Development of Hairy Root Cultures for Biomass and Triterpenoid Production in Centella asiatica

Abstract

Centella asiatica (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 C. asiatica from leaf and petiole explants. Leaves and petioles collected from C. asiatica plants were suspended in agro-stock for 30 min and co-cultured with Agrobacterium rhizogenes 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 rol 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−1), respectively. Although triterpenoid concentration was >2-fold higher in leaves than in petioles of C. asiatica 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 C. asiatica could be scaled up to enable the mass production of triterpenoids in bioreactors for the pharmaceutical and cosmetic industries.

Effect of a suitable treatment period on the genetic transformation efficiency of the plant leaf disc method

“…Studies have shown that among the many factors that determine the efficiency of genetic transformation by the leaf disc method, plant species and genotypes within the same species may have greater influence [ 31 – 34 , 73 ]. The use of different explants and explants at different development stages has a great impact on transformation efficiency [ 6 , 7 , 19 , 35 , 37 – 40 ]. This indicates that the optimal genetic transformation period must be different for different species, different genotypes of the same species, different explants of the same genotype, and receptor materials of the same explant at different developmental stages when transformation is applied.…”

“…Agrobacterium tumefaciens -mediated leaf disc transformation is a plant genetic transformation method developed by Horsch et al [ 4 , 5 ]. The improved leaf disc method includes stem segments, petioles, hypocotyls, cotyledons, and other explant materials [ 6 , 7 ]. This method introduces foreign DNA fragments or gene editors carried by Ti plasmids, Ri plasmids, and other carriers into host cells through Agrobacterium infection to study related gene functions or realize plant breeding.…”

“…Many studies have shown that different plant species, different genotypes within the same species, different explants of the same species, and different development stages of explants have a great impact on transformation efficiency, and the rate of genetic transformation can be improved when explants are cultured to a suitable state of cell development [ 6 , 7 , 19 , 31 – 34 ]. When the genetic transformation of barley was carried out, the transformation rate of barley cultured on differentiation medium for 1 day was higher than that of the uncultured, and the transformation rate dropped sharply after 1 day [ 35 ].…”

FtbZIP85 Is Involved in the Accumulation of Proanthocyanidin by Regulating the Transcription of FtDFR in Tartary Buckwheat

“…With the development of the biological industry in recent years, the cost of metabolome, transcriptome and genome research has greatly reduced. Hairy root cultures are being gradually applied to the study of metabolic pathways [ 43 , 44 , 45 , 46 , 47 ]. Nonetheless, our results indicate that ABA inhibited PA biosynthesis.…”

Induction, Flavonoids Contents, and Bioactivities Analysis of Hairy Roots and True Roots of Tetrastigma hemsleyanum Diels et Gilg

“…More than 300 species of hairy roots have been established in the culture system and used to produce bioactive compounds. Of these, 76% are herbaceous plants, such as the sunflower Helianthus annuus L. [ 14 ], as well as Urena lobata [ 15 ] and Centella asiatica [ 16 ]. Because of the difficulty in infecting woody plants and lianas, woody plants such as Taxus × media have accounted for 17% [ 17 ], while lianas such as Tripterygium wilfordii have accounted for 7% [ 18 ].…”