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小黑杨转录因子PsnbZIP1应答盐胁迫功能分析

Function Analysis of the Transcription Factor PsnbZIP1 of Populus simonii×P. nigra in Response to Salt Stress

  • 摘要: 为揭示小黑杨(Populus simonii×P. nigra)在面对非生物胁迫时,转录因子PsnbZIP1在植物体内发挥的功能,以小黑杨为试验材料,克隆得到PsnbZIP1的ORF区序列长为432 bp,并初步分析PsnbZIP1盐胁迫下的分子机制。采用q-PCR分析PsnbZIP1在150 mmol·L-1 NaCl处理小黑杨组培苗时的表达模式,发现该基因的表达量快速上升;通过生物信息学分析预测PsnbZIP1转录因子为无跨膜结构且具有信号肽的亲水性不稳定蛋白;用农杆菌(Agrobacterium)介导的烟草(Nicotiana)瞬时表达观察该基因的亚细胞定位情况,结果表明该基因为核定位蛋白;用酵母单杂交实验证明该基因编码的蛋白在酵母体内不具有转录激活功能。对PsnbZIP1基因的启动子序列进行分析,结果表明该启动子包含了生长素应答、脱落酸应答元件、光应答元件以及种子特异性调控的顺式作用调控元件,该基因可能在植物的生长发育与响应胁迫过程中发挥了重要作用;启动子还包括参与干旱诱导的MYB结合位点和MYBHv1结合位点,表明该基因有可能与一些干旱诱导相关MYB基因相互作用。

     

    Abstract: To reveal the function of transcription factor PsnbZIP1 from Populus simonii×P. nigra under abiotic stresses, P.simonii×P. nigra was used as material, and the PsnbZIP1 transcription factor gene was cloned with an ORF length of 432 bp, and the molecular mechanism of PsnbZIP1 under salt stress was analyzed. The expression pattern of PsnbZIP1 in P. simonii×P. nigra was analyzed under 150 mmol·L-1 NaCl by q-PCR, it was found that the expression of the gene increased rapidly. The PsnbZIP1 was predicted to be a hydrophilic unstable protein without trans-membrane structure but with signal peptide by bioinformatics analysis. The sub-cellular localization of the gene was observed by Agrobacterium mediated transient expression in tobacco. The results showed that the gene was a nuclear localization protein. It was confirmed by Y2HGold yeast competent cells that the gene had no transcriptional activation activity in yeast. The analysis of the promoter sequence of the PsnbZIP1 gene showed that the promoter contained auxin response elements, ABA response elements, light response elements and seed-specific cis-acting regulatory elements respectively, which might play a role in plant growth and development and response to stress.The promoter also included the MYB binding site and MYBHv1 binding site involved in drought induction, indicating that the gene was likely to interact with some of MYB involved in drought induction.

     

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