甘蔗类钙调素ScCML13与SCMV运动蛋白P3N-PIPO的互作研究  

Interaction between calmodulin-like ScCML13 of sugarcane and SCMV movement protein P3N-PIPO

作　　者：玉泉馨 杨宗桃 张海 程光远[1] 焦文迪 曾康 罗廷绪 黄国强[1] 王璐 徐景升[1] YU Quan-Xin;YANG Zong-Tao;ZHANG Hai;CHENG Guang-Yuan;JIAO Wen-Di;ZENG Kang;LUO Ting-Xu;HUANG Guo-Qiang;WANG Lu;XU Jing-Sheng(Fujian Agriculture and Forestry University/Key Laboratory of Sugarcane Biology and Genetic Breeding,Ministry of Agriculture and Rural Affairs/National Engineering Research Center for Sugarcane/Key Laboratory of Ministry of Education for Genetics,Breeding and Multiple Utilization of Crops,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/教育部作物遗传育种与综合利用重点实验室,福建福州350002

出　　处：《作物学报》2024年第7期1855-1866,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)资助。

摘　　要：类钙调素(Calmodulin-like,CML)是植物特有的钙信号感受器蛋白之一,参与植物生长发育和响应外界环境信号转导。甘蔗(Saccharum spp.hybrid)中CML应答甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)侵染尚未见报道。本研究从甘蔗热带种Badila(S.officinarum)中克隆了一个CML基因,命名为ScCML13。该基因开放读码框(open reading frame,ORF)长度为519 bp,编码172 aa。生物信息学分析表明,ScCML13属于稳定的亲水脂溶蛋白,无跨膜结构域,有4个结合Ca2+的EF-hand结构域;系统进化树分析表明,该蛋白在单子叶和双子叶植物中及单子叶C3和C_(4)植物中具有明显分化;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,ScCML13与SCMV的运动蛋白P3N-PIPO互作。亚细胞定位试验表明ScCML13定位于内质网和细胞核。共定位试验发现ScCML13会干扰SCMV-P3N-PIPO的质膜(plasma membrane)或胞间连丝(plasmodesmata)定位。实时荧光定量PCR(Real Time Quantitative PCR,RT-qPCR)分析发现,ScCML13基因主要在甘蔗叶片中表达,在节间和根中的相对表达量低;ScCML13基因在感染SCMV后2 h显著上调,随后下调至与对照组相比的水平,而在感染后期上调。Calmodulin-like(CML),one of the calcium signal receptor proteins unique to plants,is involved in plant growth and development and in response to environmental signal transduction.However,the response of CML to Sugarcane mosaic virus(SCMV)infection in sugarcane(Saccharum spp.hybrid)has not been reported.In the present study,a CML gene was cloned from a Badila(S.officinarum)and designated as ScCML13.The open reading frame(ORF)of ScCML13 gene is 519 bp in length and encodes a protein with 172 aa in length.Bioinformatics analysis showed that ScCML13 was a stable hydrophilic lipoprotein in with no transmembrane domain and contains 4 Ca2+-binding EF-hand domains.Phylogenetic tree analysis showed that the ScCML13 protein differentiated in monocotyledon and dicotyledon plants and in monocotyledonous C3 and C_(4)plants.Yeast two-hybrid(Y2H)and bimolecular fluorescence complementation(BiFC)experiments indicated that ScCML13 interacted with P3N-PIPO,the movement protein of SCMV.Subcellular localization assays demonstrated that ScCML13 was localized to the endoplasmic reticulum and nucleus,whereas the co-localization assays showed that ScCML13 interfered with the localization of SCMV-P3N-PIPO to plasma membrane or plasmodesmata.The RT-qPCR showed that ScCML13 gene was mainly expressed in sugarcane leaves,but relatively low in internodes and roots.The ScCML13 gene was significantly up-regulated at 2 h upon SCMV infection,and then down-regulated to the levels compared with the control group,while up-regulated at the later stage of infection.

关 键 词：类钙调素 甘蔗花叶病毒 P3N-PIPO 蛋白互作 

分 类 号：S435.661[农业科学—农业昆虫与害虫防治]