路易斯安那鸢尾LiPCS1基因克隆和表达分析

Cloning and qPCR Analysis of LiPCS1 Gene from Louisiana Iris

通过对路易斯安那鸢尾转录组进行分析,获得路易斯安那鸢尾络合素合酶基因(LiPCS1)的中间片段,利用RACE技术分别克隆到其全长,并对该基因进行生物信息学分析。结果表明,该基因全长1 683bp,预测编码501aa,含有半胱氨酸(Cys)、组氨酸(His)和天冬氨酸(Asp)3个必需氨基酸活性位点和重金属离子传感器基序(C368 C369 RMTC373 VRC376),与猴面花等植物同源PCS的相似位点达91.6%。表达分析结果表明,LiPCS1基因在内花瓣表达量最高,与其他组织差异显著,雄蕊、叶和茎的表达量基本在同一水平,外花瓣、根和雌蕊表达较低,雌蕊最低;随着铅处理时间增加,LiPCS1基因表达量在根系和叶片中表现先升高后下降的趋势,在铅处理3h达到最大并且差异显著。

Louisiana iris is a kind of high ornamental value of perennial plants,found to be tolerant and accumulation of heavy metal Pb.However,research on the molecular mechanism of Louisiana iris is limited.With the information of LiPCS1 iutermediate fragment form transcriptome of‘Professor Neil＇and using RACE technology,we cloned LiPCS1 for the gene bioinformatics prediction and analysis.The total length of the gene was 1 683 bp,predictive coding protein contained 501 aa.The protein contained three active essential amino acids sites,Cys,His and Asp,and base sequence of heavy metal ion sensor,C368 C369 RMTC373VRC376.The conservative analysis with other homologous PCSfound that similar loci was 91.6%.The highest expression of LiPCS1 was found in inter petals of Louisiana iris,and significant differences with other organs.The same expression level was found in stamens,leaves and stems.The expression in outer petals,roots and pistils were lower,and the expression in pistil was the lowest.