甘蔗类泛素蛋白UBL5应答SCMV侵染及其与SCMV-6K2的互作  

Sugarcane ubiquitin-like protein UBL5 responses to SCMV infection and interacts with SCMV-6K2

作　　者：杨宗桃 刘淑娴 程光远[1] 张海 周营栓 商贺阳 黄国强[1] 徐景升[1] YANG Zong-Tao;LIU Shu-Xian;CHENG Guang-Yuan;ZHANG Hai;ZHOU Ying-Shuan;SHANG He-Yang;HUANG Guo-Qiang;XU Jing-Sheng(Sugarcane Research&Development Center,China Agricultural Technology System,Fujian Agriculture and Forestry University/Key Laboratory of Sugarcane Biology and Genetic Breeding,Ministry of Agriculture and Rural Affairs/Key Laboratory of Ministry of Education for Genetics,Breeding and Multiple Utilization of Crops,Fuzhou 350002,Fujian,China)

机构地区：[1]福建农林大学国家甘蔗工程技术研究中心/农业农村部福建甘蔗生物学与遗传育种重点实验室/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》2022年第2期332-341,共10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(CXZX2018026);福建省科技厅引导性项目(2017N0003)资助。

摘　　要：泛素化修饰在蛋白功能调控、生长发育和逆境应答中具有重要作用。类泛素蛋白(ubiquitin-likeproteins,UBLs)是泛素-蛋白酶体系统(ubiquitin-proteasome system,UPS)的重要组分。本课题组前期利用酵母双杂交(yeast two-hybrid,Y2H)技术从甘蔗(Saccharumspp.hybrid)中分离鉴定了1个与甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)编码蛋白6K2互作的类泛素蛋白UBL5,命名为Sc UBL5,长度为73aa。本研究利用双分子荧光互补(bimolecular fluorescence complementation,BiFC)试验进一步证实了Sc UBL5与SCMV-6K2互作;生物信息学分析表明,Sc UBL5为稳定的亲水性非分泌蛋白,无信号肽和跨膜结构。系统进化树分析表明,Sc UBL5具有明显的种属特异性。亚细胞定位分析表明,Sc UBL5定位于细胞质和细胞核。实时荧光定量PCR分析发现,Sc UBL5基因的表达具有明显的组织特异性,在完成形态建成的正一叶、第7叶、根和第8节间中的相对表达量显著高于未成熟的心叶和第3节间;SCMV侵染对Sc UBL5基因表达影响显著,Sc UBL5基因在侵染早期显著上调,侵染后期下调但显著高于对照。Ubiquitylation plays key roles in the regulation of protein function,growth and development,and response to stress.Ubiquitin-like proteins(UBLs) are the main components of the ubiquitin-proteasome system(UPS).In our previous study,the UBL5 homologue was isolated from sugarcane(Saccharum spp.hybrid) by yeast two-hybrid(Y2 H) with the 6 K2 of Sugarcane mosaic virus(SCMV) as bait,and then designated as ScUBL5 with 73 aa in length.In the present study,the interaction of ScUBL5 with the SCMV-6 K2 was further confirmed by bimolecular fluorescence complementation assays(BiFC).Bioinformatics analysis showed that ScUBL5 is a stable hydrophilic non-secretory protein without signal peptide or transmembrane domain.Phylogenetic tree analysis showed that Sc UBL5 is species specific.Subcellular localization analysis showed that ScUBL5 is localized in cytoplasm and nucleus.Sc UBL5 gene shows obvious tissue specificity in sugarcane by real-time quantitative PCR analysis.The expression levels of ScUBL5 gene in the established morphogenesis tissues such as the 1 st leaf,the 7 th leaf,the 8 th internode and the root were significantly higher than those in the immature tissues such as leaf roll and the 3 rd internode.The expression of ScUBL5 gene is significantly affected by SCMV infection.ScUBL5 was significantly upregulated in the early stage of SCMV infection,then downregulated but significantly higher than the control at the late stage of SCMV infection.

关 键 词：甘蔗 类泛素蛋白 甘蔗花叶病毒 6K2 

分 类 号：S435.661[农业科学—农业昆虫与害虫防治]