Ruslan Komar, Selfitri Anggrahaeni Dewi, Bulan Shella A, Rukayadi Yaya
Pharmaceutical Biology Research Group, School of Pharmacy, Bandung Institute of Technology, Bandung, Indonesia.
Pharmacogn Mag. 2012 Apr;8(30):111-5. doi: 10.4103/0973-1296.96552.
Centella asiatica (L.) Urb. (Apiaceae) is an important medicinal plant, and it has been using to prepare herbal medicines. The compounds responsible for the biological activity of C. asiatica are triterpenoids such as asiaticoside. Asiaticoside is also important as a marker for standardization of C. asiatica. Due to the low content, there is a need to enhance the production of asiaticoside of C. asiatica. The biotechnological approach is one of the methods that can be used to enhance its production.
This study was designed to enhance the production of asiaticoside from C. asiatica using A. rhizogenes and elicitation experiments.
Callus cultures were initiated using Murashige and Skoog (MS) medium supplemented with 1.0 mg/L indole-3-acetic acid (IAA) and 1.0 mg/L 6-benzylaminopurin (BAP). All media were supplemented with 4% (w/w) sucrose and solidified with 0.9% agar. Elicitations were done using pectin, methyl jasmonate, and Cu(2+) ions. Transformed hairy root cultures were performed using A. rhizogenes.
Callus culture of C. asiatica was successfully initiated. Enhancement of the production of asiaticoside in the callus culture by elicitors pectin was up to 31%; methyl jasmonate (50 μM) in cell suspension cultures at day 14 was up to 171% compared to explant and 494% compared to control callus; copper ion (25 μM) at day 21 was up to 144% compared to explant, and 676% compared to control cell suspension cultures. While enhancement by genetic transformation using A. rhizogenes was 166-172% compare to untransformed roots
Elicitation and genetically transformed hairy root cultures of C. asiatica produced asiaticoside up to 172% higher than untreated callus.
积雪草（伞形科）是一种重要的药用植物，一直被用于制备草药。负责积雪草生物活性的化合物是三萜类化合物，如积雪草苷。积雪草苷也是积雪草标准化的重要标志物。由于其含量较低，有必要提高积雪草中积雪草苷的产量。生物技术方法是可用于提高其产量的方法之一。
本研究旨在利用发根农杆菌和诱导实验提高积雪草中积雪草苷的产量。
使用添加1.0 mg/L吲哚-3-乙酸（IAA）和1.0 mg/L 6-苄基氨基嘌呤（BAP）的Murashige和Skoog（MS）培养基启动愈伤组织培养。所有培养基均添加4%（w/w）蔗糖，并用0.9%琼脂固化。使用果胶、茉莉酸甲酯和铜离子进行诱导。使用发根农杆菌进行转化毛状根培养。
成功启动了积雪草的愈伤组织培养。诱导剂果胶使愈伤组织培养物中积雪草苷的产量提高了31%；在第14天，细胞悬浮培养物中的茉莉酸甲酯（50 μM）与外植体相比提高了171%，与对照愈伤组织相比提高了494%；在第21天，铜离子（25 μM）与外植体相比提高了144%，与对照细胞悬浮培养物相比提高了676%。而使用发根农杆菌进行遗传转化与未转化的根相比提高了率为166 - 172%。
积雪草的诱导和遗传转化毛状根培养物产生的积雪草苷比未处理的愈伤组织高出172%。
