以菜薹[Brassica rapa L.ssp.chinensis (L.) Mokino var.utilis Tsen et Lee]为材料,克隆得到1个WRKY转录因子,命名为BrWRKY75.氨基酸序列比对及进化树分析发现BrWRKY75与拟南芥的WRKY75同源性较高,且同属于第Ⅱ类c亚族.实时荧光定量PCR表明BrWRKY75在菜薹叶片衰老过程中表达明显增强.亚细胞定位和转录活性分析显示BrWRKY75定位于细胞核,是一种核蛋白,并且具有转录抑制活性.体外凝胶阻滞试验(EMSA)证实BrWRKY75能与W-box (TTGAC)元件特异结合.上述结果为进一步研究菜薹叶片衰老的转录调控机制奠定了基础.
Molecular Characterization of a Transcription Factor BrWRKY75 Related to Leaf Senescence of Chinese Flowering Cabbage
In the present work,a WRKY transcription factor,termed BrWRKY75 was isolated from Chinese flowering cabbage.Analysis of deduced amino acid sequence and phylogenetic tree found that BrWRKY75 showed high homology with Brassica oleracea WRKY75 and Arabidopsis thaliana WRKY75,and belong to Ⅱ c sub-group.Real-time quantitative PCR displayed that BrWRKY75 was up-regulated during leaf senescence of Chinese flowering cabbage.Sub-cellular localization and transcriptional activity analysis revealed that BrWRKY75 is a nuclear protein with transcriptional repression activity.In addition,electrophoretic mobility shift assay (EMSA) confirmed that BrWRKY75 directly bound to the W-box (TTGAC) cis-element.Taken together,these results provide basis for the further investigation of the transcriptional regulation mechanism of Chinese flowering cabbage leaf senescence.
Chinese flowering cabbageleaf senescenceWRKY transcription factortranscriptional regulation
NETL
NSTL
万方数据
维普
