Genome-wide identification of CaARR-Bs transcription factor gene family in pepper and their expression patterns under salinity stress.

In plants, ARRs-B transcription factors play a crucial role in regulating cytokinin signal transduction, abiotic stress resistance, and plant development. A number of adverse environmental conditions have caused severe losses for the pepper (Capsicum annuum L.)-a significant and economically important vegetable. Among the transcription factors of the type B-ARRs family, multiple members have different functions. In pepper, only a few members of the ARRs-B family have been reported and characterized. The current study aimed to characterize ARRs-B transcription factors in C. annuum, including phylogenetic relationships, gene structures, protein motif arrangement, and RT-qPCR expression analyses and their role in salinity stress. In total, ten genes encode CaARRs-B transcription factors (CaARR1 to CaARR10) from the largest subfamily of type-B ARRs were identified in C. annum. The genome-wide analyses of the CaARRs-B family in C. annuum were performed based on the reported ARRs-B genes in Arabidopsis. An analysis of homologous alignments of candidate genes, including their phylogenetic relationships, gene structures, conserved domains, and qPCR expression profiles, was conducted. In comparison with other plant ARRs-B proteins, CaARRs-B proteins showed gene conservation and potentially specialized functions. In addition, tissue-specific expression profiles showed that CaARRs-B genes were differentially expressed, suggesting functionally divergent. CaARRs-B proteins had a typical conserved domain, including AAR-like (pfam: PF00072) and Myb DNA binding (pfam: PF00249) domains. Ten of the CaARRs-B genes were asymmetrically mapped on seven chromosomes in Pepper. Additionally, the phylogenetic tree of CaARRs-B genes from C. annuum and other plant species revealed that CaARRs-B genes were classified into four clusters, which may have evolved conservatively. Further, using quantitative real-time qRT-PCR, the study assessed the expression patterns of CaARRs-B genes in Capsicum annuum seedlings subjected to salt stress. The study used quantitative real-time qRT-PCR to examine CaARRs-B gene expression in Capsicum annuum seedlings under salt stress. Roots exhibited elevated expression of CaARR2 and CaARR9, while leaves showed decreased expression for CaARR3, CaARR4, CaARR7, and CaARR8. Notably, no amplification was observed for CaARR10. This research sheds light on the roles of CaARRs-B genes in pepper's response to salinity stress. These findings enrich our comprehension of the functional implications of CaARRs-B genes in pepper, especially in responding to salinity stress, laying a solid groundwork for subsequent in-depth studies and applications in the growth and development of Capsicum annuum.

Genetic Diversity Analysis of Banana Cultivars (Musa sp.) in Saudi Arabia Based on AFLP Marker.

Banana plantation has been introduced recently to a temperate zone in the southeastern parts of Saudi Arabia (Fifa, Dhamadh, and Beesh, located in Jazan province). The introduced banana cultivars were of a clear origin without a recorded genetic background. In the current study, the genetic variability and structure of five common banana cultivars (i.e., Red, America, Indian, French, and Baladi) were analyzed using the fluorescently labeled AFLP technique. Nine different primer pairs combinations yielded 1468 loci with 88.96% polymorphism. Among all locations, high expected heterozygosity under the Hardy-Weinberg assumption was found (0.249 ± 0.003), where Dhamadh was the highest, followed by Fifa and Beesh, respectively. Based on the PCoA and Structure analysis, the samples were not clustered by location but in pairs in accordance with the cultivar's names. However, the Red banana cultivar was found to be a hybrid between the American and Indian cultivars. Based on ΦST, 162 molecular markers (i.e., loci under selection) were detected among cultivars. Identifying those loci using NGS techniques can reveal the genetic bases and molecular mechanisms involved in the domestication and selection indicators among banana cultivars.

DNA Barcoding of Wild Plants with Potential Medicinal Properties from Faifa Mountains in Saudi Arabia.

Wild medicinal plants are the main source of active ingredients and provide a continuous natural source for many folk medicinal products, a role that is important for society's health with an impressive record of utilization. Thus, surveying, conserving, and precisely identifying wild medicinal plants is required. The current study aimed to precisely identify fourteen wild-sourced medicinal plants from southwest Saudi Arabia, within the Fifa mountains area located in Jazan province, using the DNA barcoding technique. Two DNA regions (nuclear ITS and chloroplast rbcL) were sequenced and analyzed for the collected species using BLAST-based and phylogeny-based identification methods. Based on our analysis, ten of the fourteen species were successfully identified by DNA barcoding, five were identified as morphologically inspected, and three were morphologically indifferent. The study was able to distinguish some key medicinal species and highlight the importance of combining morphological observation with DNA barcoding to ensure the precise identification of wild plants, especially if they are medicinally relevant and associated with public health and safety usage.

Utilization of Genetic Resources, Genetic Diversity and Genetic Variability for Selecting New Restorer Lines of Rice (Oryza sativa L.).

Exploration of and understanding diversity and variability in genotypes of germplasm determines the success of rice improvement programs. One of the most important determinants of the success of breeding programs is genetic diversity and inheritance of traits. Genetic variability analysis helps breeders to determine the appropriate selection method and standards to be used to improve the preferred trait. The aim of this study was to estimate genetic components, heritability and to obtain information about genetic diversity using cluster analysis and principal component analysis. Twenty rice genotypes with three replicates in a randomized complete block design were analyzed at the Experimental Farm at Sakha Agricultural Research Station, Sakha, Kafr El-Sheikh, Egypt, during the period from 2017 to 2020. The results of the analysis of variance showed that highly significant variations were recorded between the studied genotypes for all traits. The genotypic coefficient of variation (GCV%) and phenotypic (PCV%) coefficient of variation were moderate for plant height, panicles/plant, panicle weight, spikelets/panicle, filled grains/panicle, grain yield/plant and amylose content percentage for the first-year, second-year and combined data. This indicates the existence of beneficial genetic variability that can be exploited to improve these traits. The broad-sense estimates of heritability were high and recorded values higher than 60% for all the studied traits for the two-year and combined data, except for hulling percentage. This indicates that the selection of traits that have high heritability and are less affected by the environment leads to an acceleration of the improvement of these traits. The results from the cluster analysis and principal component analysis revealed a high level of genotypic variation among the studied genotypes and genetic diversity between them. One of the most important outcomes of this study is the successful utilization of genetic resources (germplasm) from ancient varieties and lines of rice in selecting and identifying 17 new restoration lines of rice, which have various improvement purposes in rice and hybrid rice breeding programs.

Genetic Characterization of Some Saudi Arabia's Accessions from Commiphora gileadensis Using Physio-Biochemical Parameters, Molecular Markers, DNA Barcoding Analysis and Relative Gene Expression.

Commiphora gileadensis L. is a medicinal plant, known as balsam, with pharmaceutical potential for its phytochemical activities and chemical constituents. Genetic diversity is a genetic tool used in medicinal plant evolution and conservation. Three accessions from C. gileadensis were collected from three localities in Saudi Arabia (Jeddah, Jizan and Riyadh). Genetic characterization was carried out using physio-biochemical parameters, molecular markers (inter-simple sequence repeat (ISSR) and start codon targeted (SCoT)), DNA barcoding (18 S rRNA and ITS rDNA regions), relative gene expressions (phenylalanine ammonia-lyase 1 (PAL1), defensin (PR-12)) and pathogenesis-related protein (AFPRT). The results of this study showed that C. gileadensis accession C3, collected from Riyadh, had the highest content from the physio-biochemical parameters perspective, with values of 92.54 mg/g and 77.13 mg/g for total phenolic content (TPC) and total flavonoid content (TFC), respectively. Furthermore, the highest content of antioxidant enzyme activity was present in accession C3 with values of 16.87, 60.87, 35.76 and 27.98 U mg-1 for superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) (mol/min/mg FW) and ascorbate peroxidase (APX) (U mg-1 protein), respectively. The highest total number of bands and number of unique bands were 138 and 59, respectively, for the SCoT marker. The SCoT marker was the most efficient for the genetic diversity of C. gileadensis by producing the highest polymorphism (75.63%). DNA barcoding using 18 S and ITS showed the nearby Commiphora genus and clustered C. gileadensis accessions from Jeddah and Jizan in one clade and the C. gileadensis accession from Ryiadh in a separate cluster. Moreover, relative gene expression of the PAL1, defensin (PR-12) and AFPRT (PR1) genes was upregulated in the C. gileadensis accession from Ryiadh. In conclusion, ecological and environmental conditions in each locality affect the genomic expression and genetic diversity, which can help the evolution of important medicinal plants and improve breeding and conservation systems.

Green Synthesized of Thymus vulgaris Chitosan Nanoparticles Induce Relative WRKY-Genes Expression in Solanum lycopersicum against Fusarium solani, the Causal Agent of Root Rot Disease.

Fusarium solani is a plant pathogenic fungus that causes tomato root rot disease and yield losses in tomato production. The current study's main goal is testing the antibacterial efficacy of chitosan nanoparticles loaded with Thyme vulgaris essential oil (ThE-CsNPs) against F. solani in vitro and in vivo. GC-MS analysis was used to determine the chemical constituents of thyme EO. ThE-CsNPs were investigated using transmission electron microscopy before being physicochemically characterized using FT-IR. ThE-CsNPs were tested for antifungal activity against F. solani mycelial growth in vitro. A pot trial was conducted to determine the most effective dose of ThE-CsNPs on the morph/physiological characteristics of Solanum lycopersicum, as well as the severity of fusarium root rot. The relative gene expression of WRKY transcript factors and defense-associated genes were quantified in root tissues under all treatment conditions. In vitro results revealed that ThE-CsNPs (1%) had potent antifungal efficacy against F. solani radial mycelium growth. The expression of three WRKY transcription factors and three tomato defense-related genes was upregulated. Total phenolic, flavonoid content, and antioxidant enzyme activity were all increased. The outfindings of this study strongly suggested the use of ThE-CsNPs in controlling fusarium root rot on tomatoes; however, other experiments remain necessary before they are recommended.

Improving Winter Wheat Photosynthesis, Nitrogen Use Efficiency, and Yield by Optimizing Nitrogen Fertilization.

Wheat is the third most producing crop in China after maize and rice. In order to enhance the nitrogen use efficiency (NUE) and grain yield of winter wheat, a two-year field experiment was conducted to investigate the effect of different nitrogen ratios and doses at various development stages of winter wheat (Triticum aestivum L.). A total of five N doses (0, N75, N150, N225, and N300 kg ha-1) as main plots and two N ratios were applied in split doses (50%:50% and 60%:40%, referring to 50% at sowing time and 50% at jointing stage, 50% at sowing time + 50% at flowering stage, 50% at sowing time + 50% at grain filling stage, and 60% + 40% N ratio applied as a 60% at sowing time and 40% at jointing stage, 60% at sowing time and 40% at flowering stage, and 60% at sowing time and 40% at grain filling stage in subplots). The results of this study revealed that a nitrogen dose of 225 kg ha-1 significantly augmented the plant height by 27% and above ground biomass (ABG) by 24% at the grain filling stage, and the leaf area was enhanced by 149% at the flowering stage under 60 + 40% ratios. Furthermore, the N225 kg ha-1 significantly prompted the photosynthetic rate by 47% at the jointing and flowering stages followed by grain filling stage compared to the control. The correlation analysis exhibited the positive relationship between nitrogen uptake and nitrogen content, chlorophyll, and dry biomass, revealing that NUE enhanced and ultimately increased the winter wheat yield. In conclusion, our results depicted that optimizing the nitrogen dose (N225 kg/ha-1) with a 60% + 40% ratio at jointing stage increased the grain yield and nitrogen utilization rate.

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.

In-silico predicting as a tool to develop plant-based biomedicines and nanoparticles: Lycium shawii metabolites.

INTRODUCTION AND PURPOSE: In silico approach helps develop biomedicines and is useful for exploring the pharmacology of potential therapeutics using computer-simulated models. In vitro assays were used to determine the anti-microbial and cytotoxic efficacies of silver nanoparticles (AgNPs) synthesized with the shrub Lycium shawii. METHODS: In silico predicting was performed to assess the L. shawii metabolites identified using QTOF-LCMS for their pharmacological properties. L. shawii mediated AgNPs were synthesized and characterized (FTIR, TEM, SEM, DLS and EDX). The anti-bacterial efficacies of L. shawii extract, AgNPs, and penicillin-conjugated AgNPs (pen-AgNPs) were determined. The cytotoxicity of the AgNPs was measured against colorectal cancer cell line (HCT116), normal breast epithelium (MCF 10 A), and breast cancer cell line (MDA MB 231). RESULTS AND DISCUSSION: Five molecules (costunolide, catechin, emodin, lyciumaside, and aloe emodin 11-O-rhamnoside) were detected in the L. shawii extract. AgNPs (69 nm) were spherical with crystallographic structure. All three agents prepared showed inhibitory activity against the tested bacteria, the most efficacious being pen-AgNPs. High cytotoxicity of AgNPs (IC50 62 µg/ml) was observed against HCT116, IC50 was 78 µg/ml for MCF 10 A, and 250 µg/ml for MDA MB 231, of which cells showed apoptotic features under TEM examination. The in silico approach indicated that the carbonic anhydrase IX enzyme was the target molecule mediating anti-cancer and anti-bacterial activities and that emodin was the metabolite in action. CONCLUSIONS: Combining in vitro studies and in silico molecular target prediction helps find novel therapeutic agents. Among L. shawii metabolites, emodin is suggested for further studies as an agent for drug development against pathogenic bacteria and cancer.

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.

Quinoa (Chenopodium quinoa Willd.): Genetic Diversity According to ISSR and SCoT Markers, Relative Gene Expression, and Morpho-Physiological Variation under Salinity Stress.

Quinoa (Chenopodium quinoa Willd.) is a halophytic crop that can withstand a variety of abiotic stresses, including salt. The present research examined the mechanisms of salt tolerance in five different quinoa genotypes at four different salinity levels (control (60), 80, 120, and 160 mM NaCl). ISSR and SCoT analysis revealed high polymorphism percentages of 90.91% and 85.26%, respectively. Furthermore, ISSR 1 and SCoT 7 attained the greatest number of polymorphic amplicons (27 and 26), respectively. Notably, LINE-6 and M-28 genotypes demonstrated the greatest number of unique positive and negative amplicons (50 and 42) generated from ISSR and SCoT, respectively. Protein pattern analysis detected 11 bands with a polymorphism percentage 27.27% among the quinoa genotypes, with three unique bands distinguishable for the M-28 genotype. Similarity correlation indicated that the highest similarity was between S-10 and Regeolone-3 (0.657), while the lowest similarity was between M-28 and LINE-6 (0.44). Significant variations existed among the studied salinity treatments, genotypes, and the interactions between them. The highest and lowest values for all the studied morpho-physiological and biochemical traits were recorded at 60 and 160 mM NaCl concentrations, respectively, except for the Na and proline contents, which exhibited the opposite relationship. The M-28 genotype demonstrated the highest values for all studied characteristics, while the LINE-6 genotype represented the lowest in both seasons. On the other hand, mRNA transcript levels for CqSOS1 did not exhibit differential expression in roots and leaf tissues, while the expression of CqNHX1 was upregulated more in both tissues for the M-28 genotype than for the LINE-6 genotype, and its maximum induction was seen in the leaves. Overall, the genotypes M-28 and LINE-6 were identified as the most and least salinity-tolerant, respectively.