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

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; ...

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.