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为了验证百合鳞茎休眠解除过程中基因表达的差异性,以细叶百合休眠鳞茎和休眠解除鳞茎为材料,利用抑制消减杂交技术构建了百合鳞茎休眠及解除正、反向抑制消减文库。文库富集了差异基因,消减效率符合要求,插入片段集中于250~1 000 bp之间。对正、反向文库随机挑选阳性克隆测序,各获得100个表达序列标签(EST),在NCBI上进行BLASTx分析,并用KOBAS系统将获得的unigene定位到Pathways中。分析结果表明,休眠文库ESTs功能主要涉及胁迫响应方面,如苯丙氨酸代谢途径、促分裂素原活化蛋白激酶途径等与休眠有关。休眠解除文库在糖代谢、激素响应及信号转导方面表达量较高,如亚油酸代谢途径、淀粉及蔗糖代谢途径等参与了休眠解除过程。
In order to verify the difference of gene expression during lily dormancy release in lilium, the lily bulbs and dormancy lifted bulbs were used as material to construct lily bulbs dormancy and lysis suppression subtractive libraries by suppression subtractive hybridization. The library enriched the difference genes, and the efficiency of abatement met the requirements. The inserted fragments were concentrated in the range of 250-1 000 bp. Positive and negative libraries were randomly selected positive clones sequenced to obtain 100 expressed sequence tags (ESTs), BLASTx analysis on NCBI and localization of unigene to Pathways using the KOBAS system. The results showed that ESTs function of dormancy library mainly involved in stress response, such as phenylalanine metabolic pathway, mitogen-activated protein kinase pathway and dormancy. Dormancy release library in the high level of glucose metabolism, hormone response and signal transduction, such as linoleic acid metabolic pathway, starch and sucrose metabolic pathway involved in the dormancy release process.