Wang Jian-Ping, Zhou Ya-Ming, Zhang Yong-Hui
Key Laboratory of Natural Medicinal Chemistry and Resources Evaluation of Hubei Province, Department of Traditional Chinese Medicine, College of Pharmacy, Huazhong University of Science and Technology, Wuhan, People's Republic of China.
Pharmacogn Mag. 2012 Apr;8(30):149-55. doi: 10.4103/0973-1296.96569.
Despite the excellent anti-inflammatory and anti-rheumatic efficacy associated with kirenol generation, the content of kirenol in Siegesbeckea orientalis is quite low.
This study was designed to establish a reliable kirenol production protocol by transformed root cultures of S. orientalis and to investigate the antimicrobial activities of kirenol, hairy root, and S. orientalis.
Transformed root cultures of S. orientalis were established by the transformation of Agrobacterium rhizogenes A4. Transgenic status of the roots was confirmed by polymerase chain reaction (PCR) using rolB specific primers. The biomass and kirenol accumulation of hairy root clones were assessed using four different culture media: MS, MS/2, B5, and white. The antimicrobial activities of kirenol, hairy root, and S. orientalis were evaluated by the disc diffusion method.
The optimum media for kirenol synthesis was MS. The content of kirenol in transformed hairy roots made up about 80% of that observed in natural leaves of S. orientalis (1.6 mg/g dry weight). All tested samples displayed antimicrobial activity against Gram-positive pathogens including Staphylococcus epidermidis, Staphylococcus aureus, and Acinetobacter baumannii, with MIC ranging from 78 to 625 μg/mL.
The high level of kirenol contents was obtained from hairy roots of S. orientalis. Kirenol was effective against gram-positive bacteria. Interestingly, the extract from hairy roots showed a diverse antimicrobial effect from that of kirenol and S. orientalis.
尽管奇壬醇具有出色的抗炎和抗风湿功效，但东方菝葜中奇壬醇的含量相当低。
本研究旨在通过东方菝葜的发根培养建立可靠的奇壬醇生产方案，并研究奇壬醇、毛状根和东方菝葜的抗菌活性。
通过发根农杆菌A4转化建立东方菝葜的发根培养。使用rolB特异性引物通过聚合酶链反应（PCR）确认根的转基因状态。使用四种不同的培养基（MS、MS/2、B5和White）评估毛状根克隆的生物量和奇壬醇积累。通过纸片扩散法评估奇壬醇、毛状根和东方菝葜的抗菌活性。
奇壬醇合成的最佳培养基是MS。转化毛状根中奇壬醇的含量约占东方菝葜天然叶片中观察到的含量（1.6毫克/克干重）的80%。所有测试样品对包括表皮葡萄球菌、金黄色葡萄球菌和鲍曼不动杆菌在内的革兰氏阳性病原体均显示出抗菌活性，最低抑菌浓度范围为78至625微克/毫升。
从东方菝葜的毛状根中获得了高水平的奇壬醇含量。奇壬醇对革兰氏阳性菌有效。有趣的是，毛状根提取物显示出与奇壬醇和东方菝葜不同的抗菌效果。
