Abstract
BackgroundGloriosa superba L. belongs to Colchicaceae, which is an important medicinal plant containing high-priced alkaloid colchicines and other potent phytochemicals. Due to its extensive importance at the industrial level, this plant is overexploited. Moreover, indiscriminate harvesting for raw material leads to a decline in the population of this plant in the natural environment. Thus, the present study deals with the optimization of colchicine accumulation from the different intervals of in vitro and in vivo tubers of Gloriosa.ResultTo obtain in vitro tuberization, shoot tip explants were inoculated on Murashige and Skoog medium prepared with 3 mg/l BA and 0.5 mg/l Kn + 1 mg/l 2, 4-D followed by 2 mg/l BA + 0.2 mg/l NAA. In the high-performance thin-layer chromatography study (HPTLC), the linearity range of colchicine was set at a concentration range of 100–1000 ng/spot with a regression value (r) of 0.99. Its Rf value (0.25) was recorded at 254 nm. The colchicine amount in the in vivo tuber was 7.75 ± 0.25% dry weight, while the nearby amount of 7.7 ± 0.40% dry weight of colchicine was produced from 2 weeks in vitro old tuber. This value was followed by the 4th-week old tuber with 6.35 ± 0.17% dry weight and then a gradual decrease in its accumulation.ConclusionThe significant results for the accumulation of colchicine at different stages were observed. Hence, this strategy of colchicine production creates a new possibility for improved production of colchicine under in vitro conditions which will be helpful to various pharmaceutical industries without damaging the plants from the natural environment.Graphical abstract
Highlights
Gloriosa superba L. belongs to Colchicaceae, which is an important medicinal plant containing highpriced alkaloid colchicines and other potent phytochemicals
The significant results for the accumulation of colchicine at different stages were observed. This strategy of colchicine production creates a new possibility for improved production of colchicine under in vitro conditions which will be helpful to various pharmaceutical industries without damaging the plants from the natural environment
The full-strength Murashige and Skoog (MS) [18] medium was prepared by supplementing various concentrations and combinations of plant growth regulators (PGRs) like BA (N6-benzyl adenine) alone (1–3 mg/l), in combination with α-naphthalene acetic acid (NAA). (NAA 0.2 mg/l), combination of Kinetin + 2,4-Dichloro phenoxy acetic acid (2,4-D) (0.2 + 0.5, 0.5 + 1 and 1 + 1.5 mg/l)
Summary
Gloriosa superba L. belongs to Colchicaceae, which is an important medicinal plant containing highpriced alkaloid colchicines and other potent phytochemicals. It is commonly known as vachhnag, kalahari, flame lily, and glory lily Individual parts of this plant contain important phytochemicals including colchicine and are used for the treatment of various diseases like cancer, swelling, Rathod et al Futur J Pharm Sci (2021) 7:177 been carried out by the pharmaceutical industries. Due to multiple uses of colchicine, in vitro technologies would be the best option to elevate the production of colchicine along with its improved properties This technique provides a continuous, reproducible, and reliable source of the plant to meet the ever-increasing demand for pharmaceutical industries. High-performance thin-layer chromatography (HPTLC) is proven as a powerful tool for qualitative and quantitative analysis of vital bioactive compounds which are produced in various parts of plants It is considered as sophisticated among all other analytical techniques in terms of reliability, simplicity, speed, time, and cost per analysis [11, 12]. In the present investigation, an attempt was made for (1) in vitro tuber production and multiplication, (2) determination of colchicine in it and (3) optimization of the in vitro stage(s) for maximum accumulation of colchicines by using the HPTLC technique in both in vivo tuber and in vitro developed tuber at different developmental stages after method validation
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