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为揭示烟草茄尼基焦磷酸合酶(Nicotiana tabacum solanesyl diphosphate synthase,NtSPS)在烟草茄尼醇生物合成中的作用,分离了烟草品种红花大金元茄尼醇生物合成关键基因NtSPS1和NtSPS2,并对其进行了序列比对、进化分析和亚细胞定位分析,研究了其在烟草植株不同器官的表达水平以及烟草植株不同器官的茄尼醇、叶绿素含量与NtSPS表达水平的相关性。结果表明,NtSPS1和NtSPS2开放读码框(ORF)大小分别为1209、1206 bp,分别编码402、401个氨基酸;NtSPS1和NtSPS2均存在2个保守的DDxxD结构域,这与NtSPS功能发挥相关;NtSPS1、NtSPS2与番茄SPS同源性较高,这与烟草、番茄均为茄科作物有关;NtSPS1、NtSPS2均定位于叶绿体中;烟草植株不同器官NtSPS1和NtSPS2表达水平排序为:叶>茎>根,这与茄尼醇、叶绿素在烟草植株中的分布规律一致。本研究可为NtSPS调控烟草茄尼醇生物合成机制的深入研究奠定基础。
In order to reveal the role of Nicotiana tabacum solanesyl diphosphate synthase (NtSPS) in the biosynthesis of solanesol in tobacco, the key genes NtSPS1 and NtSPS2 of solanesol biosynthesis were isolated, The sequence alignment, evolutionary analysis and subcellular localization analysis were carried out. The expression level of NtSPS in different organs of tobacco plants and the expression of solanesol and chlorophyll in different organs of tobacco plants were studied. The results showed that the size of open reading frame (ORF) of NtSPS1 and NtSPS2 were 1209 and 1206 bp, encoding 402 and 401 amino acids respectively. There were two conserved DDxxD domains in NtSPS1 and NtSPS2, which were related to NtSPS function. NtSPS1 , NtSPS2 and SPS have higher homology with SPS, which is related to tobacco and tomato are Solanaceae crops; NtSPS1 and NtSPS2 are located in the chloroplast; The order of NtSPS1 and NtSPS2 expression in different organs of tobacco plant is leaf> stem> This is consistent with the distribution of solanesol and chlorophyll in tobacco plants. This study can lay the foundation for the further study on the mechanism of NtSPS regulating tobacco solanesol biosynthesis.