烟草HD-ZIP转录因子HD20的克隆及纤维分化功能的研究  

Cloning of a HD-ZIP Transcription Factor Gene HD20 and A Preliminary Study on Function of Fiber Differentiation in Tobacco

作　　者：高洁[1] 佟硕秋 钟杰[1] 贡献 吴拥军[1] GAO Jie;TONG Shuo-qiu;ZHONG Jie;GONGXian;WU Yong-jun(College of Life Science, Guizhou University, Guiyang 55002)

机构地区：[1]贵州大学生命科学学院,贵阳550025

出　　处：《生物技术通报》2018年第6期84-89,共6页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31071755/C1408);中国烟草总公司/贵州省烟草科学研究院开放课题(中烟办2014-01号]

摘　　要：旨在揭示烟草HD20在植物纤维分化中的作用。基于前期烟草基因组数据库设计特异性引物,通过RT-PCR克隆获得基因cDNA序列,将其命名为HD20,NCBI登录号为KC339003。该基因全长762 bp,编码253个氨基酸,亚细胞定位于细胞核,其编码的HD-ZIP蛋白由2个不同的结构域组成,包括N末端含有60个氨基酸的HD结构域和其后28个氨基酸组成的ZIP基序。构建植物表达载体,通过农杆菌蘸花法遗传转化拟南芥,经筛选获得阳性转基因植株,其石蜡切片显示,茎部木质部较对照发达。HD20对烟草及拟南芥的纤维分化具有调控作用。To understand the role of HD20 in the specification of fiber differentiation,we cloned the HD20 gene from tobacco by RT-PCR method. The cDNA sequence was cloned according to specific primers designed by the genomic database in tobacco and named as HD20 （GenBank accession number ：KC339003）. Sequence analysis showed that the cDNA sequence of HD20 was 762 bp and encoded 253 amino acids. Subcellular localization assays showed that it appeared in the nuclei. The HD-ZIP protein contained two significant domains,including the HD domain with 60 amino acids at the N-terminus and a ZIP matrix with 28 amino acids. We introduced HD20 into the expression vector pSH-737,transferred it by Agrobacterium mediated-dip flower method,and obtained positive transgenic plants for paraffin sections. The results of paraffin section showed that the xylem of stem was more developed than the control group. In conclusion,the result provided evidence that HD20 plays a role in the spatial control of fiber differentiation. The analysis of the regulatory effect of HD20 on tobacco and Arabidopsis helps to create a theoretical base for studying HD20 function.

关 键 词：烟草 HD20 石蜡切片 纤维分化 

分 类 号：Q943.2[生物学—植物学]