榛子甜菜碱醛脱氢酶基因BADH的克隆及在越冬过程中的表达特性分析  

Cloning and Expression Analysis of Betaine Aldehyde Dehydrogenase BADH Gene from Hazelnut(Corylus heterophylla Fisch.) during Overwintering Period

作　　者：陈新[1,2] 王贵禧[2] 徐丽[1] 张力思[1] 刘庆忠[1] 崔海金[2] 张学寅[2] 

机构地区：[1]山东省果树研究所,山东泰安271000 [2]林木遗传育种国家重点实验室/中国林业科学研究院林业研究所,北京100091

出　　处：《山东农业科学》2013年第5期6-12,共7页Shandong Agricultural Sciences

基　　金：国家林业公益性行业科研专项(201304710);中俄国际合作项目(2012DFR30700);科技部科技基础工作资助项目‘国家核桃板栗种质资源平台’(2012-048);山东省农业良种工程(鲁科农字[2011]086))

摘　　要：甜菜碱是高等植物体内一种重要的渗透调节物,甜菜碱醛脱氢酶(Betaine aldehyde dehydrogen-ase,BADH)是甜菜碱合成的关键酶之一,本文基于Solexa技术对平榛花芽转录组测序结果的分析采用RACE-PCR技术克隆到一个平榛BADH基因,命名为ChBADH(HQ700873)。ChBADH cDNA全长1 691 bp,具有一个1 512 bp的潜在编码区,编码503个氨基酸,预测其蛋白质相对分子量54.7 ku,理论等电点为5.44。Ch-BADH具有BADH家族保守的VSLELGGKSP功能活性位点和特征多肽序列,序列比较和生物信息学分析结果显示ChBADH在保守结构域和其他植物来源的BADH享有高度的一致性。以ACTIN为内参,对ChBADH基因在四个时期平榛的表达模式进行了初步的研究,荧光定量结果表明,ChBADH在寒冬时期被强烈的诱导表达,是非冷适应时期的3.5倍,推测该基因可能与榛子花芽越冬有紧密关系。Glycinebetaine is an important osmoregulation substance in higher plant and betaine aldehyde dehydrogenase（BADH） is one of the key enzymes in betaine biosynthesis.A novel BADH gene,named ChBADH（HQ700873）,was isolated by RACE-PCR based on solexa sequencing of floral bud of hazelnut（Corylus heterophylla Fisch.）.Sequence analysis revealed that ChBADH contained 1 691 nucleotides and had an opening reading frame（ORF） of 1 512 nucleotides encoding 503 amino acids.The relative molecular weight of predicted protein was 54.7 ku and its isoelectric point was 5.44.ChBADH possessed a conserved VSLELGGKSP domain and specific multi-peptide sequences.Sequence alignment and phylogenetic analysis showed that the amino acid sequence of ChBADH had high concordance with BADH from other plants.Using ACTIN as an internal reference,the expression of ChBADH in bud in 4 stages was studied by qRT-PCR.The results indicated that the transcripts of ChBADH were elevated by 2.5 fold in mid-winter than that in non-cold acclimation.It suggested that ChBADH might involve in natural overwintering of hazelnut.

关 键 词：平榛 甜菜碱醛脱氢酶 QRT-PCR 寒冬 

分 类 号：Q785[生物学—分子生物学]