换锦花LsMYB4基因的克隆与表达分析

Cloning and Expression Analysis of Ls MYB4 Gene in Lycoris sprengeri

以换锦花(Lycoris sprengeri)为材料,采用RT-PCR方法和RACE技术相结合的方法,从花瓣中克隆了MYB基因的c DNA全长序列,命名为Ls MYB4。该序列全长793 bp,包含606 bp完整开放阅读框,编码201个氨基酸,具有明显的R2R3-MYB结构域,与中国水仙Nt MYB相似性达96%。实时荧光定量PCR分析结果表明,Ls MYB4在换锦花不同组织和不同花期均有表达,其中在花瓣中的相对表达量比叶中多9~10倍,在盛花期的相对表达量比花苞期多20倍。不同花色无性系花瓣中表达量存在差异,花色较浅的无性系高于其它无性系,推测Ls MYB4在调控换锦花花色形成的过程中起着负调控作用。

A MYB gene named Ls MYB4 was cloned by RT-PCR and RACE methods from petals of Lycoris sprengeri. Sequences analysis showed that the full-length MYB c DNA was 793 bp and contained a ORF of 606 bp encoding 201 amino acids. The Ls MYB4 protein had a conserved R2R3-MYB domain and the amino acids sequence shared up to 96% homelogies with anthycyanin biosynthesis-related R2R3-MYB transcription factor in Narcissus tazetta. The expression analysis by real time q RT-PCR showed that Ls MYB4 was expressed in different tissues and flowering periods. The expression level in petal was almost ten times higher than in leaf. And in florescence period the expression of Ls MYB4 was twenty times higher than in early budding period. The Ls MYB4 expressed discrepantly in four clones of L. sprengeri,and results of it implicated that the lighter flower color,the higher expression level. This showed that Ls MYB4 played a possibly negative role in anthocyanin biosynthesis of Lycoris sprengeri.