甘蔗PsbS亚基应答甘蔗花叶病毒侵染及其与6K2蛋白的互作研究  

Sugarcane PsbS subunit response to Sugarcane mosaic virus infection and its interaction with 6K2 protein

作　　者：张海 刘淑娴 杨宗桃 王彤 程光远[1] 商贺阳 徐景升[1] ZHANG Hai;LIU Shu-Xian;YANG Zong-Tao;WANG Tong;CHENG Guang-Yuan;SHANG He-Yang;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》2020年第11期1722-1733,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(CXZX2018026)资助。

摘　　要：非光化学猝灭(non-photochemical quenching,NPQ)是高等植物主要的光保护调节机制,光合系统Ⅱ(PhotosystemⅡ,PSⅡ)的Psb S亚基在NPQ起关键作用。甘蔗(Saccharum spp.hybrid)中PSⅡPsb S亚基应答甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)的侵染尚未见报道。本课题组在前期研究中克隆了甘蔗的Psb S亚基编码基因,命名为Sc Psb S,该基因开放读码框(open reading frame, ORF)长度为798 bp,编码长度为265个氨基酸的蛋白。生物信息学分析表明, Sc Psb S为稳定的疏水性蛋白,具有叶绿体定位信号,有4个跨膜结构域;具有典型的Psb S亚基结构域。系统进化树分析表明, Sc Psb S在单子叶、双子叶以及单子叶的C3和C4植物中存在明显的分化。亚细胞定位分析表明,Sc Psb S定位于叶绿体且与SCMV-6K2部分共定位于叶绿体。双分子荧光互补(bimolecular fluorescence complementation, BiFC)试验进一步证实了ScPsbS与SCMV-6K2互作。实时荧光定量PCR分析发现, ScPsbS基因的表达具有明显的组织特异性,在成熟叶片中相对表达量最高,未成熟叶片和初衰叶中次之,茎和根中几乎不表达;SCMV侵染对ScPsbS基因表达影响显著, ScPsbS基因在侵染早期显著上调,侵染后期没有明显的差异。Non-photochemical quenching(NPQ) is the main mechanism of photoprotective regulation in higher plants. The PsbS subunit of Photosystem Ⅱ(PSⅡ) plays a key role in NPQ. The involvement of PSⅡ Psb S subunit in Sugarcane mosaic virus(SCMV) infection of sugarcane(Saccharum spp. hybrid) has not been reported. In the previous research, we cloned the coding sequence of the Psb S subunit from sugarcane and designated it as Sc Psb S. Sc Psb S had an open reading frame(ORF) length of 798 bp and encoded a protein of 265 aa. Bioinformatics analysis showed that Sc Psb S was a stable hydrophobic protein with chloroplast localization signals and four transmembrane domains. The Sc Psb S protein possesses a typical domain of Psb S protein. Phylogenetic tree analysis showed that Sc Psb S was divergent between monocotyledons and dicotyledons, or C3 plants and C4 plants. Subcellular localization analysis showed that Sc Psb S was located in chloroplasts and partially colocalized with SCMV-6 k2 in chloroplasts. The interaction of Sc Psb S with the SCMV-6K2 was further confirmed by bimolecular fluorescence complementation assays(Bi FC). Sc Psb S gene showed obvious tissue specificity in sugarcane tested by real-time quantitative PCR analysis. Sc Psb S gene had highest expression in mature leaves, followed by immature leaves and leaves beginning to senesce, and hardly expressed in stems and roots. The expression of Sc Psb S gene was significantly affected by SCMV infection, with significant upregulation in the early stage of SCMV infection, and no significant affection in the later stage of SCMV infection.

关 键 词：甘蔗 PsbS亚基 甘蔗花叶病毒 6K2 光保护 

分 类 号：S435.661[农业科学—农业昆虫与害虫防治]