Hairy Root Cultures of Gymnema sylvestre R. Br. to Produce Gymnemic Acid.
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.
Department of Biochemistry, Davangere University, Shivagangothri, Davangere, 577002, Karnataka, India.
J
J
Rajashekar
Department of Biochemistry, Davangere University, Shivagangothri, Davangere, 577002, Karnataka, India. vadlapudikumar@rediffmail.com.|Department of Biochemistry, Kuvempu University - P.G. Centre, Shivagangothri, Davangere, 577002, Karnataka, India. vadlapudikumar@rediffmail.com.
Vadlapudi
V
Kumar
Department of Biochemistry, Kuvempu University - P.G. Centre, Shivagangothri, Davangere, 577002, Karnataka, India.
V
V
Veerashree
Department of Biochemistry, Davangere University, Shivagangothri, Davangere, 577002, Karnataka, India.
D V
DV
Poornima
Department of Biochemistry, Davangere University, Shivagangothri, Davangere, 577002, Karnataka, India.
Torankumar
T
Sannabommaji
Department of Biochemistry, Davangere University, Shivagangothri, Davangere, 577002, Karnataka, India.
Hari
H
Gajula
Department of Biochemistry, Davangere University, Shivagangothri, Davangere, 577002, Karnataka, India.
B
B
Giridhara
D060054:Agrobacterium / Q000235:genetics / Q000502:physiology; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002855:Chromatography, Thin Layer / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics; D031104:Gymnema sylvestre / Q000235:genetics / Q000254:growth & development* / Q000382:microbiology / Q000502:physiology; D000069600:Plant Breeding / Q000379:methods*; D018517:Plant Roots / Q000235:genetics / Q000254:growth & development* / Q000382:microbiology / Q000502:physiology; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D012503:Saponins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic; D014315:Triterpenes / Q000032:analysis / Q000378:metabolism*
D004269:DNA, Bacterial; D012503:Saponins; C036483:T-DNA; D014315:Triterpenes; C007898:gymnemic acid
Agrobacterium rhizogenes; Elicitors; Gymnema sylvestre; Gymnemic acid; HPLC; Hairy root cultures