Method for preparing silkworm eggs for transgene of eggs of bivoltine silkworm varieties

A technology of transgenic and silkworm eggs, which is applied in the field of insect bioengineering, can solve the problems of large difference in hatching effect, difficult breeding operation, long greening process, etc., to overcome poor cocoon silk quality, short greening incubation process, low volume effect

Inactive Publication Date: 2010-05-12
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The impact of this technology on diapause is unstable. The hatching rate of silkworm eggs of some varieties can reach about 50% after transgenic operation, but the hatching effect varies greatly among varieties. The hatching rate of most Japanese dimorphic varieties is very low, and due to The greening process is as long as 35d-40d, and the hatched silkworms are weak
[0008] (3) Publication No. 101503704 (Application No.: 200910103397.6) Chinese invention patent publication discloses a method for making transgenic silkworms of silkworm diapause species, using 15°C, 75% relative humidity, and dark environment during the hatching period of parental eggs Protection: After hatching, the 1-3 instar larvae are fed in a dark environment at 23°C, 85% relative humidity, and protected by absolute light conditions at the pupal stage, so that the produced offspring silkworm eggs become non-diapause eggs. This technology also There are problems such as long greening process and weak constitution of hatched silkworms, and it is difficult to carry out rearing operation in the dark. The emergence of adults under the protection of this technology is irregular, which adds difficulties to the transgenic operation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034]Use spring-made diapause eggs of the silkworm dimorphic variety Dazao as the parent silkworm eggs, and use 22.5°C, 75%-85% relative humidity, and natural light conditions in the early stage to protect the 19th embryonic development stage; then use 20 ℃, relative humidity 75%-85%, and light intensity lower than 0.1lux under dark conditions to promote green protection until silkworm eggs hatch; after hatching, larvae are kept at 20°C-22°C, relative humidity 80%-90%, and natural light conditions. Use mulberry leaves to feed; silkworms from the third age start to use mulberry leaves to cocoon at 25°C-26°C, 75%-80% relative humidity, and natural light conditions; then at 24°C-26°C, 75%-80 %relative humidity, LD=18h: 6h under long-light conditions to protect until eclosion; collect the female moths after eclosion, and mate with male moths for 5 hours under 24.5°C and weak light conditions below 10lux. ;Mate the male and female moths separately, put the female moths on the ovip...

Embodiment 2

[0036] The spring diapause silkworm eggs of the dimorphic silkworm variety Haoyue were used as the parent silkworm eggs, and the whole greening incubation process used dark conditions of 20°C, relative humidity of 75%-85%, and light intensity lower than 0.1lux; 20°C-21°C, relative humidity 85%-90%, and natural light conditions, use mulberry leaves to feed; 3rd instar silkworms start to use mulberry leaves in 25.5°C-26°C, 75%-80% relative humidity, natural light conditions Breed until cocooning; then protect until eclosion under the conditions of 25°C-26°C, 75%-80% relative humidity, LD=18h:6h long-light system; collect eclosion female moths, 25°C-26°C, below 10lux Mate with male moths for 4 hours under weak light conditions, and use ordinary Haoyue male moths for male moths; mating male and female moths separately, put female moths on oviposition paper for production, and lay eggs in a dark environment below 0.1lux at 25°C; collect eggs produced within 10 minutes Eggs are used...

Embodiment 3

[0038] Use the diapause eggs of the silkworm dimorphic variety 75 in spring as the parent silkworm eggs, and use 24.5°C-25°C, 75%-80% relative humidity, and natural light to protect them until the 19th stage of embryonic development; Then use 20°C, relative humidity of 80%-85%, and light intensity lower than 0.1lux to protect the dark conditions until hatching; after hatching, the larvae are kept at 21°C-22°C, relative humidity of 80%, and natural light conditions, using mulberry leaves Starting from feeding the third instar silkworms, use mulberry leaves to cocoon at 25°C-26°C, 75%-80% relative humidity, and natural light conditions; then raise them at 25°C-26°C, 75%-80% relative humidity, LD=18: 6h under long-light conditions from protection to eclosion; collect the eclosion female moths, and mate with male moths for 5 hours under 25°C-26°C, 10lux weak light conditions. Moths and female moths are placed on the production oviposition paper, and the eggs are laid in a dark env...

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PUM

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Abstract

The invention relates to the field of insect bioengineering, in particular to a method for transgene of eggs of bivoltine silkworm varieties. In the method, early activated eggs are obtained by utilizing combination of protection temperature and light system in a parent generation silkworm egg hatching period, and artificial hatching treatment on the silkworm eggs before offspring fertilization; and then the early activated eggs are subjected to micro-injection and screening so as to obtain transgenic silkworm eggs. The method can solve the problems that the artificial hatching treatment after the diapaused eggs of the bivoltine silkworm varieties are subjected to transgenic injection causes damage to the silkworm eggs and the silkworm eggs cannot be hatched, also can solve the problem of low hatching rate due to transgenic injection after the early hydrochloric acid hatching treatment on the diapaused eggs of the bivoltine silkworm varieties, and also solves the problems that embryo robustness is poor and larva survival rate is low for offspring non-diapause eggs of the bivoltine silkworm varieties obtained by the hydrochloric acid hatching treatment on the diapaused eggs of the bivoltine silkworm varieties and low-temperature and dark hatching conditions for the parent generation silkworm eggs.

Description

technical field [0001] The invention relates to the field of insect bioengineering, in particular to a method for transgenic eggs of dimorphic silkworm species. Background technique [0002] Bombyx mori, also known as mulberry silkworm, scientific name Bombyx mori Linnaeus, is oviparous and is a model insect of Lepidoptera. An important physiological phenomenon of silkworm is metamorphosis. The so-called metamorphosis refers to the number of generations that occur in a year. There are differences in the silkworm varieties such as monophyletic, dimorphic, polymorphic without diapause, and polymorphic with diapause. Monomorphic silkworm species have one generation in one year, larvae have a long growth period, and the cocoons they form are large, but their bodies are weak; polymorphic silkworm species have more than 3 generations in a year, larvae have a short growth period, and the cocoons they form are small. Cocoon silk properties are poor, but the constitution is strong; ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/04C12N15/85
Inventor 徐世清司马杨虎袁红霞牛艳山甘丽萍
Owner SUZHOU UNIV
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