甘蔗谷胱甘肽硫转移酶ScGSTF1与P3N-PIPO互作应答甘蔗花叶病毒侵染的研究  

Interaction of sugarcane glutathione S-transferase ScGSTF1 with P3N-PIPO in response to SCMV infection

作　　者：杨宗桃 焦文迪 张海 张克闽 程光远[1] 罗廷绪 曾康 周营栓 徐景升[1] YANG Zong-Tao;JIAO Wen-Di;ZHANG Hai;ZHANG Ke-Ming;CHENG Guang-Yuan;LUO Ting-Xu;ZENG Kang;ZHOU Ying-Shuan;XU Jing-Sheng(Key Laboratory of Sugarcane Biology and Genetic Breeding,Ministry of Agriculture and Rural Affairs,Fujian Agriculture and Forestry University/National Engineering Research Center for Sugarcane,Fuzhou 350002,Fujian,China)

机构地区：[1]福建农林大学农业农村部福建甘蔗生物学与遗传育种重点实验室/国家甘蔗工程技术研究中心,福建福州350002

出　　处：《作物学报》2023年第10期2665-2676,共12页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31971991);福建农林大学科技创新基金项目(CXZX2019132G);广西甘蔗生物学重点实验室开放课题(GXKLSCB-202003)资助。

摘　　要：谷胱甘肽S-转移酶(glutathione S-transferase, GST, EC2.5.1.18)广泛分布于具有细胞结构的生物中。在植物中,GST参与调控生长发育、异生物质解毒及逆境应答。本课题组以甘蔗花叶病毒(Sugarcanemosaicvirus,SCMV)编码蛋白P3N-PIPO为诱饵,从甘蔗(Saccharumspp.hybrid)叶片cDNA酵母文库中筛选到1个GST基因,并从甘蔗原始栽培种Badila中克隆了该基因,其开放读码框(open reading frame, ORF)长度为645 bp,编码214个氨基酸。序列比对及进化树分析表明,该基因编码蛋白属于GST家族的Phi (F)类型(GSTF),因此命名为ScGSTF1。进一步的生物信息学分析表明,ScGSTF1不存在跨膜区域,为稳定的亲水性脂溶蛋白;GST蛋白在单子叶和双子叶植物中及C3和C4植物中存在明显的分化。酵母双杂交(yeast two-hybrid, Y2H)和双分子荧光互补(bimolecular fluorescence complementation, BiFC)试验表明, ScGSTF1与SCMV-P3N-PIPO蛋白互作。亚细胞定位试验表明ScGSTF1定位于内质网。实时荧光定量PCR分析表明, ScGSTF1在甘蔗各个组织中显著差异表达,在第3节间中的表达量最高,在心叶及根中的表达量最低;SCMV侵染显著性影响ScGSTF1基因的表达水平,侵染早期上调表达,随后下调,但仍保持显著高于对照的水平。The Glutathione S-transferases(GSTs)are widely distributed in cellular organisms and participate in the regulation of growth and development,detoxification of xenobiotics,and response to stress in plants.In the present study,we screened a sugar-cane(Saccharum spp.hybrid)cDNA library and obtained a GST gene from sugarcane original cultivar Badila using Sugarcane mosaic virus(SCMV)-encoded protein P3N-PIPO as bait.The open reading frame(ORF)of the cloned GST gene was 645 bp in length,encoding a protein with 214 amino acids.Sequence alignment and phylogenetic tree showed that this GST belonged to the Phi(F)class(GSTF)designated as ScGSTF1 in GST family.Further bioinformatics revealed that ScGSTF1 protein was a stable hydrophilic lipoprotein without transmembrane domain.Phylogenetic tree demonstrated that ScGSTF1 was divergent between monocotyledons and dicotyledonous,as well as in C3 and C4 plants.Yeast two-hybrid(Y2H)and biomolecular fluorescence com-plementation(BiFC)assays confirmed the interaction of ScGSTF1 with SCMV-P3N-PIPO.Subcellular localization assays indi-cated that ScGSTF1 was localized in the endoplasmic reticulum.The qRT-PCR analysis showed that ScGSTF1 gene was signifi-cantly differently expressed in different tissues of sugarcane plants with the highest expression level in the 3rd internode and the lowest expression level in leaf roll or roots.SCMV infection affected the relative expression level of ScGSTF1 gene,which was up-regulated at the early stage of infection and down-regulated later,but still maintained a significantly higher level than the con-trol.

关 键 词：甘蔗 甘蔗花叶病毒 谷胱甘肽硫转移酶 P3N-PIPO 蛋白互作 

分 类 号：S435.661[农业科学—农业昆虫与害虫防治]