In the present study phytosociological, ethnobotanical, pharmacognostic, physicochemical and pharmacological study of Iphiona grantioides (Boiss.) Anderb. and Pluchea arguta (Boiss.) subsp. glabra Qaiser of family asteraceae was carried out. Seven different localities in District Karak of Khyber Pukhtunkhwah, Pakistan were selected for phytosociological study of Iphiona grantioides and Pluchea arguta subsp. glabra Qaiser (Fig. 4.6). Both the plants were widely growning in the highly saline area of district Karak i.e. Andai (along road side), Sour Daag, Noshpa, Bahader Khail (Near old tunnel), Naripanoos (Near algada), Malgeen (Salt Deposit) and Karat (Salt Deposit), Karak at an elevation ranging from 450 m to 900 m. In the present study Iphiona grantioides and Pluchea arguta subsp. glabra, the two research plants, were present in close association of each other along with other 80 plants, in all the seven localities. Iphiona grantioides was found dominant at two sites, while it was second dominant at Karat ghar site and third dominant at Naripanoos site , with average importance value (IV) 21.2, and maximum IV 49.9, at Chashmai area of Andai, while minimum IV was 7.1 at Malgeen area. Important value index (IVI) of Iphiona grantioides was 7.05 with maximum IVI 16.6 and minimum IVI 2.3. Pluchea arguta subsp. glabra was found in all the seven sites, mostly growing in close association with Iphiona grantioides. It was 1st dominant at Bahader Khail sit near the old tunnel, and second dominant at Chasmai and third dominant at Malgeen, Karat and Noshpa area, having the average IV of 13.2, maximum IV of 28.3 and minimum IV of 7.1. Important value index (IVI) of Pluchea arguta subsp. glabra was 4.4, maximum IVI 9.4 and minimum IVI 1.76. Soil analysis revealed that all the soil samples analysed from different areas of District Karak were alkaline in reaction and strongly calcareous in nature. Organic matter, soil N and P were found deficient in most of area sampled. The sampled area soil had permanent salt with semi-arid climate. Pharmacognostic study of Iphiona grantioides and Pluchea arguta subsp. glabra Qaiser was carried out ,which includedmacroscopical description of leaf,stem and root of Iphiona grantioides and Pluchea arguta subsp. glabra, microscopic study which includes anatomical studies of leaf, stem and root, leaf suface study , determination of leaf constant values(Stomatal index,vein islet no. and Palisad ratio), physiochemical characteristics (Moisture contents and ash values),flourecence study, phytochemical investigation and extractive values of powder drugs of leaf,stem and roots of both the plants were determined, while ethanolic extracts of different parts of the two plants were screened out for various bioassays. The leaf of Iphiona grantioides is sucuulent in nature, pubescent, thickly covered with glandular trichomes and nonglandular hairs on both the surfaces. The leaf is amphistomatic, stomata being present on both the epidermises and the stomatal apparatus is of anomocytic type. Anatomical studies showed that both the (upper and lower) epidermises are are covered with waxy cuticle and multicellular non glandular and glandular trichomes. Pallisade cells have chloroplast and oil droplets while spongy mesophyll cells have cuboid crystals of calcium oxalate. In the midrib region the midvein is very prominent, having 1-2 vascular bundles, with sclerenchymatic cells on the lower side. Similarly leaf of Pluchea arguta subsp. glabra Qaiser was sessile, spirally arranged, almost persistant and green, herbaceous, glabrous. Lamina was simple with serrate (dentate) margins. Leaf was bitter; unplasant in taste with strong, pungent smell. Leaf anatomy of Pluchea arguta subsp.glabra. Qaiser revealed the presence of single layered upper and lower epidermises and glandular trichomes, which are sessile as well as with multicellular stalk (2-5 cells). The leaf has typical bifacial structure , with vascular system and mesophyll tissue comprises of palisade and spongy parenchymas cells containg the latter is oil globules. In the midrib region collenchymatous tissues is present below the both upper and lower epidermises and there are 3 vascular bundles in the midribVascular bundles are colletral and closed. Xylem vessels are with spiral wall thickening. Palisade ratio of Iphiona grantioides, in the present study, ranged from 5 to 6 to 6.75 .Vein iselet and vein termination number were 8 to 10 (9) and 7 to 10 (8) per mm2 respectively. The vein-islets were quite distinct as squaresh, elongated or polygonal shaped, internally provided with many forked and vascular branches, while stomatal number and stomatal index value for Iphiona grantioides was 120 to 150 (130) and 12 to 15 (13) per mm2 respectively. Pluchea arguta subsp.glabra leaf has the Palisade ratio from 6 to 7 (7.5) .Vein iselet and vein termination numbers were 10 to 12 (11) and 6 to 9 (8) per mm2 respectively while stomatal number and stomatal index value was in the range of 110 to 160 (130) and 10 to 12 (11) per mm2, respectively. Ash analysis for crude powder drug of leaf, stem, root and flower of Iphiona grantioides was carried out. Highest value of total ash was recorded for stem (7.06%) and least for leaves (3.33%) ,while roots has 4.25% total ashes and flower has 3.38% total ash value. Acid insoluble ash was in the range of 1.54% (leaf), 1.425% (root), 1.27% (stem) and 0.35% (flower). Water soluble ash was in the range of 4.36 % (leaves), 3.34% (stem), 2.23 %( roots) and 1.82% (flower). Moisture contents was heighst (9.16%) in root, 9.01 % ( stem), 7.80% (leaf) and 6.12% (flower). Similarly Ash analysis of Pluchea arguta subsp. glabra for crude powder drug of leaf, stem and root was carried out. Stem has total ash value of 7.82 %, 6.12 % to 5.6%. Acid insoluble ash was in the range of 1.66%, 1.09 % and 1.49 %. Water soluble ash was in the range of 2.56%, 3.38% and 6.5%, while moisture contents was heighst in leaves (8.87%), root (8.41 %) and lowest in stem (7.5%). Qualitative phytochemical screening of Iphiona grantioides and Pluchea arguta subsp. glabra revealed that proteins, carbohydrates, tannins, alkaloids, flavonoids, saponins, glycosides, phytosterol and tri-terphenoids and volatile oil were detected in aqueous and ethanolic extracts of various parts of both the plants. In the present study seven different solvents including ethanol, methanol, chloroform, acetone, distilled water, butanol and n-hexane were used for percent extractive values determination of different parts ( leaf , flower, root and shoot f Iphiona grantioides and leaf, shoot and root of Pluchea arguta subsp. glabra). Highest extractive values were given by Iphiona grantioides leaf i.e., (Aqueous extract (46.2%), Ethanol (37%), Butane (36%), Hexane (22.8%), Ethyl acetate (13.75) followed by root extracts i.e., Acetone (20%), Chloroform (20%) , Ethanol (19 %) and aqueous extract (16.6%). Similarly highest extractive values were given by leaves of Pluchea arguta subsp.glabra. The values are as follow, Ethyl acetate (30%), Aqueous (27.2%), Ethanol (22.2 %), and Hexane (20.75%). In the present study leaf, shoot, root and flower of Iphiona grantioides and Pluchea arguta subsp. glabra were analyzed to determine the concentration of various heavy and toxic metals including Mn, Fe, Cu, Zn, Cr, Pb, Co, Ni and Cd in these plants. Obtained results showed that both the plants have all the elements within safer and permissible limits. In the present study various bioassays were carried out to investigate the Iphiona grantioides and Pluchea arguta subsp. glabra for their curative values. Crude ethanolic extracts of Iphiona grantioides (flower, stem,leaf,root) and Pluchea arguta subsp. glabra (stem,leaf,root) were tested against 4 human pathogenic fungal strains (Trichophyton longifusis, Candida albicans, Microsporum canis, Candida glaberata), 2 animal pathogenic fungi (Aspergilus flavus , Aspergilus niger) and the 3 Plant pathogenic types (Fusarium solani, Mucor spp. and Alternaria spp.) for antifungal activity. The flower and leaf extract of Iphiona grantioides was the most effective extracts ,as shown antifungal against all the nine species at all the three (125, 250 and 500 ug/ml) doses. Similarly the crude ethanolic extract (500 ug /ml) of stem, leaves and roots had inhibitory effects against the fungal strains in the order of leaves (43.6%) ˃ stem (35%) ˃ root (30%). Antibacterial activity of ethanolic extract of various parts of Iphiona grantioides and Pluchea arguta subsp. glabra was tested against Escherichia coli, Staphylococcus viridans, Klebsiela pneumonia. B.licheniformis, Pasteurella multocida, Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella typhi and Micrococcus luteus. The results of antibacterial activity of different parts of the two plants showed that ethanolic extract of Iphiona grantioides flower was most effective against the tested microorgainism, with the highest inhibition zone of 21. 23 mm (Klebsiela pneumoniae) followed by 19.67 mm (Staphylococcus aureus ), 17.47 mm (Pasteurella multocida), 16.76 mm (Bacillus licheniformis), 15.67 mm (Micrococcus luteus), 15.00 mm (Staphylococcus viridans) while it exhibited moderate antibacterial effects against by producing zone of inhibition of 12.33 mm against Bacillus Subtilis and Pseudomonas aeroginosa. Ethanolic extract of leaves of Iphiona grantioides showed significant antibacterial effect against Escherichia coli (14 mm) while other extracts including leaf, stem and root, showed moderate antibacterial potential, in the range from 8.32 - 11.67 mm. Ethanolic extract of Pluchea arguta subsp. glabra leaf was comparatively more potent than the other two extracts i.e., stem and root, in producing prominent zone of inhibitions that is 15.33 mm (Escherichia coli), 13.33 mm (Pseudomonas aeroginosa) and 12.00 mm (Micrococcus luteus) while the lowest zone of inhibition (10.33mm) was observed in Micrococcus luteus and Pasteurella multocida by root and stem extracts , respectively. leaf ethanolic extract of Iphiona grantioides has produced an excellent phytotoxic activity against L. minor at the highest tested concentration (1000 ug/ml ) by causing (91.5%) inhibition of Lemna minor. Similarly flower extract also caused 87.2% inhibition. Pluchea arguta subsp. glabra leaf extract caused 73. 2 % inhibition, while the stem caused 70.2% inhibition. Moderate inhibitions (61.7%) were observed for the ethanolic extracts of Iphiona and Pluchea roots. Lowest inhibition (59.6%) was caused by stem extract of Iphiona grantioides at concentration 1000 ug/ml). Ethanolic extracts of flower, leaf of Iphiona grantioides showed excellent insecticidal (90%) activity against Callosobruchus analis and Rhyzopertha dominica at 10% dose concentration, respectively. Besides this, flower extract also significantly expressed insecticidal activity against Sitophilus oryzae (82%) and Tribolium casteneum (80%). Moderate lethal activity was shown by stem, leaf, flower and root extracts (in the range of 60% to 66%) against Tribolium casteneum, Sitophilus oryzae, Rhyzopertha dominica, Trogoderma granarium and Callosobruchus analis.Week inhibitory ativity (in the range of 22%-48%) was exhibited by stem, root and leaves extract against almost all the tested species except.Stem and roots of Pluchea arguta subsp. glabra displayed week inhibitory ativity against Tribolium casteneum in the order of 28% and 62%, respectively. Ethanolic extract of Pluchea arguta subsp. glabra leaf exhibited good mortality activity against Rhyzopertha dominica , Tribolium casteneum , Sitophilus oryzae , Trogoderma granarium and Callosobruchus analis in the order of 100%, 90%, 80%, 80% and 70%, respectively. The extract obtained from the stem and root exhibited a moderate (50%- 60%) inhibitory activity against Rhyzopertha dominica and Callosobruchus analis and weak activity (10% - 40%) was shown by rest of the extracts against all tested insects. Arerial parts of two plants were used for cytotoxic bioassay. The extracts displayed significant toxicity against brine shrimps. The results of the present study showed that roots of Pluchea arguta subsp. glabra the showed highest DPPH scavenging activity (82.89%) followed by ethanol extracts of leaf (77.3%) and flower (70.94 %) of Iphiona grantioides. The anthelmintic activity of Iphiona grantioides and Pluchea arguta subsp. glabra were very significant against the tested worms. Leaf extracts of Iphiona grantioides and P. arguta subsp. glabra at dose of 100 mg/ml caused death of the worm in 3.33 ±0.57 and 2.16±0.28 min, respectively. Moderate mortality time of root and stem of Iphiona grantioides was 12.43 ±0.60 and 13.83 ±0.76, respectively. In addition, weak nematocidal activity of death time of 50 ±5 was exhibited by flower extract of Iphiona grantioides. The present study has exposed the enormous and varied medicinal properties of Iphiona grantioides and Pluchea arguta subsp. glabra and both of these plants can be exploited for curative preparations on marketable level." xml:lang="en_US
