Callus Induction, Proliferation, Enhanced Secondary Metabolites Production and Antioxidants Activity of Salvia moorcroftiana L. as Influenced by Combinations of Auxin, Cytokinin and Melatonin

Abstract: Tissue culture technique is one of the best methods to reproduce salvia plant Therefore, the aim of this research was to enhance the in-vitro callus proliferation and production of secondary metabolites of S. moorcroftiana using different combinations of auxin, cytokinin and melatonin. Initially, callus induction was optimized using indole acetic acid (IAA), 2, 4-dichlorophenoxy acetic acid (2,4-D), and naphthalene acetic acid (NAA) applied at different concentrations in combination with 1 mg L-1 of 6-benzylaminopurine (BAP). The results indicates that earliest days to callus induction (14.67 days) was occurred in the media fortified with 2, 4-D+BAP (2.0+1.0 mgL-1). Whereas the highest callus initiation (100%) was induced on MS medium incorporated with 2,4-D+BAP (1+1mgL-1). Furthermore, maximum fresh weight was obtained when 2,4- D + BAP at the rate of (1+ 1mg L-1) was incorporated and dry weight was attained when 2,4- D + BAP at the rate of (2+1 mg L-1) was added to MS media. The maximum fresh and dry weight was obtained when melatonin at rate of 1.5 mg L-1 was supplemented with MS media including 2,4-D + BAP (1+1mg L-1), moreover the maximum DPPH scavenging activity, total phenolic and flavonoid content was noted when supplemented with melatonin at rate of 1.5 mg L-1. In conclusion, among various concentrations of plant growth regulators, 2,4- D + BAP at the rate of (1+ 1mg L-1) along with 1.5 g L-1 melatonin was the best for callus growth and production of secondary metabolites of S. moorcroftiana.

Palyno-morphological characterization of selected allergenic taxa of family Poaceae from Islamabad-Pakistan using microscopic techniques.

Statistics around the world show an increased risk of high allergies associated with plants. The role of allergenic pollen is now well known. The most serious allergenic diseases are asthma and rhinitis, which affect 5-30% of the population of industrialized countries. This work focuses on the exploration of allergenic grass species of Islamabad, Pakistan and use microscopic techniques to correctly identify them. This study contains 11 species of grasses which showed variations in pollen morphological features. The dominant pollen shape was prolate, prolate-spheroidal and sub oblate. The largest polar diameter was calculated for Echinochloa crus-galli (56.00 µm) while the largest equatorial diameter was found in Cenchrus ciliaris (45.00 µm). Exine ornamentation varied from Gemmate-verrucate, Scabrate-reugulate, Verrucate-reticulate, Gemmate-Scabrate, Fine-Gemmate, Gemmate-rugulate, Gemmate-reticulate, Gemmate-scabrate, Scabrate-verrucate, and Scabrate. Maximum exine thickness was found in Eleusine indica (4.90 µm) while in C. ciliaris (1.50 µm) the minimum thickness of exine was measured. In E. crus-galli (1.54 µm), the largest P to E value was measured and the smallest value was noticed in Setaria verticillata (0.86 µm). The study was performed with an aim to identify the allergenic plant species in Islamabad-Pakistan using pollen morphological studies. This study provides significant information, to arrange the pollen types within their defined classes and to discuss various pollen features for developing an allergenic pollen atlas.

Phytochemical analysis and wound healing studies on ethnomedicinally important plant Malva neglecta Wallr.

ETHNOPHARMACOLOGICAL RELEVENCE: The use of herbal medicines is increasing in developed countries as alternative and/or supportive therapy to conventional health care medicines. Malva neglecta Wallr. (Family: Malvaceae) has been reported as wound healing remedy in traditional medicines, however no experimental data is available on its wound healing potentials. The aim of this study was to explore phytochemistry and validate wound healing potentials of the plant using animal models. MATERIALS AND METHODS: M. neglecta crude methanolic extract (Mn.Cme) was chemically characterized using HPLC-DAD and GCMS analysis. Acute dermal toxicity was determined in albino rats following Organization of Economic Co-operation and Development (OECD) 402 established standards. Wound healing potentials were evaluated in rats using excision wound model. Wounds (177 mm2) were made by an excision on the skin of rats which were placed individually in cages. Mn.Cme was formulated in ointment form and was applied topically onto the wound area once daily for 14 days. The wound area was measured with translucent paper and thereafter estimated on a 1 mm2 graph sheet every 3rd day until epithelialization and complete wound closure was recorded. Wound contraction was calculated as a percentage of the original wound size. Antioxidant potentials were also evaluated via FRAP, DPPH and H2O2 free radicals scavenging assays. RESULTS: HPLC-DAD analysis revealed 25 phenolic compounds with higher amounts of hydrotyrosol (109.3 mg/g), coumaroylhexoside (97.4 mg/g), kaempferol-3-(p-coumaroyldiglucoside)-7-glucoside (37.2 mg/g), quercetin-3-O-rutinoside (31.5 mg/g) and epicatechin-3-O-(4-O-methyl)-gallate (31.3 mg/g). In GC-MS analysis, oleic acid (19.67%), taurine (17.60%), ethylene dimercaptan (14.67%), isoeugenol (14.61%), patchoulane (10.36%), methyl 12-methyltetradecanoate (8.47%) and isopropyl myristate (7.02%) were highly abundant compounds. No sign of toxicity was observed in the acute dermal toxicity test. Our test sample (Mn.Cme) exhibited considerable wound healing tendency at all doses (1 g, 1.5 g, 2 g per 10 g of ointment base) with reduced epithelialization period in a dose-related manner. Absolute healing was observed after application of 2 g of Mn.Cme ointment. Further, Mn.Cme exhibited considerable anti-radical potential in all assays. CONCLUSION: It may be concluded that M. neglecta possess very potent secondary metabolites which are previously reported for wound healing potentials. The plant has considerable antioxidant and wound healing properties and thus warrant further studies to uncover the molecular mechanism its wound healing potentials.

Lawsonia Inermis Markedly Improves Cognitive Functions in Animal Models and Modulate Oxidative Stress Markers in the Brain.

Background and Objective: Medicinal plants represent an important source of alternative medicine for the management of various diseases. The present study was undertaken to assess the potential of Lawsonia inermis ethanol (Li.Et) and chloroform (Li.Chf) extracts as memory-enhancing agents in experimental animals. Materials and Methods: Li.Et and Li.Chf were phytochemically characterized via gas chromatography-mass spectroscopy (GC-MS). Samples were tested for nootropic potentials at doses of 25, 50, 100, 200 mg/kg (per oral in experimental animals (p.o.)). Swiss albino mice of either sex (n = 210) were divided into 21 × 10 groups for each animal model. Memory-enhancing potentials of the samples were assessed using two methods including "without inducing amnesia" and "induction of amnesia" by administration of diazepam (1 mg/kg, intraperitoneally. Piracetam at 400 mg/kg (i.p.) was used as positive control. Cognitive behavioral models including elevated plus maze (EPM) and the passive shock avoidance (PSA) paradigm were used. Biochemical markers of oxidative stress such as glutathione (GSH), catalase (CAT), superoxide dismutase (SOD) levels were analyzed in the brain tissue of treated mice. Results: In 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals scavenging assay, Li.Et and Li.Chf exhibited 70.98 ± 1.56 and 66.99 ± 1.76% inhibitions respectively at 1.28 mg/mL concentration. GCMS results revealed the presence of important phytochemicals. Both samples (Li.Et and Li.Chf) at 25 mg/kg (p.o.) dose significantly (p < 0.05) improved learning and memory as indicated by decline in transfer latency and increase in step down latency in EPM and PSA models respectively. Li.Et and Li.Chf at 25 mg/kg (p.o.) showed considerable increase in GSH (2.75 ± 0.018 ***), SOD (2.61 ± 0.059 ***) and CAT (2.71 ± 0.049 ***) levels as compared to positive and negative control groups. Conclusions: This study provides the preliminary clue that L. inermis may be a potential source of memory-enhancing and anti-oxidant compounds and thus warrant further studies.