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采用RT-PCR结合RACE技术从蕙兰(Cymbidium faberi)中分离到3个B类MADS-box基因CfGLO、CfDEF1和CfDEF2,分别编码210、222和227个氨基酸。系统进化树分析显示,CfGLO属于PI/GLO类基因,CfDEF1和CfDEF2分别属于PaleoAP3组基因的PeMADS2类基因和PeMADS3类基因。RT-PCR和实时荧光定量表达分析表明,CfDEF1在2、3轮花器官侧瓣、唇瓣和蕊柱中强烈表达,在萼片中不表达;CfDEF2在1、2、3轮花器官中都表达,在营养组织中不表达;而CfGLO在所有组织中都表达,说明3个基因在蕙兰花器官的形成过程中可能扮演着不同的角色。此外,3个基因都在蕙兰幼嫩子房中有表达,显示其可能在蕙兰子房的形成过程中起一定作用。
Three B-type MADS-box genes, CfGLO, CfDEF1 and CfDEF2, were isolated from Cymbidium faberi by RT-PCR and RACE techniques and encoded 210, 222 and 227 amino acids, respectively. Phylogenetic tree analysis showed that CfGLO belonged to PI / GLO gene and CfDEF1 and CfDEF2 belonged to PeMADS2 gene and PeMADS3 gene of PaleoAP3 gene respectively. The results of RT-PCR and real-time fluorescence quantitative analysis showed that CfDEF1 was strongly expressed in the flanks, labiums and pistils of 2, 3 round organs, but not in sepals. CfDEF2 was expressed in 1, 2, , But not in vegetative tissues. However, CfGLO was expressed in all tissues, indicating that three genes may play different roles in the formation of Cymbidium orchid organ. In addition, all three genes were expressed in Cymbidium young ovary, indicating that they may play a role in the formation of Cymbidium ovary.