芜菁NAC转录因子BcNAC2基因的分离及其表达  

Isolation and Expression Analysis of BcNAC2,a NAC Transcription Factor Gene in Turnip

作　　者：张海娟[1] 吴剑锋[1] 胡帅[1] 余小林[1] 

机构地区：[1]浙江大学蔬菜科学研究所农业部园艺植物生长发育与品质调控重点开放实验室,杭州310029

出　　处：《园艺学报》2011年第6期1089-1096,共8页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(30771377);浙江省科技计划项目(2009C32029);浙江省自然科学基金项目(Y3090294);教育部留学回国人员科研启动基金项目(2009-1001);浙江省人事厅留学回国基金项目(J20090028)

摘　　要：根据拟南芥NAC2的全长序列设计引物,从芜菁中成功分离了NAC转录因子BcNAC2基因,该基因最大开放阅读框为1683bp,编码560个氨基酸,共包含6个外显子和5个内含子。将BcNAC2与其他30个NACs蛋白进行序列比对以及重构系统进化树分析发现,BcNAC2与十字花科的拟南芥的同源性最高,芜菁BcNAC2与拟南芥NAC2蛋白功能结构域内的特征序列有显著的差异。半定量RT-PCR的结果表明,BcNAC2呈部分组成型表达特征,在授粉8d的嫩角果中表达丰度最高,而且组织原位杂交结果显示,BcNAC2在胚囊中有较高的表达,由此推测,BcNAC2与芜菁的种子或胚的发育相关。BcNAC2,a NAC transcription factor gene,has been successfully isolated from turnip[Brassica campestris L.ssp.rapifera（Matzg）Sinsk,syn.B.rapa L.]using the primers designed from the sequence of the NAC2 gene in Arabidopsis thaliana.Sequence analysis results showed that the full length of genomic DNA contained six extrons and five introns,and the open reading frame（ORF）was 1 683 bp coding 560 amino acids.In addition,the sequence of BcNAC2 was much conserved comparing with those of other 30 NAC proteins,and it shared the highest homology with A.thaliana among them.In addition,there are significant difference for the motif numbers between the BcNAC2 and AtNAC2 functional domains.Moreover,RT-PCR results indicated that BcNAC2 gene presented the character of constitutive expression partially,and highest expression at 8 days after pollination.In situ hybridization result also proves that it is one of the high expression abundance genes during embryo sac development.Therefore,BcNAC2 probably involved in the development of seed or embryo in the turnip.

关 键 词：芜菁 NAC转录因子 基因分离 半定量RT-PCR 组织原位杂交 

分 类 号：S631.3[农业科学—蔬菜学]