Kayani Sadaf-Ilyas, Shen Qian, Ma Yanan, Fu Xueqing, Xie Lihui, Zhong Yijun, Tiantian Chen, Pan Qifang, Li Ling, Rahman Saeed-Ur, Sun Xiaofen, Tang Kexuan
Joint International Research Laboratory of Metabolic and Developmental Sciences, Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
Front Plant Sci. 2019 Sep 10;10:1084. doi: 10.3389/fpls.2019.01084. eCollection 2019.
Artemisinin is an effective antimalarial sesquiterpene lactone synthesized in . Various transcription factors have been previously reported that can influence the biosynthesis of artemisinin; however, the effect of YABBY family transcription factors on artemisinin biosynthesis was unknown. In the present study, we cloned and characterized AaYABBY5: a homolog of MsYABBY5 in which is involved in modulating the monoterpenes, as a positive regulator of artemisinin biosynthesis in . AaYABBY5 was found localized to the nucleus, and its expression was found to be induced by exogenous methyl jasmonic acid (MeJA) treatment. In the dual-luciferase reporter assay, it was found that AaYABBY5 significantly increased the activities of promoters of amorpha-4,11-diene synthase (), cytochrome P450 monooxygenase (), double-bond reductase 2 (), and aldehyde dehydrogenase 1 () genes. Yeast one hybrid assay showed that AaYABBY5 directly bonds to the promoters of and genes. Quantitative real-time polymerase chain reaction (qPCR) of overexpression and antisense plants revealed a significant increase in the expression of , , , and in overexpression plants and a significant decrease in the expression of these genes in antisense , respectively. Furthermore, the results of high-performance liquid chromatography (HPLC) showed that the artemisinin and its precursor dihydroartemisinic acid were significantly increased in the overexpression plants while downregulation resulted in a significant decrease in the concentration of artemisinin. Taken together, these results explicitly represent that AaYABBY5 is a positive regulator of artemisinin biosynthesis in .
青蒿素是一种在[具体植物]中合成的有效的抗疟倍半萜内酯。先前已报道多种转录因子可影响青蒿素的生物合成；然而，YABBY家族转录因子对青蒿素生物合成的影响尚不清楚。在本研究中，我们克隆并鉴定了AaYABBY5：它是[具体植物]中MsYABBY5的同源物，参与调节单萜类化合物，作为[具体植物]中青蒿素生物合成的正调控因子。发现AaYABBY5定位于细胞核，并且其表达被外源茉莉酸甲酯（MeJA）处理所诱导。在双荧光素酶报告基因测定中，发现AaYABBY5显著增加了紫穗槐-4,11-二烯合酶（[具体基因名称]）、细胞色素P450单加氧酶（[具体基因名称]）、双键还原酶2（[具体基因名称]）和醛脱氢酶1（[具体基因名称]）基因启动子的活性。酵母单杂交试验表明AaYABBY5直接与[具体基因名称]和[具体基因名称]基因的启动子结合。对[具体基因名称]过表达和反义植物的定量实时聚合酶链反应（qPCR）显示，在[具体基因名称]过表达植物中，[具体基因名称]、[具体基因名称]、[具体基因名称]和[具体基因名称]的表达显著增加，而在[具体基因名称]反义植物中这些基因的表达显著降低。此外，高效液相色谱（HPLC）结果表明，在[具体基因名称]过表达植物中青蒿素及其前体二氢青蒿酸显著增加，而[具体基因名称]下调导致青蒿素浓度显著降低。综上所述，这些结果明确表明AaYABBY5是[具体植物]中青蒿素生物合成的正调控因子。
