Wang Feng-Qing, Zhou Yan, Huang Yong, Li Ming-Jie, Tian Yun-He, Feng Fa-Jie, Chen Xin-Jian
Yao Xue Xue Bao. 2015 Feb;50(2):233-40.
Using cDNA from Rehmannia glutinosa leaf as template, a 972 bp fragment of expansin gene which containing a 762 bp ORF that encoded 253 amino acids, was cloned, named RgEXPA10, which GenBank accession number for this gene is KF011918. A 1 207 bp genomic sequence of RgEXPA10 was amplified by PCR with leaf DNA as template, sequencing analysis revealed that three exons and two introns in RgEXPA10 genomic sequence, and which GenBank accession number is KF011919. Molecular and bioinformatic analyses indicated that RgEXPA10 protein have DPBB_1 and Pollen_allerg_1 domain, also including a 26 aa nuclear localization signal and a 19 aa transmembrane region. Phylogenetic analysis revealed that RgEXPA10 showed the highest homology with AtEXPA8 among the 26 α-expansins in Arabidopsis thaliana. However, the RgEXPA10 indicated the highest homology with the expansin from Solanum lycopersicum among 22 plant species. Expression patterns using qRT-PCR analysis showed that RgEXPA10 mainly expressed in unfolded leaf, followed by the tuberous root at stage of expanding period, and rarely expressed in senescing leaf. And RgEXPA10 showed higher expression level in tuberous root at 60 and 90 days after emergence. The transcription level of RgEXPA10 significantly reduced under all the three stresses including continuous cropping conditions, salinity and waterlogging. This study will lay foundations for molecular function in development and regulation of different stresses for R. glutinosa.
以地黄叶片的cDNA为模板,克隆得到一个972 bp的扩张蛋白基因片段,其包含一个762 bp的开放阅读框,编码253个氨基酸,命名为RgEXPA10,该基因的GenBank登录号为KF011918。以叶片DNA为模板,通过PCR扩增得到RgEXPA10的1207 bp基因组序列,测序分析显示RgEXPA10基因组序列中有3个外显子和2个内含子,其GenBank登录号为KF011919。分子和生物信息学分析表明,RgEXPA10蛋白具有DPBB_1和Pollen_allerg_1结构域,还包括一个26个氨基酸的核定位信号和一个19个氨基酸的跨膜区域。系统发育分析表明,在拟南芥的26个α-扩张蛋白中,RgEXPA10与AtEXPA8的同源性最高。然而,在22种植物中,RgEXPA10与番茄的扩张蛋白同源性最高。qRT-PCR分析显示,RgEXPA10主要在未展开的叶片中表达,其次是膨大期的块根,在衰老叶片中很少表达。RgEXPA10在出苗后60天和90天的块根中表达水平较高。在连作、盐胁迫和渍水这三种胁迫条件下,RgEXPA10的转录水平均显著降低。本研究将为地黄发育和不同胁迫调控的分子功能奠定基础。