Effect of Eco-Friendly Application of Bee Honey Solution on Yield, Physio-Chemical, Antioxidants, and Enzyme Gene Expressions in Excessive Nitrogen-Stressed Common Bean (Phaseolus vulgaris L.) Plants.

Excessive use of nitrogen (N) pollutes the environment and causes greenhouse gas emissions; however, the application of eco-friendly plant biostimulators (BSs) can overcome these issues. Therefore, this paper aimed to explore the role of diluted bee honey solution (DHS) in attenuating the adverse impacts of N toxicity on Phaseolus vulgaris growth, yield quality, physio-chemical properties, and defense systems. For this purpose, the soil was fertilized with 100, 125, and 150% of the recommended N dose (RND), and the plants were sprayed with 1.5% DHS. Trials were arranged in a two-factor split-plot design (N levels occupied main plots × DH- occupied subplots). Excess N (150% RND) caused a significant decline in plant growth, yield quality, photosynthesis, and antioxidants, while significantly increasing oxidants and oxidative damage [hydrogen peroxide (H2O2), superoxide (O2•-), nitrate, electrolyte leakage (EL), and malondialdehyde (MDA) levels]. However, DHS significantly improved antioxidant activities (glutathione and nitrate reductases, catalase, ascorbate peroxidase, superoxide dismutase, proline, ascorbate, α-tocopherol, and glutathione) and osmoregulatory levels (soluble protein, glycine betaine, and soluble sugars). Enzyme gene expressions showed the same trend as enzyme activities. Additionally, H2O2, O2•-, EL, MDA, and nitrate levels were significantly declined, reflecting enhanced growth, yield, fruit quality, and photosynthetic efficiency. The results demonstrate that DHS can be used as an eco-friendly approach to overcome the harmful impacts of N toxicity on P. vulgaris plants.

Genome-Wide Analysis and Expression Profiling of DUF668 Genes in Glycine max under Salt Stress.

The DUF668 gene performs a critical role in mitigating the impact of abiotic stress factors. In this study, we identified 30 DUF668 genes in a soybean genome, distributed across fifteen chromosomes. The phylogenetic analysis classified the DUF668 genes into three groups (group I, group II, and group III). Interestingly, gene structure analysis illustrated that several GmDUF668 genes were without introns. Furthermore, the subcellular localization results suggested that GmDUF668 proteins were present in the nucleus, mitochondria, cytoplasm, and plasma membrane. GmDUF668 promoters were analyzed in silico to gain insight into the presence of regulatory sequences for TFs binding. The expression profiling illustrated that GmDUF668 genes showed expression in leaves, roots, nodules, and flowers. To investigate their response to salt stress, we utilized the RNA sequencing data of GmDUF668 genes. The results unveiled that GmDUF668-8, GmDUF668-20, and GmDUF668-30 genes were upregulated against salt stress treatment. We further validated these findings using qRT-PCR analysis. These findings provide a scientific basis to explore the functions of GmDUF668 genes against different stress conditions.

Evaluating the genotoxicity of salinity stress and secondary products gene manipulation in lime, Citrus aurantifolia, plants.

Salinity is a significant abiotic stress that has a profound effect on growth, the content of secondary products, and the genotoxicity of cells. Lime, Citrus aurantifolia, is a popular plant belonging to the family Rutaceae. The interest in cultivating this plant is due to the importance of its volatile oil, which is included in many pharmaceutical industries, but C. aurantifolia plants are affected by the NaCl salinity levels. In the present study, a comet assay test has been applied to evaluate the genotoxic impact of salinity at 0, 50, 100, and 200 mM of NaCl on C. aurantifolia tissue-cultured plants. Furthermore, terpene gene expression was investigated using a semi-quantitative real-time polymerase chain reaction. Results from the two analyses revealed that 200 mM of NaCl stress resulted in high levels of severe damage to the C. aurantifolia plants' DNA tail 21.8%, tail length 6.56 µm, and tail moment 3.19 Unit. The relative highest expression of RtHK and TAT genes was 2.08, and 1.693, respectively, when plants were exposed to 200 mM of NaCl, whereas pv4CL2RT expressed 1.50 in plants subjected to 100 mM of NaCl. The accumulation of transcripts for the RTMYB was 0.951 when plants were treated with NaCl at 50 mM, and RtGPPS gene was significantly decreased to 0.446 during saline exposure at 100 mM. We conclude that the comet assay test offers an appropriate tool to detect DNA damage as well as RtHK, TAT, and pv4CL2RT genes having post-transcriptional regulation in C. aurantifolia plant cells under salinity stress. Future studies are needed to assess the application of gene expression and comet assay technologies using another set of genes that show vulnerability to different stresses on lime and other plants.

Omics-assisted characterization of two-component system genes from Gossypium Raimondii in response to salinity and molecular interaction with abscisic acid.

The two-component system (TCS) genes are involved in a wide range of physiological processes in prokaryotes and eukaryotes. In plants, the TCS elements help in a variety of functions, including cell proliferation, response to abiotic and biotic stresses, leaf senescence, nutritional signaling, and division of chloroplasts. Three different kinds of proteins make up the TCS system in plants. These are known as HKs (histidine kinases), HPs (histidine phosphotransfer), and RRs (response regulators). We investigated the genome of Gossypium raimondii and discovered a total of 59 GrTCS candidates, which include 23 members of the HK family, 8 members of the HP family, and 28 members of the RR family. RR candidates are further classified as type-A (6 members), type-B (11 members), type-C (2 members), and pseudo-RRs (9 members). The GrTCS genes were analyzed in comparison with the TCS components of other plant species such as Arabidopsis thaliana, Cicer arietinum, Sorghum bicolor, Glycine max, and Oryza sativa. This analysis revealed both conservation and changes in their structures. We identified 5 pairs of GrTCS syntenic homologs in the G. raimondii genome. All 59 TCS genes in G. raimondii are located on all thirteen chromosomes. The GrTCS promoter regions have several cis-regulatory elements, which function as switches and respond to a wide variety of abiotic stresses. RNA-seq and real-time qPCR analysis showed that the majority of GrTCS genes are differentially regulated in response to salt and cold stress. 3D structures of GrTCS proteins were predicted to reveal the specific function. GrTCSs were docked with abscisic acid to assess their binding interactions. This research establishes the groundwork for future functional studies of TCS elements in G. raimondii, which will further focus on stress resistance and overall development.

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.

Study of genetic variability, heritability, and genetic advance for yield-related traits in tomato (Solanum lycopersicon MILL.).

Tomato is one of the most significant vegetable crops, which provides several important dietary components. Pakistan has a significant low tomato yield compared to other countries because of low genetic diversity and the absence of improved cultivars. The present study aimed to investigate the genetic variability, heritability, and genetic advance for yield and yield-related traits in tomato. For this purpose, eight tomato parents and their 15 crosses or hybrids were evaluated to study the relevant traits. Significant variation was observed for all studied traits. Higher values of the genotypic coefficient of variability (GCV) and phenotypic coefficient of variability (PCV) were recorded for yield per plant (YP) (kg) (37.62% and 37.79%), as well as the number of fruits per cluster (NFRC) (31.52% and 31.71%), number of flowers per cluster (24.63 and 24.67), and single fruit weight (g) (23.49 and 23.53), which indicated that the selection for these traits would be fruitful. Higher heritability (h2) estimates were observed for the number of flowers per cluster (NFC) (0.99%), single fruit weight (SFW) (g) (0.99%), and yield per plant (YP) (kg) (0.99%). Single fruit weight (SFW) (g) exhibited higher values for all components of variability. High genetic advance as a % of the mean (GAM) coupled with higher heritability (h2) was noted for the yield per plant (YP) (kg) (52.58%) and the number of fruits per cluster (NFRC) (43.91). NFRC and SFW (g) had a highly significant correlation with YP (kg), while FSPC had a significant positive association with YP (kg), and these traits can be selected to enhance YP (kg). Among the 15 hybrids, Nagina × Continental, Pakit × Continental, and Roma × BSX-935 were selected as high-yielding hybrids for further evaluation and analysis. These findings revealed that the best performing hybrids could be used to enhance seed production and to develop high-yielding varieties. The parents could be further tested to develop hybrids suitable for changing climatic conditions. The selection of YP (kg), SFW (g), NFC, and NFRC would be ideal for selecting the best hybrids.

Comparative Gene Enrichment Analysis for Drought Tolerance in Contrasting Maize Genotype.

Drought stress is a significant abiotic factor influencing maize growth and development. Understanding the molecular mechanism of drought tolerance is critical to develop the drought tolerant genotype. The identification of the stress responsive gene is the first step to developing a drought tolerant genotype. The aim of the current research was to pinpoint the genes that are essential for conserved samples in maize drought tolerance. In the current study, inbred lines of maize, 478 and H21, a drought-tolerant and susceptible line, were cultivated in the field and various treatments were applied. The circumstances during the vegetative stage (severe drought, moderate drought and well-watered environments) and RNA sequencing were used to look into their origins. In 478, 68%, 48% and 32% of drought-responsive genes (DRGs) were found, with 63% of DRGs in moderate drought and severe drought conditions in H21, respectively. Gene ontology (GO) keywords were explicitly enriched in the DRGs of H21, which were considerably over-represented in the two lines. According to the results of the GSEA, "phenylpropanoid biosynthesis" was exclusively enriched in H21, but "starch and sucrose metabolism" and "plant hormone signal transduction" were enhanced in both of the two lines. Further investigation found that the various expression patterns of genes linked to the trehalose biosynthesis pathway, reactive oxygen scavenging, and transcription factors, may have a role in maize's ability to withstand drought. Our findings illuminate the molecular ways that respond to lack and offer gene resources for maize drought resistance. Similarly, SNP and correlation analysis gave us noticeable results that urged us to do the same kind of analysis on other crops. Additionally, we isolated particular transcription factors that could control the expression of genes associated to photosynthesis and leaf senescence. According to our findings, a key factor in tolerance is the equilibrium between the induction of leaf senescence and the preservation of photosynthesis under drought.

Antibacterial, Antioxidant Activities, GC-Mass Characterization, and Cyto/Genotoxicity Effect of Green Synthesis of Silver Nanoparticles Using Latex of Cynanchum acutum L.

Green synthesis of nanoparticles is receiving more attention these days since it is simple to use and prepare, uses fewer harsh chemicals and chemical reactions, and is environmentally benign. A novel strategy aims to recycle poisonous plant chemicals and use them as natural stabilizing capping agents for nanoparticles. In this investigation, silver nanoparticles loaded with latex from Cynanchum acutum L. (Cy-AgNPs) were examined using a transmission electron microscope, FT-IR spectroscopy, and UV-visible spectroscopy. Additionally, using Vicia faba as a model test plant, the genotoxicity and cytotoxicity effects of crude latex and various concentrations of Cy-AgNPs were studied. The majority of the particles were spherical in shape. The highest antioxidant activity using DPPH was illustrated for CAgNPs (25 mg/L) (70.26 ± 1.32%) and decreased with increased concentrations of Cy-AGNPs. Antibacterial activity for all treatments was determined showing that the highest antibacterial activity was for Cy-AgNPs (50 mg/L) with inhibition zone 24 ± 0.014 mm against Bacillus subtilis, 19 ± 0.12 mm against Escherichia coli, and 23 ± 0.015 against Staphylococcus aureus. For phytochemical analysis, the highest levels of secondary metabolites from phenolic content, flavonoids, tannins, and alkaloids, were found in Cy-AgNPs (25 mg/L). Vicia faba treated with Cy-AgNPs- (25 mg/L) displayed the highest mitotic index (MI%) value of 9.08% compared to other Cy-AgNP concentrations (50-100 mg/L) and C. acutum crude latex concentrations (3%). To detect cytotoxicity, a variety of chromosomal abnormalities were used, including micronuclei at interphase, disturbed at metaphase and anaphase, chromosomal stickiness, bridges, and laggards. The concentration of Cy-AgNPs (25 mg/L) had the lowest level of chromosomal aberrations, with a value of 23.41% versus 20.81% for the control. Proteins from seeds treated with V. faba produced sixteen bands on SDS-PAGE, comprising ten monomorphic bands and six polymorphic bands, for a total percentage of polymorphism of 37.5%. Eight ISSR primers were employed to generate a total of 79 bands, 56 of which were polymorphic and 23 of which were common. Primer ISSR 14 has the highest level of polymorphism (92.86%), according to the data. Using biochemical SDS-PAGE and ISSR molecular markers, Cy-AgNPs (25 mg/L) showed the highest percentage of genomic template stability (GTS%), with values of 80% and 51.28%, respectively. The findings of this work suggest employing CyAgNPs (25 mg/L) in pharmaceutical purposes due to its highest content of bioactive compounds and lowest concentration of chromosomal abnormalities.

Utilization of a recombinant defensin from Maize (Zea mays L.) as a potential antimicrobial peptide.

The search for effective and bioactive antimicrobial molecules to  encounter the medical need for new antibiotics is an encouraging area of research. Plant defensins are small cationic, cysteine-rich peptides with a stabilized tertiary structure by disulfide-bridges and characterized by a wide range of biological functions. The heterologous expression of Egyptian maize defensin (MzDef) in Escherichia coli and subsequent purification by glutathione affinity chromatography yielded 2 mg/L of recombinant defensin peptide. The glutathione-S-transferase (GST)-tagged MzDef of approximately 30 kDa in size (26 KDa GST + ~ 4 KDa MzDef peptide) was immunodetected with anti-GST antibodies. The GST-tag was successfully cleaved from the MzDef peptide by thrombin, and the removal was validated by the Tris-Tricine gel electrophoresis. The MzDef induced strong growth inhibition of Rhizoctonia solani, Fusarium verticillioides, and Aspergillus niger by 94.23%, 93.34%, and 86.25%, respectively, whereas relatively weak growth inhibitory activity of 35.42% against Fusarium solani was recorded. Moreover, strong antibacterial activities were demonstrated against E. coli and Bacillus cereus and the moderate activities against Salmonella enterica and Staphylococcus aureus at all tested concentrations (0.1, 0.2, 0.4, 0.8, 1.6, and 3.2 µM). Furthermore, the in vitro MTT assay exhibited promising anticancer activity against all tested cell lines (hepatocellular carcinoma, mammary gland breast cancer, and colorectal carcinoma colon cancer) with IC50 values ranging from 14.85 to 29.85 µg/mL. These results suggest that the recombinant peptide MzDef may serve as a potential alternative antimicrobial and anticancer agent to be used in medicinal application.