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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 analyze the variation of total mRNA expression in tuber tuberization of Pinellia ternata, and provide information for studying the molecular mechanism of tuber formation in tuber. Methods: The differential display of mRNA (DDRT-PCR) was used to isolate and clone the related gene fragments of tuber development in vitro. RESULTS AND CONCLUSION: The cDNA fragments specifically expressed during the development of small tubers in vitro of 15 Pinellia ternata samples were isolated and compared with 6 homologous sequences in GenBank. The deduced amino acid sequence of the fragment has high homology with eukaryotic promoter, MADS-box protein and ethylene signal transduction factor respectively. Semi-quantitative RT-PCR further confirmed their presence and was highly expressed in tuberous tubers at different days.