菊花叶绿素a/b结合蛋白基因CmLhcb1及其启动子的克隆和表达分析

Cloning of Chlorophyll a/b Binding Protein CmLhcb1 and Promoter from Chrysanthemum morifolium and Expression Analysis

以切花菊品种‘公子’cDNA为模板克隆出叶绿素a/b结合蛋白同源基因cab,其开放阅读框为798bp,编码266个氨基酸。经多物种间比对分析,确认其属于cab基因家族的Lhcb1类,命名为CmLhcb1。同源克隆菊花品种‘清露’Lhcb1基因,氨基酸序列与‘公子’完全相同。CmLhcb1在叶片中的表达量比在茎、花和根中高,弱光和GA3处理使CmLhcb1表达上调,多效唑处理后CmLhcb1表达量受到抑制。CmLhcb1的表达受昼夜节律调节,白天表达量显著高于夜间。通过high-efficiencyTAIL-PCR(hiTAIL-PCR)方法克隆到‘公子’切花菊CmLhcb1起始密码子上游序列715bp和‘清露’起始密码子上游序列716bp,序列经PLACE数据库的比对分析,发现有很多与非生物和生物胁迫相关的元件,主要与光照、GA、ABA、水分、水杨酸和病毒相关,CmLhcb1启动子是光诱导型启动子,具有GT1-box和Z-box。

The cDNA of cut chrysanthemum ＇Gongzi＇ was used to clone homologous gene cab, which had 798 bp ORF and 266 amino acid. After blast analysis, we confirmed that the gene was ranked as Lhcbl, and was named as CmLhcbl. Using the cDNA of ＇Puma Sunny＇ chrysanthemum to homologously clone gene Lhcbl, we got the same amino sequence with that of ＇Gongzi＇ . The expression of CmLhcbl was the higher in leaf than that in stem, flower and root. Low light and GA3 treatment increased CmLhcbl expression. Paclobutrazol treatment inhibited CmLhcbl expression. The circadian clock regulated theexpression of CmLhcbl. The gene expression in day was enormously higher than that in night. The promoter sequence 715 bp of Gongzi＇ cut chrysanthemum and 716 bp of＇Puma Sunny＇ were cloned using high-efficiency TAIL-PCR （hiTAIL-PCR）, and many biologic and abiotic stress responsive elements related to light, GA, ABA, water, SA and virus were found by PLACE Databank. The promoter was light responsive, and it had the GTl-box and Z-box element.