马尾松纤维素合成酶基因PmCesA1的克隆及其分析  

Cloning and Analysis on PmCesA1 Gene Encoding Pinus massoniana Cellulose Synthase

作　　者：阮维程 潘婷[1] 季孔庶[1] 

机构地区：[1]南京林业大学南方现代林业协同创新中心,南京210037

出　　处：《分子植物育种》2015年第4期861-870,共10页Molecular Plant Breeding

基　　金：国家科技支撑项目(2012BAD01B0202);林业公益性行业科研专项经费(201104010);江苏高校优势学科建设工程(PAPD);南方现代林业协同创新项目共同资助

摘　　要：植物纤维素合成酶(Ces A)是林木中纤维素合成过程中的重要酶,与木材形成密切相关。本研究以马尾松嫩枝的总RNA反转录得到的c DNA为模板,利用反转录PCR和RACE技术克隆得到马尾松Ces A1基因的全长c DNA序列,命名为Pm Ces A1。序列长度为3 142 bp,包含有一个长为2 955 bp的开放阅读框(open reading frame,ORF),编码984个氨基酸。序列分析表明:Pm Ces A1序列与已报道的火炬松(Pinus taeda)Ces A1基因(AY789650.1)的相似性达99%。对Pm Ces A1所编码蛋白的氨基酸序列聚类分析,结果显示马尾松与松科植物具有较近的亲缘关系。采用实时定量PCR测定了Pm Ces A1在马尾松的嫩枝、叶和根中的表达量,发现以嫩枝中的表达量最高(2.53)。本研究获得的全长基因可以用来构建正义表达载体,再通过遗传转化得到转基因植株,可进一步分析该基因在植株内的表达情况和作用机制,为马尾松纤维素合酶的研究和获得高纤维得率的马尾松良种提供帮助。Plant cellulose synthase （CesA） is a family of enzymes playing an important role in the process of cell-ulose biosynthesis and closely related to the formation of forest timber. In this study, total RNA extracted from twigs of Pinus massoniana was reverse transcribed into cDNA and used as PCR template. With the strategies of RT-PCR and RACE, we cloned full-length CesA 1 gene cDNA （named PmCesA 1） that is 3 142 bp in length, and has a 2 955 bp open reading frame （ORF）, encoding 984 amino acids. Sequence analysis showed that PmCesA 1 has a 99%similarity with reported CesA1 gene （AY789650.1） of P. taeda. The amino acid sequence clustering analysis showed P. masso-niana had a closer kinship with Pinaceae. Based on the PmCesA 1 expression levels of twigs, leaves and roots in P. massoniana measured by RT-PCR, it was found that the twigs had the highest expression level of PmCesA 1 （2.53）. The full length gene obtained in this study can be used to construct sense expression vectors and get transgenic plants. Then the expression and mechanism of action about PmCesA 1 in P. massoniana can be further analyzed, which will help for theoretical study of cellulose synthase and breeding improved P. massoniana with high content of cellulose.

关 键 词：马尾松 纤维素合成酶基因 CDNA克隆 PmCesA1 序列分析 

分 类 号：Q943.2[生物学—植物学]