半夏试管小块茎发育过程中相关基因差异表达的研究

Differential display of mRNA from microtubers of Pinellia ternata in vitro

目的:分析半夏试管块茎形成过程中总mRNA差异表达的变化规律,为研究半夏试管块茎形成的分子机制提供了信息。方法:利用mRNA差异显示技术(DDRT-PCR),分离和克隆半夏试管小块茎发育的相关基因片段。结果和结论:分离到了15个半夏试管小块茎发育过程中特异表达的cDNA片段,在GenBank中进行同源性比较发现6个没有同源序列与之匹配,推测为新的cDNA片断;3个片段推导的氨基酸序列分别与真核生物启动子,MADS-box蛋白和乙烯信号转导因子有高度同源性。半定量RT-PCR进一步证实它们存在,并在半夏块茎形成不同天数高度特异表达。

Objective： To study mRNA differential expression from microtubes of Pinellia ternata in vitro, and give more infor- mation of the molecular mechanism in the formation of microtubers. Method： DDRT-PCR was used to identify the expressed gene frag- ments related to the microtubers development of P. ternata in vitro. Results and Conclusion： Fifteen cDNA fragments differentially expressed in the induction of microtubers of P. ternata in vitro were identified and characterized by using mRNA differential display DDRT-PCR. The deduced amino-acid sequences of six fragments of cDNA showed no significant homology with ESTs and genes in the Genbank databases, and they could be new cDNA fragments. However, the remaining three showed significant homologies with se- quences encoding components of eukaryotic translation initiation factor 3 subunit H, MADS-box protein and ethylene signal transcription factor, respectively. Their differential expression patterns were confirmed by semi-quantitative RT-PCR analysis. And the expression level of the induction of microtubers of P. ternata in vitro was different.