Li Meiya, Jiang Fusheng, Yu Xiangli, Miao Zhiqi
Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China ; Institute of Biotechnology, College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Institute of Biotechnology, College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Biomed Res Int. 2015;2015:504932. doi: 10.1155/2015/504932. Epub 2015 Feb 1.
Taxadiene is the first committed precursor to paclitaxel, marketed as Taxol, arguably the most important anticancer agent against ovarian and breast cancer. In Taxus, taxadiene is directly synthesized from geranylgeranyl diphosphate (GGPP) that is the common precursor for diterpenoids and is found in most plants and microbes. In this study, Artemisia annua L., a Chinese medicinal herb that grows fast and is rich in terpenoids, was used as a genetic engineering host to produce taxadiene. The TXS (taxadiene synthase) gene, cloned from Taxus and inserted into pCAMBIA1304, was transformed into Artemisia annua L. using the Agrobacterium tumefaciens-mediated method. Thirty independent transgenic plants were obtained, and GC-MS analysis was used to confirm that taxadiene was produced and accumulated up to 129.7 μg/g dry mass. However, the high expression of TXS did not affect plant growth or photosynthesis in transgenic Artemisia annua L. It is notable that artemisinin is produced and stored in leaves and most taxadiene accumulated in the stem of transgenic Artemisia annua L., suggesting a new way to produce two important compounds in one transgenic plant: leaves for artemisinin and stem for taxadiene. Overall, this study demonstrates that genetic engineering of the taxane biosynthetic pathway in Artemisia annua L. for the production of taxadiene is feasible.
紫杉二烯是紫杉醇（商品名为泰素）的首个关键前体物质，紫杉醇可谓是对抗卵巢癌和乳腺癌最重要的抗癌药物。在红豆杉属植物中，紫杉二烯直接由香叶基香叶基二磷酸（GGPP）合成，GGPP是二萜类化合物的常见前体，存在于大多数植物和微生物中。在本研究中，生长迅速且富含萜类化合物的中国药用植物黄花蒿被用作基因工程宿主来生产紫杉二烯。从红豆杉中克隆并插入到pCAMBIA1304中的紫杉二烯合酶（TXS）基因，采用根癌农杆菌介导的方法转化到黄花蒿中。获得了30株独立的转基因植株，气相色谱 - 质谱联用（GC - MS）分析证实了紫杉二烯的产生，其积累量高达129.7μg/g干重。然而，TXS的高表达并未影响转基因黄花蒿的植物生长或光合作用。值得注意的是，青蒿素在叶片中产生和储存，而大多数紫杉二烯在转基因黄花蒿的茎中积累，这表明了在一种转基因植物中生产两种重要化合物的新途径：叶片用于生产青蒿素，茎用于生产紫杉二烯。总体而言，本研究表明通过基因工程改造黄花蒿中的紫杉烷生物合成途径来生产紫杉二烯是可行的。
