摘要
苎麻是中国的传统纤维作物,能够生产最长的自然纤维。本研究旨在克隆苎麻纤维素合酶基因BnCesA1全长编码序列,对其表达模式进行分析。以已知的苎麻纤维素合酶基因序列(DQ077190)为基础,设计5'RACE引物,以湘苎三号为材料,得到了BnCesA1的5'端,拼接后得到了BnCesA1的全长序列,并从湘苎三号的cDNA中成功克隆到包括BnCesA1全部编码序列的cDNA序列。扩增得到的BnCesA1基因cDNA为3253bp,编码区3246bp,编码含1082个氨基酸的多肽。通过对这个基因进行核酸序列和蛋白结构域分析表明,BnCesA1和毛果杨、欧美山杨、巨桉、大叶相思等其他物种的纤维素合酶基因都有很高的同源性,根据得到的BnCesA1的5'端设计特异性表达检测引物,分析其在湘苎三号苎麻品种各组织中的表达情况,结果显示BnCesA1在所检测的各组织中均有表达,表达量为茎皮>叶>顶芽>根。本研究首次克隆到苎麻中编码全长蛋白的纤维素合酶基因,并且苎麻BnCesA1在茎皮中高表达提示该基因可能在苎麻韧皮纤维合成中有重要作用。
Boehmeria nivea（ramie）is a traditional Chinese fibre crop,and provide the longest nature fibre.This study aimed to clonethe complete coding sequences of CesA1 gene from ramie and analyze its express pattern among different ramie tissues.According to the known sequence of BnCesA1（DQ077190）,5′RACE PCR groove primers were designed to amplify the 5′terminal fragment of BnCesA1 using cDNA from ramie selection ＂Xiangzhu 3＂.Then the full length gene sequence was assembled and cloned.The whole jointed sequence spans 3253bp and has an open reading frame of 3246bp,which codes for a protein of 1082 amino acids.Nuclear acid sequence analysis and protein domain comparison indicate that BnCesA1 has a very high homology with Populus trichocarpa,Populus tremuloides,Eucalyptus camaldulensis,Acacia mangium.Specific primers were designed according 5′sequence of BnCesA1 for semi-quantitative RT-PCR analysis to detect the BnCesA1 expression pattern among different ramie tissues.The result demonstrated that BnCesA1 are expressed in various tissues of ＂Xiangzhu 3＂.And the expression level in tissues from high to low is stem,leaf,bud and root.Altogether,our research cloned the first full-length coding sequence of ramie cellulose synthesase,and the higher expression of BnCesA1 in bark suggest that this gene may play an important role in bast fibre synthesis of ramie.
出处
《植物遗传资源学报》
CAS
CSCD
北大核心
2012年第5期851-857,共7页
Journal of Plant Genetic Resources
基金
湖南省作物种质创新与资源利用国家重点实验室培育基地一般重点项目(09KFXM02)
省科技计划项目博士后专项基金(S2010R1040)
湖南省应用基础重点项目(2010TP4004-1
2010FJ2013)
关键词
苎麻
纤维素合酶基因
RACE
表达分析
Rmaie（Boehmeria nivea）
Cellulose synthase
RACE
Express analysis
