In Vitro Propagation of Cannabis sativa L. and Evaluation of Regenerated Plants for Genetic Fidelity and Cannabinoids Content for Quality Assurance.
Cannabis sativa L. (Marijuana; Cannabaceae), one of the oldest medicinal plants in the world, has been used throughout history for fiber, food, as well as for its psychoactive properties. The dioecious and allogamous nature of C. sativa is the major constraint to maintain the consistency in chemical profile and overall efficacy if grown from seed. Therefore, the present optimized in vitro propagation protocol of the selected elite germplasm via direct organogenesis and quality assurance protocols using genetic and chemical profiling provide an ideal pathway for ensuring the efficacy of micropropagated Cannabis sativa germplasm. A high frequency shoot organogenesis of C. sativa was obtained from nodal segments in 0.5 μM thidiazuron medium and 95 % in vitro rhizogenesis is obtained on half-strength MS medium supplemented with 500 mg/L activated charcoal and 2.5 μM indole-3-butyric acid. Inter Simple Sequence Repeats (ISSR) and Gas Chromatography-Flame Ionization Detection (GC-FID) are successfully used to monitor the genetic stability in micropropagated plants up to 30 passages in culture and hardened in soil for 8 months.
National Center for Natural Product Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, University of Mississippi, Mississippi, MS, 38677, USA. hlata@olemiss.edu.
Hemant
H
Lata
National Center for Natural Product Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, University of Mississippi, Mississippi, MS, 38677, USA.
Suman
S
Chandra
National Center for Natural Product Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, University of Mississippi, Mississippi, MS, 38677, USA.|Department of Pharmacognosy, School of Pharmacy, University of Mississippi, Mississippi, MS, 38677, USA.
Ikhlas A
IA
Khan
National Center for Natural Product Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, University of Mississippi, Mississippi, MS, 38677, USA.|Department of Pharmaceutics, School of Pharmacy, University of Mississippi, Mississippi, MS, 38677, USA.
Mahmoud A
MA
ElSohly
D000064:Acclimatization; D002186:Cannabinoids / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D002188:Cannabis / Q000235:genetics* / Q000254:growth & development* / Q000502:physiology; D002849:Chromatography, Gas / Q000379:methods; D003470:Culture Media / Q000378:metabolism; D046508:Culture Techniques / Q000379:methods; D018744:DNA, Plant / Q000235:genetics; D018895:Microsatellite Repeats; D063246:Organogenesis, Plant; D000069600:Plant Breeding / Q000379:methods; D010937:Plant Growth Regulators / Q000378:metabolism; D018520:Plant Shoots / Q000235:genetics / Q000254:growth & development / Q000502:physiology; D013242:Sterilization / Q000379:methods
D002186:Cannabinoids; D003470:Culture Media; D018744:DNA, Plant; D010937:Plant Growth Regulators
Cannabis sativa L.; GC-FID; Genetic fidelity; ISSR markers; Micropropagation; Nodal explants