普通烟草钙依赖蛋白激酶NtCDPK15的基因克隆及表达分析  

Cloning and Expression of Calcium-dependent Protein Kinase 15 (NtCDPK15) in Nicotiana tabacum

作　　者：康乐[1,2] 张丽[1,2] 张磊[1,2] 吴清章[1,2] 丁安明[1,2] 宗鹏[1,2] 刘贯山[1] 

机构地区：[1]中国农业科学院烟草研究所,烟草行业烟草基因资源利用重点实验室,青岛266101 [2]中国农业科学院研究生院,北京100081

出　　处：《中国烟草科学》2013年第3期48-54,共7页Chinese Tobacco Science

摘　　要：钙依赖蛋白激酶(CDPK)对Ca2+信号转导通路下游元件的调控具有重要作用。本实验采用同源克隆技术从普通烟草中分离得到1个CDPK基因,命名为NtCDPK15(GenBank登录号为JN662020),cDNA全长为1963 bp,ORF大小为1569 bp,编码522个氨基酸。多序列比对结果表明,NtCDPK15的编码产物具有典型的CDPK保守序列,C末端调控区有4个EF手性结构。系统进化树分析显示,NtCDPK15可能来源于绒毛状烟草,预测NtCDPK15与NtCDPK1这对旁系同源基因在功能上可能非常保守。实时荧光定量PCR分析结果表明,NtCDPK15在根和叶中的表达量显著高于茎中的表达量,但经盐胁迫诱导后NtCDPK15在茎中的表达量显著增加,同时,盐胁迫还使基因在叶中的表达量显著上调,ABA胁迫可诱导基因在根中的表达,而干旱胁迫可使基因在根和叶中的表达量下调,这些结果说明NtCDPK15在普通烟草中的表达受盐胁迫及ABA胁迫诱导,但受干旱胁迫抑制。Calcium-dependent protein kinases（CDPKs） play important roles in regulating downstream components in calcium signaling pathways. NtCDPK15 （GenBank Accession No. JN662020）, was isolated from common tobacco （Nicotiana tabacum） by rapid amplification of cDNA ends （RACE）. The NtCDPK15 cDNA is 1963 bp long and contains an open reading frame （ORF） of 1569 bp encoding 522 amino acids. Sequence alignments indicated that NtCDPK15 was a typical CDPK which had well-conserved C-terminal calmodulin-like structure with 4-EF hand motifs for calcium-binding. Pbylogenetic analysis indicated the NtCDPK15 might originate from Nicotiana tomentosiformis, and NtCDPK15 and NtCDPK1 might correspond to conserved functions. The results of real-time quantitative reverse transcription-PCR （qRT-PCR） showed that the relative expression of NtCDPK15 in roots and leaves was significantly higher than those in stems, but highly increased in stems treated with high-salt. Salt stress and ABA treatment made the relative expression of the gene increased in leaves and roots, but the gene expression was lowered subjected to drought stress in roots and leaves. These indicate that in common tobacco NtCDPK15 is induced by high-salt and ABA stresses, and suppressed by drought stress.

关 键 词：普通烟草 CDPK RACE 非生物胁迫 基因表达 

分 类 号：S572[农业科学—烟草工业]