烟草转录因子基因NtGRAS-R1的克隆与功能分析  

Cloning and Function Analysis of Transcription Factor Gene NtGRAS-R1 from Nicotiana tabacum L.

作　　者：李富欣[1,2] 许芳芳[1] 李素敏[1] 肖万福[1] 孙艳敏 刘卫群[1] 郭红祥[1] 

机构地区：[1]河南农业大学生命科学学院,河南郑州450002 [2]河南省烟草公司济源市公司,河南济源454650 [3]濮阳市农业科学院,河南濮阳457000

出　　处：《河南农业科学》2015年第5期58-62,共5页Journal of Henan Agricultural Sciences

基　　金：河南省烟草公司重点项目(HYKJ201308)

摘　　要：为了分析Nt GRAS-R1的生物学功能,在NCBI上BLAST植物的EST数据库拼接获得NtGRAS-R1的全长序列;根据该序列设计特异引物,利用PCR方法从烟草根系c DNA中扩增NtGRAS-R1,将其连接到p S1300表达载体上,采用农杆菌介导的花序侵染法转化拟南芥,采用RTPCR法检测转基因拟南芥植株;获得稳定转基因拟南芥后观察生长性状,并用q PCR方法检测AtCLV3基因的表达情况。结果显示,Nt GRAS-R1基因属于HAM亚家族,编码508个氨基酸;观察发现,转基因拟南芥植株根长和根体积明显大于野生型;q PCR结果表明,转基因拟南芥At CLV3的表达量明显低于野生型拟南芥。初步表明Nt GRAS-R1参与根系生长发育调控过程。The aim of this research is to explore the function of NtGRAS-R1 . The full-length sequence of NtGRAS-R1 was acquired with in silico cloning method. The ORF fragment of NtGRAS-R1 was obtained with PCR amplification method, and then pS1300-NtGRAS-R1 expression vector was constructed. Transgenic Arabidopsis with NtGRAS-R1 was obtained with Agrobacterium tumefaciens-mediated floral dip transformation method. RT-PCR was used to identify transgenic plants. The phenotypic of transgenic Arabidopsis was observed, and the expression of AtCLV3 was detected with qPCR method. The results showed that NtGRAS-R1 belonged to HAM subfamily,having an ORF of 1 527 bp to encode a protein of 508 amino acids. Compared to wild type plants, the root length and volume of transgenic Arabidopsis obviously increased. The expression of AtCLV3 in transgenic Arabidopsis was lower than wild type plant. In the present study, NtGRAS-R1 was successfully cloned, and the phenotypic analysis of transgenic Arabidopsis shows that NtGRAS-R1 has a role in the process of tobacco roots growth and development.

关 键 词：烟草 根系 NT GRAS-R1 打顶 发育调控 

分 类 号：S572[农业科学—烟草工业]