Abstract:The full-length cDNA sequences of DEFICIENS(DEF) and GLOBOSA(GLO)-like genes were cloned from Paphiopedilum concolor Pfitz.by reverse transcriptase-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE).They were named PcDEF and PcGLO,respectively.Expression transcript levels of PcDEF and PcGLO in different flower bud developmental stages and different tissues were analyzed by semi-quantitative RT-PCR and real-time PCR.The results revealed that the full-length cDNA sequences of PcDEF and PcGLO were 1 039 bp and 934 bp,respectively,and encoded 224 and 210 amino acids,respectively.Protein alignment analysis indicated that both PcDEF and PcGLO proteins had typical structure domains of MADS-box protein,containing a MADS domain and a K domain.Protein sequence identity search showed that PcDEF shared 75%~96% similarities with other known DEF/AP3-like MADS-box proteins of Orchidaceae and PcGLO shared 87%~98% similarities with other known GLO/PI-like MADS-box proteins of Orchidaceae.Phylogenetic analysis clearly showed that PcDEF and PcGLO proteins were classed into AP3 and PI subgroups of B group of MADS-box proteins’ family,respectively.Expression analyses revealed that PcDEF and PcGLO were detected in all six flower bud developmental stages,but showed different expression patterns in tissues examined.Compared with the transcripts of PcDEF in columns,sepals,bracts and roots,higher levels in mature flowers,lips and petals,lower levels in flower stems and leaves,and only low transcript levels were detected in ovaries.PcGLO was expressed in all tested tissues,including leaves,roots,flower stems,ovaries,bracts,sepals,petals,lips and columns,but at different abundance.