Indra D. Bhatt

Optimization extraction conditions for improving phenolic content and antioxidant activity in Berberis asiatica fruits using response surface methodology (RSM).

This study for the first time designed to optimize the extraction of phenolic compounds and antioxidant potential of Berberis asiatica fruits using response surface methodology (RSM). Solvent selection was done based on the preliminary experiments and a five-factors-three-level, Central Composite Design (CCD). Extraction temperature (X1), sample to solvent ratio (X3) and solvent concentration (X5) significantly affect response variables. The quadratic model well fitted for all the responses. Under optimal extraction conditions, the dried fruit sample mixed with 80% methanol having 3.0 pH in a ratio of 1:50 and the mixture was heated at 80 °C for 30 min; the measured parameters was found in accordance with the predicted values. High Performance Liquid Chromatography (HPLC) analysis at optimized condition reveals 6 phenolic compounds. The results suggest that optimization of the extraction conditions is critical for accurate quantification of phenolics and antioxidants in Berberis asiatica fruits, which may further be utilized for industrial extraction procedure.

Assessment of Antioxidant Properties in Fruits of Myrica esculenta: A Popular Wild Edible Species in Indian Himalayan Region

Crude extract of Myrica esculenta fruits, a wild edible species of Indian Himalayan Region, was evaluated for phenolic compounds and antioxidant properties. Results revealed significant variation in total phenolic and flavonoid contents across populations. Among populations, total phenolic content varied between 1.78 and 2.51 mg gallic acid equivalent/g fresh weight (fw) of fruits and total flavonoids ranged between 1.31 and 1.59 mg quercetin equivalent/g fw. Antioxidant activity determined by 2,2′-azinobis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging, 1,1-diphenyl-2-picrylhydrazyl radical scavenging and ferric reducing antioxidant power (FRAP) exhibited considerable antioxidant potential and showed significant positive correlation with total phenolic and total flavonoids content. High performance liquid chromatography analysis revealed significant variation (P <  .01) in phenolic compounds (i.e., gallic acid, catechin, hydroxybenzioc acid and ρ-coumaric acid) across populations. This study provides evidences to establish that consumption of M. esculenta fruits while providing relished taste would also help in reduction of free radicals. Therefore, this wild edible species deserves promotion in the region through horticulture and forestry interventions.

Micropropagation of Indian wild strawberry.

An efficient method of micropropagation based on an increased percentage survival of explants and reduced phenol-induced browning in wild strawberry has been developed. Serial transfer of nodal explants was carried out at 24-, 48- and 96-h intervals. Nodal segments cultured on Murashige and Skoog medium supplemented with 6-benzyladenine (4.0 μM) and α-naphthalene acetic acid (0.1 μM) gave the best (94.4%) explant establishment and shoot number (22.3) per explant. Of the cytokinins tested, 6-benzyladenine was found more effective than kinetin and N6-(γ,γ dimethylallyamino) purine. Excised shoots rooted on half-strength agar-gelled medium with 1.0 μM α-naphthalene acetic acid. Rooted shoots with fully expanded leaves acclimatized successfully and about 70% of plantlets survived ex vitro.

Factors controlling micropropagation of Myrica esculenta buch. – Ham. ex D. Don: a high value wild edible of Kumaun Himalaya

Various factors such as browning, season, media type and plant growth regulators influence the micropropagation of female trees of Myrica esculenta . These factors have successfully been addressed after addition of some media additives, collection of the explant at right season, standardizing the media type and use of plant growth regulators at varying concentrations. Polyvinylpyrollidone (PVP - 0.5%) was found effective for successful partial removal of phenolic compounds and obtaining maximum percent survival of explants. The explant collection season played an important role in reducing phenol induced browning and winter season was found best for explant establishment. The maximum number of shoots (4-5/explant) was obtained in Woody Plant Medium (WPM) supplemented with 10µM kinetin and 0.1µM naphthalene acetic acid (NAA). Of all the media types tried, Woody Plant Medium was found to be the best. Kinetin was found superior to benzyl amino purine and N 6 (γ,γ-dimethylallyamino)purine (2iP) for explant establishment and multiplication. NAA induced rooting to 45.8% of explants in 1/2 strength Woody Plant Medium. However, ex vitro survival percentage was low. Key Words: Browning, in vitro, plant regeneration, tissue culture, woody plant medium. African Journal of Biotechnology Vo l.3(10) 2004: 534-540

Habitat-dependent variations in berberine content of Berberis asiatica Roxb. ex. DC. in Kumaon, Western Himalaya.

The variation of the berberine content in roots and stem bark of Berberis asiatica with altitude and edaphic conditions in the western Himalaya was estimated by HPLC. The comparative assessment revealed a significantly higher berberine content in roots than in stem barks. Moreover, the berberine content varied significantly with altitude and edaphic conditions both in root and stem bark samples. The populations growing at low altitude contained significantly more berberine than the ones growing at high altitude. Also the moisture and potassium (K) percentage of the soil significantly influenced the berberine content.

Micropropagation of Pittosporum napaulensis (DC.) Rehder & Wilson - a rare, endemic Himalayan medicinal tree.

An in vitro propagation protocol has been developed from mature trees of Pittosporum napaulensis. The best bud proliferation (83.1%), shoot number (21 axillary shoots/ explant) and shoot length (5.5 cm) was achieved in Murashige and Skoog (MS) medium supplemented with 5.0 μM N−6 benzyladenine and 0.1 μM α- naphthalene acetic acid. Of the three cytokinins tested (N−6 benzyladenine, kinetin and thidiazuron), N−6 benzyladenine proved to be the best for shoot induction. Shoot regeneration potential varied among genotypes. Regenerated shoots rooted after 48 hours treatment on half-strength MS liquid medium supplemented with 20 μM indole-3-butyric acid. Rooted shoots transferred to 120 g (w/v) soilrite + sand + soil (1:1:1) mixture showed 70% survival. Twenty-one plantlets are growing well in green house conditions.

Assessment of Nutritional and Antioxidant Potential of Selected Vitality Strengthening Himalayan Medicinal Plants

Three Himalayan medicinal plants (Habenaria intermedia, H. edgeworthii, and Roscoea procera), widely used in vitality strengthening Ayurvedic formulations in India, were assessed for nutritional phytochemical constituents, and antioxidant activity. These target species emerged as a good source of minerals and possessed important micro elements. Individually, H. intermedia contained a high content of total phenols, thiamins, tannins, and calcium; R. procera was rich in potassium and iron content; and H. edgeworthii emerged as a good source of sodium. While various antioxidant assays provided evidences on the antioxidant potential of target species, greater antioxidant potential of H. intermedia as compared to the other two species was revealing. This study, therefore, highlighted the possibilities of harnessing nutritional and antioxidant potential of these species.

The availability, fruit yield, and harvest of Myrica esculenta in Kumaun (West Himalaya), India.

Abstract Myrica esculenta Buch.-Ham. ex D. Don is a popular, potentially income-generating wild edible in the Indian Himalaya. The species prefers Pinus roxburghii Sarg., Quercus leucotrichophora A. Cam., and mixed Quercus forests, contributing 15–26% of total tree density in the forests. It performs best in Pinus roxburghii forests, where its density correlates with Pinus tree biomass. The regeneration of Myrica is poor in all the habitats. However, recruitment of species increases consistently from abundant Myrica to no-Myrica stands. The fruit yield increases with tree size category and differs between habitats. The potential yield at different sites is 2.0–4.2 tonnes/ha, of which 2.8–7.2% is harvested for income generation. The income generated from Myrica fruit is significant, considering the regional annual per capita income. The possible impact of fruit harvesting and other disturbance factors on the regeneration of the species is discussed. There are significant options for enhancing the income-generating potential through value addition.

Combined effect of cytokinins on multiple shoot production from cotyledonary node explants of Bauhinia vahlii

Using seedling explants, an improved regeneration protocol was developed for Bauhinia vahlii. A combination of thidiazuron and kinetin (1.0 μM each) increased the number of shoots significantly (p > 0.05) up to four successive subculture cycles. Over 83% shoots rooted on one-fourth strength Murashige and Skoog (MS) medium supplemented with 1.0 μM α-naphthaleneacetic acid (NAA). Fifty percent of plantlets (15 No.) successfully acclimatized in 90 g (w/v) soilrite + sand + soil (2:1:1) in the shed house. Preconditioning at different sucrose concentrations prior to acclimatization showed no effect on percent survival but improved plant quality.

Characterization of Agro-diversity by Seed Storage Protein Electrophoresis: Focus on Rice Germplasm from Uttarakhand Himalaya, India

Abstract The characteristics of 48 rice varieties from Uttarakhand Himalaya, India were detected by morphological and biochemical markers. The grains of the selected rice varieties varied in their morphological (grain length, grain width and grain weight) and biochemical characters (sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE). Based on the presence of 70, 65, 60, 57, 37–39, 22–23, 13 and 10 kDa protein bands in the 48 rice varieties, seven types of profiles were identified. An unweighted pair group average method with arithmetic mean (UPGMA) dendrogram based on cluster analysis of genetic similarity of the protein bands showed two distinct groups with 1%–78% similarity coefficients. The presence of characteristic bands in selected varieties is a useful parameter for identification of rice germplasm.