title: Plant regeneration, genetic fidelity, and active ingredient content of encapsulated hairy roots of Picrorhiza kurrooa Royle ex Benth.
abstract: Among five hairy root lines of Picrorhiza kurrooa that were established through Agrobacterium rhizogenes, one (H7) was selected for encapsulation due to high accumulation of picrotin and picrotoxinin (8.3 and 47.6 μg/g DW, respectively). Re-grown encapsulated roots induced adventitious shoots with 73 % frequency on MS medium supplemented with 0.1 μM 6-benzylaminopurine, following 6 months of storage at 25 °C. Regenerated plantlets had 85 % survival after 2 months. Regenerants were of similar morphotype having increased leaf number and branched root system as compared to non-transformed plants. The transformed nature of the plants was confirmed through PCR and Southern blot analysis. Genetic fidelity analysis of transformed plants using RAPD and ISSR showed 5.2 and 3.6 % polymorphism, respectively. Phytochemical analysis also showed that picrotin and picrotoxinin content were similar in hairy root line and its regenerants.
authors: Molecular Taxonomy Lab, Botany Division, Forest Research Institute (FRI), P.O. New Forest, Dehradun, Uttarakhand, India. janhvi_mishra@rediffmail.com
authors: Janhvi Mishra
authors: JM
authors: Rawat
authors: Balwant
authors: B
authors: Rawat
authors: Shakti
authors: S
authors: Mehrotra
mesh_terms: D060054:Agrobacterium / Q000235:genetics; D015139:Blotting, Southern; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry; D016172:DNA Fingerprinting; D005838:Genotype; D032261:Picrorhiza / Q000033:anatomy & histology / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D010852:Picrotoxin / Q000031:analogs & derivatives* / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D018520:Plant Shoots / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D016133:Polymerase Chain Reaction; D019105:Random Amplified Polymorphic DNA Technique; D012038:Regeneration; D054830:Sesterterpenes; D013696:Temperature; D014170:Transformation, Genetic
chemical_list: D003470:Culture Media; D010852:Picrotoxin; D054830:Sesterterpenes; C022961:picrotoxinin; C411705:picrotin