萜类化合物是植物中最大的代谢物群组之一,在植物应对生物和非生物胁迫过程中发挥着重要作用。萜烯合成酶(Terpene synthases, TPSs)是植物萜类化合物合成的关键酶,然而目前关于油菜萜烯合成酶研究很少报道。为探究TPSs在油菜中的作用,...萜类化合物是植物中最大的代谢物群组之一,在植物应对生物和非生物胁迫过程中发挥着重要作用。萜烯合成酶(Terpene synthases, TPSs)是植物萜类化合物合成的关键酶,然而目前关于油菜萜烯合成酶研究很少报道。为探究TPSs在油菜中的作用,本文对甘蓝型油菜萜类合酶基因BnaTPSA08-1 (BnaA08T0101600Z)进行了功能研究。在克隆得到甘蓝型油菜BnaTPSA08-1基因CDS序列基础上,成功构建了BnaTPSA08-1基因的过表达双元载体pFGC5941-35s::BnaTPSA08-1。利用农杆菌介导的甘蓝型油菜下胚轴遗传转化方法成功获得5株稳定遗传的BnaTPSA08-1基因过表达转基因植株。通过收集过表达转基因植株释放的有机挥发性气体,并用GC-MS测定气体组分,结果显示:与对照植株的有机挥发性组分比较,过表达植株产生25种不同类型的挥发性气体,其中十一烷及其衍生物占比最多,这说明BnaTPSA08-1基因产物的生化功能是促进十一烷及其衍生物的合成,这为进一步研究BnaTPSA08-1基因在甘蓝型油菜中的生物学功能打下了良好基础。Terpenoids are one of the largest groups of metabolites in plants and play an important role in plant response to biotic and abiotic stresses. Terpene synthases (TPSs) are key enzymes in the synthesis of terpene compounds in plants. However, there are few reports on terpene synthases in rapeseed. In order to explore the role of TPSs in rapeseed, the function of the terpene synthase gene BnaTPSA08-1 (BnaA08T0101600Z) in Brassica napus was studied. Based on the cloned CDS sequence of BnaTPSA08-1 gene in Brassica napus, the over-expression vector pFGC5941-35s::BnaTPSA08-1 was successfully constructed. Five transgenic plants with stable BnaTPSA08-1 gene over-expression were successfully obtained by Agrobacteria-mediated hypocotyl genetic transformation of Brassica napus. The organic volatile gases released by overexpressed transgenic plants were collected and the gas components were determined by GC-MS. The results showed that: Compared with the organic volatile components of control plants, over-expressed plants produced 25 different types of volatile gases, among them, undecane and undecane derivatives account for the most, indicating that the biochemical function of BnaTPSA08-1 gene product is to promote the synthesis of undecane and undecane derivatives. This provides a good basis for further study on the biological function of BnaTPSA08-1 gene in Brassica napus.展开更多
文摘萜类化合物是植物中最大的代谢物群组之一,在植物应对生物和非生物胁迫过程中发挥着重要作用。萜烯合成酶(Terpene synthases, TPSs)是植物萜类化合物合成的关键酶,然而目前关于油菜萜烯合成酶研究很少报道。为探究TPSs在油菜中的作用,本文对甘蓝型油菜萜类合酶基因BnaTPSA08-1 (BnaA08T0101600Z)进行了功能研究。在克隆得到甘蓝型油菜BnaTPSA08-1基因CDS序列基础上,成功构建了BnaTPSA08-1基因的过表达双元载体pFGC5941-35s::BnaTPSA08-1。利用农杆菌介导的甘蓝型油菜下胚轴遗传转化方法成功获得5株稳定遗传的BnaTPSA08-1基因过表达转基因植株。通过收集过表达转基因植株释放的有机挥发性气体,并用GC-MS测定气体组分,结果显示:与对照植株的有机挥发性组分比较,过表达植株产生25种不同类型的挥发性气体,其中十一烷及其衍生物占比最多,这说明BnaTPSA08-1基因产物的生化功能是促进十一烷及其衍生物的合成,这为进一步研究BnaTPSA08-1基因在甘蓝型油菜中的生物学功能打下了良好基础。Terpenoids are one of the largest groups of metabolites in plants and play an important role in plant response to biotic and abiotic stresses. Terpene synthases (TPSs) are key enzymes in the synthesis of terpene compounds in plants. However, there are few reports on terpene synthases in rapeseed. In order to explore the role of TPSs in rapeseed, the function of the terpene synthase gene BnaTPSA08-1 (BnaA08T0101600Z) in Brassica napus was studied. Based on the cloned CDS sequence of BnaTPSA08-1 gene in Brassica napus, the over-expression vector pFGC5941-35s::BnaTPSA08-1 was successfully constructed. Five transgenic plants with stable BnaTPSA08-1 gene over-expression were successfully obtained by Agrobacteria-mediated hypocotyl genetic transformation of Brassica napus. The organic volatile gases released by overexpressed transgenic plants were collected and the gas components were determined by GC-MS. The results showed that: Compared with the organic volatile components of control plants, over-expressed plants produced 25 different types of volatile gases, among them, undecane and undecane derivatives account for the most, indicating that the biochemical function of BnaTPSA08-1 gene product is to promote the synthesis of undecane and undecane derivatives. This provides a good basis for further study on the biological function of BnaTPSA08-1 gene in Brassica napus.
