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为了深入研究桃花芽休眠的分子机制,利用抑制差减杂交技术(SSH),以解除休眠期的花芽的RNA为实验方Tester,以休眠期花芽的RNA为驱动方Driver,构建休眠解除cDNA文库,测定分析了180个上调表达的cDNA片段,测序成功128个,除去冗余序列,得到28个单一序列。对序列进行BLAST比对及功能注释,发现这些序列分别属于新陈代谢、氧化还原、信号转导、抗性相关等类别基因。运用qRT-PCR技术对HisH3、Dhn和Metallothionein基因进行检测,结果表明它们在花芽休眠解除过程中均上调表达。
In order to further study the molecular mechanism of dormancy in peach buds, RNA was used to drive dormant flower buds to construct a dormancy-relieved cDNA library using subtractive hybridization (SSH) A total of 180 cDNA fragments were amplified and sequenced. 128 sequences were successfully sequenced and 28 redundant sequences were obtained. BLAST alignment and functional annotation of the sequences revealed that these sequences belong to the categories of metabolism, redox, signal transduction and resistance-related, respectively. The results of qRT-PCR analysis of HisH3, Dhn and Metallothionein genes showed that they all up-regulated in flower bud dormancy release.