Enhanced production of berberine in In vitro regenerated cell of Tinospora cordifolia and its analysis through LCMS QToF

Abstract

Tinospora cordifolia is a prioritized medicinal plant and having an immense medicinal importance especially in Indian medicinal system. But this plant needs a regeneration protocol for its rapid propagation. An efficient regeneration protocol was developed for T. cordifolia using nodal explants. High frequency of multiple shoot formation was induced when the nodal segments were cultured on MS medium supplemented with BAP (1.0 mg L−1) and 2-iP (0.5 mg L−1). The highest mean number of shoots per nodal explant (7.9 ± 0.45) with highest shoot length (9.3 ± 0.48 cm) and 86% response were achieved on this media and hormonal concentration. The optimum rooting was obtained on ½ strength of MS medium augmented with IBA (0.5 mg L−1) with 8.3 ± 0.46 cm root length and 89% response. Micropropagated plantlets were found to be identical with the mother plant when clonal fidelity of these plantlets were analyzed with inter simple sequence repeat (ISSR) marker. The berberine content was analyzed through LCMS QToF and the highest amount was found in in vitro callus (19.8 µg/gm) followed by stem (9.3 µg/gm) and leaves of field-grown plants (8.4 µg/gm). Further, presence of berberine was confirmed by ESI–MS spectra with protonated molecular ions ([M + H]+) at m/z 336. Furthermore, MS–MS fragmentation pattern confirmed for the presence of berberine in both the samples. Both the spectra (standard and samples) showed common peaks for berberine in the form of protonated molecular ions ([M + H]+) at m/z 320, m/z 304, m/z 292, m/z 278 in MS/MS mode. The study revealed that developed protocol is potent for rapid mass propagation of this plant species with high accumulation of important secondary metabolite berberine.

Microplants production of Eucalyptus cloeziana from indirect organogenesis

“…Hypocotyl and cotyledon of in vitro cultured seedlings were the most responsive tissues as explants for the regeneration of new E. cloeziana plants via indirect organogenesis, showing a response similar to that of other studies (Mittal and Sharma 2017). A possible cause for this tissue differentiation, considering that these events may vary according to the plant genotype (Salla et al 2018), is related to the regulation of physiological processes that favor the formation of new tissues because they contain cells with high juvenility and cellular competence (Wendling et al 2015), leading to increased organ production and development (Gupta and Karmakar 2017).…”

“…Thus, the influence of the incubation conditions has been studied in several Eucalyptus species (Fernando et al 2016) during organogenesis. These changes may occur due to the composition of the culture medium, nutrition, balance between the plant growth regulators, type and ontogenetic age of the explant and determination of the tissues (Brondani et al 2012, Mittal and Sharma 2017, Souza et al 2019.…”

A review on analytical methods for natural berberine alkaloids

“…A Waters Acquity UPLC instrument system equipped with Q-TOF-MS was used for analysis to provide a good foundation for future research on chemical structure characteristics. The enhanced production of berberine in in vitro regenerated cells of Tinospora cordifolia was analysed through UHPLC-Q-TOF-MS equipped with a HSS T3 column, and the eluted compounds were detected from m/z 100 to 1000 by using a Q-TOF mass spectrometer full scan equipped with ESI in positive ion mode [81]. An efficient regeneration protocol was developed for Tinospora cordifolia using nodal explants in this paper and the study revealed that the developed protocol was potent for rapid mass propagation of this plant species with high accumulation of important secondary berberine metabolites.…”

Optimization of salicylic acid and chitosan treatment for bitter secoiridoid and xanthone glycosides production in shoot cultures of Swertia paniculata using response surface methodology and artificial neural network

“…Advancement in plant biotechnology holds great assurance for the conservation and production of improved variety of medicinally important rare and endangered flora and large-scale production of plant derived secondary metabolites. In a variety of medicinal plants, tissue culture technique has been successfully proven as a potential alternative strategy for the production of valuable bio-active compounds with clinical significance [9]. In Swertia spp., very few reports are available on the in vitro strategies that are used to enhance growth and secondary metabolite production utilizing hairy/adventitious root cultures, rhizobacteria etc.…”