摘要
【目的】研究LdATPaseE通过类胰岛素信号通路影响保幼激素和蜕皮激素通路,调节马铃薯甲虫幼虫化蛹的分子机制。【方法】LdATPaseE cDNA序列通过马铃薯甲虫转录组数据分析和RT-PCR克隆获得;qPCR检测其在马铃薯甲虫各发育阶段,以及四龄幼虫不同组织的相对表达量;采用喂食幼虫dsRNA的方法,分析该基因在马铃薯甲虫幼虫的生长发育过程中的影响,并测定类胰岛素、保幼激素和蜕皮激素通路基因对该基因的表达响应机制。【结果】喂食二龄幼虫ds LdATPaseE后,成功敲低了靶标基因,阻止二龄幼虫的生长,显著增加了二龄幼虫的死亡率。三龄和四龄幼虫喂食ds LdATPaseE-1和ds LdATPaseE-2,在极低浓度下即敲低了靶标基因,阻止了幼虫生长,显著降低马铃薯甲虫幼虫的化蛹率。敲低LdATPaseE还显著升高了LdInR与Ld4EBP的表达量,抑制一个蜕皮激素合成基因的转录,降低20E滴度并降低了一个20E响应基因的表达。敲低LdATPaseE还抑制了一个JH合成酶基因的表达,降低了JH滴度,下调了一个JH早期响应基因的表达量。【结论】沉默LdATPaseE后,其可能通过抑制IIS信号途径,降低20E和JH的滴度、下调20E和JH信号从而影响幼虫生长和发育。
【Objective】Previous experiments showed that feeding different doses of LdATPaseE to Colorado Potato Beetles'larvae resulted in a gradual decline in the feeding amount,mobility,digging ability,pupation rate,and emergence rate of the larvae.This experiment attempts to explain the LdATPaseE gene expression pattern and how the insulin-like signaling pathway affects juvenile hormone and ecdysone hormone,thereby regulating the pupa and emergence of Coloradopotato beetle larvae.【Methods】The LdATPaseEc DNA was obtained by transcriptome data and RT-PCR cloning from Colorado Potato Beetles.Sequence alignment and phylogenetic analysis were used to determine the integrity and phylogeny of the gene's cDNA.The expression of larvae in different tissues of Colorado Potato Beetles at different developmental stages was detected by qPCR.The effect of the gene on the growth and development of Colorado Potato Beetle larvae was observed by feeding the larvae dsRNA,and the effects of juvenile hormone and ecdysone on the expression of the gene were determined.【Results】ds LdATPaseE-1 and ds LdATPaseE-2 were fed to second-instar larvae,leading to successfully knocking down of the target gene's expression level,and prevented the growth of second-instar larvae and significantly increased the mortality of second-instar larvae.Feeding ds LdATPaseE-1 and ds LdATPaseE-2 at the third and fourth instar stage of larvae,the target gene was knocked down at even very low concentrations,preventing larval growth and significantly reducing the pupation rate of potato beetle larvae.In addition,knockdown of LdATPaseE also significantly increased the expression of LdInR and Ld4EBP,inhibited the transcription of an ecdysone synthesis gene,and decreased the 20E titer and the expression of a 20E response gene.Silence of LdATPaseE also inhibited the expression of a JH synthase gene,reduced the JH titer,and down-regulated the expression of a JH early response gene.【Conclusion】Upon silencing LdATPaseE,it may affect larval growth and development by inhibiting its suppressioninsulin/insulin-like pathways,thereby reducing the titer of 20E and JH,down-regulating 20E and JH signals.
作者
廖兰兰
王俊
付开赟
丁新华
何江
吐尔逊·阿合买提
郭文超
LIAO Lanlan;WANG Jun;FU Kaiyun;DING Xinhua;HE Jiang;Tursun Ahmat;GUO Wenchao(College of Plant Science,Tarim University,Alar Xinjiang 843300,China;Key Laboratory of Integrated Crop Pest Management in Northwestern Oasis of China,MOARA,Institute of Plant Protection,Xinjiang Academy of Agricultural Sciences,Urumqi 830091,China;Plant Protection Station of Xinjang Uygur Autoncmous Regiou,Urumqi 830049;Post-Doctoral Mobile Station of Xinjiang Agricultural University/Post-Doctoral Workstation of Xinjiang Academy of Agricultural Sciences,Urumqi 830091,China;Institute of Applied Microbiology,Xinjiang Academy of Agricultural Sciences,Urumqi 830091,China)
出处
《新疆农业科学》
CAS
CSCD
北大核心
2021年第6期1124-1137,共14页
Xinjiang Agricultural Sciences
基金
新疆维吾尔自治区自然科学基金青年科学基金(2016D01B043)。
