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类黄酮-3’5’-羟化酶(F3’5’H)基因是合成蓝色飞燕草色素类花色苷的关键酶基因。本研究采用RT-PCR法从彩色马铃薯品种‘转心乌’中克隆到了F3’5’H基因的c DNA,并进行了生物信息学和组织表达模式分析,希望能探明F3’5’H基因在彩色马铃薯花色苷合成中的作用及表达方式。克隆到的F3’5’H c DNA序列全长1 720 bp,编码509个氨基酸残基,同源比对表明F3’5’H与茄科植物聚在一起,其次是其它双子叶植物。F3’5’H具有信号肽和明显的跨膜结构域,属于分泌蛋白且为稳定的亲水蛋白,定位于细胞质。说明F3’5’H在细胞质中的粗糙型内质网上合成前体后,跨膜运输到其它部位或细胞器中发挥作用。α螺旋和无规则卷曲是F3’5’H的主要二级结构元件。F3’5’H具有细胞色素P450的“PPGP”、“AGTDT”、“FGAGRRICAG”三段基序,且只有一个功能结构域,与细胞色素P450的功能结构域相匹配,属于细胞色素P450家族的一员。组织特异性表达结果表明:F3’5’H相对表达量和花色苷含量均是块茎高于叶片和地上茎,它们的变化趋势基本一致,花色苷含量较高的器官,其F3’5’H的相对表达量也高,说明花色苷的积累与F3’5’H的表达正相关。
The flavonoid 3’5’-hydroxylase (F3’5’H) gene is a key enzyme gene for the synthesis of blue delphinium anthocyanins. In this study, c-DNA of F3’5’H gene was cloned from color potato ’Zhuanwuwu’ by RT-PCR, and bioinformatics and tissue expression patterns were analyzed in the hope of exploring F3’5’H The Role of Genes in the Synthesis of Anthocyanins in Colored Potato and Its Expression. The cloned F3’5’H cDNA sequence was 1 720 bp in length and encoded 509 amino acid residues. The homologous alignment indicated that F3’5’H was clustered with solanaceous plants, followed by other dicotyledonous plants. F3’5’H has a signal peptide and a distinct transmembrane domain, belongs to the secreted protein and is a stable hydrophilic protein, localized in the cytoplasm. This shows that F3’5’H plays a role in transmembrane transport to other sites or organelles after it is synthesized in the rough endoplasmic reticulum in the cytoplasm. Alpha helix and random curl are the main secondary structural elements of F3’5’H. F3’5’H has the “PPGP ”, “AGTDT ”, “FGAGRRICAG ” three-segment motifs of cytochrome P450 and has only one functional domain matching the functional domain of cytochrome P450, Belongs to a member of the cytochrome P450 family. Tissue-specific expression results showed that: F3’5’H relative expression levels and anthocyanin content are tubers higher than leaves and aboveground stems, their trend is basically the same, higher anthocyanin content of the organ, the F3’5’H Relative expression level is also high, indicating that anthocyanin accumulation and F3’5’H expression is positively correlated.