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Callus induction and Organogenesis in Sugarcane (Saccharum officinarum L.) var 93v297

100%

Rao A. S.

tom 48

An efficient protocol for induction of callus and regeneration of a sugar cane var 93v297 has been developed and reported here. Callus induction from immature young leaf explants derived from 2-3-month-old plants was achieved on Murashige and Skoog’s (MS) medium supplemented with different auxins viz, 2,4-D, NAA and IAA. Among different auxins, 2, 4-D at 3.5mg/l + 0.5mg/l BAP was found favourable in inducing callus. Addition of coconut milk and BAP further enhanced the growth of callus maximum being on MS medium supplemented with 0.5mg/l BAP (3602.33±0.88mg). Calli were further evaluated for regeneration. MS medium supplemented with 1.0 mg/l BAP was found suitable where 100% calli regenerated with maximum number of multiple shoots per callus mass (41.40±0.89). Highest number of root emergence (28.33±1.16) and maximum root length (3.40±0.67cm) was achieved on MS medium supplemented with 3mgl/l NAA. The in vitro grown plants were transferred to polycups containing a mixture of sterilized sand, soil and cocopeet (1:1:1) for hardening. The hardened plants were transferred to green-house conditions where they survived with 90% frequency.

HPTLC/HPLC and gravimetric methodology for the identification and quantification of gymnemic acid from Gymnema sylvestre methanolic extracts

51%

Ahmed A. B. A. , Rao A. S. , Rao M. V. , Taha R. M.

2013

tom Vol. 25, no. 2

339--361

Gymnemic acid (GA), a well known anti-diabetic compound has been detected in methanol extracts of intact leaves and in vitro callus cultures derived from leaf explants of Gymnema sylvestre. Callus biomass was developed in MS medium with optimum plant growth regulators (OPGRs) of 2,4-D (1.5 mg L-1) + KN (0.5 mg L-1) under abiotic stress conditions at 45 days determined by growth curve analysis. GA detection and quantification were carried out using thin-layer chromatography (TLC), highperformance thin-layer chromatography (HPTLC), high-performance liquid chromatography (HPLC), and gravimetric techniques. GA detection peak area and their absorption spectra were evaluated through HPTLC and HPLC with the standard GA. Quantification of GA had showed the linearity, accuracy, robustness and precision by HPLC. GA content was significantly higher in gravimetric method than HPLC. All these methods were found to be simple, accurate, selective and rapid and could be successfully applied for the determination of GA. It could have potential as a pharmaceutical drug for Type 1 diabetes mellitus (IDDM) and obesity.