杨树ARGONAUTE基因的克隆及序列分析  

Cloning and Characterization of ARGONAUTE Genes in Populus

作　　者：胥猛[1] 谢雯凡[1] 潘惠新[1] 苏晓华[2] 张守攻[2] 黄敏仁[1] 

机构地区：[1]南京林业大学江苏省杨树种质创新与品种改良重点实验室,南京210037 [2]中国林业科学研究院林业研究所,北京100091

出　　处：《林业科学》2011年第3期46-51,共6页Scientia Silvae Sinicae

基　　金：国家"973"项目(2009CB119107);国家林业公益性行业科研专项(201004004)

摘　　要：作为RNA诱导沉默复合体的核心元件,Argonaute(AGO)蛋白在植物生长发育过程中发挥着重要作用。以欧美杨和美洲黑杨不定根cDNA为模板,采用RT-PCR技术分离得到PeAGO5和PdAGO5基因的cDNA克隆,测序和序列分析结果表明2个目的cDNA片段均为2809bp,基因内部含有完整的开放阅读框,可编码907个氨基酸,预测蛋白质分子量分别为102.31ku和102.34ku,理论等电点(pI)分别为9.34和9.7。所推导的2个氨基酸序列之间仅有7个差异位点,含有3个保守的结构域,DUF1785、PAZ和Piwi结构域。它们与拟南芥AtAGO5蛋白的相似性分别为64.8%和64.3%,故将其命名为PeAGO5和PdAGO5。系统进化分析发现,PeAGO5和PdAGO5,与拟南芥AtAGO1,AtAGO5和AtAGO10蛋白同属一个进化分支。此外,采用实时定量PCR技术检测PeAGO5基因在杨树硬枝插穗不定根发育过程中的表达模式,推测该基因参与不定根发育调控。As the core component of RNA induced silencing complex （RISC）,Argonaute （AGO） proteins play an essential role in plant development and cellular metabolism.Our previous studies suggested that a predicted gene from Populus trichocarpa genome sequence,POPTR _ 0001s22120.1,was preferentially expressed in adventitious root development of Populus cuttings.In this study,the target cDNA clones were isolated from the cDNA library prepared from adventitious roots of P.× euramericana and P.deltoides using RT-PCR technique,respectively.The two cDNAs were both 2 809 bp,and would encode for polypeptides of 907 amino acid residues with the molecular mass around 102.31 ku and 102.34 ku,the theoretical isoelectric point （pI） of 9.34 and 9.7,respectively.There is difference in seven distinct amino acid residues between the two deduced proteins which contain 3 conserved domains： DUF1785,PAZ and Piwi,and share 64.8% and 64.3% identify with AtAGO5 protein of Arabidopsis thaliana,respectively.They were,therefore,named as PeAGO5 and PdAGO5.Phylogenetic tree revealed that PeAGO5,PdAGO5,AtAGO1,AtAGO5 and AtAGO10 were assigned to the same clade.Quantitative real-time PCR analysis showed that PeAGO5 was differentially expressed during adventitious root development in hardwood cuttings of the poplars,indicating that PeAGO5 would be involved in adventitious root development.

关 键 词：杨树 硬枝插穗 Argonaute蛋白 表达模式 QRT-PCR 

分 类 号：S718.46[农业科学—林学] Q943.2[生物学—植物学]