Yu Han-Song, Ma Lan-Qing, Zhang Ji-Xing, Shi Guang-Lu, Hu Yao-Hui, Wang You-Nian
Food Science and Engineering College, Jilin Agricultural University, Changchun 130118, People's Republic of China.
Key Laboratory of Urban Agriculture (North) of Ministry of Agriculture PR China, Beijing University of Agriculture, Beijing 102206, People's Republic of China.
Phytochemistry. 2011 Jun;72(9):862-70. doi: 10.1016/j.phytochem.2011.03.020. Epub 2011 Apr 16.
Salidroside, the 8-O-β-D-glucoside of tyrosol, is a novel adaptogenic drug extracted from the medicinal plant Rhodiola sachalinensis A. Bor. Due to the scarcity of R. sachalinensis and its low yield of salidroside, there is great interest in enhancing production of salidroside by biotechnological manipulations. In this study, two putative UDP-glycosyltransferase (UGT) cDNAs, UGT72B14 and UGT74R1, were isolated from roots and cultured cells of methyl jasmonate (MeJA)-treated R. sachalinensis, respectively. The level of sequence identity between their deduced amino acid sequences was ca. 20%. RNA gel-blot analysis established that UGT72B14 transcripts were more abundant in roots, and UGT74R1 was highly expressed in the calli, but not in roots. Functional analysis indicated that recombinant UGT72B14 had the highest level of activity for salidroside production, and that the catalytic efficiency (Vmax/Km) of UGT72B14 was 620% higher than that of UGT74R1. The salidroside contents of the UGT72B14 and UGT74R1 transgenic hairy root lines of R. sachalinensis were also ∼420% and ∼50% higher than the controls, respectively. UGT72B14 transcripts were mainly detected in roots, and UGT72B14 had the highest level of activity for salidroside production in vitro and in vivo.
红景天苷是酪醇的8 - O - β - D - 葡萄糖苷,是从药用植物高山红景天中提取的一种新型适应原性药物。由于高山红景天资源稀缺且红景天苷产量低,人们对通过生物技术手段提高红景天苷产量产生了浓厚兴趣。在本研究中,分别从茉莉酸甲酯(MeJA)处理的高山红景天的根和培养细胞中分离出两个推定的尿苷二磷酸糖基转移酶(UGT)cDNA,即UGT72B14和UGT74R1。它们推导的氨基酸序列之间的序列同一性水平约为20%。RNA凝胶印迹分析表明,UGT72B14转录本在根中更为丰富,而UGT74R1在愈伤组织中高表达,但在根中不表达。功能分析表明,重组UGT72B14对红景天苷生产的活性水平最高,并且UGT72B14的催化效率(Vmax/Km)比UGT74R1高620%。高山红景天UGT72B14和UGT74R1转基因毛状根系的红景天苷含量也分别比对照高约420%和约50%。UGT72B14转录本主要在根中检测到,并且UGT72B14在体外和体内对红景天苷生产的活性水平最高。