摘要
WRKY蛋白是近年来在植物中发现的一类转录因子家族,本研究从拟南芥中克隆得到转录因子WRKY18的DNA序列,利用荧光体式显微镜观察酵母单杂交菌落,确定了该转录因子与DNA序列TTGACT结合。分别通过半定量、实时荧光定量PCR技术检测其在不同组织及花药发育不同时期的表达,发现该转录因子为组成型表达。该基因在成熟拟南芥根、茎、叶、花的各个组织中均有表达,而在花药发育的9、10时期表达量显著高于其它时期。分别使用扫描电子显微镜和光学显微镜观察比较突变体与野生型植株的花粉形态差异和花粉萌发情况,发现WRKY18对拟南芥花粉发育有重要影响,WRKY18缺失造成拟南芥花粉外形异常、花粉萌发率降低。本研究对进一步分析转录因子WRKY18在拟南芥花粉发育过程中的表达与调控作用具有一定的基础意义。
WRKY proteins have been found in plants in recent years, which belong to a class of transcription factor family. In this study, the transcription factor WRKY18 was cloned from A. thaliana. By fluorescence stereo microscope, a DNA sequence, TTGACT, is identified as binding to WRKY18 in vitro using one-hybrid experiment. The expression of WRKY18 in roots, stems, leaves and flowers of A. thaliana was detected using semi-quantitative RT-PCR. The expression of WRKY18 during different developmental stages of the A. thaliana anther was also detected using quantitative RT-PCR. The results show that WRKY18 is constitutively expressed in A. thaliana. And the expression of WRKY18 in the 9th and 10th stages of anther development is significantly higher than that in other stages. In addition, the pollen morphology and pollen germination of T-DNA insertion mutant and wide type were comparatively observed by scanning electron microscopy and optical microscope. The results show that the partial infertility mutant pollens have an abnormal shape. This research indicates that WRKY18 acts as a transcription factor and plays an important role during the development of pollen, and the study is of significance for the research of the regulation and expression of WRKY18 in pollen development of A. thaliana.
出处
《电子显微学报》
CAS
CSCD
2014年第2期156-162,共7页
Journal of Chinese Electron Microscopy Society
基金
国家自然科学基金资助项目(No.31170574
No.31370590)
关键词
拟南芥
转录因子
WRKY18
表达分析
花粉发育
Arabidopsis thaliana （ L. ） Heynh.
transcription factor
WRKY18
expression
pollen development
