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本研究利用兼并-PCR及RACE技术克隆了兴安落叶松(Larix gmelinii)的咖啡酸-O-甲基转移酶基因(命名为Lg COMT),并对其基因结构和表达模式进行了分析。所克隆的Lg COMT c DNA全长为1 173 bp,包含1 092 bp的开放阅读框,编码364个氨基酸;基因组DNA全长为1 400 bp,包含3个外显子和2个内含子。系统进化分析结果显示,Lg COMT与日本落叶松、挪威云杉和北美云杉等裸子植物COMT的亲缘关系较近。表达模式分析结果显示,Lg COMT基因主要在兴安落叶松的幼茎中表达,与其功能相一致。本研究为进一步分析兴安落叶松咖啡酸-O-甲基转移酶基因的功能,表达特性及其在木质素单体生物合成中的调控机理奠定了基础。
In this study, the caffeic acid-O-methyltransferase gene (named as Lg COMT) of Larix gmelinii was cloned by using combination-PCR and RACE techniques. The gene structure and expression pattern of Lg COMT were also analyzed. The cloned Lg COMT c DNA was 1 173 bp in length and contained a 1 092 bp open reading frame encoding 364 amino acids. The full-length genomic DNA was 1 400 bp in length and contained 3 exons and 2 introns. Phylogenetic analysis showed that Lg COMT had closer genetic relationship with COMT, such as Japanese larch, Norwegian spruce and North American spruce. The expression pattern analysis showed that the Lg COMT gene was mainly expressed in the young stem of Larix gmelinii, which was consistent with its function. This study laid the foundation for the further analysis of the function, expression characteristics and regulation of L-caffeic acid-O-methyltransferase gene in lignin monomeric biosynthesis.