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目的在厚朴转录组测序基础上,对厚朴1-脱氧-D-木酮糖-5-磷酸合成酶(DXS)基因DXS1(MoDXS1)和厚朴DXS2(MoDXS2)基因进行全面的生物信息学分析。方法利用生物信息学方法对厚朴MoDXS1和MoDXS2基因编码氨基酸序列的理化特性、亲/疏水性、功能域、二级结构、三级结构和系统发育进化等进行分析和预测;采用实时荧光定量PCR分析厚朴MoDXS1和MoDXS2基因的相对表达量。结果 MoDXS1和MoDXS2均为不稳定的亲水蛋白,蛋白结构域分析显示MoDXS1、MoDXS2与其他植物的DXS基因有很高的同源性:二级结构均为混合型结构的蛋白质,α-螺旋是所有基因多肽链中大量的结构元件,利用同源建模法对三级空间结构进行了预测;同源序列对比显示MoDXS家族与烟草、丹参和拟南芥的DXS蛋白具有较高的同源性:系统进化树分析表明MoDXS1与其他被子植物亲缘关系较近,但MoDXS2单独聚为一支;实时荧光定量PCR结果显示DXS1在厚朴与凹叶厚朴间无显著差异,但DXS2在凹叶厚朴中的表达量要明显高于厚朴;GC-MS检测结果表明,3种指标性成分β-石竹烯、β-氧化石竹烯和β-桉叶油醇在凹叶厚朴皮或叶中的量均高于厚朴。结论本研究结果为解析厚朴萜类次生代谢调控机制奠定了理论基础。
OBJECTIVE: To conduct a comprehensive bioinformatics study on the genes of Magnaporthe grisea 1-deoxy-D-xylulose-5-phosphate synthase (DXS) genes DXS1 (MoDXS1) and Magnolia officinalis DXS2 (MoDXS2) analysis. Methods Bioinformatics methods were used to analyze and predict the physicochemical properties, hydrophilicity / hydrophobicity, functional domain, secondary structure, tertiary structure and phylogenetic evolution of the encoded amino acid sequences of Magnolia officinalis MoDXS1 and MoDXS2 genes. Real-time fluorescence quantitative PCR The relative expression levels of MoDXS1 and MoDXS2 in Magnolia officinalis were analyzed. Results Both MoDXS1 and MoDXS2 were unstable hydrophilic proteins. The analysis of protein domain showed that MoDXS1 and MoDXS2 had high homology with DXS genes of other plants. The secondary structures were both mixed-structure proteins. The α-helix was All the structural elements in the polypeptide chain of all genes were predicted by using the homology modeling method. The homologous sequence comparison showed that the MoDXS family had high homology with the DXS protein of tobacco, Salvia miltiorrhiza and Arabidopsis Phylogenetic tree analysis showed that MoDXS1 was closely related to other angiosperms but MoDXS2 was clustered together. The results of real-time PCR showed that there was no significant difference between DXS1 and Magnolia officinalis, but the expression of DXS2 in Magnolia officinalis The content of β-Caryophyllene, β-Oxidation Caryophyllene and β-Eucalyptol were higher than those of Magnolia officinalis by GC-MS. Park. Conclusion The results of this study laid the theoretical foundation for the analysis of the regulation mechanism of hops terpenoids.