Zhang Gao-Hua, Wang He, Wang Xu-Da, Feng Ming, Li Huai-Mei, Li Shu-Ying
Institute of Dalian Biotechnology, Liaoning Academy of Agricultural Sciences, Dalian 116024, China.
Yi Chuan. 2012 Jun;34(6):742-8. doi: 10.3724/sp.j.1005.2012.00742.
The AlHAK1 gene encoding a high-affinity K+ transporter was isolated from Aeluropus littoralis (Gouan) Parl, a graminaceous halophyte, and plays a crucial role in nutrition and ion homeostasis in plant cell. To investigate the regulation role of AlHAK1 on the transcriptional level, an about 1.3 kb 5'-flanking region of the AlHAK1 gene containing a putative promoter was cloned by genome walking method. Cis-regulatory elements analysis showed AlHAK1-promoter region contained typical TATA and CAAT boxes, and some growth and development relative motifs, as well as environmental re-sponsive elements. To reveal the function and regulating role, the AlHAK1 promoter was fused to the β-glucuronidase (GUS) reporter gene in the pCAMBIA1301 vector and introduced into rice via Agrobacterium-mediated transformation. Histo-chemical staining indicated that the GUS expression directed by AlHAK1 promoter was observed in leaves, stems, roots, anther, lemma, and palea. GUS quantitative fluorometric analysis indicated that GUS activity directed by AlHAK1 promoter was lower than CaMV35S and Ubiquitin constitutive promoters; however, in the roots and stems the GUS activity was rela-tively high and displayed a tissue-specific expression pattern. Under ABA, high temperature or drought stress, the GUS activity directed by AlHAK1 promoter was inducible in the roots and stems, suggesting the elements of HSE (-682 bp) and MybBS (-1 268 bp) might play a role in the inducible regulation.
从禾本科盐生植物海滨碱蓬中分离出编码高亲和力钾离子转运体的AlHAK1基因,该基因在植物细胞的营养和离子稳态中起关键作用。为了研究AlHAK1在转录水平上的调控作用,采用基因组步移法克隆了AlHAK1基因约1.3 kb的5′侧翼区域,其中包含一个假定的启动子。顺式作用元件分析表明,AlHAK1启动子区域含有典型的TATA盒和CAAT盒,以及一些与生长发育相关的基序和环境响应元件。为了揭示其功能和调控作用,将AlHAK1启动子与pCAMBIA1301载体中的β-葡萄糖醛酸酶(GUS)报告基因融合,并通过农杆菌介导的转化导入水稻。组织化学染色表明,在叶片、茎、根、花药、内外稃中均观察到AlHAK1启动子指导的GUS表达。GUS定量荧光分析表明,AlHAK1启动子指导的GUS活性低于CaMV35S和泛素组成型启动子;然而,在根和茎中GUS活性相对较高,并呈现出组织特异性表达模式。在脱落酸、高温或干旱胁迫下,AlHAK1启动子指导的GUS活性在根和茎中可诱导,表明热激元件(-682 bp)和MybBS元件(-1268 bp)可能在诱导调控中起作用。