In vitro regeneration approaches for restoration of Ceropegia mohanramii-an endemic and critically endangered asclepiad.

The study aimed to develop an efficient, rapid, and large-scale in vitro regeneration system for propagation, conservation, and restoration of an endemic and critically endangered herb, Ceropegia mohanramii. The cultures were established using nodal explants on Murashige and Skoog's (MS) medium supplemented with 6-benzylaminopurine (BAP: 1.0 mg/l). Nodal buds cultured on MS medium supplemented with BAP (2.0 mg/l) along with indole-3-butyric acid (IBA, 0.5 mg/l) resulted with production of maximum number of shoots (17.1 ± 1.2) in hundred percent of the cultures. MS medium supplemented with BAP (2.0 mg/l) along with diverse concentrations of indole-3acetic acid (IAA) promoted the in vitro flowering. In vitro regenerated shoots were transferred to one-half MS medium fortified with singular supplementation of auxins, where IBA (1.5 mg/l) served optimal for production of maximum number of roots (5.7 ± 0.6). In vitro derived plantlets were hardened under controlled conditions in a glasshouse and subsequently transferred to soil. Over 1200 saplings were transplanted to eight different localities of the Western Ghats where over 76% survival is recorded after 1 year of transplantation.

Highly efficient in vitro regeneration, establishment of callus and cell suspension cultures and RAPD analysis of regenerants of Swertia lawii Burkill.

Highly efficient in vitro regeneration system has been developed for Swertia lawii Burkill, an important herb used as substitute for Swertia chirayita. Shoot tips explants were cultured on MS medium with various phytohormones for multiple shoot production. The best shoot production frequency (100%) and maximum shoots (10.4 ± 0.8) were obtained on MS media containing TDZ (3.0 mg l-1) in combination with IBA (0.3 mg l-1). Maximum callus induction (95 ± 4.8%) and callus growth (1.7 ± 0.4 gm) was achieved on MS medium with 2, 4-D (3.0 mg l-1). Cell suspension cultures were established and studied for their growth kinetics. Shoots were rooted best (22.1 ± 2.5) in 1/2 MS medium with IAA (3.0 mg l-1). The genetic uniformity of the micropropagated clones was assessed using RAPD markers. Out of 405 bands, 400 (98.76%) were monomorphic and rest 5 (1.24%) were polymorphic. High multiplication frequency and low risk of genetic instability ensures the efficacy of this protocol.

Phytochemical composition, antioxidant activity and HPLC profiles of Swertia species from Western Ghats.

Swertia chirayita is one of the potential medicinal plants of the family Gentianaceae in traditional medicine. Due to its high demand and scarcity, trade of chirayita is affected by adulterants. Swertia species from Western Ghats were compared with S. chirayita for phytochemical characterisation and antioxidant activities by using different extracts. This study revealed that acetone is the best extraction solvent of phenolic and flavonoid compounds with antioxidant properties as compared with other extracts. S. chirayita showed better antioxidant activity than other species with highest content of phenolics and flavonoids. Among the species from Western Ghats, Swertia minor has better antioxidant properties with higher content of phenolics and flavonoids when compared with S. chirayita. Gallic acid was detected in all species under study by using HPLC analysis. The Swertia species under study showed similar phytochemical properties and antioxidant potential and hence their use as substitute to S. chirayita needs to be further investigated.

Seasonal variation in content of camptothecin from the bark of Nothapodytes nimmoniana (Grah.) Mabb., using HPLC analysis.

OBJECTIVE: To study and compare seasonal variation in camptothecin (CPT) content from bark samples of Nothapodytes nimmoniana obtained from geographically and climatologically isolated populations. METHODS: A standard High Performance Liquid Chromatography methodology was used to analyze and quantify CPT from bark samples of N. nimmoniana. RESULTS: Sample collected from Amboli yielded highest CPT content 1.337 g/100 g dry bark powder during the monsoon compared to other localities in study. Monsoon (August) showed to accumulate higher levels of CPT in barks of N. nimmoniana as compared to summer (May). Amboli averaged highest accumulation of CPT compared to other localities under study. CONCLUSION: These findings indicate season to have control over accumulation of CPT. Locality Amboli has highest CPT content in all seasons and were the elite population during the study. The study also suggests the need for further investigation in lights of biosynthesis in the plant.

Free amino acid profiling in grain amaranth using LC-MS/MS.

Reliable and reproducible techniques for identification and quantification of amino acids usually require derivatization. However, techniques such as LC-MS/MS may perhaps sideline the derivatization with significant accuracy. Understanding the interaction between free amino acids and several physiological processes apparently requires that the pool be studied in the seed itself. Data on free amino acid content of grain amaranth is very inadequate. The present analysis revealed a higher concentration of essential amino acids especially Threonine and Tryptophan than non essential amino acids. The study constitutes first report of use of LC-MS/MS method for analysing the free amino acid composition of amaranth grain.