RESUMO
BACKGROUND: Gymnema sylvestre is a medicinal woody perennial vine known for its sweetening properties and anti-diabetic therapeutic uses in the modern and traditional medicines. Its over-exploitation for the therapeutic uses and to meet the demand of pharmaceutical industry in raw materials supply for the production of anti-diabetic drugs has led to considerable decline in its natural population. RESULTS: An efficient system of shoot bud sprouting from nodal segment explants and indirect plant regeneration from apical meristem-induced callus cultures of G. sylvestre have been developed on Murashige and Skoog (MS) medium amended with concentrations of cytokinins. Of the three growth regulators tested, N6-benzylaminopurine (BAP) was the most efficient and 2.0 mg L-1 gave the best shoot formation efficiency. This was followed by thidiazuron (TDZ) and kinetin (Kin) but, most of the TDZ-induced micro shoots showed stunted growth. Multiple shoot formation was observed on medium amended with BAP or TDZ at higher concentrations. The produced micro shoots were rooted on half strength MS medium amended with auxins and rooted plantlets acclimatized with 87% survival of the regenerates. CONCLUSIONS: The developed regeneration system can be exploited for genetic transformation studies, particularly when aimed at producing its high yielding cell lines for the anti-diabetic phytochemicals. It also offers opportunities for exploring the expression of totipotency in the anti-diabetic perennial vine.
Assuntos
Gymnema sylvestre/crescimento & desenvolvimento , Morfogênese/efeitos dos fármacos , Reguladores de Crescimento de Plantas/farmacologia , Brotos de Planta/crescimento & desenvolvimento , Regeneração/efeitos dos fármacos , Compostos de Benzil/farmacologia , Gymnema sylvestre/efeitos dos fármacos , Cinetina/farmacologia , Compostos de Fenilureia/farmacologia , Brotos de Planta/efeitos dos fármacos , Purinas/farmacologia , Tiadiazóis/farmacologiaRESUMO
BACKGROUND: Gymnema sylvestre is a medicinal woody perennial vine known for its sweetening properties and antidiabetic therapeutic uses in the modern and traditional medicines. Its over-exploitation for the therapeutic uses and to meet the demand of pharmaceutical industry in raw materials supply for the production of anti-diabetic drugs has led to considerable decline in its natural population. RESULTS: An efficient system of shoot bud sprouting from nodal segment explants and indirect plant regeneration from apical meristem-induced callus cultures of G. sylvestre have been developed on Murashige and Skoog (MS) medium amended with concentrations of cytokinins. Of the three growth regulators tested, N6-benzylaminopurine (BAP) was the most efficient and 2.0 mg L-1 gave the best shoot formation efficiency. This was followed by thidiazuron (TDZ) and kinetin (Kin) but, most of the TDZ-induced micro shoots showed stunted growth. Multiple shoot formation was observed on medium amended with BAP or TDZ at higher concentrations. The produced micro shoots were rooted on half strength MS medium amended with auxins and rooted plantlets acclimatized with 87% survival of the regenerates. CONCLUSIONS: The developed regeneration system can be exploited for genetic transformation studies, particularly when aimed at producing its high yielding cell lines for the anti-diabetic phytochemicals. It also offers opportunities for exploring the expression of totipotency in the anti-diabetic perennial vine.
Assuntos
Reguladores de Crescimento de Plantas/farmacologia , Regeneração/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Gymnema sylvestre/crescimento & desenvolvimento , Morfogênese/efeitos dos fármacos , Compostos de Fenilureia/farmacologia , Purinas/farmacologia , Tiadiazóis/farmacologia , Compostos de Benzil/farmacologia , Brotos de Planta/efeitos dos fármacos , Gymnema sylvestre/efeitos dos fármacos , Cinetina/farmacologiaRESUMO
Gymnema sylvestre R. Br. (Asclepiadaceae) is an endangered species extensively used in the management of diabetes, obesity, and treatment of various diseases. Uncontrolled exploitation to meet the increasing demand and low seed viability hastens the disappearance of the plant from its natural habitat. Hairy root culture provides a suitable alternative for the enhanced production of active principles. The current protocol provides the optimized culture conditions for the establishment of hairy root cultures and elicitation studies and also confirmation of stable integration of A. rhizogenes plasmid T-DNA into host genetic material by PCR and RT-PCR. Furthermore, it also discusses the suitable methods for the extraction procedures, and qualitative and quantitative analysis of gymnemic acid by HPTLC and HPLC.
Assuntos
Gymnema sylvestre/crescimento & desenvolvimento , Melhoramento Vegetal/métodos , Raízes de Plantas/crescimento & desenvolvimento , Saponinas/metabolismo , Triterpenos/metabolismo , Agrobacterium/genética , Agrobacterium/fisiologia , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia em Camada Delgada/métodos , DNA Bacteriano/genética , Gymnema sylvestre/genética , Gymnema sylvestre/microbiologia , Gymnema sylvestre/fisiologia , Raízes de Plantas/genética , Raízes de Plantas/microbiologia , Raízes de Plantas/fisiologia , Plasmídeos/genética , Reação em Cadeia da Polimerase/métodos , Saponinas/análise , Saponinas/genética , Transformação Genética , Triterpenos/análiseRESUMO
Gymnema sylvestre R. Br. is an important medicinal plant known for its antidiabetic potential. In the present study, phytosterols from G. sylvestre was identified and quantified in vivo and in vitro. Maximum callus induction was observed in MS medium supplemented with 0.5 mg L(-1) of 2, 4-D. The protein content was significantly high both in aerial plant parts and callus tissue. Phytosterols were identified using chromatographic and spectral studies. beta-sitosterol, campesterol and stigmasterol were identified both in vivo and in vitro. Lanosterol was identified only in callus culture. Phytosterols have reported for the first time in callus culture of G. sylvestre.
