Taxonomic study and diversity of Postia s.lat. in Swat, Pakistan: addition of five brown rot Polypores to the country.

Brown rot Polypores are ecologically significant as they play a crucial role in maintaining the carbon cycle and contribute to humus formation in forest ecosystems through their lignocellulose degradation ability. It is important to note that some species can significantly impact timber, potentially causing decay in economically valuable wood. Many Asian countries including Pakistan are still under the exploratory phase and have undocumented species diversity in Polypore fungi. In the current study, collections representing five different species belonging to two families, Postiaceae and Adustoporiaceae, were subjected to detailed morphoanatomical and molecular analyses. A combined matrix of two gene datasets (ITS and nrLSU) was analyzed using three different phylogenetic methods viz. Maximum Parsimony (MP), Maximum Likelihood (ML), and Bayesian inference (BI). Our study presents descriptions of five previously undocumented brown rot Polypore species from the country including Fuscopostia fragilis (Fr.) B.K. Cui, L.L. Shen & Y.C. Dai, Amaropostia stiptica (Pers.) B.K. Cui, L.L. Shen & Y.C. Dai, Cyanosporus piceicola B.K. Cui, L.L. Shen & Y.C. Dai, Spongiporus balsameus (Peck) A. David, Rhodonia placenta (Fr.) Niemelä, K.H. Larss. & Schigel. Regarding the molecular data, nodes of our subject sequences were substantially supported and fell under their respective species clades with high ML bootstrap values (≥ 95), MP bootstrap ≥ 74 and BI probabilities ≥ 0.98. Findings of the study will not only contribute to our understanding of local Polypores species diversity but also enhance knowledge of geographical distribution in global context.

Biochemical and phenological characterization of diverse wheats and their association with drought tolerance genes.

Drought is one of the most important wheat production limiting factor, and can lead to severe yield losses. This study was designed to examine the effect of drought stress on wheat physiology and morphology under three different field capacities (FC) viz. 80% (control), 50% (moderate) and 30% (severe drought stress) in a diverse collection of wheat germplasm including cultivars, landraces, synthetic hexaploid and their derivatives. Traits like grain weight, thousand grain weight and biomass were reduced by 38.23%, 18.91% and 26.47% respectively at 30% FC, whereas the reduction rate for these traits at 50% FC were 19.57%, 8.88% and 18.68%. In principal component analysis (PCA), the first two components PC1 and PC2 accounted for 58.63% of the total variation and separated the cultivars and landraces from synthetic-based germplasm. Landraces showed wide range of phenotypic variations at 30% FC compared to synthetic-based germplasm and improved cultivars. However, least reduction in grain weight was observed in improved cultivars which indicated the progress in developing drought resilient cultivars. Allelic variations of the drought-related genes including TaSnRK2.9-5A, TaLTPs-11, TaLTPs-12, TaSAP-7B-, TaPPH-13, Dreb-B1 and 1fehw3 were significantly associated with the phenological traits under drought stress in all 91 wheats including 40 landraces, 9 varieties, 34 synthetic hexaploids and 8 synthetic derivatives. The favorable haplotypes of 1fehw3, Dreb-B1, TaLTPs-11 and TaLTPs-12 increased grain weight, and biomass. Our results iterated the fact that landraces could be promising source to deploy drought adaptability in wheat breeding. The study further identified drought tolerant wheat genetic resources across various backgrounds and identified favourable haplotypes of water-saving genes which should be considered to develop drought tolerant varieties.

Water stress effects on stay green and chlorophyll fluorescence with focus on yield characteristics of diverse bread wheats.

MAIN CONCLUSION: The study provided an insight toward better understanding of stay-green mechanisms for drought tolerance improvement and identified that synthetic-derived wheats proved as a promising germplasm for improved tolerance against water stress. Stay-green (SG) trait is considered to be related with the ability of wheat plants to maintain photosynthesis and CO2 assimilation. The present study explored the interaction of water stress with SG expression through physio-biochemical, agronomic and phenotypic responses among diverse wheat germplasm comprising of 200 synthetic hexaploids, 12 synthetic derivatives, 97 landraces and 16 conventional bread wheat varieties, for 2 years. The study established that variation of SG trait existed in the studied wheat germplasm and there was positive association between SG trait and tolerance to water stress. The relationship of SG trait with chlorophyll content (r = 0.97), ETR (r = 0.28), GNS (r = 0.44), BMP (r = 0.34) and GYP (r = 0.44) was particularly promising under water stress environment. Regarding chlorophyll fluorescence, the positive correlation of фPSII (r = 0.21), qP (r = 0.27) and ETR (r = 0.44) with grain yield per plant was noted. The improved ΦPSII and Fv/Fm of PSII photochemistry resulted in the high photosynthesis activity in SG wheat genotypes. Regarding relative water content and photochemical quenching coefficient, synthetic-derived wheats were better by maintaining 20.9, 9.8 and 16.1% more RWC and exhibiting 30.2, 13.5 and 17.9% more qP when compared with landraces, varieties and synthetic hexaploids, respectively, under water stress environment. Synthetic derived wheats also exhibited relatively more SG character with good yield and were more tolerant to water stress in terms of grain yield, grain weight per plant, better photosynthetic performance through chlorophyll fluorescence measurement, high leaf chlorophyll and proline content, and hence, may be used as novel sources for breeding drought tolerant materials. The study will further facilitate research on wheat leaf senescence and will add to better understanding of SG mechanisms for drought tolerance improvement.

A Novel Based Synthesis of Silver/Silver Chloride Nanoparticles from Stachys emodi Efficiently Controls Erwinia carotovora, the Causal Agent of Blackleg and Soft Rot of Potato.

In recent years, the biological synthesis of silver nanoparticles has captured researchers' attention due to their unique chemical, physical and biological properties. In this study, we report an efficient, nonhazardous, and eco-friendly method for the production of antibacterial silver/silver chloride nanoparticles utilizing the leaf extract of Stachys emodi. The synthesis of se-Ag/AgClNPs was confirmed using UV-visible spectroscopy, DPPH free radical scavenging activity, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and X-ray diffraction (XRD). An intense peak absorbance was observed at 437 nm from the UV-visible analysis. The Stachys emodi extract showed the highest DPPH scavenging activity (89.4%). FTIR analysis detected various bands that indicated the presence of important functional groups. The SEM morphological study revealed spherical-shaped nanoparticles having a size ranging from 20 to 70 nm. The XRD pattern showed the formation of a spherical crystal of NPs. The antibacterial activity performed against Erwinia carotovora showed the maximum inhibition by centrifuged silver nanoparticles alone (se-Ag/AgClNPs) and in combination with leaf extract (se-Ag/AgClNPs + LE) and leaf extract (LE) of 98%, 93%, and 62% respectively. These findings suggested that biosynthesized NPs can be used to control plant pathogens effectively.

Green Synthesis of Silver Nanoparticles Using Euphorbia wallichii Leaf Extract: Its Antibacterial Action against Citrus Canker Causal Agent and Antioxidant Potential.

Biologically synthesized silver nanoparticles are emerging as attractive alternatives to chemical pesticides due to the ease of their synthesis, safety and antimicrobial activities in lower possible concentrations. In the present study, we have synthesized silver nanoparticles (AgNPs) using the aqueous extract of the medicinal plant Euphorbia wallichii and tested them against the plant pathogenic bacterium Xanthomonas axonopodis, the causative agent of citrus canker, via an in vitro experiment. The synthesized silver nanoparticles were characterized by techniques such as UV-Vis spectroscopy, Fourier transform infrared spectroscopy, energy-dispersive X-ray spectroscopy, X-ray diffraction analysis and transmission electron microscopy. Moreover, the plant species were investigated for phenolics, flavonoids and antioxidant activity. The antioxidant potential of the extract was determined against a DPPH radical. The extract was also evaluated for phenolic compounds using the HPLC technique. The results confirmed the synthesis of centered cubic, spherical-shaped and crystalline nanoparticles by employing standard characterization techniques. A qualitative and quantitative phytochemical analysis revealed the presence of phenolics (41.52 mg GAE/g), flavonoids (14.2 mg QE/g) and other metabolites of medicinal importance. Different concentrations (1000 µg/mL to 15.62 µg/mL-2 fold dilutions) of AgNPs and plant extract (PE) alone, and both in combination (AgNPs-PE), exhibited a differential inhibition of X. axanopodis in a high throughput antibacterial assay. Overall, AgNPs-PE was superior in terms of displaying significant antibacterial activity, followed by AgNPs alone. An appreciable antioxidant potential was recorded as well. The observed antibacterial and antioxidant potential may be attributed to eight phenolic compounds identified in the extract. The Euphorbia wallichii leaf-extract-induced synthesized AgNPs exhibited strong antibacterial activity against X. axanopodis, which could be exploited as effective alternative preparations against citrus canker in planta in a controlled environment. In addition, as a good source of phenolic compounds, the plant could be further exploited for potent antioxidants.

Occurrence and Distribution of Heavy Metals in Mining Degraded Soil and Medicinal Plants: A Case Study of Pb/Zn Sulfide Terrain Northern Areas, Pakistan.

Mining activities have serious environmental impacts, thus releasing heavy metals (HMs) such as cadmium (Cd), lead (Pb), chromium (Cr), zinc (Zn) and nickel (Ni) into the surrounding environment. The current paper investigated the impacts of mining activities of Pb-Zn sulfide on soil and medicinal plants. Hence, soil samples (n = 36) and medicinal plants (n = 36) samples were collected, acid extracted and were analyzed through Inductively Coupled Plasma Mass Spectrometry (ICP-MS) for HMs quantification. Our results showed that mineralized zones showed high HMs enrichment levels as compared to non-mineralized zones. Various Indices for HMs assessment revealed that the contaminated soil of the study area had low to extreme level. The mean concentrations of HMs in mining degraded soil and medicinal plants were significantly higher (p ≤ 0.01) and were found in order of Zn > Pb > Cr > Ni > Cd and Cr > Zn > Pb > Ni > Cd respectively. Similarly, some widely consumable medicinal plants showed good metal accumulation for Cd, Cr and Pb i.e., 3.57 mg kg1, 350 mg kg-1 and 335 mg kg-1 in C. sativa, while 5.9 mg kg-1, 276.9 mg kg-1 and 188.7 mg kg-1 in R. hestatus respectively. Hence, the present study recommended that medicinal plants grown in mining areas should be analyzed for HMs concentration before consumption.

Prevalent fecal contamination in drinking water resources and potential health risks in Swat, Pakistan.

Fecal bacteria contaminate water resources and result in associated waterborne diseases. This study assessed drinking water quality and evaluated their potential health risks in Swat, Pakistan. Ground and surface drinking water were randomly collected from upstream to downstream in the River Swat watershed and analyzed for fecal contamination using fecal indicator bacteria (Escherichia coli) and physiochemical parameters (potential of hydrogen, turbidity, temperature, electrical conductivity, total dissolved solid, color, odor and taste). The physiochemical parameters were within their safe limits except in a few locations, whereas, the fecal contaminations in drinking water resources exceeded the drinking water quality standards of Pakistan Environmental Protection Agency (Pak-EPA), 2008 and World Health Organization (WHO), 2011. Multivariate and univariate analyses revealed that downstream urbanization trend, minimum distance between water sources and pit latrines/sewerage systems, raw sewage deep well injection and amplified urban, pastures and agricultural runoffs having human and animal excreta were the possible sources of contamination. The questionnaire survey revealed that majority of the local people using 10-20years old drinking water supply schemes at the rate of 73% well supply, 13% hand pump supply, 11% spring supply and 3% river/streams supply, which spreads high prevalence of water borne diseases including hepatitis, intestinal infections and diarrhea, dysentery, cholera, typhoid fever, jaundice and skin diseases in children followed by older and younger adults.

Health risk assessment of heavy metals and bacterial contamination in drinking water sources: a case study of Malakand Agency, Pakistan.

Human beings are frequently exposed to pathogens and heavy metals through ingestion of contaminated drinking water throughout the world particularly in developing countries. The present study aimed to assess the quality of water used for drinking purposes in Malakand Agency, Pakistan. Water samples were collected from different sources (dug wells, bore wells, tube wells, springs, and hand pumps) and analyzed for different physico-chemical parameters and bacterial pathogens (fecal coliform bacteria) using standard methods, while heavy metals were analyzed using atomic absorption spectrophotometry (AAS-PEA-700). In the study area, 70 % of water sources were contaminated with F. coliform representing high bacterial contamination. The heavy metals, such as Cd (29 and 8 %), Ni (16 and 78 %), and Cr (7 %), exceeded their respective safe limits of WHO (2006) and Pak-EPA (2008), respectively, in water sources, while Pb (9 %) only exceeded from WHO safe limit. The risk assessment tools such as daily intake of metals (DIMs) and health risk indexes (HRIs) were used for health risk estimation and were observed in the order of Ni > Cr > Mn > Pb > Cd and Cd > Ni > Pb > Mn > Cr, respectively. The HRI values of heavy metals for both children and adults were <1, showing lack of potential health risk to the local inhabitants of the study area.

Population exposure to emerging perfluoroalkyl acids (PFAAs) via drinking water resources: Application of multivariate statistics and risk assessment models.

This study assessed the occurrence, origins, and potential risks of emerging perfluoroalkyl acids (PFAAs) for the first time in drinking water resources of Khyber Pakhtunkhwa, Pakistan. In total, 13 perfluoroalkyl carboxylic acids (PFCAs) with carbon (C) chains C4-C18 and 4 perfluoroalkyl sulfonates (PFSAs) with C chains C4-C10 were tested in both surface and ground drinking water samples using a high-performance liquid chromatography system (HPLC) equipped with an Agilent 6460 Triple Quadrupole liquid chromatography-mass spectrometry (LC-MS) system. The concentrations of ∑PFCAs, ∑PFSAs, and ∑PFAAs in drinking water ranged from 1.46 to 72.85, 0.30-8.03, and 1.76-80.88 ng/L, respectively. Perfluorobutanoic acid (PFBA), perfluorohexanoic acid (PFHxA), and perfluoropentanoic acid (PFPeA) were the dominant analytes in surface water followed by ground water, while the concentration of perfluorobutane sulfonate (PFBS), perfluorooctanoic acid (PFOA), perfluoroheptanoic acid (PFHpA), perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHxS), perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluoroundecanoic acid (PFUnDA), and perfluorododecanoic acid (PFDoDA) were greater than long-chain PFOA and PFOS. The correlation statistics, which showed a strong correlation (p < 0.05) between the PFAA analytes, potentially indicated the fate of PFAAs in the area's drinking water sources, whereas the hierarchical cluster analysis (HCA) and principal component analysis (PCA) statistics identified industrial, domestic, agricultural, and commercial applications as potential point and non-point sources of PFAA contamination in the area. From risk perspectives, the overall PFAA toxicity in water resources was within the ecological health risk thresholds, where for the human population the hazard quotient (HQ) values of individual PFAAs were < 1, indicating no risk from the drinking water sources; however, the hazard index (HI) from the ∑PFAAs should not be underestimated, as it may significantly result in potential chronic toxicity to exposed adults, followed by children.

Antibacterial Efficacy of Green Synthesized Silver Nanoparticles Using Salvia nubicola Extract against Ralstonia solanacearum, the Causal Agent of Vascular Wilt of Tomato.

Ralstonia solanacearum is a phytopathogen causing bacterial wilt diseases of tomato and affecting its productivity, which leads to prominent economic losses annually. As an alternative to conventional pesticides, green synthesized nanoparticles are believed to possess strong antibacterial activities besides being cheap and ecofriendly. Here, we present the synthesis of silver nanoparticles (Sn-AgNPs) from medicinally important aqueous plant extracts of Salvia nubicola. Characterization of biologically synthesized nanoparticles was performed through UV-vis spectrophotometry, Fourier transform infrared spectroscopy (FTIR), energy-dispersive X-ray spectroscopy, X-ray diffraction, scanning electron microscopy, transmission electron microscopy (TEM), and thermogravimetric analysis. The antibacterial activity of the biosynthesized silver nanoparticles was tested against the phytopathogen R. solanacearum through in vitro experiments. Preliminary phytochemical analysis of the plant extracts revealed the presence of substantial amounts of flavonoids (57.08 mg GAE/g), phenolics (42.30 mg GAE/g), tannins, and terpenoids. The HPLC phenolic profile indicated the presence of 25 possible bioactive compounds. Results regarding green synthesized silver nanoparticles revealed the conformation of different functional groups through FTIR analysis, which could be responsible for the bioreduction and capping of Ag ions into silver NPs. TEM results revealed the spherical, crystalline shape of nanoparticles with the size in the range of 23-63 nm, which validates SEM results. Different concentrations of Sn-AgNPs (T1 (500 µg/mL) to T7 (78.1 µg/mL)) with a combination of plant extracts (PE-Sn-AgNPs) and plant extracts alone exhibited an efficient inhibition of R. solanacearum. These findings could be used as an effective alternative preparation against the bacterial wilt of tomato.

A comprehensive review on traditional uses, phytochemistry and pharmacological properties of Paeonia emodi Wall. ex Royle: current landscape and future perspectives.

Paeonia emodi Wall. ex Royle is commonly known as Himalayan paeony has great importance as a food and medicine. The practice of Paeonia emodi Wall. ex Royle is very ancient and it is conventionally used for a wide range of illnesses in the folk system of medicine because of its wide beneficial phytochemical profile. The main purpose of the current review was the synthesis of recent data on botany, ethnopharmacology, phytochemistry and potential pharmacological mechanisms of action of Paeonia emodi Wall. ex Royle, thus offering new prospects for the development of new adjuvant natural therapies. Using scientific databases such as PubMed/MedLine, Scopus, Web of Science, ScienceDirect, Google Scholar, Springer, and Wiley, a comprehensive literature search was performed for Paeonia emodi Wall. ex Royle. For searching, we used the next MeSH terms: "Biological Product/isolation and purification", "Biological Products/pharmacology", "Drug Discovery/methods", "Ethnopharmacology, Medicine", "Traditional/methods", "Paeonia/chemistry", "Plant Extracts/pharmacology", "Phytochemicals/chemistry", "Phytochemicals/pharmacology", "Plants, Medicinal". The results of the most recent studies were analyzed and the most important data were summarized in tables and figures. Phytochemical research of Paeonia emodi Wall. ex Royle has led to the isolation of triterpenes, monoterpenes, phenolic acids, fatty acids, organic compounds, steroids, free radicals and some other classes of primary metabolites. In addition, diverse pharmacological activities like antibacterial, antifungal, anticoagulant, airway relaxant lipoxygenase and beta-glucuronidase inhibiting activity, radical scavenging activity, phytotoxic and insecticidal activities have been reported for Paeonia emodi Wall. ex Royle. Different bioactive compounds of Paeonia emodi Wall. ex Royle has proven their therapeutic potential in modern pharmacological and biomedical research to cure numerous gastrointestinal and nervous disorders. In future, further in vitro and in vivo therapeutic studies are required to identify new mechanisms of action, pharmacokinetics studies, and new pharmaceutical formulations for target transport and possible interaction with allopathic drugs. Also, new research regarding quality evaluation, toxicity and safety data in humans is needed.

Comparative Effects of Hydropriming and Iron Priming on Germination and Seedling Morphophysiological Attributes of Stay-Green Wheat.

Seed priming is considered to play an essential role in the overall improvement of agricultural crops. The current research work was carried out in order to investigate the comparative effects of hydropriming and iron priming on the germination behavior and morphophysiological attributes of wheat seedlings. The experimental materials consisted of three wheat genotypes including a synthetically derived wheat line (SD-194), stay-green wheat genotype (Chirya-7), and conventional wheat variety (Chakwal-50). The treatments included hydro (distilled and tap water)- and iron priming (10 and 50 mM) of wheat seeds for 12 h duration. Results indicated that both priming treatment and wheat genotypes exhibited highly different germination and seedling characteristics. These included germination percentage, root volume, root surface, root length, relative water content, chlorophyll content, membrane stability index, and chlorophyll fluorescence attributes. Furthermore, the synthetically derived line (SD-194) was the most promising in majority of the studied attributes by exhibiting a high germination index (2.21%), root fresh weight (7.76%), shoot dry weight (3.36%), relative water content (19.9%), chlorophyll content (7.58%), and photochemical quenching coefficient (2.58%) when compared with stay-green wheat (Chirya-7). The study also found that hydropriming with tap water and priming wheat seeds with low concentrations of iron yielded better results when a comparison was made with wheat seeds primed at high concentrations of iron. Therefore, wheat seed priming with tap water and iron solution for 12 h is recommended for optimum wheat improvement. Furthermore, current findings suggest that seed priming may have the prospect of an innovative and user-friendly approach for wheat biofortification with the aim of enhanced iron acquisition and accumulation in grains.

High throughput biochemical profiling, and functional potential analysis for valorization of grape peduncles.

Vitis vinifera L., commonly known as grape is a major fruit crop in the world. Grapes seem to confer health benefits due to their chemical components, biological and antioxidant activities. The present study is conducted to evaluate the biochemical constituents, antioxidant, and antimicrobial potential of ethanolic grape peduncles (EGP) extract. The result of phytochemical analysis revealed the presence of various phytochemicals such as flavonoid, tannin, carbohydrates, alkaloids, cardiac glycoside, phenol, steroid, terpenoids, quinones and anthraquinones. Furthermore, total phenolic content (TPC) and total flavonoid contents (TFC) were 7.35 ± 0.25 mg GAE/g (Gallic Acid Equivalent per gram) and 29.67 ± 0.13 mg QE/g (Quercetin Equivalent per gram) respectively. DPPH (2, 2-diphenyl-1-picrylhydrazyl) free radical scavenging assay revealed IC50 = 159.3 µg/mL. The antibacterial and antifungal study disclosed that the extract was highly potent against Salmonella typhi with maximum zone of inhibition of 27.2 ± 1.60 mm and Epidermophyton floccosum with 74 ± 1.81% inhibition. The extract was analyzed for its cytotoxicity and antileishmanial activity and showed no activity against HeLa cell line and promastigotes of Leishmania major. Elements Fe, Mn, Ni, Pb and Cd were determined by atomic absorption spectroscopy and approximately 50 compounds were identified by Gas Chromatography-Mass Spectrometry (GC-MS). Current work suggest that grape peduncles can be a promising source of bioactive medicinal component.

Traditional Uses of Wild Edible Mushrooms among the Local Communities of Swat, Pakistan.

Mushrooms play a crucial role in human life as well as in nature, providing food, medicine, and carrying out vital processes of decomposition, nutrient recycling, and developing mycorrhizal association with plants. The traditional system of knowledge about identification, collection, and usage of mushrooms has been accumulated through the shared experiences of many generations. Unfortunately, there have been continuous threats to the folk knowledge of mushrooms mainly due to habitat degradation, urbanization, and contemporary medication. The current research was, therefore, aimed to document an ethnomycological knowledge possessed by the ethnic communities of Swat, Pakistan. The purposive randomized sampling was carried out using chain referral method. Ethno-mycological information was collected from 62 informants using free listing, preference ranking, and use totaled methods. In total, 34 species of mushrooms belonging to 31 genera and 21 families were reported. About 85% of the reported species belong to Basidiomycetes, and 12.5% to Ascomycetes are used as food and for medicinal purposes. Morchella angusticeps, M. esculenta, Pleurotus sp., Auricularia sp., Flammulina velutipes, Agaricus bisporus, Ganoderma lucidum, and Sanghuangporus sanghuang were among the most cited edible and medicinal mushrooms. The current study revealed that district Swat is rich in wild edible and medicinal mushrooms (WEMs), and the local communities possess rich traditional knowledge about their collection, storage, and utilization. The diversity of WEMs of this region could contribute substantially to the socio-economic uplifting of the local communities through appropriate domestication and commercialization. Anthropogenic factors, coupled with depletion of traditional knowledge, threaten the diversity of WEMs in the region; therefore, in situ and ex situ conservation strategies are highly recommended.

Simplified floral dip transformation method of Arabidopsis thaliana.

Being a model plant, Arabidopsis transformation is a frequent routine in genetics and molecular laboratories. Here, we explain a modified floral dip method routinely used in our laboratory for Arabidopsis transformation by applying drought stress after plant treatment with Agrobacterium. By using this method, hundreds of individual transgenic plants are produced from just 30 infiltrated plants by increasing transformation frequency from 1% to 10% within 2-3 months.

The poly(A) polymerase PAPS1 mediates pollen maturation by regulating sperm cell differentiation in plants.

In flowering plants, a haploid microspore undergoes an asymmetric division to produce the male germline that encounters a mitotic division to produce two germ cells. The resulting germ cells undergo a series of specialization events to produce the two sperm cells required for double fertilization. These events include to upregulate male germline-specific while downregulating male germline-nonspecific regulon, but how these specializations events are regulated, are still unresolved. To know how plant sperm cell is specialized, we mutagenized Arabidopsis double homozygous transgenic line (MGH3p-MGH3::eGFP and ACTIN11p-H2B::mRFP) by an ethyl methane sulfonate (EMS) treatment and isolated a mutant with sperms identity loss, resulting in a completely male defective plant. Second-generation sequencing identified a point mutation G/A causing premature stop codon TGG/TGA in the poly(A) polymerase PAPS1 that is linked with phenotype. Further, we found that paps1 mutant fails to upregulate male germline-specific regulon and to downregulate male germline-nonspecific factors required for sperm cell differentiation and attaining pollen maturation. Previously, polyadenylation of pre-mRNAs by PAPS1 has been found crucial for both RNA-based silencing processes and the processing of pre-mRNAs into mature mRNAs ready for translation. This study concludes that PAPS1 mediates sperm cell differentiation through upregulating specific while silencing the nonspecific factors of male germlines.

Immunoinformatic-based design of immune-boosting multiepitope subunit vaccines against monkeypox virus and validation through molecular dynamics and immune simulation.

Monkeypox virus is the causative agent of monkeypox disease, belonging to an orthopoxvirus genus, with a disease pattern similar to that of smallpox. The number of monkeypox cases have robustly increased recently in several countries around the world, potentially causing an international threat. Therefore, serious measures are indispensable to be taken to mitigate the spread of the disease and hence, under these circumstances, vaccination is the best choice to neutralize the monkeypox virus. In the current study, we used immunoinformatic approaches to target the L1R, B5R, and A33R proteins of the monkeypox virus to screen for immunogenic cytotoxic T-lymphocyte (CTL), helper T-lymphocyte (HTL), and B-cell epitopes to construct multiepitope subunit vaccines. Various online tools predicted the best epitope from immunogenic targets (L1R, B5R, and A33R) of monkeypox virus. The predicted epitopes were joined together by different linkers and subjected to 3D structure prediction. Molecular dynamics simulation analysis confirmed the proper folding of the modeled proteins. The strong binding of the constructed vaccines with human TLR-2 was verified by the molecular docking and determination of dissociation constant values. The GC content and codon adaptation index (CAI) values confirmed the high expression of the constructed vaccines in the pET-28a (+) expression vector. The immune response simulation data delineated that the injected vaccines robustly activated the immune system, triggering the production of high titers of IgG and IgM antibodies. In conclusion, this study provided a solid base of concept to develop dynamic and effective vaccines that contain several monkeypox virus-derived highly antigenic and nonallergenic peptides to control the current pandemic of monkeypox virus.

Wild Vicia Species Possess a Drought Tolerance System for Faba Bean Improvement.

Faba bean (Vicia faba L.), a drought-sensitive crop, is drastically affected by drought stresses compromising its growth and yield. However, wild relatives of faba bean are considered a reservoir of potential genetic resources for tolerance against abiotic stresses. This study was conducted to characterize wild relatives of faba bean for identification of a specific tolerance system required for its improvement against drought stress. The study focused on physiological, biochemical, and anatomical responses of wild Vicia species under drought stress conditions. The experiment was carried out under various levels of drought stress imposed through different field capacities (FC) which included 80% FC ie (well-watered condition), 55% FC (moderate stress), and 30% FC (severe stress). When compared to plants grown in a control environment, drought stress significantly reduced the studied physiological attributes including soluble sugars (21.3% and 15.8%), protein contents (14.7 and 14.6%), and chlorophyll (8.4 and 28.6%) under moderate (55% FC) and severe drought stress (30% FC), respectively. However, proline content increased by 20.5% and 27.6%, peroxidase activity by 48.5% and 57.1%, and superoxide dismutase activity by 72.6% and 64.8% under moderate and severe stress, respectively. The studied anatomical attributes were also affected under drought stress treatments, including diameter of stem xylem vessels (9.1% and 13.7%), leaf lower epidermal thickness (8.05% and 13.34%), and leaf phloem width (5.3% and 10.1%) under moderate and severe stress, respectively. Wild Vicia spp. showed better tolerance to water-deficit conditions as compared to cultivated Vicia L. The observed potential diversity for drought tolerance in wild Vicia spp. may assist in improvement of faba bean and may also help in understanding the mechanisms of adaptations in drought-prone environments.

Bio-Functional Potential and Biochemical Properties of Propolis Collected from Different Regions of Balochistan Province of Pakistan.

Propolis is a well-known resinous natural substance collected by honeybees (Apis mellifera L.) from plants exudations. Variations in chemical composition of propolis are due to different sources from which it is collected and change in climate and geographical location. In this study, different propolis samples were collected from different regions of Balochistan and examined for its chemical composition, total phenolics and total flavonoid contents, and antioxidant potential by using DPPH radical scavenging assay and antimicrobial activity. Bioactive components analysis revealed the presence of steroids, carbohydrates, flavonoids, coumarins, cardiac glycosides, quinones, anthraquinones, terpenoids, tannins, and phlobatannins at different levels. The total phenolics contents were ranged from 2.9343 ± 1.247 to 6.0216 ± 2.873 mg GAE g-1, and flavonoid contents were found to be 0.1546 ± 0.087 to 0.6586 ± 0.329 mg QE g-1, respectively. The antioxidant ability of each extract was analyzed by their concentration having 50% inhibition (IC 50). The propolis sample P3 possessed lower IC 5027.07 ± 0.73 mg mL-1 with higher % inhibition of DPPH radical, and P8 showed lower % inhibition by having IC 5084.43 ± 2.07 mg mL-1. The antibacterial activity of all samples was analyzed against a wide group of bacteria including Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Klebsiella pneumonia and propolis extract (P4) was highly active against Klebsiella pneumoniae with the maximum diameter of zone of inhibition 20.33 ± 1.52 mm, and propolis extract (P3) showed maximum zone of inhibition against Escherichia coli19.06 ± 1.90, while propolis extract (P2) was found less active with minimum diameter of zone of inhibition 7.46 ± 1.50 mm. The antifungal activity of extract was considered as active against the fungal species. Propolis extract (P3) showed 82% of zone of inhibition against Aspergillus Niger, and propolis extract (P1) was highly active against Aspergillus parasiticus with 80% of zone of inhibition. By comparing the vibration frequencies in wave numbers of the sample spectrograph acquired from an FTIR spectrophotometer, the functional groups present in the extracts were identified. The presence of seven elements (Fe, Zn, Mn, Ni, Pb, Cd, and Cr) was analyzed through atomic absorption spectrophotometer. The obtained concentrations were within the permissible ranges established by the World Health Organization. The GC-MS analysis revealed the presence of 80 different compounds belonged to different classes. The obtained results confirmed the imperative potential of propolis which can be used in various biological applications.

Exploiting the drought tolerance of wild Elymus species for bread wheat improvement.

Crop wild resources are excellent sources of new genetic variation for resilience against climate extremes. However, detailed characterization of the desirable phenotypes is essential before using these crop wild resources in breeding programs. This current study was, therefore, conducted to investigate the water stress responses of eight wild Elymus species and two wheat cultivars. The experiment was carried out under varying levels of osmotic stress induced by polyethylene glycol and progressive water stress through different field capacities. Water stress significantly reduced both physiological and biochemical traits compared to control, ranging from 7.1% (protein content) to 34.5% (chlorophyll) under moderate stress and 9.1-45.8% under severe stress. The anatomical features were also affected under progressive water stress, including a reduction in xylem vessel diameter (7.92 and 16.50%), phloem length (4.36 and 7.18%), vascular bundle length (3.09 and 6.04%), and ground tissue thickness (2.36 and 5.52%), respectively. Conclusively, Elymus borianus (endemic to Swat, Pakistan), E. russelli, E. caninus, E. longioristatus, and E. dauhuricus outperformed the check wheat cultivar, Pirsabak 2005, which is a rainfed variety. The results revealed that Elymus species belonging to the tertiary gene pool of bread wheat could be an excellent drought tolerance source for use in a breeding program.