多花水仙花瓣抑制消减杂交cDNA文库的构建及分析

Construction and analysis of the suppression subtractive hybridization cDNA libraries between yellow and white perianth of Narcissus tazetta

采用抑制消减杂交(SSH)技术,以多花水仙的栽培品种‘黄花水仙2号’黄色花瓣和‘金盏银台’白色花瓣的cDNA分别作为测试子和驱动子,建立正、反向抑制消减杂交cDNA文库.随机挑选正、反向文库各216个阳性克隆测序,对已知功能的uniESTs进行基因本体论(GO)分类分析,荧光定量PCR检测部分可能参与色素形成的uniESTs.结果表明:水仙花色差异表达正、反向消减文库经检验质量可靠,为分离花色形成相关基因奠定了基础;GO功能分类分析显示,水仙花色差异可能受到多个代谢途径的影响;经初步筛选得到11个与水仙花色形成相关的uniESTs,为进一步克隆基因全长和功能验证提供参考.

The cDNA libraries derived from forward and reverse suppression subtractive hybridization （SSH） were constructed be- tween yellow and white perianth of Narcissus tazetta. A total of 216 clones randondy selected from each library were used for sequen- cing. The uniESTs of known function was annotated by gene ontology （GO） function classification. The expression of uniESTs in- volved in the color formation was verified by fluorescence quantitative PCR. The forward and reverse SSH cDNA libraries were tested to he of reliable quality and were good for separation flower color formation related genes. GO function classification results showed that the flower color difference of narcissus might be influenced by multiple metabolic pathway. Eleven uniESTs were obtained for further investigation of their roles in the flower color formation of narcissus.