铁皮石斛AP1 MADS-box基因克隆与表达分析

Cloning and Expression Analysis of AP1 MADS-Box Gene from Dendrobium candidum

铁皮石斛是重要的兰科植物,具有较高的观赏和药用价值。AP1(APETALA1)MADS-box是影响铁皮石斛成花的关键基因。本研究以铁皮石斛为试验材料,采用PCR和RACE技术对铁皮石斛花器官中的两个同源基因AP1-A MADS-box和AP1-B MADS-box进行克隆,并对该成花基因进行表达分析。结果表明,这两个基因分别编码253个和213个氨基酸,OFR长度分别为762 bp和642 bp,与春兰、建兰等物种的同源性较高。采用RT-PCR和Real-Time PCR技术对这两个基因进行表达分析,发现AP1-A MADS-box和AP1-B MADS-box基因主要在唇瓣和合蕊柱中表达,除此之外,AP1-B MADS-box基因在萼片中的表达量也较高。

Dendrobium candidum is an important orchid plant with high ornamental and medicinal value, and AP1(APETALA1) MADs-box is a key gene affecting flowering of D. candidum. In this study, two homologous genes AP1-A MADS-box and AP1-B MADS-box were cloned from D. candidum by PCR and RACE techniques, and the expression of these homologous genes was analyzed. The two genes encoded 253 and 213 amino acids, and their OFR lengths were 762 bp and 642 bp, respectively, indicating high homology with Cymbidium goeringii and Cymbidium ensifolium. RT-PCR and real-time PCR were used to analyze the expression of AP1-A MADS-box and AP1-B MADs-box genes, and it was found that AP1-A MADS-box and AP1-B MADS-box genes were mainly expressed in the labellum and staminal column. Additionally, AP1-B MADS-box gene was also highly expressed in sepals.