摘要
【目的】以前期通过穗发育芯片筛选到一个水稻Dof家族转录因子OsDof6为研究对象,进一步探究OsDof6的表达模式与生物学功能。【方法】对OsDof6的基因、蛋白及启动子序列进行生物信息学分析,利用CRISPR/Cas9技术对该基因进行定点编辑,通过实时定量PCR和亚细胞定位技术分析该基因的表达模式。【结果】对该基因的启动子序列分析表明,该基因启动子中存在大量与光响应、激素响应及胁迫响应相关的顺式调控元件。利用CRISPR/Cas9技术获得了2种不同突变类型的功能缺失突变体9522Dof6-3和9522Dof6-4。对突变体T1株系进行表型观察发现,相较于对照,两种突变体在营养生长阶段分蘖数明显降低,转入生殖生长阶段后,两种突变体的抽穗时间推迟约3 d。实时定量PCR结果显示OsDof6在水稻根、茎、叶和穗中均有不同程度的表达,在穗发育后期相对表达量明显提高;通过亚细胞定位分析发现OsDof6定位于细胞核。【结论】初步判断OsDof6基因会影响水稻分蘖数与抽穗期。
【Objective】Our goal is to further explore the expression pattern and biological function of OsDof6,a rice Dof family transcription factor screened with the panicle development chip in the previous study.【Method】Bioinformatics approaches were applied to analyze OsDof6 gene,protein and protein promoter sequences.The gene was edited using CRISPR/Cas9 technology.Real-time quantitative PCR and subcellular localization technology was used to reveal the gene expression pattern.【Result】The gene promoter contains a large number of photo-,hormone-,and stress-responses and tissue expression elements.Then,two mutants of different mutation types 9522Dof6-3 and 9522Dof6-4 were obtained by using CRISPR/Cas9 technology.Through observation of the T1 generation of the mutant materials,we found that the mutants had significantly less tillers and longer heading dates(delayed by 3 days)compared to the control materials.Real-time quantitative PCR results showed that OsDof6 was expressed in varying degrees in the roots,stems,leaves and panicles,and the relative expression was significantly increased at late stages of panicle development.Subcellular localization analysis showed that OsDof6 belongs to a nuclear localization gene.【Conclusion】OsDof6 gene affects the tiller number and heading date of rice.
作者
张立成
李懿星
王天抗
邱牡丹
宋书锋
董皓
李磊
刘建丰
李莉
ZHANG Licheng;LI Yixing;WANG Tiankang;QIU Mudan;SONG Shufeng;DONG Hao;LI Lei;LIU Jianfeng;LI Li(College of Agronomy,Hunan Agricultural University,Changsha 410128,China;State Key Laboratory of Hybrid Rice,Hunan Hybrid Rice Research Center,Changsha 410125,China;Longping Branch,Graduate School of Hunan University,Changsha 410125,China)
出处
《中国水稻科学》
CAS
CSCD
北大核心
2020年第5期397-405,共9页
Chinese Journal of Rice Science
基金
国家自然科学基金资助项目(31671669)
国家重点研发计划资助项目(2016YFD0101103)
国家自然科学基金资助项目(31901530)。
作者简介
通信联系人:刘建丰,liujf501@aliyun.com;通信联系人:李莉,E-mail:lili@hhrrc.ac.cn。
