棉花微管结合蛋白基因GhCLASP1的克隆与表达分析  

Cloning and Expression Analysis of GhCLASP1 Gene in Gossypium hirsutum L.

作　　者：朱守鸿 薛飞[1] 赵兰杰[2] 刘永昌[1] 李艳军[1] 熊显鹏[1] 孙杰[1] 

机构地区：[1]石河子大学农学院/新疆生产建设兵团绿洲生态农业重点实验室,新疆石河子832003 [2]石河子大学生命科学学院,新疆石河子832003

出　　处：《西北植物学报》2015年第10期1941-1948,共8页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(U1128301);国家"十二五"科技支撑计划(2011BAD35B05);石河子大学高层次人才专项(RCZX201212)

摘　　要：CLASP蛋白(CLASPs)是一种能够特异地与微管结合,参与调节微管结构与功能的微管结合蛋白(MAPs),在植物细胞形成、细胞伸长等方面起着关键作用。该研究利用电子克隆结合RT-PCR技术从陆地棉(Gossypium hirsutum L.)中克隆获得1个CLASP基因,命名为GhCLASP1(NCBI登录号为KP742966)。序列分析显示,GhCLASP1属于CLASP基因家族,开放阅读框长度为4 188bp,编码含1 395个氨基酸残基的蛋白;GhCLASP1蛋白含有1个HEAT重复结构域和2个CLASP-N端结构域。进化分析表明,GhCLASP1与可可树(Theobroma cacao)CLASP蛋白的亲缘关系最近。qRT-PCR分析表明,GhCLASP1在棉花的根、茎、叶、花及纤维发育的不同时期均有表达,且GhCLASP1基因表达量最大值出现在纤维发育次生壁加厚期(开花后27d),推测GhCLASP1基因可能对棉花纤维次生壁的形成起着重要的作用。利用本氏烟草(Nicotiana benthamiana)叶片瞬时表达系统对GhCLASP1基因编码的蛋白进行亚细胞定位结果显示,GhCLASP1蛋白定位在细胞膜上。上述结果为进一步研究CLASP基因在棉花中的功能奠定了基础。CLIP-Associated Proteins（CLASPs）is specifically binding to microtubules of microtubule associated proteins（MAPS）,involved in mediating the structure and function of microtubules,and play key roles during cell formation and cell elongation.A new CLASPgene was obtained fromGossypium hirsutum L.by electronic cloning and reverse-transcription polymerase chain reaction（RT-PCR）technology.Sequence analysis showed that GhCLASP1 belongs to the CLASP gene family,the full length ORF of GhCLASP1is4 188 bp,encoding 1 395 amino acids.GhCLASP1 has one HEAT domain and two CLASP-N structures.The phylogenetic analysis of homologue CLASP proteins of other plants showed that GhCLASP1 had a close relationship with TcCLASP of Theobroma cacao.Real-time quantitative reverse transcription PCR（qRT-PCR）showed that GhCLASP1 was expressed in root,stem,leaf,petal and fiber at different development periods.The highest expression of the GhCLASP1 was observed in the secondary period of the cotton fiber wall thick（27day post anthesis）and suggesting that GhCLASP1 might play an important role in the formation of secondary wall synthesis of cotton fiber.Transient expression system of the Nicotiana benthamianaleaves was used to analyze subcellular localization,and showed that GhCLASP1 might locatein the cell membrane.This study proved a foundation for studying the functions of CLASPgenes in fiber development.

关 键 词：棉花 纤维发育 CLASP 表达分析 亚细胞定位 

分 类 号：Q785[生物学—分子生物学] Q786