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采用RT-PCR结合RACE法,分离克隆龙眼(Dimocarpus longan Lour.)胚性愈伤组织生长素受体基因TIR1(Transport inhibitor response 1,即DL-TIR1)和生长素结合蛋白基因ABP1(Auxin biding protein 1,即DL-ABP1),运用生物信息学方法对序列进行分析,并通过实时荧光定量PCR法研究其在龙眼体细胞胚胎(以下简称龙眼体胚)发生过程中的表达。结果表明:DL-TIR1为2328bp的mRNA全长序列(GenBank,GQ870264),包含1个长为1761bp的开放阅读框,5′非编码区为118bp,3′非编码区为449bp,推定的氨基酸序列含586个氨基酸,与其它植物TIR1具有85%～57%同源性;DL-TIR1在龙眼体胚的各阶段均有表达,整个变化趋势呈近似“W”状,在子叶形胚中的表达量最高。DL-ABP1为869bp的mRNA全长序列(GenBank,GQ900592),包含1个长为567bp的开放阅读框,5′非编码区为43bp,3′非编码区为259bp;推定的氨基酸序列含188个氨基酸,与其它植物ABP1具有87%～72%同源性,DL-ABP1在龙眼体胚中整个表达变化趋势也呈现近似“W”状。生长素受体DL-TIR1和“可能的”生长素受体DL-ABP1在龙眼体胚中的变化趋势极为相似。
RT-PCR and RACE were used to isolate Dimocarpus longan Lour. The expression of auxin receptor 1 (TIR1) and auxin-binding protein ABP1 (Auxin biding protein) 1, ie, DL-ABP1). The sequences were analyzed by bioinformatics methods and their expression in somatic embryos of longan (hereinafter referred to as Longan somatic embryos) was studied by real-time fluorescence quantitative PCR. The results showed that DL-TIR1 was full-length sequence of 2328bp in length (GenBank, GQ870264), containing an open reading frame of 1761bp. The 5 ’untranslated region was 118bp and the 3’ non-coding region was 449bp. The putative amino acid sequence Which contains 586 amino acids and has 85% -57% homology with other plants TIR1. DL-TIR1 is expressed in all stages of longan somatic embryos, and the whole variation trend is similar to “W” shape in cotyledons The highest expression. The full-length cDNA sequence of 869bp was obtained from DL-ABP1 (GenBank, GQ900592), containing 567bp open reading frame (ORF) with 43bp in 5 ’untranslated region and 259bp in 3’ untranslated region. The putative amino acid sequence contained 188 Amino acids, which shared 87% -72% homology with other plant ABP1. The trend of the expression of DL-ABP1 in longan somatic embryos also showed similar “W” shape. The trend of the changes of auxin receptor DL-TIR1 and “possible ” auxin receptor DL-ABP1 in longan somatic embryos is very similar.