Micromorphological Sculptural Diversity in Foliar Epidermis and Trichomes Features among Invasive Species.

This study examines the role of light microscopic (LM) and scanning electron microscopic (SEM) micromorphological traits of the epidermis in identifying and classifying invasive plants. SEM was conducted to increase our understanding of microscopic qualities that are not visible in light microscopy and to elucidate unclear affinities among invasive species. The study examines invasive species' morphological and anatomical characteristics from the Pothohar Plateau of Pakistan for the first time. The results showed that various micromorphological features are very useful for species' accurate identification. Adaxial and abaxial surfaces of leaves showed variations in subsidiary cells, glands, anticlinal wall patterns, stomata, and epidermal cells. Epidermal cell shapes observed were irregular, elongated, rectangular, and polygonal. Epidermal cells having maximum length were calculated in Stellaria media (126.3 µm) on adaxial side. On the abaxial surface, the minimum length was noticed in Eucalyptus camaldulensis (28.5 µm). Both glandular and nonglandular trichomes were examined, ranging from unicellular to multicellular. Most of the investigated specimens of leaves were amphistomatic, while some were hypostomatic, like Alternanthera pungens, Calotropis procera, Cannabis sativa, Lantana camara, and Thevetia peruviana. Leaf epidermal morphology contains numerous useful systematic features for accurate identifications of plant species. The micromorphological attributes under observation provide a standard criterion to the researcher for identifications of invasive flora in future morpho-taxonomic studies.

Distinguishing features of Lycium L. species (family Solanaceae) distributed in Egypt based on their anatomical, metabolic, molecular, and ecological characteristics.

Among the 70-80 species of the genus Lycium (family Solanaceae) disjunctly distributed around the world, only three are frequently distributed in different locations in Egypt. Due to the morphological similarities between these three species, there is a need for alternative tools to distinguish them. Thus, the objective of this study was to revise the taxonomic features of Lycium europaeum L., Lycium shawii Roem. & Schult., and Lycium schweinfurthii var. aschersonii (Dammer) Feinbrun in consideration of their anatomical, metabolic, molecular, and ecological characteristics. In addition to analysis of their anatomical and ecological features, DNA barcoding was performed for molecular characterization through internal transcribed spacer (ITS) sequencing and start codon targeted (SCoT) markers. Furthermore, metabolic profiling of the studied species was conducted based on gas chromatography-mass spectrometry (GC-MS). The observed anatomical features of the adaxial and abaxial epidermal layers, type of mesophyll, crystals, number of palisade and spongy layers, and the vascular system showed variations between the studied species. Beyond this, the anatomy of the leaves showed an isobilateral structure in the studied species, without distinct differences. Species were molecularly identified in terms of ITS sequences and SCoT markers. The ITS sequences were deposited in GenBank with accession numbers ON149839.1, OP597546.1, and ON521125.1 for L. europaeum L., L. shawii, and L. schweinfurthii var. aschersonii, respectively. The sequences showed variations in GC content between the studied species; this was 63.6% in L. europaeum, 61.53% in L. shawii, and 63.55% in L. schweinfurthii var. aschersonii. A total of 62 amplified fragments, including 44 polymorphic fragments with a ratio of 70.97%, were obtained in the SCoT analysis, as well as unique amplicons in L. europaeum L., shawii, and L. schweinfurthii var. aschersonii of 5, 11, and 4 fragments, respectively. Through GC-MS profiling, 38 compounds were identified with clear fluctuations in the extracts of each species. Of these, 23 were distinguishing chemicals that could help in chemical identification of the extracts of the studied species. The present study succeeds in identifying alternative clear and diverse characteristics that can be used to distinguish between L. europaeum, L. shawii, and L. schweinfurthii var. aschersonii.

Membrane-processed honey samples for pollen characterization with health benefits.

Better processing techniques must be utilized widely due to the rising demand for honey. The most common honey processing techniques are applied to melissopalynomorphs to check the quality and quantity of valuable honey using microporous ultrafiltration membranes. It is essential to have the ability to selectively filter out sugars from honey using ultrafiltration. This study authenticated 24 honey samples using membrane reactors ultrafiltration protocol to describe the pollen spectrum of dominant vegetation. The purpose of this study was also to explore nutritional benefits as well as the active phytochemical constituents of honey samples. Honey samples were collected and labeled Acacia, Eucalyptus, and Ziziphus species based on plant resources provided by local beekeepers. A variety of honeybee flora was collected around the apiaries between 2020 and 2021. Honey analysis revealed that the pollen extraction of 24 bee foraging species belonging to 14 families. The honey membrane technology verified the identities of honey and nectar sources. Also, pollen identified using honey ultrafiltration membranes revealed dominant resources: Acacia spp. (69%), Eucalyptus spp. (52%) and Ziziphus spp. Honey filtration using a membrane technology classified 14 samples as unifloral, represented by six dominant pollen types. The absolute pollen count in the honey sample revealed that 58.33% (n = 14) belong to Maurizio's class I. Scanning ultrasculpturing showed diverse exine patterns: reticulate, psilate, scabrate-verrucate, scabrate-gemmate, granulate, perforate, microechinate, microreticulate, and regulate to fossulate for correct identification of honey pollen types. Honey ultrafiltration should be utilized to validate the botanical sources of honey and trace their biogeographic authenticity. Thus, it is imperative to look at the alternative useful method to identify the botanical origin of filtered honey. It is critical to separate honey from adulteration by a standardized protocol. Membrane technology has yielded significant outcomes in the purification of honey.

Tendril Anatomy: A Tool for Correct Identification among Cucurbitaceous Taxa.

This research examined the histological micro-structure of tendril vasculature in cucurbitaceous taxa. In this research, the tendril anatomy of 17 taxa of Cucurbitaceae categorized into seven genera, including Cucumis (five species), Cucurbita and Luffa (three species each), Citrullus and Momordica (two species each) while Lagenaria and Praecitrullus (one species each), collected from different areas of the Thal desert were examined via microscopic imaging to explore its taxonomic significance. Tendril transverse sections were cut with a Shandon Microtome to prepare slides. The distinctive characteristics of taxonomic value (qualitative and quantitative) include tendril and vascular bundle shape, variation in the number of vascular bundles, tendril diameter length, layers of sclerenchyma, and shape of collenchyma and epidermal cells. Tendril shapes observed are irregular, slightly oval-shaped, slightly C shaped, angular (4-angled, 6-angled, or polygonal), and star shaped. Quantitative measurements were taken to analyze the data statistically using SPSS software. Cucurbita pepo had a maximum tendril diameter length of 656.1 µm and a minimum in Momordica balsamina of 123.05 µm. The highest number of vascular bundles (12) were noticed in Luffa acutangula var.amara. Angular type was prominent in collenchyma, and irregular shape was dominant in sclerenchyma cells. A maximum of seven to nine sclerenchyma layers were present in Lagenaria siceraria and a minimum of two or three layers in Cucumis melo subsp. agrestis, Cucumis melo var. flexuosus, and Cucumis melo var.cantalupensis. Epidermis cells also show great variations with a rectangular shape being dominant. Statistical UPGMA dendrogram clustering of tendril vasculature traits shows that histological sections studied with microscopic techniques can be used to identify species and will play a vital role in future taxonomic and phylogenic linkages.

Effect of Potassium Deficiency on Physiological Responses and Anatomical Structure of Basil, Ocimum basilicum L.

The aim of this study was to investigate the effect of a variable supply of potassium to culture medium on physiological and anatomical parameters (histological sections at the third internode) in basil, Ocimum basilicum. Thirty-four-day-old plants grown on basic nutrient medium were divided into four batches and grown on media with varying doses of potassium: 0.375 mM, 0.250 mM, 0.125 mM and 0 mM K+. After 64 days of culture, a final harvest was performed. The results showed that root and shoot growth in basil was decreased with decreased K+ concentration. This restriction was associated with a reduction in root elongation and leaf expansion, which was coupled with a decrease in chlorophyll and carotenoid contents. The estimation of electrolyte leakage reveals that this parameter was increased by potassium deficiency. With respect to total polyphenol and flavonoid contents, only the third leaf-stage extracts exhibited a decrease under low-K+ conditions. However, variability in response of phenolic compounds was recorded depending on the organ and the K+ concentration in the medium. Stem cross sections of potassium-deficient basil plants revealed a decrease in the diameter of these organs, which can be attributed to a restriction of the extent of different tissue territories (cortex and medulla), as well as by a reduction in cell size. These effects were associated with a decrease in the number of conducting vessels and an increase in the number of woody fibers.

Palyno-Morphological Characteristics as a Systematic Approach in the Identification of Halophytic Poaceae Species from a Saline Environment.

Pollen morphology of 12 salt-tolerant grasses (Calamagrostis pseudophragmites, Cenchrus biflorus, Cenchrus ciliaris, Cenchrus pennisetiformis, Cymbopogon jwarancusa, Dactyloctenium aegyptium, Echinochloa colona, Echinochloa crus-galli, Saccharum griffithii, Saccharum ravennae, Saccharum spontaneum, and Urochloa ramosa) from the Salt Range was studied using scanning electron microscopy (SEM) and light microscopy (LM). The main aim was the elucidation of taxonomic significance of pollen morphology, which might be useful for taxonomists in the identification of halophytic Poaceae taxa. The described pollen morphology is a good source of taxonomic characteristics that can help in species and genera differentiation. The investigated variations in the pollen micromorphological features and exploration of new palynological traits support and strengthen the systematics of Poaceae. The pollen shape of all studied species was sub-spheroidal, and exine ornamentation includes microechinate-areolate (11 spp.) and microechinate (1 spp.). Two types of pollen apertures were reported, i.e., monoporate (11 spp.) and diporate (Cenchrus pennisetiformis). The annulus was found in all species while operculum was observed only in three species. The univariate and multivariate analyses were used to analyze the quantitative data. The highest pollen viability values (%) depicted the adaptability of Poaceae taxa in the salt region. Overall, the pollen characteristics in polar and equatorial view, apertures, annulus, operculum, and surface ornamentation of pollen grains of the family Poaceae are of great taxonomic significance for the species identification.

Genetic Diversity, Analysis of Some Agro-Morphological and Quality Traits and Utilization of Plant Resources of Alfalfa.

Alfalfa (Medicago sativa L.) is one of the most important perennial forage crops to build effective diets for livestock producers. Forage crop improvement depends largely on the availability of diverse germplasms and their efficient utilization. The present investigation was conducted at Ismailia Agricultural Research Station to assess twenty-one alfalfa genotypes for yield components, forage yield and quality traits during 2019/2020 and 2020/2021. The genotypes were evaluated in field experiments with three replicates and a randomized complete block design, using analysis of variance, estimate of genetic variability, estimate of broad sense heritability (hb2) and cluster analysis to identify the inter relationships among the studied genotypes as well as principal component analysis (PCA) to explain the majority of the total variation. Significant differences were found among genotypes for all studied traits. The general mean of the studied traits was higher in the second year than the first year. Moreover, the combined analysis showed highly significant differences between the two years, genotypes and the year × gen. interaction for the traits studied. The genotype F18 recorded the highest values for plant height, number of tiller/m2, total fresh yield and total dry yield, while, the genotype F49 ranked first for leaf/stem ratio. The results showed highly significant variation among the studied genotypes for crude protein %, crude fiber % and ash %. Data revealed that the genotypes P13 and P5 showed the highest values for crude protein %, whereas, the genotype F18 recorded the highest values for crude fiber % and ash content. The results revealed high estimates of genotypic coefficient and phenotypic coefficient of variation (GCV% and PCV%) with high hb2, indicating the presence of genetic variability and effective potential selection for these traits. The cluster analysis exhibited considerable genetic diversity among the genotypes, which classified the twenty one genotypes of alfalfa into five sub-clusters. The genotypes F18, F49, K75, S35, P20, P5 and P13 recorded the highest values for all studied traits compared with other clusters. Furthermore, the PC analysis grouped the studied genotypes into groups and remained scattered in all four quadrants based on all studied traits. Ultimately, superior genotypes were identified can be utilized for crop improvement in future breeding schemes.

Sorghum Allelopathy: Alternative Weed Management Strategy and Its Impact on Mung Bean Productivity and Soil Rhizosphere Properties.

The reduction of herbicide use and herbicide-resistant weeds through allelopathy can be a sustainable strategy to combat the concerns of environmental degradation. Allelopathic crop residues carry great potential both as weed suppressers and soil quality enhancers. The influence of sorghum crop residues and water extracts on the weed population, soil enzyme activities, the microbial community, and mung bean crop productivity was investigated in a two-year experiment at the Student Research Farm, University of Agriculture Faisalabad. The experimental treatments comprised two levels of sorghum water extract (10 and 20 L ha-1) and two residue application rates (4 and 6 t ha-1), and no sorghum water extract and residues were used as the control. The results indicated that the incorporation of sorghum water extract and residue resulted in significant changes in weed dynamics and the soil quality indices. Significant reduction in weed density (62%) and in the dry weight of weeds (65%) was observed in T5. After the harvest, better soil quality indices in terms of the microbial population (72-90%) and microbial activity (32-50%) were observed in the rhizosphere (0-15 cm) by the same treatment. After cropping, improved soil properties in terms of available potassium, available phosphorus soil organic matter, and total nitrogen were higher after the treatment of residue was incorporated, i.e., 52-65%, 29-45%, 62-84%, and 59-91%, respectively. In the case of soil enzymes, alkaline phosphatase and dehydrogenase levels in the soil were 35-41% and 52-77% higher, respectively. However, residue incorporation at 6 t ha-1 had the greatest effect in improving the soil quality indices, mung bean productivity, and reduction of weed density. In conclusion, the incorporation of 6 t ha-1 sorghum residues may be opted to improve soil quality indices, suppress weeds, harvest a better seed yield (37%), and achieve higher profitability (306 $ ha-1) by weed suppression, yield, and rhizospheric properties of spring-planted mung beans. This strategy can provide a probable substitute for instigating sustainable weed control and significant improvement of soil properties in the mung bean crop, which can be a part of eco-friendly and sustainable agriculture.

Biological Applications of Ball-Milled Synthesized Biochar-Zinc Oxide Nanocomposite Using Zea mays L.

Nanotechnology is one of the vital and quickly developing areas and has several uses in various commercial zones. Among the various types of metal oxide-based nanoparticles, zinc oxide nanoparticles (ZnO NPs) are frequently used because of their effective properties. The ZnO nanocomposites are risk-free and biodegradable biopolymers, and they are widely being applied in the biomedical and therapeutics fields. In the current study, the biochar-zinc oxide (MB-ZnO) nanocomposites were prepared using a solvent-free ball-milling technique. The prepared MB-ZnO nanocomposites were characterized through scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) spectroscopy, X-ray powder diffraction (XRD), and thermogravimetric analysis (TGA), Fourier-transform infrared spectroscopy (FTIR), and ultraviolet-visible (UV) spectroscopy. The MB-ZnO particles were measured as 43 nm via the X-ray line broadening technique by applying the Scherrer equation at the highest peak of 36.36°. The FTIR spectroscope results confirmed MB-ZnO's formation. The band gap energy gap values of the MB-ZnO nanocomposites were calculated as 2.77 eV by using UV-Vis spectra. The MB-ZnO nanocomposites were tested in various in vitro biological assays, including biocompatibility assays against the macrophages and RBCs and the enzymes' inhibition potential assay against the protein kinase, alpha-amylase, cytotoxicity assays of the leishmanial parasites, anti-inflammatory activity, antifungal activity, and antioxidant activities. The maximum TAC (30.09%), TRP (36.29%), and DPPH radicals' scavenging potential (49.19%) were determined at the maximum dose of 200 µg/mL. Similarly, the maximum activity at the highest dose for the anti-inflammatory (76%), at 1000 µg/mL, alpha-amylase inhibition potential (45%), at 1000 µg/mL, antileishmanial activity (68%), at 100 µg/mL, and antifungal activity (73 ± 2.1%), at 19 mg/mL, was perceived, respectively. It did not cause any potential harm during the biocompatibility and cytotoxic assay and performed better during the anti-inflammatory and antioxidant assay. MB-ZnO caused moderate enzyme inhibition and was more effective against pathogenic fungus. The results of the current study indicated that MB-ZnO nanocomposites could be applied as effective catalysts in various processes. Moreover, this research provides valuable and the latest information to the readers and researchers working on biopolymers and nanocomposites.

ZnO Nanoparticle-Mediated Seed Priming Induces Biochemical and Antioxidant Changes in Chickpea to Alleviate Fusarium Wilt.

Chickpea (Cicer arietinum L.) is one of the main pulse crops of Pakistan. The yield of chickpea is affected by a variety of biotic and abiotic factors. Due to their environmentally friendly nature, different nanoparticles are being synthesized and applied to economically important crops. In the present study, Trichoderma harzianum has been used as a stabilizing and reducing agent for the mycosynthesis of zinc oxide nanoparticles (ZnO NPs). Before their application to control Fusarium wilt of chickpea, synthesized ZnO NPs were characterized. X-ray diffraction (XRD) analysis revealed the average size (13 nm) of ZnO NPs. Scanning electron microscopy (SEM) indicated their spherical structure, and energy dispersive X-ray analysis (EDX) confirmed the oxide formation of ZnO NPs. Transmission electron microscopy (TEM) described the size and shape of nanoparticles, and Fourier transform infrared (FTIR) spectroscopy displayed the presence of reducing and stabilizing chemical compounds (alcohol, carboxylic acid, amines, and alkyl halide). Successfully characterized ZnO NPs exhibited significant mycelial growth inhibition of Fusarium oxysporum, in vitro. In a greenhouse pot experiment, the priming of chickpea seeds with ZnO NPs significantly increased the antioxidant activity of germinated plants and they displayed 90% less disease incidence than the control. Seed priming with ZnO NPs helped plants to accumulate higher quantities of sugars, phenol, total proteins, and superoxide dismutase (SOD) to create resistance against wilt pathogen. These nanofungicides were produced in powder form and they can easily be transferred and used in the field to control Fusarium wilt of chickpea.