continuously purchased. These exotic resources not only cost high capital, but also cannot be purchased at any time as needed, as opposed to the common silkworm species. These problems generally exist in present silkworm breeding production, if operation units can reserve seeds by themselves, a large amount of purchase funds can be saved, and the requirement of annual display in own garden can be met. Because silkworm eggs have a diapause phenomenon, the conventional production method for removing the diapause mainly comprises low-temperature treatment, pickling treatment, hydrogen peroxide treatment and the like. The low-temperature treatment generally requires low-temperature refrigeration at 5 ℃ for more than 3 months, so that the requirement of hatching at any time cannot be met. The immediate pickling and hydrogen peroxide treatment need professional equipment, operation standards and strict technical requirements, and how to utilize a simple method to immediately relieve diapause becomes a difficult problem. Meanwhile, due to the special display function of the park, the silkworm is required to exist at each period, so that the diapause of the silkworm eggs is required to be relieved immediately, and how to delay the relief of the hatching of the diapause silkworm eggs is required to be researched, so that the silkworm eggs are hatched in batches, and the display effect of the simultaneous existence of the silkworm of different silkworm ages is met.
disclosure of Invention
Aiming at the defects of the prior art, the invention provides a method for instantly removing diapause and delaying incubation by refrigeration through heat shock of silkworms, which adopts a silkworm diapause variety as an original material, instantly removes the diapause through a heat shock technology, delays incubation through a low-temperature refrigeration technology, realizes the incubation of the silkworms at any time, obtains living things of all periods from silkworm eggs-newly-hatched silkworms-1 age-2 age-3 age-4 age-5 age-cocoons, is used for exhibiting in a park and achieves a good exhibition effect.
the invention is realized by the following steps:
A method for heat shock immediate diapause release and refrigeration delayed incubation of silkworms comprises the following steps:
S1, hatching the silkworm eggs after pickling in a climate box environment with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ until hatching and feeding the silkworm eggs to cocoons;
s2, collecting male and female moths of the synchronized moths at a protection temperature of 25 ℃, mating for 4-5 hours at 25 ℃ in indoor light, then splitting, placing the male and female moths on silkworm paper, laying eggs in a dark environment, and laying eggs for 6-8 hours;
s3, after spawning, protecting the silkworm excrement paper with silkworm eggs in a thermostat at 25 ℃, thermally exciting the silkworm excrement paper for 5 seconds by hot water at a preset temperature after a preset time, and drying the silkworm excrement paper in the air and then dividing the dried silkworm excrement paper into three parts;
S4, placing the first silkworm egg subjected to instant heat shock treatment in a climate box with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ for hatching, and then collecting ants for feeding; placing the second part of the silkworm eggs subjected to instant heat shock treatment in a climate box with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ for hatching, placing the silkworm eggs in an intermediate warehouse for 2 hours after the silkworm eggs are spotted, wrapping the silkworm eggs with a film, placing the silkworm eggs in a cold storage, taking out a preset amount of silkworm eggs in batches every 3 days, placing the taken-out silkworm eggs in the intermediate warehouse for 2 hours, and placing the silkworm eggs in an environment at 25 ℃ for collecting and feeding; and placing the third silkworm eggs subjected to instant heat shock treatment in an intermediate storage for 6 hours, refrigerating in a refrigerator, wrapping the silkworm eggs in the refrigerator by using a film, taking out a preset amount of silkworm eggs in batches every 5 days after 30 days, placing the taken-out silkworm eggs in the intermediate storage for 24 hours, then placing in a climate box with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ for hatching, and collecting and feeding the ants.
Preferably, in step S4, the second immediately heat-shocked silkworm egg is spotted and then stored in the refrigerator for a total storage time of 3 to 20 days.
Preferably, in step S4, the third immediately heat-shocked silkworm egg is stored in the refrigerator for a total time of 30 to 50 days.
preferably, in step S4, the second instant heat-shock-treated silkworm eggs are taken out of the refrigerator once every 3 days for a total of 6 times, wherein the amount of the taken-out silkworm eggs is 1/6 of the total amount of the second instant heat-shock-treated silkworm eggs; the third immediately heat-shocked egg was taken out of the refrigerator every 5 days for a total of 4 times, each time the total amount of the third immediately heat-shocked egg was 1/4.
Preferably, the intensity of the indoor light is 50-300 Lux.
preferably, the preset time is 12-24h, and the preset temperature is 53.1 +/-0.5 ℃.
Preferably, the temperature of the intermediate reservoir is 15-16 ℃.
Preferably, the temperature of the refrigerator is 5 ± 0.5 ℃.
Preferably, the refrigerator is a refrigerator.
Preferably, the film is a polyethylene film.
Compared with the prior art, the invention has the following beneficial effects:
The invention provides a simple and easy method for immediately removing the diapause of silkworm eggs, which realizes the acid-immersion-free immediate diapause removal through heat shock without professional acid-immersion diapause removal equipment and operation technology, has no special requirements on equipment and technology, and is simple and convenient and easy to operate;
The method adopts a low-temperature refrigeration delayed incubation technology to realize the incubation of the silkworms at any time and the batch and fractional incubation after the heat shock immediately relieves the diapause of the silkworm seeds, and simultaneously obtains the living matters of each period from the silkworm eggs, namely the silkworm eggs, 1 age, 2 age, 3 age, 4 age, 5 age and cocoons, is used for displaying in a garden, achieves a good display effect, solves the problem that the silkworms with different silkworm ages are displayed at the same time in the current agricultural kingdom and the theme of the silkworms in the agricultural garden, saves high seed purchasing cost, can realize the annual uninterrupted display effect of the silkworm seeds, and is suitable for wide and effective popularization;
the method for instantly releasing diapause and refrigerating delayed incubation by heat shock of the silkworms is simple and easy to operate, production and management of the silkworms can be artificially adjusted according to the requirements of a display plan, specific germplasm resources do not need to be continuously purchased, purchasing and storing work of the resources is simplified, and cost is reduced.
drawings
FIG. 1 is a flow chart of the method for heat shock immediate diapause release and refrigeration delayed hatching of silkworms of the present invention;
FIG. 2 shows the state change of silkworm during heat shock immediate diapause release and cold storage delayed hatching of silkworm.
Detailed Description
Exemplary embodiments, features and aspects of the present invention will be described in detail below with reference to the accompanying drawings. In the drawings, like reference numbers can indicate functionally identical or similar elements. While the various aspects of the embodiments are presented in drawings, the drawings are not necessarily drawn to scale unless specifically indicated.
The invention provides a method for thermally shocking, immediately removing diapause and refrigerating and delaying incubation of silkworms, which is characterized in that the heat shock is adopted to immediately remove diapause and then the low-temperature refrigerating and delaying incubation are utilized to obtain living things of silkworm eggs, namely the silkworm larvae, namely the silkworms, 1 to 2 to 3 to 4 to 5 to cocoon, in each period, and the living things are used for exhibiting in a garden area, and the method comprises the following steps:
s1, hatching the silkworm eggs after pickling in a climate box environment with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ until hatching and feeding the silkworm eggs to cocoons;
S2, collecting male and female moths of the synchronized moths at a protection temperature of 25 ℃, mating for 4-5 hours at 25 ℃ in indoor light, then splitting, placing the male and female moths on silkworm paper, laying eggs in a dark environment, and laying eggs for 6-8 hours;
s3, after spawning, protecting the silkworm egg-containing paper in a thermostat at 25 ℃, thermally exciting the silkworm egg-containing silkworm egg;
S4, directly putting the first silkworm egg subjected to instant heat shock treatment in a climate box with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ for hatching, and then collecting ants for feeding; placing the second silkworm egg subjected to instant heat shock treatment in a climate box with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ for hatching, placing the silkworm egg in an intermediate warehouse with the temperature of 15-16 ℃ for 2 hours after the silkworm egg is spotted, wrapping the silkworm egg by using a polyethylene film, placing the silkworm egg in a cold storage with the temperature of 5 +/-0.5 ℃, taking out the silkworm egg in batches, taking out a part of silkworm egg every 3 days, placing the taken-out silkworm egg in the intermediate warehouse with the temperature of 15-16 ℃ for 2 hours, and then placing the silkworm egg in the environment with the temperature of 25 ℃ for collecting and feeding the silkworm; placing the third instant heat-shock treated silkworm in an intermediate storage at 15-16 deg.C for 6h, refrigerating in a refrigerator at 5 + -0.5 deg.C, and wrapping with polyethylene film in the refrigerator; after 30 days, the silkworm eggs are taken out in batches, a part of silkworm eggs are taken out every 5 days, the taken-out silkworm eggs are firstly placed in an intermediate storage for 24 hours at the temperature of 15-16 ℃, and then are placed in a climate box for hatching and collecting ants for feeding, wherein the temperature of the climate box is 24-25.5 ℃, and the difference between the humidity and the dryness is 1.5-2 ℃.
in this embodiment, the total storage time of the second instantly heat-shocked silkworm in the refrigerator is 3-20 days, and the total storage time of the third instantly heat-shocked silkworm in the refrigerator is 30-50 days.
Example 1
S1, hatching and hatching the silkworm diapause variety q-quail spotted silkworm eggs which are purchased from the silkworm research institute of Chinese academy of agricultural sciences in a climate box environment with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ and feeding the silkworm diapause variety q-quail spotted silkworm eggs to cocoons;
S2, collecting male and female moths of the moths at the cocoon protection temperature of 25 ℃, mating for 4-5h at the temperature of 25 ℃ in room light, then separating, placing the male moths on silkworm paper, laying eggs in the dark environment for 6-8 h;
s3, after spawning, placing the silkworm egg paper in a thermostat at 25 ℃, after 12-24 hours, thermally stimulating the silkworm egg paper with hot water at 53.1 ℃ for 5 seconds, immediately removing diapause, and drying in the air and then dividing into three parts for treatment;
s4, directly putting the first silkworm egg subjected to instant heat shock treatment in a climate box with the temperature of 24-25.5 ℃ and the humidity difference of 1.5-2 ℃ for hatching, and then harvesting and feeding the ants, wherein the hatching rate is about 62%; directly placing the second silkworm eggs subjected to instant heat shock treatment in a climate box with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ for hatching, placing the silkworm eggs in an intermediate warehouse with the temperature of 15-16 ℃ for 2 hours after the silkworm eggs are hatched, wrapping the silkworm eggs with a polyethylene film, placing the silkworm eggs in a refrigerator with the temperature of 5 ℃, taking out 1/6 silkworm eggs every 3 days, taking out the silkworm eggs for 6 times in total, placing the taken-out silkworm eggs in the intermediate warehouse with the temperature of 15-16 ℃ for 2 hours, placing the silkworm eggs in an environment with the temperature of 25 ℃ for preparing ant collection and feeding, wherein the hatching rate is reduced along with the prolonging of the storage time in the refrigerator, and the sixth hatching rate is 45.6 percent as shown in table 1; placing the third instant heat-shock treated silkworm egg in an intermediate storage at 15-16 deg.C for 6h, cooling in a refrigerator at 5 deg.C, and wrapping with polyethylene film in the refrigerator; after 30 days, 1/4 silkworm eggs are taken out every 5 days, the silkworm eggs are taken out in 4 batches in total, the taken silkworm eggs are firstly placed in an intermediate storage at the temperature of 15-16 ℃ for 24 hours and then placed in a climate box at the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ for hatching and feeding the collected silkworms, the hatching rate is as shown in table 1, the hatching rate is slightly increased along with the prolonging of the time of a refrigerating chamber, and the highest hatching rate is 88.5%.
TABLE 1 incubation rates of three different silkworm species after heat shock treatment of silkworm diapause variety q-Coturnix silkworm eggs
Example 2
S1, hatching and hatching the diapause silkworm Ze-tiger spot silkworm eggs which are purchased from the silkworm research institute of Chinese agricultural academy of sciences in a climate box environment with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ and feeding the silkworm eggs to cocoons;
S2, collecting male and female moths of the simultaneous moth transformation under the protection temperature of the seed cocoons at 25 ℃, mating for 4-5h at 25 ℃ under indoor light, then splitting, placing the male moths on silkworm paper, and laying eggs in a dark environment for 6-8 h;
S3, after spawning, protecting the breeding silkworms in a thermostat with the temperature of 25 ℃, thermally stimulating the silkworm connecting paper with hot water with the temperature of 53.1 ℃ for 5S after 12-24h, immediately releasing diapause, drying in the air and then processing in three parts;
s4, directly putting the first silkworm egg subjected to instant heat shock treatment in a climate box with the temperature of 24-25.5 ℃ and the humidity difference of 1.5-2 ℃ for hatching, and then harvesting and feeding the ants, wherein the hatching rate is about 85%; the second part of the silkworm eggs subjected to instant heat shock treatment is directly placed in a climate box with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ for hatching, after the silkworm eggs are hatched, the silkworm eggs are placed in a 15-16 ℃ intermediate warehouse for 2 hours, the silkworm eggs are wrapped by a polyethylene film, the silkworm eggs are placed in a 5 ℃ refrigerator, 1/6 silkworm eggs are taken out every 3 days, the silkworm eggs are taken out in 6 batches in total, the taken silkworm eggs are placed in the 15-16 ℃ intermediate warehouse for 2 hours, and then the silkworm eggs are placed in a 25 ℃ environment for collecting and feeding, wherein the hatching rate is shown in table 2, the hatching rate is reduced along with the prolonging of the storage time in the refrigerator, and the sixth hatching rate is 42.5%; placing the third instant heat-shock treated silkworm egg in an intermediate storage at 15-16 deg.C for 6h, cooling in a refrigerator at 5 deg.C, and wrapping with polyethylene film in the refrigerator; after 30 days, 1/4 silkworm eggs are taken out every 5 days, the silkworm eggs are taken out in 4 batches in total, the taken silkworm eggs are firstly placed in an intermediate storage at the temperature of 15-16 ℃ for 24 hours and then placed in a climate box at the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ for hatching and feeding the collected silkworms, the hatching rate is slightly increased along with the extension of the time of a refrigerating chamber, and the highest hatching rate is 90.5%.
TABLE 2 incubation rates of three different silkworm species after heat shock treatment of silkworm diapause variety Ze-tiger spot silkworm eggs
the conventional method comprises the steps of placing silkworm eggs in a refrigerator for refrigeration, taking the silkworm eggs out of the refrigerator for pickling to remove diapause when the silkworm eggs are subjected to cold soaking until the specified refrigeration period is reached, carrying out incubation and incubation at the natural temperature after the silkworm eggs subjected to the acid soaking are dried, and feeding the silkworm eggs to cocoons. The invention can meet the requirements of garden exhibition by simple artificial adjustment of production and management of silkworm eggs according to the requirements of plans through simple management of silkworm eggs, and saves manpower and material resources.
Finally, it should be noted that: the above-mentioned embodiments are only used for illustrating the technical solution of the present invention, and not for limiting the same; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (9)
1. a method for heat shock immediate diapause release and refrigeration delayed incubation of silkworms is characterized by comprising the following steps: which comprises the following steps:
S1, hatching the silkworm eggs after pickling in a climate box environment with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ until hatching and feeding the silkworm eggs to cocoons;
S2, collecting male and female moths of the synchronized moths at a protection temperature of 25 ℃, mating for 4-5 hours at 25 ℃ in indoor light, then splitting, placing the male and female moths on silkworm paper, laying eggs in a dark environment, and laying eggs for 6-8 hours;
S3, after spawning, protecting the silkworm egg-containing paper in a thermostat at 25 ℃, after 12-24 hours, thermally exciting the silkworm egg-containing paper for 5 seconds by using hot water at 53.1 +/-0.5 ℃, and carrying out air drying and treatment in three parts;
s4, placing the first silkworm egg subjected to instant heat shock treatment in a climate box with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ for hatching, and then collecting ants for feeding; placing the second part of the silkworm eggs subjected to instant heat shock treatment in a climate box with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ for hatching, placing the silkworm eggs in an intermediate warehouse for 2 hours after the silkworm eggs are spotted, wrapping the silkworm eggs with a film, placing the silkworm eggs in a cold storage, taking out a preset amount of silkworm eggs in batches every 3 days, placing the taken-out silkworm eggs in the intermediate warehouse for 2 hours, and placing the silkworm eggs in an environment at 25 ℃ for collecting and feeding; and placing the third silkworm eggs subjected to instant heat shock treatment in an intermediate storage for 6 hours, refrigerating in a refrigerator, wrapping the silkworm eggs in the refrigerator by using a film, taking out a preset amount of silkworm eggs in batches every 5 days after 30 days, placing the taken-out silkworm eggs in the intermediate storage for 24 hours, then placing in a climate box with the temperature of 24-25.5 ℃ and the dry-wet difference of 1.5-2 ℃ for hatching, and collecting and feeding the ants.
2. the method for heat shock immediate diapause release and cold storage delayed hatching of silkworms of claim 1, wherein: in step S4, the second instant heat-shock treated silkworm egg is spotted and then stored in a refrigerator for 3-20 days.
3. The method for heat shock immediate diapause release and cold storage delayed hatching of silkworms of claim 1, wherein: in step S4, the third instant heat shock treated silkworm egg is stored in the refrigerator for 30-50 days.
4. The method for heat shock immediate diapause release and cold storage delayed hatching of silkworms of claim 2 or 3, wherein: in step S4, the second instantaneous heat-shock-treated silkworm eggs are taken out of the refrigerator every 3 days for 6 times, wherein the number of the taken-out silkworm eggs is 1/6 of the total number of the second instantaneous heat-shock-treated silkworm eggs; the third immediately heat-shocked egg was taken out of the refrigerator every 5 days for a total of 4 times, each time the total amount of the third immediately heat-shocked egg was 1/4.
5. The method for heat shock immediate diapause release and cold storage delayed hatching of silkworms of claim 1, wherein: the intensity of the indoor light is 50-300 Lux.
6. the method for heat shock immediate diapause release and cold storage delayed hatching of silkworms of claim 1, wherein: the temperature of the intermediate storage is 15-16 ℃.
7. The method for heat shock immediate diapause release and cold storage delayed hatching of silkworms of claim 1, wherein: the temperature of the refrigerator is 5 +/-0.5 ℃.
8. the method for heat shock immediate diapause release and cold storage delayed hatching of silkworms of claim 5, wherein: the refrigerator is a refrigerator.
9. The method for heat shock immediate diapause release and cold storage delayed hatching of silkworms of claim 1, wherein: the film is a polyethylene film.
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| CN112106738A (en) * | 2020-09-11 | 2020-12-22 | 河南省蚕业科学研究院 | Method for nursing one or two-release autumn tussah to relieve diapause |
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| CN113481207B (en) * | 2021-07-15 | 2023-04-07 | 西南大学 | Essential gene Bmtret1 for silkworm egg diapause and application thereof |
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