【摘要】 【目的】探明NtChlH在烟草叶绿素代谢中的作用，为利用该基因选育烟叶集中成熟的品种提供基因资源和理论依据。【方法】采用基因克隆、生信分析以及实时荧光定量等方法研究烟草NtChlH的CDs序列、组织表达模式、蛋白表征及其在叶绿素合成中的作用。【结果】通过克隆获得烟草镁离子螯合酶H亚基基因NtChlH的CDs序列，同时克隆NtChlH起始密码子ATG上游2101 bp启动子序列，对基因序列和启动子序列进行分析以及对NtChlH进行亚细胞定位，随后对NtChlH进行组织特异性分析，再研究两种烟草品种在暗处理下对光的敏感性。NtChlH的CDs序列长度为3342 bp,编码1114个氨基酸，亚细胞定位分析结果表明，NtChlH蛋白定位于细胞膜上；启动子顺式作用元件分析表明，NtChlH含有响应光照、厌氧诱导、伤口、茉莉酸甲酯和生长素等多种信号的顺式作用元件；NtChlH基因的组织表达分析显示，NtChlH在富含叶绿素的源组织中表达量高，黄叶K326在叶绿素合成过程中对光的敏感性低于K326。【结论】研究明确NtChlH的序列、组织表达模式、蛋白表征以及在叶绿素合成中的作用，为利用该基因选育烟叶集中成熟的品种提供基因资源和理论依据。更多还原

【Abstract】 【Objective】The role of NtChlH in the chlorophyll metabolism of Nicotiana tabacum was explored, which provided the genetic resources and theoretical basis for selecting tobacco varieties with concentrated maturity by using this gene.【Method】Gene cloning, bioinformatics analysis and real-time fluorescence quantification were used to study the CDs sequence, tissue expression pattern, protein characterization of tobacco NtChlH and its role in chlorophyll synthesis.【Result】The CDs sequence of Mg-chelatase H subunit gene NtChlH was obtained by cloning, and the 2101 bp promoter sequence in the upstream of NtChlH start codon ATG were cloned, the gene sequence and promoter sequence were analyzed, and NtChlH was subcellularly localized, followed by tissue-specific analysis of NtChlH,and then the sensitivity of the two tobacco varieties to light under dark treatment was studied.The CDs sequence length of NtChlH was 3342 bp and encoded 1114 amino acids.The results of subcellular localization analysis showed that NtChlH protein was localized on the cell membrane, while the cis-acting element analysis of the promoter showed that NtChlH contained cis-acting elements such as in response to light, anaerobic induction, wound, methyl jasmonate and auxin.The tissue expression analysis of NtChlH gene showed that NtChlH was highly expressed in chlorophyll-enriched source tissues.Yellow leaf K326 was less sensitive to light during chlorophyll synthesis than that of K326.【Conclusion】The sequence, tissue expression pattern, protein characterization of NtChlH and its role in chlorophyll synthesis are clarified, which provide the genetic resources and theoretical basis for selecting tobacco varieties with concentrated maturity by using this gene.更多还原