Cao Xue, Qin Zhenfen, Fan Panhui, Wang Sifan, Meng Xiangxiao, Wan Huihua, Yang Wei, Chen Shilin, Yao Hui, Chen Weiqiang, Sun Wei
State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Acta Pharm Sin B. 2025 Aug;15(8):4300-4305. doi: 10.1016/j.apsb.2025.03.038. Epub 2025 Mar 18.
Genetic transformation is a fundamental tool in molecular biology research of medicinal plants. Tailoring transgenic technologies to each distinct medicinal plant would necessitate a substantial investment of time and effort. Here, we present a simple hairy root transformation method that does not require sterile conditions, utilizing strain K599 and the visible RUBY reporter system. Transgenic hairy roots were obtained for six tested medicinal plant species, roots or rhizomes of which have recognized medicinal value, spanning four botanical families and six genera (, , , , , and ). Furthermore, two previously identified UGTs that convert liquiritigenin into liquiritin in heterologous systems were studied using the method. Our results indicate that overexpression of but not and Cas9-mediated knockout of profoundly influenced the accumulation of liquiritin and isoliquiritin in licorice roots. Therefore, the method described here represents a simple, rapid and widely applicable hairy root transformation method that enables fast gene functional study in medicinal plants.
遗传转化是药用植物分子生物学研究中的一项基本工具。针对每一种独特的药用植物量身定制转基因技术将需要投入大量的时间和精力。在此，我们展示了一种简单的毛状根转化方法，该方法不需要无菌条件，利用K599菌株和可见的红宝石报告系统。我们获得了六种受试药用植物的转基因毛状根，这些植物的根或根茎具有公认的药用价值，涵盖四个植物科和六个属（、、、、、和）。此外，还使用该方法研究了两个先前在异源系统中将甘草素转化为甘草苷的UGT。我们的结果表明，的过表达而非和Cas9介导的的敲除对甘草根中甘草苷和异甘草苷的积累有深远影响。因此，本文所述方法代表了一种简单、快速且广泛适用的毛状根转化方法，能够在药用植物中快速进行基因功能研究。
