白茶萎凋过程中cDNA文库筛选与EST分析

Screening and EST Analysis of cDNA Library during Withering of White Tea(Camellia sinensis)

萎凋是白茶品质形成的关键工序。以国家级审定茶树良种‘福鼎大白茶’鲜叶—萎凋叶作为供试材料,利用双链特异性核酸酶(DSN)介导,进行抑制差减杂交技术,构建鲜叶—萎凋叶正向差减文库;利用反向印迹杂交(Northern-blot)技术进行筛选与评价文库质量,获得具有差异基因的阳性克隆,并将阳性克隆的测序结果进行ESTs分析和归类,分离出白茶加工过程中重要功能基因。从分子生物学角度分析白茶萎凋过程中品质形成的关键代谢途径与其基因调控的具体情况,证实白茶萎凋过程中的基因对品质形成具有重要影响。

Withering is a key process in the formation of white tea quality.Using the national-level certified tea plant variety'Fuding Dabai'tea fresh leaf-withered leaf as the test material,double-strand specific nuclease(DSN)mediation was used to carry out suppression subtractive hybridization technology to construct the fresh leaf-withered leaf positive difference library;Northern blot technology was used to screen and evaluate the quality of the library,obtain positive clones with different genes,and ESTs analysis and classification of the sequencing results of positive clones were performed to isolate genes with important functions in the processing of white tea.From a molecular biology perspective,the key metabolic pathways involved in quality formation during the white tea withering process and the specific conditions of gene regulation were analyzed,confirming that genes involved in the withering process have a significant influence on quality formation.