Maintaining the artemisinin content through direct and indirect in vitro regeneration and their assessment of variations with the field grown mother plants of Artemisia annua L.

An efficient protocol for maintaining the artemisinin content in tissue culture and high frequency of in vitro direct and indirect regenerations of multiple shoots of high artemisinin yielding genotypes of Artemisia annua has been developed and their comparison with field grown mother plant has been carried out.The eleven elite genotypes(containing more than 1%artemisinin)were tested on seven different modified media formulations.Modified half MS(Murashige and Skoog’s)media containing 100 mg L^(-1) myo-inositol,0.5 g L^(-1) casine hydrolysate,5 mg L^(-1) biotin,2 mL L^(-1) RT(Revised Tobacco)vitamin stock,0.5 mg L^(-1) BAP and 0.01 mg L^(-1) NAA showed best regeneration while,above modified MS medium containing 0.2 mg L^(-1) BAP and 0.2 mg L^(-1) NAA showed maximum shoot multiplication with maintained artemisinin content.Based on the chemical profiles of both the systems,minor difference was observed in their artemisinin content.A large scale culture of these plants maintained the normal growth index along with the artemisinin content and could be a better alternative to maintain the high artemisinin yielding genotypes with their genetic constraint in specific media combinations and also used as base material for further genetic improvement.

Genetic Variability and Elite Line Selection for High Essential Oil and Nepetalactone Content in Catmint (<i>Nepeta cataria</i>L.)

Nepeta cataria L., commonly known as catmint or catnip, belongs to the family “Lamiaceae” and is indigenous to Europe and Asia. The essential oil of this species is known for the richness and diversity of nepetalactones (NPL) which are used as mosquito/insect repellents in perfumery and cosmetic industries. Reports on Indian catmint germplasm are very meager and warrants exploration of its commercial potential as a natural, non-toxic source of insect repellents. With this objective, commercial open-pollinated seeds of catmint collected from its native, temperate habitat in Himalayas were introduced in the tropical plains. Subsequent to adaptation to a new zone we were able to isolate nineteen individual plants based on plant growth. Hydrodistillation of the fresh herb yielded essential oil in the range of 0.01% to 0.2%. Gas Chromatography (GC) and GC-Mass Spectrometry (GC-MS) analyses of the oil revealed the dominance of monoterpene hydrocarbon, namely, 4aα,7α,7aα NPL (1) isomer (84%). The other two isomers of nepetalactone, 4aα,7α,7aβ NPL (2) and 4aα,7β,7aα NPL (3) were also present, although in very less amounts (1.0% and 1.6%, respectively). Sesquiterpenes identified were α-humulene (traces), (E)-caryophyllene (0.6%) and caryophyllene oxide (1.7%). We compared the identified Indian catmint chemotype with the other oils from temperate, sub-tropical and tropical locations based on literature search. The Indian chemotype was found to be similar to the oils from Burundi, France, Turkey, UK and USA, mainly due to more accumulation of 4aα,7α,7aα NPL (1) isomer. These oils grouped together in Principal Component Analysis. Breeding lines are presently being developed to improve yield related traits in this plant. Multidisciplinary R&D efforts along with setting up industry related guidelines are required to successfully commercialize catmint cultivation. Several species of Nepeta genus have high nepetalactone content too and their potential as a commercial source of these isomers still needs to be explored.

Insect feeding deterrent and growth inhibitory activities of scopoletin isolated from Artemisia annua against Spilarctia obliqua （Lepidoptera： Noctuidae）

Artemisia annua （Asteraceae） is well known for its antimalarial activities due to presence of the compound artemisinin. We isolated a methoxy coumarin from the stem part of A. annua and confirmed its identity as scopoletin through mass spectral data. The structure was established from 1H-nuclear magnetic resonance （NMR）, 13C-NMR. The compound scopoletin was evaluated for its feeding deterrence and growth inhibitory potential against a noxious lepidopteran insect, Spilartctia obliqua Walker. Scopoletin gave FD50（feeding deterrence of 50%） value of 96.7 μg/g diet when mixed into artificial diet. S. obliqua larvae （12-day-old） exposed to the highest concentration （250μg/g diet） of scopoletin showed 77.1% feeding-deterrence. In a growth inhibitory assay, scopoletin provided 116.9% growth inhibition at the highest dose of 250μg/g diet with a GI50 （growth inhibition of 50%） value of 20.9μg/g diet. Statistical analysis showed a concentrationdependent dose response relationship toward both feeding deterrent and growth inhibitory activities. Artemisinin is found mainly in the leaves of A. annua and not in the stems, which are typically discarded as waste. Therefore identification of scopoletin in stems of A. annua may be important as a source of this material for pest control.

Comparative assessment for removal of anionic dye from water by different waste-derived biochar vis a vis reusability of generated sludge

In this study,four biochars prepared from different crop residue waste i.e.sugarcane bagasse(SBB),coconut shell(CNB),paddy straw(PDB),and distilled waste of lemongrass(LGB)were evaluated for removal of Remazol Brilliant Blue R from the aqueous system.The RBBR adsorption capacities of biochar were 97-79%for SBB,99.9-99.47%for CNB,66.1-48%for PDB,and 78-68%for LGB,dominantly controlled by their aromaticity and mineral content.The Langmuir and Freundlich isotherms and pseudo-second-order kinetic models have described the chemisorption of RBBR on biochar surfaces.The thermodynamic data suggested that adsorption was spontaneous and endothermic.These biochars demonstrated excellent reusability(till four cycles with 50-61%regeneration).The purified water and biochar dye sludge demonstrated no phytotoxicity.The findings obtained in this study may provide supports for the potential of biochars for anionic dye removal from water and utilization of generated sludge for zero waste-producing technologies in the future.