文献类型：期刊文献

中文题名：穗花杉中不存在紫杉醇的化学及分子生物学证据

英文题名：Chemical and Molecular Biological Evidence Reveals That There Is No Taxol in Amentotaxus argotaenia(Hance)Pilger

作者：王帅[1] 邵芬娟[1] 李论[2] 芦强[1] 邱德有[1]

第一作者：王帅

机构：[1]中国林业科学研究院林业研究所;[2]广西桂林小庐山生态农业发展有限公司

年份：2016

卷号：36

期号：4

起止页码：661-666

中文期刊名：西北植物学报

外文期刊名：Acta Botanica Boreali-Occidentalia Sinica

收录：CSTPCD;;北大核心:【北大核心2014】；CSCD:【CSCD2015_2016】；

基金：中国林业科学研究院林业研究所中央级公益性科研院所基本科研业务费专项(RIF2014-01)

语种：中文

中文关键词：穗花杉;紫杉醇;紫杉二烯合成酶;LC-MS;原核表达

外文关键词：Amentotaxus argotaenia;taxol; taxadiene synthase; LC-MS; prokaryotic expression

分类号：Q785;Q789

摘要：穗花杉(Amentotaxus argotaenia(Hance)Pilger)是红豆杉科穗花杉属的一个种。由于穗花杉与红豆杉属植物具有较近的亲缘关系,有研究者认为穗花杉也可以合成紫杉醇,并利用HPLC法测到其含有紫杉醇。但这仅仅是依靠HPLC中出峰时间来判断,并没有质谱结果。该研究利用LC-MS对穗花杉茎叶的化学提取物分析结果显示,穗花杉的茎叶中均未发现紫杉醇。为了进一步证明穗花杉不能合成紫杉醇,该研究利用欧洲红豆杉(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,TbTS)的序列TBLASTN比对穗花杉本地转录组,从中找出并克隆得到与TbTS同源性最高的AarTSL1基因;利用原核表达分析AarTSL1基因的编码蛋白,结果发现该基因不具有紫杉二烯合成酶编码基因的功能。该研究从化学和生物学两个方面均证明穗花杉无法合成紫杉醇,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。
Amentotaxus argotaenia （Hance） Pilger is one member of Amentotaxus in Taxaceae. Due to the close genetic relationship between A. argotaenia and Taxus sp. , some researchers thought that taxol could be synthesized in A. argotaenia and testified this hypothesis using HPLC method. This conclusion was just reached based on the retention time of the peak in the extract of A. argotaenia corresponding to that of the taxol standard, but not from any mass spectra data. In this study, we analyzed the extraction of the stems and leaves of A. argotaenia with LC-MS, and found that there was no taxol in the extraction. In order to prove our hypothesis, AarTSL1 gene was isolated and cloned by using the taxadiene synthase of Taxus baccata to TBLASTN the transcriptome of A. argotaenia. Functional analysis of AarTSL1 with prokaryotic expression showed that the AarTSL1 protein cannot catalyze GGPP （geranylgeranyl pyrophos- phate） to taxadiene, key intermedia in taxol biosynthesis pathway. Taking together, our chemical and mo- lecular biology evidence proved that A. argotaenia cannot product taxol. Our results will be helpful to the research work on the taxonomic study of A. argotaenia.