摘要
【目的】比较高浓度氧处理法(高氧法)与传统HCl处理法(HCl法)阻断蚕卵滞育效果的差异,探讨高氧法阻断滞育在家蚕Bombyx mori转基因技术体系上的应用。【方法】采用高氧法和HCl法在产后20 h处理二化性蚕品种‘大造’,比较孵化率、孵化时间,调查高氧法的最佳处理条件;将高氧法应用于一化性蚕品种‘土耳其’和转基因二化性蚕品种‘大造’,调查其对一化性蚕品种蚕卵阻断滞育效果及转基因二化性蚕品种显微注射后蚕卵阻断滞育效果。【结果】高氧法对二化性蚕卵在孵化率、孵化时间上都与HCl法有相近的阻断滞育的效果,其最佳条件为产卵后20 h用体积分数为70%的O2处理40 h;该方法也能成功阻断一化性蚕品种,孵化率达(71±20)%;高氧法处理的显微注射转基因家蚕卵孵化率达到了(49±9)%,而对照组的孵化率为0。【结论】高氧法为今后生产上替代HCl法,采用安全环保的蚕卵阻断滞育技术,解决转基因蚕卵阻断滞育和提高转基因家蚕育种进度的关键问题提供了可行的新方法。
【Objective】To compare the differences between the effects of high oxygen concentration treating mehod(hyperoxia method)and traditional HCl treating method(HCl method)on prevention of diapause in silkworm(Bombyx mori)eggs,and investigate the application of hyperoxia method in transgenic technology system of silkworm.【Method】The bivoltine silkworm‘Dazao’were treated by hyperoxia method and HCl method on the 20th hour after spawning.The hatching rate and incubation period were compared,and the best condition of hyperoxia method was investigated.The univoltine silkworm‘Tuerqi’and transgenic bivoltine silkworm‘Dazao’were treated by hyperoxia method.The effects of hyperoxia method and microinjection on blocking univoltine silkworm diapause were investigated.【Result】Hyperoxia method had similar dispause prevention effects with HCl method considering hatching rate and incubation duration.The best condition was treating 40 hours by 70%(φ)O2 after spawning for 20 hours.Hyperoxia method could successfully prevent diapause in univoltine silkworm eggs.The hatching rate was(71±20)%.The hatching rate of micro-injected transgenic silkworm eggs treated by hyperoxia method approached(49±9)%,while that of control group was 0.【Conclusion】Hyperoxia method is a feasible new method for replacing HCl method in the future production,enables safe and environmental friendly prevention of diapause in silkworm eggs,solves key problems of diapause prevention in transgenic silkworm eggs,and improves breeding progress of transgenic silkworms.
作者
龚竞
尚杉
张育浩
颜嘉敏
朱勇
GONG Jing;SHANG Shan;ZHANG Yuhao;YAN Jiamin;ZHU Yong(Key Laboratory of Sericultural Biology and Genetic Breeding,Ministry of Agriculture and Rural Affairs/College of Biotechnology,Southwest University,Chongqing 400715,China)
出处
《华南农业大学学报》
CAS
CSCD
北大核心
2020年第1期61-68,共8页
Journal of South China Agricultural University
基金
国家自然科学基金面上项目(31872429)
国家自然科学基金(31802139)
重庆市基础与前沿研究计划(cstc2017jcyjAC0475)
关键词
家蚕
阻断滞育
高氧法
转基因
扫描电镜
silkworm
diapause prevention
hyperoxia method
transgene
scanning electron microscopy