青杄CYP1基因的克隆与表达  

Cloning and Expression Analysis of CYP1 from Picea wilsonii

作　　者：罗朝兵[1] 鞠丹[1] 刘中帅 张凌云[1] 

机构地区：[1]省部共建森林培育与保护教育部重点实验室(北京林业大学),北京100083

出　　处：《东北林业大学学报》2015年第10期14-20,共7页Journal of Northeast Forestry University

基　　金：转基因生物新品种培育重大专项(2014ZX08009-003-002);国家自然科学基金项目(31270663)

摘　　要：以青杄(Picea wilsonii)的c DNA文库为模板,根据青杄基因Pw CYP1的EST序列设计引物,通过RACE-PCR方法获取Pw CYP1的全长c DNA序列。生物信息学分析表明:Pw CYP1全长c DNA为962 bp,编码框为516 bp组成,编码一个171氨基酸的多肽,Protparam工具预测蛋白的分子量约为17.98 k Da,理论等点为8.34。组织特异性试验表明,Pw CYP1在青杄花粉中的表达量最高,种子中最少。花粉萌发试验显示,Pw CYP1在青杄花粉萌发中表达量先下降,在后期表达量重新上调。在青杄种子萌发试验中,Pw CYP1表达量先下降,在第4天达到最低值后表达量被上调。同时,Pw CYP1受脱落酸(abscisic acid,ABA)、茉莉酸甲酯(methyl jasmonate,Me JA)、赤霉素(Gibberellin,GA)、干旱和盐胁迫的诱导且表达量上调,这些结果显示Pw CYP1参与了发育及多种逆境胁迫和对激素的响应过程。The full length c DNA of Pw CYP1 was obtained by RACE PCR method based on the full length c DNA library of Picea wilsonii and EST fragment of Pw CYP1. Bioinformatic tools were applied for predicting Pw CYP1. The full length c DNA of Pw CYP1 was 962 bp and the ORF was 516 bp which encoded 171 aa. The theoretical molecular weight of Pw CYP1 was 17. 98 k Da with isoeletric point of 8. 34. By the tissue-specific expression experiment,the expression level Pw CYP1 in pollen was the highest among all tissues and the expression level in seed was the lowest. By pollen germination assays,the expression of Pw CYP1 was dropped down to a nadir at the 24 th hour and then increased. During seed germination,the expression of PwCYP1 was decreased,peaked on the 4th day and then increased. The expression of Pw CYP1 was significantly induced by abscisic acid,methyl jasmonate,gibberellin,drought and salt stress. Pw CYP1 has a potential function in development and responding to stresses and phytohormone treatments.

关 键 词：青杄 亲环素蛋白 CYP1基因 植物激素 

分 类 号：S791[农业科学—林木遗传育种]