RESUMEN
The efficient production of plant-derived medicinal compounds (PDMCs) from in vitro plants requires improvements in knowledge about control of plant or organ development and factors affecting the biosynthesis pathway of specific PDMCs under in vitro conditions, leading to a realistic large-scale tool for in vitro secondary metabolite production. Thus, this study aimed to develop an in vitro technique, through the induction and proliferation of calli, for production of plant fresh weight, and to compare the PDMC profile obtained from the plants versus in vitro calli of Phyllanthus amarus. It was successfully possible to obtain and proliferate two types of calli, one with a beige color and a friable appearance, obtained in the dark using Murashige and Skoog (MS) medium plus 2,4-dichlorophenoxyacetic acid (2,4-D), and a second type with a green color, rigid consistency, and nonfriable appearance obtained under light conditions and MS medium plus 6-benzyladenine (6-BA). In vitro micropropagated plants that gave rise to calli were also acclimatized in a greenhouse and cultivated until obtaining the mass for PDMC analysis and used as a control. While the micropropagated-derived plants concentrated the lignans niranthin, nirtetralin, and phyllanthin, the Phyllanthus amarus calli proliferated in vitro concentrated a completely different biochemical profile and synthesis of compounds, such as betulone, squalene, stigmasterol, and ß-sitosterol, in addition to others not identified by GC-MS database. These results demonstrate the possibility of applying the calli in vitro from Phyllanthus amarus for production of important PDMCs unlike those obtained in cultures of differentiated tissues from field plants.
Asunto(s)
Phyllanthus/química , Extractos Vegetales/aislamiento & purificación , Botánica/métodos , Cámbium/metabolismo , Proliferación Celular , Citocininas , Oscuridad , Técnicas In Vitro , Células Vegetales , Extractos Vegetales/química , Plantas Medicinales/químicaRESUMEN
Abstract A precise and accurate method for the identification and authentication of Phyllanthus niruri L. from P. debilis Klein ex Willd. and P. urinaria L., Phyllanthaceae, was developed using high-performance liquid chromatography. Chromatographic fingerprint analysis was combined with simultaneous quantification of phyllanthin and hypophyllanthin for the developed method. Phyllanthin and hypophyllanthin were successfully separated and quantified under this proposed method. The highest amount of phyllanthin and hypophyllanthin was found in P. niruri compared to P. debilis and P. urinaria. Fingerprint chromatogram of the three Phyllanthus species showed distinct profiles that these may be used to identify and authenticate each Phyllanthus species, which improved by marker compounds present in each species. The combination of chromatographic fingerprint analysis and discriminant analysis was successfully discriminated all three species, including P. niruri adulterated with P. debilis or P. urinaria. The method can be used for the identification and authentication of P. niruri from related species, such as P. debilis and P. urinaria.