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Rhodiola imbricata is a rare medicinal plant of the trans-Himalayan region of Ladakh. It is used for the treatment of numerous health ailments. Compact callus aggregate (CCA) suspension cultures of Rhodiola imbricata were established to counter extinction threats and for production of therapeutically valuable phenolic compounds to meet their increasing industrial demands. The present study also investigated the effect of jasmonic acid (JA) on production of phenolic compounds and bioactivities in CCA suspension cultures. CCA suspension cultures established in an optimized Murashige and Skoog medium supplemented with 30 g/l sucrose, 3 mg/l NAA, and 3 mg/l BAP showed maximum biomass accumulation (8.43 g/l DW) and highest salidroside production (3.37 mg/g DW). Upon 100 μM JA treatment, salidroside production (5.25 mg/g DW), total phenolic content (14.69 mg CHA/g DW), total flavonoid content (4.95 mg RE/g DW), and ascorbic acid content (17.93 mg/g DW) were significantly increased in cultures. In addition, DPPH-scavenging activity (56.32%) and total antioxidant capacity (60.45 mg QE/g DW) were significantly enhanced upon JA treatment, and this was positively correlated with increased accumulation of phenolic compounds. JA-elicited cultures exhibited highest antimicrobial activity against Escherichia coli. This is the first report describing the enhanced production of phenolic compounds and bioactivities from JA-elicited CCA suspension cultures of Rhodiola imbricata.

Study was conducted during 2013-14 to examine the role of various dormancy breaking treatments, viz. hot water treatment, scarification, stratification, concentrated acids (H 2 SO 4 , HNO 3 and HCl), gibberellic acid, potassium nitrate, alcohol, and acetone and gamma-rays irradiation on the percentage germination, seedling growth and seed vigour. Dried seeds were incubated in the plant growth chambers for 20-28 days at constant temperature of 25±2 °C under continuous light (16 hrs) photoperiod after its treatments. Maximum percent germination 97.2% was obtained in Innula racemosa followed by Rheum webbianum (95.1%), Carum carvi (93.4%), Saussurea lappa (90.01%) and Bunium persicum (81.4%) when seeds were pretreated with acid (H 2 SO 4 for 5 minutes). According to results obtained in present study, all studied species found best germination with H 2 SO 4 for 5 minutes in duration of 30 days. The seedlings derived from seeds exposed to the various treatments performed well when grown in a green house. Maximum length of seedlings were found in 24.3 cm in S. lappa followed by R. webbianum (23.8 cm) and C. carvi (22.2 cm) when seeds were pretreated with H 2 SO 4 for 5 minutes, on the other side B. persicum (19.4cm) in hot water treatment at 80°C for 20 minutes and I. racemosa (17.4 cm) in 0.2 KNO 3 for 10 minutes. Highest value of seed vigour index (2263) and lowest seed vigour index (390) was found in R. webbianum and B. persicum. The well developed seedlings were observed in 90 days and transplanted it for further developments. The data have implications for conservation and cultivation of the species studied.

Aim: The present report aimed to study the therapeutic and phytochemical properties of stems and roots of trans-himalayan plant Codonopsis clematidea. Material and Methods: The crude samples of stems and roots were explored for their chlorophyll, carotenoid and carbohydrate content in order to understand the matrix of these two plant parts. The extraction of phytochemicals was carried out by three different methods viz. sonication, maceration and soxhlet in methanol to identify the best extraction method. Further, analysis of Total Flavonoid Content (TFC) and Total Polyphenolic Content (TPC) were carried out using rutin trihydrate and gallic acid as a reference standard. Antioxidant capacity was estimated using three methods viz. 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging assay and Ferric Reducing Antioxidant Power (FRAP) assay. In addition to this, GC-MS analysis was also performed for the identification of volatile constituents of Methanol Extract of Stems (MES) and Methanol Extract of Roots (MER). Results: The MES and Dimethylsulfoxide Extract of Stems (DES) were found to have higher Chlorophyll a (Ch a) content in comparison to Chlorophyll b (Ch b) and Carotenoids (C ca). Carbohydrate profile showed that stems and roots have the highest content of fructose in comparison to other sugar moieties. The soxhlet method showed the highest percentage extractive yield in the stems as well as roots. Results revealed that the MES showed higher antioxidant potential as compared to the MER. A correlation has also found to exist between the results of TPC, TFC and antioxidant assays, since TPC and TFC are key constituents responsible for the antioxidant potential. Conclusion: These results have been found to suggest the richness of MES in natural phenols, flavonoids and antioxidants. Further, study should be conducted over identification and characterization of compounds present in the extract.

4This stress condition helps in increasing secondary metabolite production. 5 These plants have been used for medico-religious purposes which in turn help in generating income for local people.6 Out of several medicinal plants found in trans-Himalayas, Rhodiola imbricata frames to be a distinctive herb due to the presence of various phytopharmaceutical constituents which can be helpful for human kind. High Efficiency in vitro Plant Regeneration and Secondary Metabolite Quantification from Leaf Explants of Rhodiola imbricataABSTRACT Aim: Rhodiola imbricata is an endangered medicinal plant of the trans-Himalayan Leh-Ladakh region belonging to the family Crassulaceae. An efficient propagation and regeneration system via direct shoot organogenesis from leaf explant and evaluation of cinnamyl alcohol (Secondary metabolite) was established in this study. Material and Methods: In vitro grown leaves were inoculated using Murashige and Skoog (MS) medium supplemented with (alpha)-naphtalene acetic acid (NAA) in combination with 6-benzyladenine (BAP) for callus proliferation and regeneration. Results: The highest percentage of rhizogenous callus was induced in medium containing NAA (10.0-15.0 µM). The highest percentage of shoot formation from leaf derived callus was obtained in the medium containing of NAA (5.0 µM) and BAP (2.5 µM) as well as in NAA (1.0 µM), BAP (5.0 µM) (38.88% and 37.49%) respectively. Rooting of regenerated shoots were effective when a lower concentration of NAA (0.5 μM) was used alone. A maximum number of roots (22.0) and higher length (0.6 cm) was observed. The in-vitro plantlets with well-developed shoots and roots were acclimatized successfully to natural field conditions with a survival rate of over 80%. Cinnamyl alcohol (Secondary metabolite) evaluation was also done for the very first time and an upregulation of 49.6% and 30.6% were observed in in-vitro roots and shoots when compared with mother root and shoot respectively. Hence, it was proved that the content of secondary metabolites obtained from in-vitro raised plants is higher than mother plant. Conclusion : These results may lay a foundation for genetic improvement and can be used to determine sequential level of targeted secondary metabolites through cell culture in Rhodiola imbricata.