关键词: 龙眼, 胚性愈伤组织, 生长素受体基因, 生长素结合蛋白基因, 克隆, 序列分析, 实时荧光定量PCR法

Abstract: The RT-PCR with RACE method was used to clone the complete mRNA sequences of the transport inhibitor response 1 gene（DL-TIR1） and auxin binding protein 1 gene（DL-ABP1）from embryogenic calli of longan（Dimocarpus longan Lour.）. And bioinformatics methods were used to analyze obtained sequences and putative amino acid sequences. Then qRT-PCR（real-time reverse transcription PCR）method was used to determine the mRNA transcription level of two genes in the process of somatic embryogenesis in longan. The results were as follows：The full length TIR1（DL-TIR1，GenBank，GQ870264） mRNA，about 2 328 bp，consisting of an open reading frame of 1 761 bp ，and 5′ and 3′ untranslated regions of 118 bp and 449 bp，respectively. The putative protein had 586 amino acids，and the identity with the other polypeptides varied between 85%–57%. DL-TIR1 expressed in 8 different stages during somatic embryogenesis，which showed approximately a“W”curve. The full length ABP1（DL-ABP1，GenBank，GQ900592）mRNA，about 869 bp，consisting of an open reading frame of 567 bp，and 5' and 3' untranslated regions of 43 bp and 259 bp，respectively. The putative protein had 188 amino acids，and the identity with the other polypeptides varied between 87%–72%. The expression of DL-ABP1 in 8 different stages during somatic embryogenesis also showed approximately a“W”curve. The mRNA transcription level of the auxin receptor gene DL-TIR1 and the potential auxin receptor gene DL-ABP1 were very similar.