关键词: 龙眼, 体胚发生, 磷酸丙糖异构酶, 基因克隆, 实时荧光定量PCR

Abstract: The complete cDNA sequence of triosephosphate isomerase gene（TPI）of longan embryogenic callus was obtained by reverse transcription polymerase chain reaction（RT-PCR）with rapid amplification of cDNA ends（RACE）. And the mRNA transcription level of this gene was surveyed by real-time reverse transcription PCR（qRT-PCR）during somatic embryogenesis in longan. The results showed that the full-length cDNA sequences of two type longan embryogenic callus TPI were cloned，namely TPIⅠ（GenBank accession number FJ595244）and TPIⅡ（GenBank accession number GU073294）. The complete nucleotide sequences of two TPI were 1 084 bp and 1 113 bp respectively. And they all contained a 765-nucleotides-long open reading frame（ORF）which encoded a protein of 254 amino acid residues. The results showed that the mRNA transcription levels of longan TPI were different at the different stages of longan somatic embryogenesis. In conclusion，it was preliminarily revealed that two type TPI genes and their encoded functional proteins may play an important role during the early somatic

Key words: longan, somatic embryogenesis, triosephosphate isomerase, gene cloning, real-time PCR