水虻HiACC和HiFAS的克隆与表达模式分析

Cloning and Experssion Pattern of HiACC and HiFAS in Black Soldier Fly

通过实时荧光定量PCR(RT-qPCR)技术克隆获得了4个水虻脂肪酸合成关键基因乙酰辅酶A羧化酶(ACC)基因HiACC和脂肪酸合成酶(FAS)基因HiFAS1、HiFAS2、HiFAS3,对其进行序列分析和系统进化树构建,并利用RT-qPCR检测分析了其在水虻不同发育阶段、组织、营养条件下的表达模式。结果表明,HiACC、HiFAS1、HiFAS2、HiFAS3等4个基因在双翅目昆虫中高度保守;4个基因均在卵期、4~5龄幼虫期表达量较高,在蛹期表达量较低;4个基因主要集中在脂肪体和前中肠表达;与基础饲喂对照组相比,饥饿组、高脂饲喂组的4个基因表达量均显著下调,其中在饥饿胁迫下表达量最低。为后续研究ACC和FAS的表达调控机制,提高水虻油脂积累效率提供了理论依据。

We cloned four key genes of fatty acid synthesis,namely acetyl-coenzyme A carboxylase(ACC)gene HiACC and fatty acid synthase(FAS)genes HiFAS1,HiFAS2,and HiFAS3 by real-time quantitative fluorescent PCR(RT-qPCR)technique,and carried out sequence analysis and phylogenetic tree construction.Moreover,we analyzed the expression patterns of the four genes in different growth stages,tissues,and nutritional conditions by RT-qPCR.The results show that HiACC,HiFAS1,HiFAS2,and HiFAS3 genes are highly conserved in Diptera insects.The four genes are highly expressed in the egg stage and 4th to 5th instar larvae,but are lower expressed in pupal stage.The four genes are mainly expressed in fat body and the anterior and middle intestine.Compared with the expression level of basic feeding control group,the expression levels of the four genes in starvation treatment group and high fat feeding group are significantly down-regulated,and the expression levels are the lowest under starvation stress.This study provides a theoretical basis for the further research on the regulation mechanism of ACC and FAS expression and the improvement of oil accumulation efficiency in black soldier fly.