细叶百合休眠与解除SSH文库构建及其调控基因的初步筛选

 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。

Preliminary Screening of Dormancy Regulation Related Genes from SSHLibrary Constructed Using Dormant and Dormancy Breaking Bulbs ofLilium pumilum

In order to verify the differences of gene expression in the process of lily bulb dormancybreaking，two SSH-cDNA libraries were constructed using dormant and dormancy breaking bulbs. Theforward library contained genes preferentially expressed in dormancy breaking bulbs and the reverselibrary contained genes preferentially expressed in dormant bulbs. The results showed that differentialexpress genes were enriched effectively in two libraries，and subtraction efficiency reached the demand.Most of the length of inserted fragments was 250–1 000 bp. One hundred positive clones were randomlyselected for sequencing from the two libraries. We analyzed ESTs in BLASTx of NCBI and unigenespositioning in the pathways using KOBAS system. Results indicated that in dormant bulb library，thesefunctionally known genes were detected in stress response. Phenylalanine metabolism and MAPKsignaling pathway were associated with dormancy. In bulb dormancy breaking library，the function of ESTs involved in sugar metabolism，hormone response and signal transduction，linoleic acid metabolismand starch and sucrose metabolic pathway was involved in dormancy breaking process.