丹参ERF转录因子SmERF1基因的表达分析和亚细胞定位  

Expression analysis and subcellular localization of an ERF transcription factor from Salvia miltiorrhiza

作　　者：吴文燕[1] 蒋喜红[1] 刘春生[1] 黄璐琦[2] 申业[2] 

机构地区：[1]北京中医药大学中药学院,北京100029 [2]中国中医科学院中药资源中心,北京100070

出　　处：《中国中药杂志》2013年第7期957-961,共5页China Journal of Chinese Materia Medica

基　　金：国家“重大新药创制”科技重大专项(2009ZX09301005)

摘　　要：目的:对前期已经克隆的丹参SmERF1基因进行聚类分析,分析SmERF1基因在不同诱导子处理后不同时间的表达情况;并对其进行亚细胞定位分析。方法:利用MEGA5软件对SmERF1基因及拟南芥ERF基因家族进行聚类分析;利用半定量RT-PCR方法,分析SmERF1基因在不同诱导子处理后不同时间的表达情况;将SmERF1与GFP融合,在洋葱表皮瞬时表达,以确定SmERF1蛋白表达部位。结果:丹参SmERF1属于ERF家族第Ⅶ亚族;YE+Ag+诱导对SmERF1基因的表达没有影响,ABA和MeJA可以抑制该基因的表达,而水杨酸诱导会出现先抑制,后随处理时间加长,SmERF1基因表达又恢复;亚细胞定位确定SmERF1基因在细胞核内特异表达。结论:SmERF1是一个AP2/ERF转录因子,属于AP2/ERF家族第Ⅶ亚族,其表达受ABA,SA,MeJA等激素调节控制。Objective： A SmERFIgene was isolated from Salvia miltiorrhiza, and expression patterns to different stress condition were analysed in the root tissues of S. miltiorrhiza. Method： The cDNA of SmERF1 gene from S. miltiorrhiza was isolated by RT- PCR, and the phylogenetic tree using the neighbour-joining tree method in Mega 5 was obtained. To confirm the protein is likely to localize in the nucleus, the SmERFI coding region was fused to the N-terminus of the GFP gene under the control of the CaMV 35S promoter and transferred into onion epidermal cells using the particle bombardment method. Semi-quantitative RT-PCR analysis revealed different expression pattern of SmERFI gene in response to exogenous ABA, MeJA and SA. Result ： The phylogenetic tree analysis revealed that SmERFI is most similar to AP2/ERF VII subgroup members. The transient expression of the SmERFI ： ： GFP fusion protein indicated that the SmERFI was exclusively localized to the nucleus. The transcript of SmERF1 highly accumulated when the plants were treated with MeJA, while accumulated slightly in response to exogenous ABA, salicylic acid. Conclusion： These results suggest hormone such as ABA, MeJA and SA signaling pathways can be involved in the activation and inhibition of the SmERFI.

关 键 词：丹参 ERF转录因子 表达分析 亚细胞定位 

分 类 号：S567.53[农业科学—中草药栽培] R282.7[农业科学—作物学]