脂肪酸合成酶是脂肪酸合成通路中的关键酶,脂肪酸的合成对昆虫的正常生长发育和繁殖至关重要,而生殖的正常关系到昆虫的种群繁衍.本研究旨在利用RNA干扰(RNA interference,RNAi)技术沉默柑橘全爪螨(Panonychus citri)脂肪酸合成酶(fatty acid synthase,FAS)基因PcFAS,通过脂质组学技术分析该基因沉默后对柑橘全爪螨雌成螨脂质代谢的影响.采用饲喂法导入dsRNA沉默柑橘全爪螨PcFAS基因,基于液相色谱-质谱联用(liquid chromatography-mass spectrometry,LC-MS)技术进行脂质组学研究,系统解析机体脂质组成与表达变化模式.运用主成分分析(principal component analysis,PCA)、偏最小二乘判别分析(partial least squares discriminant analysis,PLS-DA)和KEGG富集分析等方法分析两组的脂质代谢差异及差异脂质分子的富集通路.脂质组学分析结果显示,从柑橘全爪螨中共鉴定出40个脂质亚类,1 012个脂质分子,其中涉及5种脂肪酸、3种甘油脂、15种甘油磷脂、7种鞘脂、6种糖脂、1种固醇脂、3种衍生脂.基于变量投影重要度(variable importance in the projection,VIP)和变异倍数(fold change,FC),以 PLS-DA 模型第一主成分的 VIP＞1,FC＞1.200或＜0.833且P＜0.05为筛选标准,共筛选出150个显著性差异脂质分子(60个上调,90个下调).经KEGG通路富集分析,差异脂质代谢产物主要富集的4条通路有脂肪消化与吸收,维生素消化与吸收,脂肪细胞脂解的调节和胆固醇代谢.PcFAS沉默后影响了柑橘全爪螨雌成螨机体内脂质的组成和含量变化,干扰其正常的脂质代谢,这可能会导致其生长发育障碍或死亡.本研究可为基于脂类合成关键基因的害螨防治靶标的筛选提供重要的理论依据.
Lipidomic-based Analysis of the Effects of Silencing Fatty Acid Synthase Gene on Lipid Metabolism in Panonychus citri
Fatty acid synthase is a key enzyme in the fatty acid synthesis pathway.The synthesis of fatty acids is crucial for the normal growth,development,and reproduction of insects,and the normal reproduction is related to the population reproduction of insects.This study aims to silence the fatty acid synthase(FAS)gene PcFAS in Panonychus citri using RNA interference(RNAi)technology and the molecular mechanism of lipid metabolism in adult females of P.citri after gene silencing through lipidomics was analyzed.The dsRNA was delivered into adult female of P.citri by feeding to silence PcFAS gene.Lipidomics research was conducted based on li-quid chromatography-mass spectrometry(LC-MS)to systematically analyze the lipid composition and expression changes in P.citri.Principal component analysis(PCA),partial least squares discriminant analysis(PLS-DA),and KEGG enrichment analysis were used to explore metabolic product differences and pathways between the two groups.The results of lipidomics analysis revealed that 40 lipid subclasses and 1 012 lipid molecules were identified in P.citri,involving 5 fatty acids,3 glycerides,15 glycerophospholipids,7 sphingolipids,6 glycolipids,1 sterol lipid,and 3 derivative lipids.Basing on variable importance in the projection(VIP)and fold change(FC),using PLS-DA model with VIP＞1,FC＞1.200 or＜0.833 and P＜0.05 as the selection criteria,150 lipid molecules with significant differences were screened out(60 up-regulated and 90 down-regulated).In KEGG pathway enrichment analysis,the main enriched pathways of differential lipid metabolites were fat digestion and absorption,vitamin digestion and absorption,regulation of lipolysis in adipocytes and cholesterol metabolism.The composition and content of lipids in female adults of P.citri were disrupted by silencing of PcFAS so that P.citri would be inhibited development even death because of disordered lipid metabolism.Therefore,the present study provides the important theoretical basis for developing important agricultural pests control technology targeting on the lipids biosynthesis genes involving in the lipids metabolism.
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