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毛白杨磷酸肌醇特异性磷脂酶PtoPLC4基因的克隆与表达

发表时间:2013-10-15  浏览量:1226  下载量:578
全部作者: 张杰伟,朱丹,张中保,陈亚娟,魏建华
作者单位: 北京市农林科学院北京农业生物技术研究中心,农业基因资源与生物技术北京市重点实验室
摘 要: 磷酸肌醇特异性磷脂酶C(phosphoinositide-specific phospholipase C,PI-PLC)是一类重要的磷酸肌醇信号途径中的酶。研究利用同源克隆和实时定量PCR(real-time quantitative PCR, RT-PCR)方法,从毛白杨(Populus tomentosa)中克隆到长度为1 686 bp编码区的PtoPLC4 cDNA(GeneBank No. JX861370),预测其编码561个氨基酸,分子量为64 ku,等电点为6.28. 对预测的氨基酸序列进行结构分析,结果表明:PtoPLC4具有EF手型结构域,X,Y和C2结构域,与其他已知的植物PI-PLC结构相似。实时定量荧光PCR检测表明,该基因在毛白杨不同组织中表达丰度不同,在叶片的表达量最高,其次为茎和根。用ABA,NaCl,低温和干旱等胁迫处理的实验表明,PtoPLC4的转录主要受到NaCl和干旱的影响,而受ABA和低温胁迫影响较小,推测PtoPLC4响应高盐和干旱胁迫,可作为调控林木抗逆的目标基因用于林木基因工程改良。
关 键 词: 林木遗传育种;毛白杨;PtoPLC4;基因克隆;盐胁迫;干旱胁迫
Title: Cloning and expression analysis of phospholipase PtoPLC4 gene in Populus tomentosa
Author: ZHANG Jiewei, ZHU Dan, ZHANG Zhongbao, CHEN Yajuan, WEI Jianhua
Organization: Key Laboratory of Agricultural Genetic Resources and Biotechnology, Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences
Abstract: Phosphoinositide-specific phospholipase C (PI-PLC) is a critical enzyme in the phospholipid signaling pathway. The full length PtoPLC4 gene (GeneBank No. JX861370) was cloned from Populus tomentosa seedlings cDNA.Sequencing results revealed that the cDNA length was 1 686 bp encoding 561 amino acids. The mature protein with the predicted molecular weight of 64 ku and the isoelectric point of 6.28. The sequence analysis indicated that the polypeptide contained EF-hand, X, Y and C2 domains. The overall structure of putative PtoPLC4 protein was similar to other plant PI-PLCs. Real-time quantitative PCR (RT-PCR) showed the PtoPLC4 expression level was different in different tissues. The expression level from high to low was the following order: leaves, stems and roots. The results also indicated that the transcribe of PtoPLC4 was mainly affected by NaCl and drought, while it only responded to ABA and low temperature lightly in P.tomentosa. These results suggested that PtoPLC4 gene could be taken for target gene in cultivating new clones for salt and drought resisting.
Key words: tree genetics and breeding; Populus tomentosa; PtoPLC4; gene cloning; salt stress; drought stress
发表期数: 2013年10月第19期
引用格式: 张杰伟,朱丹,张中保,等. 毛白杨磷酸肌醇特异性磷脂酶PtoPLC4基因的克隆与表达[J]. 中国科技论文在线精品论文,2013,6(19):1869-1875.
 
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