FENG Wanqian,MA Lipeng,JIANG Xiaoxiao,et al.Cloning and Expression Analysis of a Transcription Factor Gene ZjDREB1.2 From Zoysia[J].Northern Horticulture,2018,42(18):75-85.[doi:10.11937/bfyy.20180246]
结缕草转录因子基因ZjDREB1.2的克隆及胁迫下的表达分析
- Title:
- Cloning and Expression Analysis of a Transcription Factor Gene ZjDREB1.2 From Zoysia
- Keywords:
- Zoysia; transcription factor; DREB; abiotic stress
- 文献标志码:
- A
- 摘要:
- 结缕草是一类多耐逆暖季型草坪草，但不耐低温。以日本结缕草‘Meyer’‘胶东青’和沟叶结缕草‘马尼拉’为材料，克隆获得了结缕草（Zoysia）一个新的DREB类转录因子基因，命名为ZjDREB1.2（GenBank登录号：KP696367.1）。结果表明：该基因开放阅读框长702 bp，推测编码的蛋白质含233个氨基酸，平均分子量为25.2 KDa，理论等电点为4.91。保守结构域预测显示ZjDREB1.2蛋白含有一个不完整的AP2结构域，结构域的上、下游均具有DREB1亚组特异性序列。系统进化树分析结果显示ZjDREB1.2蛋白属于DREB家族A-1亚组，与C4植物的同源基因遗传距离较近。在日本结缕草‘Meyer’中，ZjDREB1.2基因在低温、高盐和干旱胁迫2~12 h内均上调表达，其中受低温诱导上调幅度最大。与沟叶结缕草‘马尼拉’相比较，低温胁迫72 h后日本结缕草中ZjDREB1.2的表达量相对较高。酵母单杂交结果显示，ZjDREB1.2蛋白具有强的转录激活功能，但不表现出与DRE元件结合功能。暖季型植物中这种AP2结构域不完整DREB1基因的进化机制及功能值得研究。
- Abstract:
- Zoysia is a type of warm season turf grass and shows high tolerance to multiple stresses except cold.A new dehydration-responsive element binding (DREB) gene,designated as ZjDREB1.2 (GenBank accession No:KP696367.1),was isolated from three cultivars of zoysiagrass,‘Meyer’ (Zoysia japonica),‘Jiaodongqing’ (Z.japonica) and ‘Manila’ (Z.matrella).The results showed that the open reading frame of ZjDREB1.2 was 702 bp in length,encoding 233 amino acids.The relative molecular mass of ZjDREB1.2 protein was predicted to be 25.2 kDa with a theoretical isoelectric point of 4.91.The protein possesses an incomplete AP2/ERF DNA-binding domain,which was flanked up- and down-stream directly by two signature sequences of DREB1 genes.Phylogenetic tree analysis indicated that ZjDREB1.2 protein belonged to the A-1 group of DREB subfamily and was a close relative of the DREB1 homologous of C4 plants.In the cultivar ‘Meyer’,the transcript of ZjDREB1.2 gene was up-regulated under low temperature,high salinity and drought stresses during the 2-12 hours,with the greatest response to cold,and then down-regulated to the normal around or even lower level after 72 hours.Compared with Zoysia matrella,Zoysia japonica showed a relative higher transcript level of ZjDREB1.2 at the 72 hours cold stress point.The yeast one-hybrid assay indicated that ZjDREB1.2 protein showed a strong transcriptional activity but no DRE-element binding activity.The evolution of DREB1 gene in warm season plants with incomplete AP2 domain and its function is worthy of further research.
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备注/Memo
第一作者简介：冯婉倩（1994-），女，硕士研究生，研究方向为生物化学与分子生物学。E-mail:s161084@muc.edu.cn.责任作者:韦善君（1975-），女，博士，讲师，研究方向为植物生理与分子生物学。E-mail:wei.s.j@163.com.基金项目:国家自然科学基金资助项目（31100507）；中央民族大学自主创新资助项目（2016SHXY05）；中央民族大学大学生创新训练资助项目（URTP2015110021，URTP2014110029）。收稿日期：2018-03-19