Diacytic Stomata Research Articles

Anatomic and Phytochemical Investigation of Herbal Tea Bags Sold as Lemon Balm (Melissa officinalis L.) in the Market

Melissa officinalis L., an ethnobotanically valuable plant, has been used for the treatment of several diseases since ancient times. However, different plants with the same name are sold instead of lemon balm in markets that sell herbal products in Turkey. For this purpose, 15 different brands of tea bags in crushed form, sold as lemon balm (Melissa officinalis L.) in markets, were analyzed. The total phenolic, flavonoid and antioxidant capacities of the hydroalcoholic extracts of these tea samples along with the stomatal structure were investigated. Additionally, chemical compositions and rosmarinic acid contents were determined by the high-performance thin layer chromatography method. Among the samples examined, S1, S2, S9 and S12 tea samples were found to meet the eligibility criteria. The leaves of these samples had diacytic stomata and the rosmarinic acid ratio in their phytochemical composition was over 2%. Furthermore, caffeic acid was detected in these samples.

STUDY ON ANATOMICAL CHARACTERIZATIONOF RHINACANTHUS NASUTUS (L.) KURZ

Rhinacanthus nasutus (L.) Kurz, belonging to the family Acanthaceae, is an herbaceous shrub. This plant holds significant value in traditional medicine. The leaves of R. nasutus are commercially available as tea bags, marketed as a natural remedy for various ailments. However, the conservation status of R. nasutus necessitates appropriate protective measures to ensure its sustainability. Anatomical studies provide various histological layers, including the epidermis, cortex, vascular bundles, and pith. The leaves feature numerous diacytic stomata and hypostomatic nature. The leaf midrib exhibits a narrow epidermis with numerous glandular hairs. The medulla contains arc-shaped vascular bundles and lateral accessory bundles, with the stele comprising linear xylem elements towards the inner side and phloem elements on the outer region. Transfusion tracheids are present between the palisade and spongy cells, with tissues containing large quantities of oil droplets. The petiole shows a narrow epidermis with numerous glandular hairs. The vascular region features a V-shaped stele. The stem reveals the stelar region is extensive and the inner region is rich in developed xylem elements with linear multiple vessels. The medullary ray cells are narrow, and the pith region displays intruded primary xylem with thick-walled parenchyma cells. Wood anatomical analysis shows xylem parenchyma cells that are paratracheal. The vessel elements possessing perforation plates and transverse walls with bordered pits. Tracheids have a narrow lumen, and the lateral walls contain simple pits. The wood demonstrates multiseriate ray parenchyma and the bark consists of phloem rays and narrow fibers. In conclusion, R. nasutus exhibits a variety of histological and anatomical characteristics that support its traditional medicinal uses. The detailed anatomical studies contribute to understanding the plants structural adaptations and potential environmental interactions, thereby facilitating its identification and utilization in herbal medicine. The conservation of this species is critical to preserving its medicinal value and ensuring its availability for future research and therapeutic applications.

Morpho-anatomical studies of family lamiaceae species of district Lahore, Punjab: a revision to flora of Pakistan

BackgroundThis study was aimed to determine the taxonomic position and delimitation of fifteen Lamiaceae taxa using leaf epidermal morpho-anatomical features in Lahore. A main objective of the study was also the revision and upgradation of Lamiaceae taxa in the flora of Pakistan, as no details of studied species are found in the flora of Pakistan.MethodsThe examination of significant anatomical parameters, such as epidermal cell shape and size, stomatal types, guard and subsidiary cells shape and size, stomatal cavity size, trichome size and shape, oil droplets, crystals, and secretory cavity characteristics were studied using light microscopic (LM) and scanning electron microscopic (SEM) techniques. Among all the studied Lamiaceae species, these anatomical features varied significantly. Principal component analysis and correlation were done to distinguish the species’ similarities.ResultsMost species had pentagonal and hexagonal epidermal cells with straight anticlinal wall thickness. On the adaxial surface, paracytic stomata were found in Ocimum basilicum L. and Rosmarinus officinalis L. Diacytic stomata was observed in Ajuga reptans L. and anisocytic stomata in Galeopsis tetrahit L. In the abaxial surface, trichomes were present in five species, i.e., Mentha suaveolens Ehrh. A. reptans, Thymus vulgaris L., M. haplocalyx, and Salvia splendens Ewat. In S. splendens, peltate and glandular trichomes were seen whereas, in other species, trichomes were long, unbranched glandular and had tapering ends. In adaxial side trichomes were present only in M. suaveolens, A. reptans, S. bazyntina, O. basciculum, S. splendens, S. officinalis, S. rosemarinus. In other species, trichomes were absent on the adaxial surface. In abaxial view, M. suaveolens had the largest length of trichomes, and O. basciculum had the smallest. S. splendens L. had the largest trichome width, while T. vulgaris had the smallest.ConclusionHence, according to these findings, morpho-anatomical traits are useful for identifying Lamiaceae taxa. Also, there is a need of upgradation and addition of studied taxa in flora of Pakistan comprehensively.

Studies on Various Taxonomic, Pharmacognostic and Phytochemical Properties of Sanchezia speciosa Leonard Acanthaceae

Sanchezia speciosa Leonard, Acanthaceae is used as ornamental plant. It used in folklore medicine in the management of pain, anti-microbial and insecticide. An investigation was carried out to evaluate the various taxonomic, pharmacognostic and phytochemical standards to ensure the identity, purity, safety and efficacy of the medicinal plant, S. speciosa. Various observations were recorded which included microscopy, micromeritics, fluorescence, chemomicroscopy, soluble extractive values, moisture contents and phytochemical analysis. The results indicated that the epidermal shapes were both polygonal on the adaxial and abaxial surfaces. Stomatal distribution was amphistomatic with anisocytic and diacytic stomata on the adaxial and abaxial surface, respectively. Stomatal index was 21.47% on the adaxial surface and 15.43% on the abaxial surface. Micromeritics results for the leaf powder were bulk volume of 25.33± 0.408, tapped volume of 19.00± 0.00, bulk density of 0.40± 0.006, tapped density of 0.53± 0.021, Hausner’s ratio of 1.34± 0.037, Carr’s index of 25.00± 2.550 and angle of repose of 43.07°. The micromeritics indicated a passable flow. The chemo microscopy indicated presence of lignin, mucilage and starch. Fluorescence properties showed different colours under different ultraviolet lights. The water-soluble, methanol-soluble, ethanol-soluble extractive values were 14.7% w/w, 4.3% w/w and 5.0% w/w, respectively. Moisture content was 8.5% w/w. Total ash, acid-insoluble and water-soluble ash values were 20.0 % w/w, 2.0% w/w and 4.2% w/w, respectively. GC-MS of methanol extract revealed the total of 39 phytochemicals with over 5 prominent peaks having higher area% which include hexadecanoic acid methyl ester (2.64%), 9,12,15-octadecatrienoic acid methyl ester (4.77%), phytol (9.98%), 9,12,15 octadecatrienoic acid (12.97%), Stigmasterol (2.00%) and Squalene (28.16%). Other compounds are glycerin (5.83%), n-hexadecanoic acid (2.85%) and 9 octadecenamide (2.18%). Many of them possess good pharmacological properties. The data generated from the present study would help to authenticate S. speciosa and also affirm its folklore use in traditional medicine which has potential for further development into drug product.

Microscopical, phytochemical, and LC/MS analysis of Ginkgo biloba leaves

Every medicinal practitioner knows Ginkgo as the plant source of an extract that is good for memory improvement. Ginkgo biloba extract is classified as one of the medicines in the treatment of dementia and social exclusion brought by vascular or neurodegenerative disorders. In such disorders, the extract was reported to be successful in improving symptoms such as depression, attention, memory disturbances, vertigo, tinnitus, and anxiety. The aerial part of Ginkgo bioloba was obtained from China, Utilizing fresh leaves allows for a microscopic inspection, the concentrated extracts by soxhlet were screened by standard methods for the qualitative investigation of secondary metabolites present in the plant, and a small quantity of extract was analyzed by LC/MS instrument. Microscopical examination shows diacytic stomata, helical vessels, fiber, and unicellular unbranched trichomes. Qualitative analysis is positive for tannin, glycoside, flavonoid, terpene, and phenolic compounds detected while saponin, coumarin, and alkaloid gave a negative result. While LC/MS shows important compounds that have important biological activities such as phenolic acids, flavonoids, and flavonoid glycosides which are reported for their different pharmacological activity, the Ginkgo plant is a promising drug that can help in the treatment of different diseases and required further studies.

Taxonomic characteristics of <i>Spinacia oleracea</i> L.

The research focused on the taxonomic study and proximate analysis of Spinacia oleracea L. The samples were fixed in formaldehyde acetic acid alcohol, dehydrated in alcohol solutions of 50%, 70% and 90%, sectioned, stained in 2% of Safranin O, counter stained in Alcian blue for 5 minutes, mounted in glycerine and photo-micrographed using Leica WILD MPS 52 camera on Leitz Draplan microscope. The proximate analysis was done using the Association of Official Analytical Chemists’ methods. The result revealed that spinach is glabrous and reach up to 4 to 8ft in length. The foliar organs are simple, ovate to ovate-triangular and alternate to rosette phyllotaxy. Margins are entire to slightly serrated, reticulate veined, up to 5 ± 4 cm in length and 3 ± 2 cm in width. The basal leaves are far larger than the ones towards the apical meristem. The inflorescence has spikelet of actinomorphic flowers pollinated by wind. The fruit is a berry/ drupe. The foliar epidermal study revealed presence of simple elongated, linear and glandular trichomes, and amphistomatic diacytic stomata. Anatomical study showcased a layer of epidermis, hypodermis of 2 to 3 rolls of collenchyma, general cortex of 7 to 10 rolls of parenchyma and the pith of parenchyma. The node is unilacunar with 2 lateral leaf traces and 1 leaf trace median. The proximate compositions of: 0.19 ± 0.00 Carbohydrate (%); 3.75 ± 0.00 Protein (%); 0.02 ± 0.00 Lipid (%); 94.74 ± 0.36 Moisture (%) content; 0.62 ± 0.32 Fiber (%) and 1.18 ± 0.03 Ash (%). These information would assist for further delimitation of the species.

Фармакогностическое исследование сырья Nepeta pannonica

Фармакогностический анализ лекарственного растительного сырья является важным этапом введения вида в официальное использование. Объектом исследования являлись надземные органы Nepetapannonica(семейство Lamiaceae), для которого выполнен макро- и микроскопический анализ. Определены следующие диагностические признаки на макроскопическом уровне: лист — форма листовой пластинки, тип жилкования; стебель — тип ветвления побега, окраска, супротивное расположение листьев, слабое опушение; соцветие — расположение цветков; чашечка цветка — форма зубцов, тип опушения, мелкая ребристость; венчик цветка — форма и размеры, окраска. В качестве микроскопических признаков сырья определены форма клеток эпидермиса верхней и нижней сторон, участков над жилками; диацитные устьица, наличие мелких многоклеточных трихом и крупных эфирномасличных железок в строении листа, стебля, чашечки и венчика цветка; разделение мезофилла на столбчатую и губчатую ткани на поперечном срезе листа.

Pharmacognostic Evaluation of the Leaves of Thurnbegia laevis Nees (Acanthaceae)

Thurnbegia laevis Nees. belong to the Acanthaceae family. It is used in folklore medicine for the management of diabetes, piles, joint pains as well as skin related problems. This study aims to evaluate the leaves of the plant by employing the quality control parameters. The leaves of T. laevis were collected, identified, air-dried, pulverized and stored in glass bottles. Standard procedures were used to carry out microscopy on the fresh leaves, micromeritics, chemomicroscopy, fluorescence analysis, soluble-extractive values, moisture contents and ash values using powdered samples of the leaf. Methanol extract of the leaf was used to conduct GC-MS analysis. The result showed that the leaf was amphistomatic with diacytic stomata on both surfaces. The micromeritic studies showed that the leaf had an angle of repose of 37.47ᵒ, Carr’s Index of 23.01% and Hausner’s ratio of 1.3. Water-soluble, ethanol-soluble and methanol-soluble extractive values were 21%w/w, 10%w/w and 11%w/w, respectively. Fluorescence analysis of the leaf showed different colours indicating the presence of different phytochemicals. The moisture content, total ash, acid-insoluble and water-soluble ash values were 5.31%w/w, 15.18%w/w, 2.7%w/w and 5.6%w/w, respectively. Chemomicroscopy of the leaf powder showed the presence of mucilage, lignin, starch, cellulose and protein while, oil and calcium oxalate crystals were absent. The GC-MS analysis showed the presence of thirty-five (35) phytochemicals with compounds like n-Hexadecanoic acid, 9,12,15-Octadecatrienoic acid, Vitamin E, Stigmasterol, Supraene and, Benzoic acid. These compounds have antioxidant, anticonvulsant, antiserum, antifungal, hypoglycaemic, antimicrobial, anti-inflammatory and larvicidal activities. The results stated could be used to establish standards for the authentication of the fresh and powdered drug leaf products of Thunbergia laevis.

Anatomical adaptation of water-stressed Eugenia uniflora using green synthesized silver nanoparticles and melatonin.

Cell and sub-cellular anatomical adjustments are adaptations utilized by plants to tolerate abiotic stress. Both melatonin and Morinda lucida-silver nanoparticles (ML-AgNPs) are recognized as bio-stimulants. The study examined the morphological changes and adaptive characteristics of these bio-stimulants under water-stress Eugenia uniflora. Twenty-four hours was spent priming the seeds with melatonin (0.06 mg/L), ML-AgNPs (0.06 mg/L), and a mixture (1:1) of the two. The seeds were sown and subjected to water stress for 7 days. The leaves, stems, and roots of water-stressed E. uniflora were sectioned, dried, and examined using a microscope. Drought stress led to the production of non-glandular trichomes on the abaxial and the transformation of paracytic stomata into diacytic stomata. During water stress, melatonin enlarges intercellular gaps and stomata, increases sponge and palisade parenchyma, and thickens epidermis (stem and root) and fibers. The ML-AgNPs diminished the size of mesophyll, intercellular gaps, stomata, and stem fiber. The ML-AgNPs increased the size of bulliform cells and activated the mechanical resistance features of sclerophyllous leaves (thick-celled epidermis and sclerieds) and ray parenchyma (root and stem). Equally, Melatonin and ML-AgNPs increased stem and root anatomical characteristics (xylem, bark, pith, cortex, epidermis, and vascular bundles). Stomata of E. uniflora are susceptible to alterations and undergo cell division into two new stomata (stomatogensis) in response to varying conditions (melatonin and ML-AgNPs). Melatonin adopted a strategy for maintaining a high plant water status, possibly by osmoregulation, whereas E. uniflora primed with ML-AgNPs survived by minimizing transpirational water loss through morphological changes.

A New Taxon of Salvia (Lamiaceae) From Türki̇Ye

Salvia divaricata Montbret & Aucher ex Benth. subsp. artvinense Eminagaoglu, Ozcan & Akyıldırım is described as a new endemic subspecies from Ardanuç (Artvin, Türkiye). It is related to Salvia divaricata Montbret & Aucher ex Benth. and S. tomentosa Mill. from which it differs in stem, leaf characters and flower color and numbers. A key is given to distinguish the new subspecies from the other species in the genus Salvia. Morphology, stem, petiole and leaf anatomy, and nutlet micromorphology were investigated. Nearly rounded stem, hemispherical petiole, bifacial leaf with diacytic stomata, and ovoid to rotund nutlet with glabrous and distinctly rough to protuberances ornamentation were determined for this subspecies. Taxonomic interpretations of the new subspecies are given using morphological, anatomical and phylogenetic analyses. Bangladesh J. Plant Taxon. 29(2): 167-181, 2022 (December)

Comparative Leaf Anatomy and Micromorphology of Thunbergia erecta (Benth.) T. Anderson and Thunbergia laurifolia Lindl. in Peninsular Malaysia.

Comparative leaf anatomy and micromorphology study was carried out on two selected species from the genus Thunbergia Retz. of Acanthaceae subfamily Thunbergioideae. These two investigated species were T. erecta and T. laurifolia from Peninsular Malaysia. The leaf anatomical study involve several methods such as cross-section using sliding microtome on the petioles, midribs, lamina and marginal, leaf epidermal peeling, leaf clearing and observation under a light microscope. The leaf micromorphology method involve the observation under a scanning electron microscope (SEM). This study aimed to investigate the taxonomic value of leaf anatomy and micromorphology characteristics of genus Thunbergia. The results have shown that there were five common characteristics present in both species studied and several variable characters that might be useful for species differentiation of T. erecta and T. laurifolia. The five common characteristics recorded were the presence of raphide, sinuous anticlinal walls, diacytic stomata, majority opened and minority closed venation in lamina and the presence of peltate glandular (unicellular terminal) trichome. The variable characteristics included were petiole, and marginal outlines, types of vascular bundles, the presence of druse, marginal venation, stomata occurrence, types of wax, cuticular sculpturing and types of trichomes. In conclusion, findings in this study showed that leaf anatomical and micromorphological characteristics possessed taxonomic value that can be used in the species identification for the genus Thunbergia specifically for T. erecta and T. laurifolia.

Pharmacognostic evaluation and coagulant analysis of the ethanol extracts of the leaves of Ocimum gratissimum L. (Lamiaceae)

Ocimum gratissimum belongs to the group of plants known as spices. The plant is an erect small plumb with many branches usually not more than 1m high. It is of the familylabiatae, genus Ocimum and species gratissimum. It is commonly known as scent leaf. This project is focused on the pharmacognostic and coagulant activities of the ethanol extract of the leaves of Ocimum gratissimum. The ethanol leaf extract of the plant was obtained using cold maceration with ethanol and then filtered and concentrated using rotary evaporator. The extract was used for coagulant evaluations, pharmacognostic evaluation and chromatographic (thin layer chromatography) analysis. The LD50 was determined in rats using Lorkes method. Microscopic result shows the presence of a covering or uniseratetrichomes, sclerenchyma none pitted with lumen and diacytic stomata. The numerical data obtained were 8.75%, 12% and 8.45% for moisture content, alcohol extractive value and total cash value. The LD50 shows that there was no death recoded because of the holistic use of the plants. The result of the coagulation research are as follow: Group 1 clotting time 1: 69 min and bleeding time 2: 62min, group 2 clotting time 1: 39 min and bleeding time 4: 79min, group 3 clotting time 1: 37 min and bleeding time 1: 99min, group 4 clotting time 1: 81mins and bleeding time is 2: 28mins, group 5 clotting time is 1: 47mins and bleeding time is 2: 49mins) This shows that the ethanol extract of Ocimum gratissimumon rats exhibited coagulant property. The phytochemical analysis of this plant reveals that it contains alkaloids, flavonoids, saponins, terpenoids, tannins, glycosides and resins. It can therefore be concluded that the ethanol extract of Ocimum gratissimum possesses coagulant activity. This study can be of great importance to the development of new coagulant drugs. Citation: Osualaet al.(2021) Pharmacognostic Evaluation and Coagulant screening of Ethanol Extract of the Leaves ofOcimumgratissimumL. (Lamiaceae) Journal

Micromorphological and Pharmacognostic Studies of Leaf and Stem of Solenostemon monostachyus P. Beauv (Lamiaceae)

Solenostemon monostachyus P. Beauv (Lamiaceae), it’s ethnomedical uses include anti-plasmodial, anti-pyretic, antiulcerogenic, antioxidant, anti-inflammatory and anti-nociceptive activities. The aim of this study was to employ the quality control parameters in the evaluation of the leaf and stem of Solenostemon monostachyus to aid in the identification and standardization of the medicinal plant. The plant leaves and stems were collected, identified, air-dried, pulverized and stored in separate glass containers. Standard procedures were employed to obtain the microscopic features of the fresh and powdered samples, micromeritic, chemomicroscopy, fluorescence properties, moisture contents, ash values and soluble extractive values. The results of the microscopic studies using fresh and powdered leaf samples revealed the presence of diacytic stomata on both the abaxial and adaxial surfaces (amphistomatic), with stomatal index of 27.9% and 14.8% respectively. The result of the micromeritics properties of the powdered leaf and stem samples showed angles of repose of 38.0o and 46.0o, Carr’s index of 23.7% and 32.5% and Hausner’s ratios of 1.3 and 1.5 respectively. Results for the moisture content, total, acid-insoluble and water-soluble ash values were 11.7%w/w, 13.7%w/w, 1.8%w/w and 9.4%w/w for the leaf and 13.3%w/w, 17.3%w/w, 1.8%w/w and 9.2%w/w for the stem respectively. Extractive values for water-soluble, methanol-soluble and ethanol-soluble were 26.5%w/w, 32.3%w/w, 14.5%w/w and 15.5%w/w, 15.8%w/w and 14.5%w/w for the leaf and stem respectively. Chemomicroscopy indicated the presence of lignin, calcium oxalate crystals and protein in the leaf and stem. The results obtained therefore could be used to establish pharmacopoeial standard for the fresh and powdered drug products of Solenostemon monostachyus, thus preventing adulteration.

Plectranthus neochilus Schltr.: Anatomic and cytogenetic analyses and chemical characterization of its essential oil

Although used for medicinal purposes, there are many inconsistencies in the literature regarding the identification of Plectranthus neochilus Schltr. due to the morphological similarities with other species belonging to the so-called “P. caninus complex”. Studies that develop tools for the correct identification of medicinal plants are of vital importance as the use of the wrong species can lead to serious health problems. Thus, the objective of the study was to determine a profile to identify individuals of P. neochilus based on anatomic, chemical and cytogenetic analyses. The cytogenetic analysis showed that all the individuals used in the present study belonged to the same species (2n = 30), P. neochilus, in agreement with literature. The presence of diacytic stomata, absence of digitiform trichome, distribution of peltate glandular trichomes on both sides of the leaves are also in agreement with previous reports for P. neochilus. However, the leaf mesophyll and the layout and morphology of glandular trichomes were different from those previously described in the literature. Histochemical tests showed the absence of phenolic compounds but presence of total and neutral terpenes and lipids. The essential oil obtained by hydrodistillation contained β-caryophyllene as the major compound (20.11%) which also coincides with other studies of P. neochilus in Brazil. These results show the cytogenetic, anatomical and the chemical profile of the essential oil of P. neochilus in Brazil and can be used as a tool for its identification.

Morphology, Anatomy and Secondary Metabolites Investigations of Premna odorata Blanco and Evaluation of Its Anti-Tuberculosis Activity Using In Vitro and In Silico Studies.

In-depth botanical characterization was performed on Premna odorata Blanco (Lamiaceae) different organs for the first time. The leaves are opposite, hairy and green in color. Flowers possess fragrant aromatic odors and exist in inflorescences of 4–15 cm long corymbose cyme-type. In-depth morphological and anatomical characterization revealed the great resemblance to plants of the genus Premna and of the family Lamiaceae, such as the presence of glandular peltate trichomes and diacytic stomata. Additionally, most examined organs are characterized by non-glandular multicellular covering trichomes, acicular, and rhombic calcium oxalate crystals. P. odorata leaves n-hexane fraction revealed substantial anti-tuberculous potential versus Mycobacterium tuberculosis, showing a minimum inhibition concentration (MIC) of 100 μg/mL. Metabolic profiling of the n-hexane fraction using gas-chromatography coupled to mass spectrometry (GC/MS) analysis revealed 10 major compounds accounting for 93.01%, with trans-phytol constituting the major compound (24.06%). The virtual screening revealed that trans-phytol highly inhibited MTB C171Q receptor as M. tuberculosis KasA (β-ketoacyl synthases) with a high fitting score (∆G = −15.57 kcal/mol) approaching that of isoniazid and exceeding that of thiolactomycin, the co-crystallized ligand. Absorption, distribution, metabolism, excretion and toxicity predictions (ADME/TOPKAT) revealed that trans-phytol shows lower solubility and absorption levels when compared to thiolactomycin and isoniazid. Still, it is safer, causing no mutagenic or carcinogenic effects with higher lethal dose, which causes the death of 50% (LD50). Thus, it can be concluded that P. odorata can act as a source of lead entities to treat tuberculosis.

STANDARDIZATION AND TOXICOLOGICAL STUDIES OF THE LEAF OF SENNA SIAMEA IRWIN & BARNEBY (FABACEAE) COLLECTED FROM AGULU IN AWKA SOUTH OF ANAMBRA STATE

Background: Pharmacognostic standards and toxicological studies of the leaf of S. siamea were carried out. Objectives: To determine the quantitative phytochemical constituents present, pharmacognostic standards and toxicological effect of the methanol leaf extract and fractions of S. siamea. Methodology: Fresh leaves of S. siamea were collected, dried, pulverizes to powder and subjected to pharmacognostic and quantitative phytochemical analysis. The pulverized leaves were cold macerated in methanol for three days with intermittent shaking and exchange of fresh solvent every 24 hrs (aliquot extraction). Fractionation was done using n-hexane, ethylacetate and butanol. Chromatographic analysis of the methanol extract and fractions were obtained using standard methods. The methanol extract and fractions where screened for phytochemical constituents and toxicological effects using standard methods. Acute toxicity (LD50) of the extract was determined using Lorke’s method. Results: Macroscopic and microscopic examinatios revealed acute apex, pinnate veination and diacytic stomata, trichomes respectively. Phytochemical screening of the extract and fractions revealed the presence of flavonoids, alkaloids, saponins, tannins etc. Proximate analysis of S. siamea revealed moisture content of 5.50 %, total ash value of 15.00 %, water soluble ash of 2.00 % etc. The LD50 was found to be > 5000 mg/kg. Increase in ALT, AST and ALP when metanol extract and fractions were administered were not statistically different from the control group, indicating no hepatotoxic potential. Increase in PCV, Hb and RBC when metanol extract and fractions were administered were not statistically different from control group. The non-significant decrease in the levels of ALP, ALT and AST indicates that the extract has possible hepatoprotective effect. The non-significant decrease in the levels of RBC, Hb and PCV indicates that the extract does not affect the heamatopoietic system when administered orally and at the doses used in the study. Conclusions: Specific standards obtained from the study may be compiled into a monograph of S.siemea plant that may constitute in herbal pharmacopoeia.

Biosystematic studies of some Egyptian species of Cestrum (Solanaceae).

Cestrum is the second largest genus of family Solanaceae, after Solanum, distributed in warm to subtropical regions. Species of genus Cestrum are one of the most ethnopharmacological relevant plants, for their broad biological and pharmacological properties. There is a scarcity to taxonomical studies and identification of these plants in Egypt, thus, the objective of this study was to implement various morphological features, chemical markers and molecular tools to emphasize the taxonomical features of the different Cestrum species. Morphologically, the epidermal cells of C. diurnum, C. elegans and C. parqui were irregular with sinuate anticlinal wall patterns for both surfaces, while, C. nocturnum has anticlinal walls, sinuolate with polygonal to irregular epidermal cells on the abaxial surface. The species of Cestrum have hypostomatic leaves, except C. parqui that has amphistomatic leaves. The experimented species of Cestrum have Anomocytic and anisocytic stomata, while, C. elegans has a diacytic stomata. The morphologically identified Cestrum spp were molecular confirmed based on their ITS sequences, the sequences of C. diurnum, C. nocturnum, C. elegans and C. parqui were deposited on genbank with accession # MT742788.1, MT749390.1, MW091481.1 and MW023744.1, respectively. From the SCOT analyses, the four species of Cestrum were grouped into 2 clusters (I, II), cluster I contains C. elegans, C. nocturnum and C. parqui, while cluster II contains only C. diurnum with 100% polymorphism for all primers. From the GC-MS profile, the C. diurnum exhibited a diverse metabolic paradigm, ensuring their richness with different metabolites comparing to other experimented Cestrum species. Among the total resolved metabolites, 15-methyltricyclo 6.5.2-pentadeca-1,3,5,7,9, 11,13-heptene was the highly incident compound in C. elegans (35.89%) followed by C. parqui (21.81%) and C. diurnum (11.28%), while it absent on C. nocturnum. The compound, 2,2',6,6'-tetra-tert-butyl-4,4'-methylenediphenol was highly detected in C. elegans and C. dirunum with minor amounts in the other Cestrum species. Cypermethrin and 3-butynyl-2,2,5-trimethyl-1,3-dioxane-5-methanol were pivotally reported in C. nocturnum. Taken together, from molecular and metabolic markers, C. diurnum, C. parqui and C. elegans have higher proximity unlike to C. nocturnum.

Taxonomic implication of leaf epidermal anatomy of selected taxa of Scrophulariaceae from Pakistan.

The family Scrophulariaceae consists of taxonomically complex genera and species. The delimitation of the taxa within this family is always challenging. In this paper, we studied leaf epidermis anatomical characteristics and its taxonomic significance of four species belonging to four genera of the family Scrophulariaceae collected from northern Pakistan. The species were examined under light and scanning electron microscopes (LM and SEM). Qualitative and quantitative foliar epidermal anatomical features were examined for both adaxial and abaxial surfaces. Qualitative characters like epidermal cell shape, epidermal cell cover, anticlinal wall, trichomes type, stomata type and stomata position were examined. Quantitative characters like the length and width of leaf epidermis, stomata, stomatal pore, subsidiary cell and trichomes for both adaxial and abaxial surfaces were studied and measured. Stomatal index within the species and between the species was found to be different on adaxial and abaxial surfaces. Diacytic stomata and glandular trichomes on epidermis were only found in Anticharis glandulosa while rest of the taxa has anomocytic type stomata and dendroid trichomes on both surfaces. Based on the micromorphological characters, we did principal component analysis (PCA), and cluster analysis for the species delimitation and identification. A taxonomic key has been provided to delimit and identify the studied taxa based on foliar epidermal characters. The aim of the present research was to elucidate the micromorphological characters to distinguish the studied taxa for taxonomic purposes.

Scanning electron microscopic screening of 20 medicinally important Asteroideae taxa.

In the present study anatomical characterization of 20 medicinally important Asteroideae species were done under light and scanning electron microscopy. Variety of qualitative and quantitative anatomical characters like epidermal cells, stomata, guard cells, subsidiary cells, trichomes and oil droplets were observed. Generally pentagonal, polygonal, irregular or hexagonal, smooth, undulating thick walled epidermal cells were observed in studied species. In abaxial surface Thymophylla tenuiloba L. possessed the largest length of epidermal cell that is, 221.6 (156.6-286.6) μm whereas Bellis perenis L. showed the smallest length that is, 46.4 (32.6-60.2) μm. Average width of epidermal cells ranged from 57 (22-92) μm to 169 (127.9-210.1) μm. Cosmos sulphureus Cav. had smallest width while Thymophylla tenuiloba L. had the largest width. In adaxial surface Artemisia absinthium L. possessed the largest length of epidermal cell that is, 269 (165.1-372.9) μm whereas Bellis perenis L. showed the smallest length that is, 61.4 (42.6-80.2) μm. Average width of epidermal cells ranged from 50.8 (32.6-69) μm to 260 (116-202) μm. Thymophylla tenuiloba L. had smallest width while Dahlia pinnata Cav. had the largest width. Among stomatal characters anisocytic, anomocytic, and diacytic stomata were observed in selected species of Asteroideae. Nonglandular uniserate, multicellular, unbranched pointed tips with bulbous base trichomes were reported in some Asteroideae members while some possessed glandular, capitates mushroom like multicellular trichomes covered with tubercle papicles. Rounded, oval, triangular shaped oil droplets were observed in some species. It is concluded that qualitative and qualitative anatomical variations in trichomes, stomata and epidermal cells are of good taxonomic value for the Asteroideae species.

TAXONOMIC IMPORTANCE OF ANTICLINAL WALLS AND STOMATA PATTERNING IN SOME MELASTOMA L. SPECIES FROM FRASER HILL

The epidermal characteristics of five selected Melastoma L. species in Fraser Hill, Pahang that belongs to Melastomataceae family had been investigated. These five species namely M. muticum Ridl., M. decemfidum Roxb., M. perakense Ridl., M. sanguineum x malabathricum and M. malabathricum var. normale. The objective of this study is to determine whether epidermal characteristics in Melastoma could be taxonomic value in systematic and diagnostic investigations. Methods of the investigation involved epidermal peel and scanning electron microscopy. Results obtained revealed that the presence of hypostomatic stomata and guard cells pairs were elliptic in shaped for all species studied. Apart, the pattern of anticlinal walls on adaxial and abaxial surfaces was straight to wavy for all species studied except for wavy to sinuous anticlinal walls on abaxial surface of M. sanguineum x malabathricum. Furthermore, two types of stomata were observed among species studied such as anomocytic and diacytic stomata. Results showed that diacytic type was only present in M. sanguineum x malabathricum therefore could be a criterion to diagnose the species. Lastly, this present study was also reported on the presence of two types of guard cell pairs such as raised or slightly raised and sunken guard cell pairs. In conclusion, the present study revealed that the anticlinal walls and stomata patterning possess as taxonomic importance in identification and classification of Melastoma either at genus or species level.