紫花针茅WRKY53基因的克隆、序列分析及功能鉴定

Molecular Cloning,Sequence Analyses,and Functional Identification the of WRKY53 Gene in Stipa purpurea

植物WRKY转录因子家族在叶片衰老中起着重要的调节作用,其成员WRKY53基因与叶片衰老密切相关。本文利用紫花针茅(Stipa purpurea Griseb.)的一条EST序列,克隆获得Sp WRKY53基因的全长CDS序列,其开放阅读框为1443 bp,编码480个氨基酸,蛋白质分子量为50.9 k D,理论等电点为7.64。序列比对结果和系统进化分析表明,Sp WRKY53基因有2个WRKY序列,与已报道的小麦(Triticum aestivem L.)的WRKY53基因结构相似。亚细胞定位结果显示该蛋白定位于细胞核上。紫花针茅WRKY53基因的过表达以及表达分析表明,该基因能够加速植物叶片衰老并在多种非生物胁迫下上调表达。本研究为WRKY53基因在牧草分子育种中的进一步研究奠定了基础。

The gene family of the WRKY transcription factors in plants play an important role in controlling leaf senescence. The WRKY53 gene is closely related to leaf senescence. Based on the EST sequence, the full-length CDS of Sp WRKY53 was cloned from Stipa purpurea.Sequence analysis showed that the opening reading frame of Sp WRKY53 was 1443 bp,which encoded a 480-amino acid peptide. Its protein molecular weight and isoelectric point were50. 9 k D and 7.64, respectively. Sequence alignment and phylogenetic analysis results indicated that the Sp WRKY53 gene with two WRKY sequences displayed similar structures with those reported for the WRKY53 gene of wheat. Sp WRKY53 was localized in the cell nucleus by subcellular analysis. Sp WRKY53 was upregulated and could promote leaf senescence under abiotic stress. This study will lay a foundation for further study of the WRKY53 gene in the molecular breeding of forage grass.