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香榧(Torreya grandis Fort.ex Lindl.cv.Merrillii)是红豆杉科榧树属的一个种。由于香榧与红豆杉属植物具有较近的亲缘关系,研究者认为香榧也可以合成紫杉醇,并利用HPLC法验证其中含有紫杉醇。但他们仅仅依靠HPLC中出峰时间来判断,并没有质谱的分析结果。本研究利用LC-MS对香榧茎叶的化学提取物进行分析发现香榧并不能合成紫杉醇。为了进一步证明香榧不能合成紫杉醇,我们利用欧洲红豆杉(Taxus baccata L.)紫杉醇生物合成关键基因-紫杉二烯合成酶基因(Taxadiene synthase gene,Tb TS)TBLASTN比对香榧本地转录组,从中找出并克隆得到与Tb TS同源性最高的Tgr TSL1基因。利用原核表达对Tgr TSL1基因进行功能分析,发现该基因不具有紫杉二烯合成酶编码基因的功能。这样,我们从化学和生物学两个方面对香榧无法合成紫杉醇这个结论进行了证明。这为以后通过植物学和化学分类学等研究穗花杉的分类学地位提供了新的有用信息。
Torreya grandis Fort.ex Lindl.cv.Merrillii is a species of the genus Taxus. Due to the close genetic relationship between Fragrant and Taxus, the researchers believe that Fragrans can also be synthesized paclitaxel, and the use of HPLC method to verify that it contains paclitaxel. But they only rely on the peak time in the HPLC to judge, there is no mass spectrometry results. In this study, LC-MS analysis of the chemical extracts of stems and leaves of sambuci showed that sambucus did not synthesize paclitaxel. In order to further prove that Taxol can not synthesize paclitaxel, we compared the taxol synthase gene (Tb TS) TBLASTN, a key gene of taxol biosynthesis in Taxus baccata L., with the local transcriptome The Tgr TSL1 gene with the highest homology to Tb TS was cloned and cloned. The functional analysis of Tgr TSL1 gene using prokaryotic expression showed that the gene does not have the function of encoding gene of taxadiene synthase. In this way, we have proved from two aspects of chemistry and biology that the taxane can not synthesize paclitaxel. This provided new and useful information for the later study of the taxonomic status of Pinus thunbergii through botany and chemical taxonomy.