Development of Hairy Root Cultures for Biomass and Triterpenoid Production in <i>Centella asiatica</i>
2022
Seungeun Baek | Jong-Eun Han | Thanh-Tam Ho | So-Young Park
<i>Centella asiatica</i> (Apiaceae) is a tropical/subtropical medicinal plant, which contains a variety of triterpenoids, including madecassoside, asiaticoside, madecassic acid, and asiatic acid. In this study, we tested the efficiency of hairy root (HR) induction in <i>C. asiatica</i> from leaf and petiole explants. Leaves and petioles collected from <i>C. asiatica</i> plants were suspended in agro-stock for 30 min and co-cultured with <i>Agrobacterium rhizogenes</i> for 3 days to induce HR formation. The transformation efficiency of leaf and petiole explants was approximately 27% and 12%, respectively. A total of 36 HR lines were identified by PCR-based amplification of <i>rol</i> genes, and eight of these lines were selected for further analysis. Among all eight HR lines, the petiole-derived lines HP4 and HP2 displayed the highest growth index (37.8) and the highest triterpenoids concentration (46.57 mg∙g<sup>−1</sup>), respectively. Although triterpenoid concentration was >2-fold higher in leaves than in petioles of <i>C. asiatica</i> plants, the accumulation of triterpenoids in petiole-derived HR cultures was 1.4-fold higher than that in leaf-derived HR cultures. Additionally, in both leaf- and petiole-derived HR cultures, terpenoid production was higher in HRs than in adventitious roots. These results demonstrate that the triterpenoid content in the explant does not affect the triterpenoid content in the resultant HRs. The HR culture of <i>C. asiatica</i> could be scaled up to enable the mass production of triterpenoids in bioreactors for the pharmaceutical and cosmetic industries.
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