Induction of root colonization by Piriformospora indica leads to enhanced asiaticoside production in Centella asiatica

Centella asiatica (Indian pennywort) has wide application in Indian and Chinese traditional medicines with documented evidence for wound healing and neuroprotective and anti-aging potential. Asiaticoside, a trisaccharide triterpene, is the most medicinally active compound in the plant. beta-Amyrin synthase and squalene synthase have been identified as the two key genes in the triterpenoid pathway which regulate the production of asiaticoside in C. asiatica. The paper reports salient findings of our study utilizing the growth-promoting endophytic fungus Piriformospora indica to successfully colonize roots of C. asiatica in vitro cultures for investigating the effect of the mutualistic association on asiaticoside production. Co-cultivation of P. indica resulted in the rapid enhancement of root and shoot biomass of host plant, which was visible after 7 days of culture and continued up to 45 days. P. indica co-cultivation also favored the synthesis of asiaticosides, as evidenced by HPLC analysis which indicated about twofold increase (0.53% (w/w) in leaves and 0.23% (w/w) in whole plant) over control (0.33% (w/w) in leaves and 0.14% (w/w) in whole plant). Real-time PCR results confirmed the strong upregulation of squalene synthase and beta-amyrin synthase transcripts in P. indica-challenged plants compared with the control. Our data demonstrate the potential use of P. indica as a means to enhance plant secondary metabolite production in planta with scope for further field evaluation.