Field response and molecular screening of European wheat germplasm against powdery mildew at the Himalayan region of Pakistan.

Wheat powdery mildew possesses a significant threat to wheat crops not only on a global scale but also in the northern region of Pakistan. Recognizing the need for effective measures, the exploration and utilization of exotic germplasm take on critical importance. To address this, a series of trials were made to investigate the response of 30 European (EU) lines, in addition to the local checks (Siran, Atta-Habib (AH) and Ghanimat-e-IBGE) against wheat powdery mildew at the Himalayan region of Pakistan. The study involved field testing from 2018 to 2022 across multiple locations, resulting in 38 different environments (location × year). In addition to field evaluations, molecular genotyping was also performed. The disease was absent on the tested lines during 2018, 2019, and 2020 whereas it ranged from 0 to 100% at Chitral location during 2021, where 100% was observed only for one EU wheat line "Matrix." The disease prevailed only at Gilgit location (0-60% for EU wheat line "F236") and at Nagar location (0-10% for EU wheat lines Substance and Nelson) during the disease season of 2022. Most of the EU wheat lines showed very low ACI values, due to an overall low disease pressure. Matrix showed the maximum ACI (1.54) followed by Ritter (1.25) and Bli_autrichion (0.87), whereas the minimum (0.1) was for Substance, JB_Asano, and KWS_Loft followed by Canon (0.19), all exhibiting partial resistance. The molecular marker-based screening revealed that Pm38 was the most prevalent and detected in 100% of wheat lines followed by Pm39 (60%) and Pm8 (30%). Six wheat lines (20%) possessed all three Pm genes (Pm8, Pm38, and Pm39) concurrently. The variability observed in this study can be utilized in future breeding efforts aimed at developing resistant wheat varieties.

Microscopic (LM and SEM) visualization of pollen ultrastructure among honeybee flora from lower Margalla Hills and allied areas.

Microscopic visualization of micro-morphological characters were analyzed using a scanning electron microscopic (SEM) tool, which has proven to be very successful to analyze the pollen surface peculiarities. The significant goal of this research was to perform microscopic examination of pollen of some of the most frequently visited honeybee floral species around apiaries. Micro-morphological characterization of frequented honeybees foraged plants were discussed. A total of 15 species, belonging to 11 different families were identified for the foraging activities of honeybees, namely, Lantana camara, Jatropha integerrima, Helianthus annuus, Tecoma stans, Lagerstroemia indica, Duranta erecta, Cosmos sulphureus, Hymenocollis littoralis, Moringa oleifera, Cestrum nocturnum, Parthenium hysterophorus, Volkameria inermis, Catharanthus roseus, Malvastrum coromandelianum, and Citharexylum spinosum. The microscopic slides were prepared using the acetolysis method, and the qualitative and quantitative features were measured and described using microscopic tools. The pollen type sculpture varies from psilate scabrate to echinate and colpi from tricolpate to tetracolpate. Quantitative parameters such as polar diameter, mesocolpium distance, equatorial dimensions, aperture size, spine diameter, and exine thickness were calculated using IBM SPSS Statistics 20. The exine thickness was measured at its maximum in C. roseus (3.85 µm), whereas it was at its minimum of 0.90 µm in L. indica and D. erecta. Pollen fertility was estimated to be highest in H. annus (88%). The current research validates scanning microscopic features of pollen of the honeybee floral species, which is helpful for the accurate identification and commercializing of honey production to generate revenue for beekeepers through the exploration of bee floral diversity. RESEARCH HIGHLIGHTS: Pollen spectrum examination using microscopic imaging techniques. Variations examined through scanning microscopy in pollen ultrastructure. Micro-morphology has taxonomic value for accurate identification of honeybee flora. Authentication of Honeybee floral species for sustainable beekeeping.

Antimicrobial activities of different solvent extracts from stem and seeds of Peganum Harmala L.

Wild medicinal herbs have been used as folk and traditional medicines all across the world since well before recorded history. This present study was designed to test the antimicrobial activities of five different solvent extracted samples (n-hexane, n-butanol, ethyl acetate, methanol, and water) of Peganum harmala using stems and seeds. Two different strains of Gram-negative bacteria (Escherichia coli and Klebsiella pneumonia), two Gram-positive bacteria (Bacillus subtilus and Staphylococcus aureus), and one fungal strain (Candida albicans) were used. The antimicrobial activities were measured using a disc diffusion assay. Two concentrations of the extracts (1 and 2mgDisc-1) were used. Ethyl acetate fraction was found more affective among the tested solvents and showed maximum activity (zone of inhibition) against S. aureus (65.53 and 81.10%), E. coli (46.22 and 61.29%) while n-butanol and water fractions gave maximum activity against S. aureus (78.86 and 70.00%) and K. pneumonia (57.00 and 61.39%) respectively. Water fraction showed maximum activity against C. albicans (60.00 and 81.88%). In the case of the stem, Ethyl acetate again showed more activity against B. subtilus (38.57 and 42.10%) and S. aureus (36.66 and 46.66%) while n-butanol showed maximum activity against K. pneumonia (24.55 and 32.44%) and E. coli (27.93 and 37.61%). Methanol was found more effective against C. albicans (25.71 and 43.80%). Seed extracted samples were found more effective compared to the stem. Ethyl acetate, butanol, and aqueous extracted samples showed good activity against the tested microbes, so these fractions are recommended for study their mechanism of actions and isolation of bioactive metabolites responsible for antimicrobial activities. The P. harmala should be evaluated for their bioactive compounds to be used in future studies. Our objective is to provide the framework for future study on the roles of P. harmala as traditional medicines.

Micro-morphological diversity of pollen among Asteraceous taxa from Potohar Plateau-Pakistan.

Microscopic pollen characters from 19 species of family Asteraceae collected from different areas of the Potohar Plateau of Pakistan have been investigated using light and scanning electron microscopy. This study uses microscopic techniques to provide detailed information on pollen and its exine structure based on qualitative and quantitative characters. The present study has observed three types of pollen grains based on their apertures, that is, trizonocolporate, tetrazonocolporate, and tricolporate. Exine sculpturing is echinate in all studied taxa. Some species also exhibit Lacunae which differ in their shape and sizes. The size of the spines and the number of spines between the colpi play a vital role in differentiating the species of the same genus. Pollen fertility ranges from 94.3 to 74.19, showing that these species are well established in the study area. For the correct identification of species, a light and scanning electron microscopy was used that provides sufficient information for a taxonomist to distinguish species taxonomically. RESEARCH HIGHLIGHTS: Light and Scanning electron microscope is comparatively used to investigate the pollen characters of some species of the family Asteraceae. A scanning electron microscopy is used to study the micro-morphological characters of pollen grains. Qualitative and quantitative characters of pollen were studied.

Biogenic Synthesis of Silver Nanoparticles Using Phagnalon niveum and Its In Vivo Anti-Diabetic Effect against Alloxan-Induced Diabetic Wistar Rats.

Background: Type-2 diabetes mellitus (T2DM) is a non-communicable, life-threatening syndrome that is present all over the world. The use of eco-friendly, cost-effective and green synthesised nanoparticles (NPs) as a medicinal therapy in the treatment of T2DM is an attractive option. Aim: The present study aimed to evaluate the anti-diabetic potential of the phyto-synthesised silver nanoparticles (AgNPs) obtained from Phagnalon niveum plant methanolic extract. Methods: The green synthesised AgNPs made from Phagnalon niveum plant methanolic extract were analysed by Ultraviolet-Visible (UV-Vis) spectroscopy, and the functional groups involved in the reduction of the silver ions (Ag+) were characterised by Fourier Transform Infrared (FTIR) spectroscopy. The size and crystallinity were assessed via X-ray Diffraction (XRD). The morphology of AgNPs was confirmed using Scanning Electron Microscopy (SEM). The amount of silver (Ag) was estimated via energy dispersive X-ray (EDX) analysis. An intraperitoneal injection of 200 mg alloxan per kg albino Wistar rats' body weight, at eight weeks old and weighing 140-150 g, was used to induce diabetes mellitus (N = 25; n = 5/group). Group C: untreated normal control rats that only received distilled water, group DAC: diabetic control rats that received alloxan 200 mg/Kg body weight, DG: diabetic rats treated with glibenclamide at 0.5 mg/kg body weight, DE: diabetic rats that received methanolic P. niveum extract at 10 mg/Kg body weight, and DAgNPs: diabetic rates that received AgNPs synthesised from P. niveum at 10 mg/kg body weight. The blood glucose levels were monitored on days 0, 7, and 14, while lipid, liver, and kidney profiles were checked after dissection at the end of treatment (day 21). On the final day of the period study (day 21), an oral glucose tolerance test was carried out by administering orally 2 g/kg body weight of glucose to the respective groups, and the blood glucose level was checked. A fasting glucose level was measured using a glucometer. Urine samples were collected from each animal and analysed using lab-made assay kits for glucose, bilirubin, pH, leukocytes, and nitrite, among other factors. For statistical analyses, a one-way ANOVA and Dunnett's test were applied. Results: The green-mediated synthesis of AgNPs using P. niveum methanolic extract produced spherical and mono-dispersed NPs with a size ranging from 12 to 28 nm (average: 21 nm). Importantly, a significant reduction of blood glucose levels and an increase in body weight, as well as a remarkable improvement in lipid, liver, and kidney profiles, were noticed. Conclusions: The biosynthesised AgNPs significantly improved the abnormalities in body weight, urine, and serum levels, indicating that it is a promising anti-diabetic agent.

Characterization of anatomical foliar epidermal features of herbaceous flora of Tilla Jogian, Pakistan by using light microscopy techniques.

In current study, 22 herbaceous plant specimens have been examined with the help of light microscope to carry out leaf epidermal anatomy. Foliar micro-morphology of herbaceous plants from Tilla Jogian, Jhelum Pakistan was held first time. The plants were collected from different parts of Tilla Jogian at different elevations. The main objective of the study is to use this information as an aid in plant taxonomy. Significant variations were observed in both qualitative and quantitative features. With the help of microscopy significant variations in characters like stomata size, number, epidermal cell size, shape and number, guard cell size, trichomes, and subsidiary cells were recorded. On the basis of these findings, light microscopy of leaf epidermal anatomical features holds great interest for taxonomists to identify complete taxa. Foliar epidermal anatomy of herbaceous flora serves as baseline data along with taxonomical data and can be helpful in demarcation and limitation of nearly associated taxa to gain better use of anatomical characters.

Implication of scanning electron microscopy and light microscopy for oil content determination and seed morphology of Verbenaceae.

Due to increase in energy catastrophe and accompanying environmental issues, the search for renewable and alternative energy resources is a fiercely disputed problem in the scientific world. Biodiesel has proven to be the finest sustainable alternative to gasoline. Because of their environment-friendly character and cost-effectiveness, nonedible oil-bearing seeds could be the possible resource for biodiesel production. Therefore, the present study uses scanning electron microscopy (SEM) to investigate and identify micromorphological features of six novel, nonedible oil-bearing seeds of Verbenaceae as potential biodiesel feed stocks. Selected seeds of Verbenaceae range in size from 1 to 5 mm in width and 1.8 to 6.4 mm in length, according to light microscopic studies. The oil content of seeds ranges from14 to 19%. Seed texture, color, shape, size, periclinal and anticlinal wall patterns, and cell outline are all more variable in SEM-mediated seed ultrastructure analyses. The findings of this work imply that SEM could be a useful method for revealing hidden micromorphological features in a variety of nonedible oil-producing seeds, which could aid to future investigation, proper identification, seed categorization, and authenticity. These seeds were hard to be identified by the native people due to their resemblance in seed color and shape, so there is a need to give a detailed description of seed characters for proper identification of these plants. This work will determine the new features of the seeds, which will be helpful in near future.

Microscopic characterization of petiole anatomy of Asteraceous taxa of Western Himalaya-Pakistan.

Petiole anatomy of 15 species of family Asteraceae was examined which aimed to investigate petiolar anatomical structures for species level identification. Shandon Microtome was used for petiole histological preparations. Both qualitative and quantitative features were studied under microscope which showed significant variation in petiole, collenchyma, parenchyma shape/size, vascular bundles arrangement/size, and vessel elements quantity. Artemisia japonica Thunb., Cirsium vulgare (Savi) Ten., Myriactis nepalensis Less., Seriphidium brevifolium Ling & Y.R.Ling, Taraxacum officinale (L.) Weber ex F.H.Wigg., and Xanthium strumarium L. showed winged petioles. Maximum length and width of upper and lower epidermis was found in Tagetes erecta L. which is 23.05 ± 0.89 µm, 24.9 ± 1.257 µm length and 21.75 ± 1.38067 µm, 22.75 ± 0.467 µm width, respectively. Petioles of Parthenium hysterophorus L. was longest one with 9.85 ± 10.45 µm while A. japonica Thunb. showed highest number of vessel elements. Maximum size of vascular bundles was found in T. erecta L. with 5.05 ± 14.25 µm. Artemisia annua L., C. vulgare (Savi) Ten, Cyanthillium cinereum (L.) H.Rob., Helianthus annus L., M. nepalensis Less., P. hysterophorus L., Senecio chrysanthemoides DC. have trichomes while Tussilago farfara L. has highest number of vascular bundles. All species have angular collenchyma type except M. nepalensis Less., P. hysterophorus L., S. brevifolium Ling & Y.R.Ling, Tagetes minuta L., T. officinale L., S. chrysanthemoides DC., and T. farfara L. Cluster analysis implemented that distinct plant species in cluster. Petiolar anatomical structures and taxonomic key will helpful for distinguishing Asteraceous taxa at genus and species level. This taxonomic significant investigation will also provide baseline to taxonomists for other Asteraceae studies and phylogenetic research.

Implication of scanning electron microscopy as a tool for identification of novel, nonedible oil seeds for biodiesel production.

Biodiesel is a promising, bio-based, renewable, nontoxic, environment friendly, and alternative fuel for petroleum derived fuels which helps to reduce dependency on conventional fossil fuels. In this study, six novel, nonedible seed oil producing feedstock were explored for their potential for sustainable production of biodiesel. It is very important to correctly identify oil yielding plant species. Scanning electron microscopy (SEM) was used as reliable tool for authentic identification of oil yielding seeds. Macromorphological characters of seeds were studied with light microscopy (LM). Outcomes of LM of seeds exposed distinctive variation in seed size from 16.3 to 3.2 mm in length and 12.4 to 0.9 mm in width, shape varied from oval to triangular, and color from black to light brown. Oil content of nonedible seed ranged from 25 to 30% (w/w). Free fatty acid content of seed oil varied from 0.32 to 2.5 mg KOH/g. Moreover, ultra structural study of seeds via SEM showed variation in surface sculpturing, cell arrangement, cell shape, periclinal wall shape, margins, protuberances, and anticlinal wall shape. Surface sculpturing varied from rugged, reticulate, varrucose, papillate, and striate. Periclinal wall arrangements confirmed variation from rough, wavy, raised, depressed, smooth, and elevated whereas, anticlinal walls pattern showed variation from profuse undulating, smooth, raised, grooved, deep, curved, and depressed. It was concluded that SEM could be a latent and advanced technique in unveiling hidden micromorphological characters of nonedible oil yielding seeds which delivers valuable information to researchers and indigenous people for precise and authentic identification and recognition.

Anatomical characterization of 18 commercially important varieties of Phoenix dactylifera L. by using microscopy.

Date palm (Phoenix dactylifera L.) is an important traditional crop having socioeconomic values. To best of literature review and our knowledge, there is a gap of taxonomic study on the role of morpho-anatomical features of date palm for correct identification of its varieties. With the advancement in the field of microscopy, anatomical features are considered very effective taxonomical markers. The present study was aimed to analyze the taxonomic variations using leaf epidermal anatomy and to highlight the importance of leaf epidermis to resolve identification ambiguities among the different varieties of date palms. The study includes analysis of leaf epidermal anatomy of 18 different commercial varieties of date palm collected from different areas of Pakistan. A diverse range of shape and size of epidermal cells, subsidiary cells, guard cells, stomata, stomatal pore was observed on abaxial as well as adaxial among studied date palm varieties via LM and found to be very helpful tool in discrimination of varieties. The shape of epidermal cells on both abaxial and adaxial surfaces varies including elongated, irregular, polygonal, curved, and spherical or sometime mixed of these. The values of stomatal indices on both surfaces showed wide variations.

Molecular detection of Cryptosporidium: an emerging parasite in different water sources of 2010 flood-affected district Nowshera, Pakistan.

Cryptosporidium is a water-borne zoonotic parasite worldwide, usually found in lakes and rivers contaminated with sewage and animal wastes, causing outbreaks of cryptosporidiosis. In this study, 300 water samples were collected from four designated places of flood-affected district Nowshera consist of different water sources to find out the prevalence of Cryptosporidium via polymerase chain reaction (PCR). The overall prevalence of Cryptosporidium was 30.33% (91/300) with more prevalent 44% in drain water and low 5% in bore/tube well water. The prevalence in open well and tap water was recorded 33% and 20%, respectively. The highest prevalence was recorded in summer (June-September). The result of this study ensures enormous contamination of drinking water that requires appropriate treatment, cleaning and filtration to provide safe drinking water. Preventing water-borne disease and proper treatment of water supplies is essential to public health.

Application and implication of scanning electron microscopy for evaluation of palyno-morphological features of Vitaceae from Pakistan.

The present study aimed to investigate the palyno-morphological features of species of family Vitaceae from Pakistan. A total of nine species, belonging to four genera were collected, pressed, identified, and then analyzed microscopically. Both quantitative and qualitative characters of the pollen grains were recorded including polar and equatorial diameter, P/E ratio, number of colpi and pores, exine thickness and shapes of the pollen in both polar and equatorial view, and exine sculpturing using Leica microscope fitted with camera Meiji Infinity 1 and then analyzed statistically using software IBM SPSS Statistics 20. The results of the present study demonstrated the variations in polar and equatorial diameter, exine thickness, P/E ratio, pollen shape, and exine sculpturing of the studied species and highlighted the significance of pollen morphology as an identification tool. The present study may contribute to better understand the classification at genus level, which will support the future phylogenetic characterization of the family.

Continentalic acid exhibited nephroprotective activity against the LPS and E. coli-induced kidney injury through inhibition of the oxidative stress and inflammation.

The present study investigated the effect of the continentalic acid (CNT) isolated from the Aralia Continentalis against the LPS and E. coli-induced nephrotoxicity. The LPS and E. coli administration markedly altered the behavioral parameters including spontaneous pain, tail suspension and survival rate. However, the treatment with CNT dose dependently improved the behavioral parameters. The CNT treatment significantly improved the renal functions test (RFTs) and hematological parameters following LPS and E. coli-induced kidney injury. Furthermore, the LPS and E. coli administration markedly compromised the anti-oxidant enzymes and enhanced the oxidative stress markers. However, the CNT treatment markedly enhanced the anti-oxidants enzymes such as GSH, GST, Catalase and SOD, while attenuated the oxidative stress markers such as MDA and POD. The MPO enzyme is widely used marker for the neutrophilic infiltration, the LPS and E. coli administration markedly increased the MPO activity. However, the CNT treatment markedly attenuated the MPO activity in both LPS and E. coli-induced kidney injury. Furthermore, the CNT treatment markedly attenuated the NO production compared to the LPS and E. coli-induced kidney injury group. Additionally, the CNT treatment improved the histological parameters markedly (H and E, PAS and Masson's trichome staining) and protect the kidney from the inflammatory insult of the LPS and E. coli evidently. The comet assay revealed marked DNA damage, however, the CNT treatment markedly prevented the LPS and E. coli-induced kidney damage. The CNT treatment markedly enhanced the expression of Nrf2, while attenuated the iNOS expression in both models of kidney injury.

Scanning electron microscopy as a tool for authentication of oil yielding seed.

Now-a-days, plant species are consumed globally for various purposes and this increasing demand leads to adulteration due to gradually exploitation in natural resources. The major causes of adulteration may be confusion in nomenclature, unawareness of authentic sources, unavailability of authentic sources, color resemblances, deficiencies in collection procedures, and misidentification. This study aims to use the microscopic techniques such as scanning electron microscopy for the authentication of the oil yielding seeds of four important and traditionally used species Prunus persica, Prunus domestica, and Eruca sativa and Argemone Mexicana from their adulterants. All of these are versatile in usage. Locally, these four plants are adulterated badly and there is need to provide a criteria and a complete monograph for correct identification. This research may prove to be helpful for quality control and as well for future studies to explore other novel aspects of these plants.

Comparative SEM and LM foliar epidermal and palyno-morphological studies of Amaranthaceae and its taxonomic implications.

Palynological features as well as comparative foliar epidermal using light and scanning electron microscope (SEM) of 17 species (10genera) of Amaranthaceae have been studied for its taxonomic significance. Different foliar and palynological micro-morphological characters were examined to explain their value in resolving the difficulty in identification. All species were amphistomatic but stomata on abaxial surface were more abundant. Taxonomically significant epidermal character including stomata type, trichomes (unicellular, multicellular, and capitate) and epidermal cells shapes (polygonal and irregular) were also observed. Pollens of this family are Polypantoporate, pores large, spheroidal, mesoporous region is sparsely to scabrate, densely psilate, and spinulose. All these characters can be active at species level for identification purpose. This study indicates that at different taxonomic levels, LM and SEM pollen and epidermal morphology is explanatory and significant to identify species and genera.

Molecular detection of Toxoplasma gondii in water sources of district Nowshehra, Khyber Pakhtunkhwa, Pakistan.

Toxoplasmosis is spread through contamination of water sources and results in morbidity globally. In the current study 300 water samples were processed by polymerase chain reaction (PCR) for detection of Toxoplasma gondii. The overall prevalence in different water sources was 6.6% (17/300). Among different water sources the highest prevalence was recorded in drain water at 7% (7/100), followed by tube well water at 7.5% (3/40) and open well water at 5% (5/100) ,and the lowest was recorded in tap water at 3.33% (2/60). The highest prevalence was recorded in summer. Evidence indicates that cleaning and filtration need to be adopted to avoid the health hazards of waterborne zoonotic parasites.

3-Hydr-oxy-1,2,3,9-tetra-hydro-pyrrolo[2,1-b]quinazolin-4-ium chloride dihydrate: (+)-vasicinol hydro-chloride dihydrate from Peganum harmala L.

The title compound, C(11)H(13)N(2)O(+)·Cl(-)·2H(2)O, the dihydrate of (+)-vasicinol hydro-chloride, is a pyrrolidinoquinazoline alkaloid. It was isolated from the ethyl acetate fraction of the leaves of Peganum harmala L. The pyrrolidine ring has an envelope conformation with the C atom at position 2 acting as the flap and the C atom at position 3, carrying the hydroxyl substituent, has an S configuration. The absolute configuration was determined as a result of the anomalous scattering of the Cl atom. In the crystal structure, mol-ecules stack along the a axis, connected to one another via inter-molecular O-H⋯Cl and N-H⋯Cl hydrogen bonds, forming approximately triangular-shaped R(2) (1)(7) rings, and O-H⋯Cl and O-H⋯O hydrogen bonds, forming penta-gonal-shaped R(5) (4)(10) rings. The overall effect is a ribbon-like arrangement running parallel to the a axis.