三疣梭子蟹蜕皮周期及眼柄切除后血淋巴甲基法尼酯浓度测定及其合成酶基因的表达分析  

Changes in the hemolymph levels of methyl farnesoate and gene expression of enzymes in its biosynthesis during the molt cycle and after eyestalk ablation of the swimming crab, Portunus trituberculatus

作　　者：邵娟 郑亮 郑宏坤 谢熙[1] 朱冬发[1] SHAO Juan;ZHENG Liang;ZHENG Hong-kun;XIE Xi;ZHU Dong-fa(School of Marine Science,Ningbo University,Ningbo 315211,China)

机构地区：[1]宁波大学海洋学院,宁波315211

出　　处：《生物学杂志》2019年第2期28-32,共5页Journal of Biology

基　　金：国家自然科学基金项目(No.41376152;41776165);宁波市科技局重大农业科技项目(No.2016C11003)

摘　　要：甲基法尼酯(MF)是昆虫保幼激素III(JH III)的非环氧化物,是甲壳动物中JH的对等物,参与调控甲壳动物的各项生理活动。为了研究MF在蜕皮过程中的可能作用,利用气相色谱-质谱联用(GC-MS)中的选择离子监测模式(SIM)对三疣梭子蟹蜕皮周期中血淋巴中MF含量变化进行了检测,并利用实时荧光定量PCR(qPCR)对MF合成途径中关键酶的基因表达变化进行了分析。结果显示,三疣梭子蟹血淋巴MF含量在蜕皮前期显著上升,至D_1亚期升至最高。而AACT、FPS和JHAMT 3个MF合成酶在蜕皮周期中的基因表达模式虽然并不相同,但基本符合它们在MF合成通路中的位置关系,表明MF合成酶的转录调控可能存在级联效应。通过眼柄切除(ESA)进一步研究了眼柄因子对MF生物合成的调控作用,结果发现ESA能够迅速提升血淋巴MF含量和PtHMGR、PtFPS、PtFAMeT和PtJHAMT等基因的表达水平,这说明眼柄因子对MF的生物合成存在转录水平的调控。Methyl farnesoate(MF)is the unepoxidized form of insect JH III,and is commonly considered as an equivalent of JH in crustaceans,which plays essential roles in many physiological processes of crustaceans.To investigate the possible role of MF in the molting process of the swimming crab,Portunus trituberculatus,the hemolymph levels were detected using the gas chromatography-mass spectrography(GC-MS)with selected ion monitoring(SIM),and the gene expression of the enzymes in MF biosynthesis pathway was detected using qPCR.The results showed that the hemolymph MF titer increased significantly in the pre-molt stage,and to the maximum in D 1 substage.The gene expression patterns of three MF biosynthesis enzymes including AACT,FPS,and JHAMT were not the same during the molt cycle,while still consistent with their location in the MF biosynthesis pathway,which indicated that the transcriptional regulation of MF synthesis enzymes might be cascaded.In addition,the regulatory role of eyestalk factor on the MF biosynthesis was investigated by eyestalk ablation(ESA)experiment.The results showed that ESA could rapidly induce the hemolymph MF titer and the gene expression of MF synthesis enzymes including PtHMGR,PtFPS,PtFAMeT and PtJHAMT,suggesting the existence of transcriptional regulation of eyestalk factors on MF biosynthesis.

关 键 词：甲基法尼酯 生物合成 蜕皮 三疣梭子蟹 基因表达 

分 类 号：Q78[生物学—分子生物学] S917.4[农业科学—水产科学]