马尾松PmPIN2基因的克隆及实时定量分析  

Cloning and Real-time Quantitative Analysis of PmPIN2 Gene from Pinus massoniana

作　　者：武星 季子钧 吴晓刚[1] 陈佩珍[1] 吴帆 孙晓波 季孔庶[1] Wu Xing;Ji Zijun;Wu Xiaogang;Chen Peizhen;Wu Fan;Sun Xiaobo;Ji Kongshu(Co-Innovation Center for Sustainable Forestry in Southern China,Key Laboratory of Forest Genetics and Biotechnology of Ministry of Education,Nanjing Forestry University,Nanjing,210037;School of Resources ＆ Environment,Anhui Agriculture University,Hefei,230036)

机构地区：[1]南京林业大学南方现代林业协同创新中心、林木遗传与生物技术省部共建教育部重点试验室,南京210037 [2]安徽农业大学,资源与环境学院,合肥230036

出　　处：《分子植物育种》2018年第15期4880-4889,共10页Molecular Plant Breeding

基　　金：“十三五”国家重点研发计划项目课题(2017YFD0600304)、江苏高校优势学科建设工程(PAPD)和南方现代林业协同创新项目共同资助

摘　　要：PIN2基因在植物生根机理中起关键作用,对马尾松PIN2基因进行研究,可为解决马尾松无性系生根困难、促根壮苗培育提供帮助。本研究以马尾松幼苗全株为试材,利用PCR技术和RACE技术克隆了马尾松PIN2基因全长,并通过相关软件和实时荧光定量进行生物信息学和组织特异性分析。结果发现PIN2基因全长3 706 bp,包括2 103 bp完整ORF序列,编码700个氨基酸。生物信息学分析表明,PIN2蛋白为疏水性蛋白,相对分子量76.43 k D,等电点(p I)为9.06,总亲水性平均数0.023,PmPIN2编码的蛋白与PIN家族具有同样典型结构域。实时荧光定量分析结果显示,PmPIN2基因在马尾松根、茎、叶、花中均有表达,其中根中的表达量最高,花中最低。PIN2基因是参与植物生根过程的重要基因,对马尾松PmPIN2基因的研究为PIN基因家族在生根机制方面的作用探究提供帮助。The PIN2 gene plays a key role in the plant rooting mechanism. The study on PIN2 gene in Pinus massoniana may provide help to solve the rooting problems of P.massoniana clones and improve the growth of seedling and rooting. In this study, the whole P.massoniana seedling was used as experimental material to clone the whole length of P. massoniana PIN2 gene by PCR and RACE techniques. Bioinformatics and tissue-specific analysis were also performed with related software and real-time fluorescence quantification. The results showed that PIN2 gene was 3706 bp in total length and encoded 700 amino acids, including 2103 bp complete ORF sequence. Bioinformatics analysis showed that PIN2 protein was a hydrophobic protein, which the relative molecular weight of protein was 76.43 kD, and the isoelectric point （pI） was 9.06. The total mean ofhydrophilicity was 0.023. the PmPIN2 encoded protein had the same typical structural domain as the PIN family. The result of real-time quantitative fluorescence analysis showed that the PmPIN2 gene was expressed in roots, stems, leaves and flowers of P.massoniana, among which the highest expression was in roots and the lowest was in flowers. The PIN2 gene was an important gene involved in the rooting process of the plant, the study on the PmPIN2 gene in P.massoniana would provide help to explore the role of PIN gene family in rooting mechanism.

关 键 词：马尾松 PmPIN2基因 实时荧光定量分析 

分 类 号：Q943.2[生物学—植物学]