Genotypic and Phenotypic Structure of the Population of Phytophthora infestans in Egypt Revealed the Presence of European Genotypes

Late blight disease of potato and tomato, caused by Phytophthora infestans, results in serious losses to Egyptian and global potato and tomato production. To understand the structure and dynamics of the Egyptian population of P. infestans, 205 isolates were collected from potato and tomato plants during three growing seasons in 2010–2012. The characterization was achieved by mating-type assay, metalaxyl sensitivity assay, and virulence pattern. Additionally, genotyping of 85 Egyptian isolates and 15 reference UK isolates was performed using 12 highly informative microsatellite (SSR) markers David E. L. Cooke and five effector (RxLR) genes. Mating-type testing showed that 58% (118 of 205) of the isolates belonged to mating type A1, 35% (71 isolates) to mating type A2, and the rest 8% (16 isolates) were self-fertile. The phenotype of metalaxyl response was represented as 45% resistant, 43% sensitive, and 12% as intermediate. Structure analysis grouped the 85 identified genotypes into two main clonal lineages. The first clonal lineage comprised 21 isolates belonging to A2 mating type and 8 self-fertile isolates. This clonal lineage was identified as Blue_13 or EU_13_A2. The second main clonal lineage comprised 55 isolates and was identified as EU_23_A1. A single isolate with a novel SSR genotype that formed a distinct genetic grouping was also identified. The effector sequencing showed good correspondence with the virulence data and highlighted differences in the presence and absence of loci as well as nucleotide polymorphism that affect gene function. This study indicated a changing population of P. infestans in Egypt and discusses the findings in the context of late blight management

Lignin-Loaded Carbon Nanoparticles as a Promising Control Agent against Fusarium verticillioides in Maize: Physiological and Biochemical Analyses

Lignin, a naturally occurring biopolymer, is produced primarily as a waste product by the pulp and paper industries and burned to produce electricity. Lignin-based nano- and microcarriers found in plants are promising biodegradable drug delivery platforms. Here, we highlight a few characteristics of a potential antifungal nanocomposite consisting of carbon nanoparticles (C-NPs) with a defined size and shape containing lignin nanoparticles (L-NPs). Spectroscopic and microscopic studies verified that the lignin-loaded carbon nanoparticles (L-CNPs) were successfully prepared. Under in vitro and in vivo conditions, the antifungal activity of L-CNPs at various doses was effectively tested against a wild strain of F. verticillioides that causes maize stalk rot disease. In comparison to the commercial fungicide, Ridomil Gold SL (2%), L-CNPs introduced beneficial effects in the earliest stages of maize development (seed germination and radicle length). Additionally, L-CNP treatments promoted positive effects on maize seedlings, with a significant increment in the level of carotenoid, anthocyanin, and chlorophyll pigments for particular treatments. Finally, the soluble protein content displayed a favorable trend in response to particular dosages. Most importantly, treatments with L-CNPs at 100 and 500 mg/L significantly reduced stalk rot disease by 86% and 81%, respectively, compared to treatments with the chemical fungicide, which reduced the disease by 79%. These consequences are substantial considering the essential cellular function carried out by these special natural-based compounds. Finally, the intravenous L-CNPs treatments in both male and female mice that affected the clinical applications and toxicological assessments are explained. The results of this study suggest that L-CNPs are of high interest as biodegradable delivery vehicles and can be used to stimulate favorable biological responses in maize when administered in the recommended dosages, contributing to the idea of agro-nanotechnology by demonstrating their unique qualities as a cost-effective alternative compared to conventional commercial fungicides and environmentally benign nanopesticides for long-term plant protection

Biological Activity of Four <i>Trichoderma</i> Species Confers Protection against <i>Rhizoctonia solani</i>, the Causal Agent of Cucumber Damping-Off and Root Rot Diseases

The cucumber (Cucumis sativa L.) is often subjected to several fungal diseases. Rhizoctonia solani-induced cucumber damping-off and root rot are the most common diseases reported from the commercial greenhouses of the eastern area of Saudi Arabia. The objective of the current study is to explore the antagonistic activity of four Trichoderma species against R. solani in vitro and in vivo. Ten R. solani isolates (eight belonging to AG-4 and two belonging to AG-A and AG2-1) were studied. AG4 isolates were pathogenic to cucumber plants, while AG-A and AG2-1 isolates were non-pathogenic. Seven isolates of Trichoderma spp., named T. hamatum KSATR8, T. harzianum (KSATR9 and KSATR10), T. asperellum (KSATR11, KSATC, and KSAT1E), and T. longibrachiatum KSATS were isolated, and the identities of both R. solani and Trichoderma isolates were confirmed based on the phylogenetic analysis of the DNA sequence of the ITS region. The dual culture findings indicated that T. asperellum KSATC and KSAT1E exhibited the most significant inhibitory activities against R. solani, with values of 79.33 and 70.89%, respectively. Scanning electron microscope (SEM) images showed a considerable degradation in the cell wall and collapsing of R. solani hyphae by all Trichoderma species. Under greenhouse conditions, the application of T. asperellum KSATC and KSAT1E at concentrations of 2 × 108 conidia/mL revealed a reduction in root rot and damping-off incidence percentages with values that did not reveal a significant (p 0.05) difference from those of Rizolex-T fungicide. Nevertheless, the efficacy of the fungicide attained 86.67%, being higher than that of T. asperellum KSATC, which reached 80%. Trichoderma asperellum KSATC and KSAT1E were the greatest in increasing peroxidase, catalase, and chitinase enzymes activities in cucumber plants. Conversely, a significant (p 0.05) elevation in polyphenol oxidase enzyme (0.762 and 0.97 U/g FW) and total phenol content (0.55 and 0.62 mg/g FW) was recorded in cucumber plants treated with T. harzianum KSATR9 and KSATR10, respectively. The statistical analysis results displayed no considerable variations among cucumber plants regarding total chlorophyll content as a response to treatments with Trichoderma species and fungicides. Therefore, we endorse using T. asperellum KSATC and KSAT1E as an alternative to fungicides to manage root rot and damping-off in cucumbers

Isolation and Characterization of a Novel Pathogenesis-Related Protein-1 Gene (AvPR-1) with Induced Expression in Oat (Avena sativa L.) during Abiotic and Hormonal Stresses

Pathogenesis-related protein-1 (PR-1) plays crucial roles in regulating plant responses to biotic and abiotic stresses. This study aimed to isolate and characterize the first PR-1 (AvPR-1) gene in oat (Avena sativa L.). AvPR-1 presented conserved signal peptide motifs and core amino acid composition in the functional protein domains as the protein sequence of AvPR-1 presented 98.28%, 97.7%, and 95.4% identity with known PR1 proteins isolated from Triticum aestivum PRB1-2-like, Triticum dicoccoides PRB1-2-like, and Aegilops tauschii subsp. tauschii, respectively. Bioinformatic analysis showed that the AvPR-1 protein belongs to the CAP superfamily (PF00188). Secondary and 3D structure analyses of the AvPR-1 protein were also conducted, confirming sequence conservation of PR-1 among studied species. The AvPR-1 protein harbors a calmodulin-binding domain located in its C-terminal part as previously shown for its wheat homolog TdPR1.2. Moreover, gene expression analysis showed that AvPR-1 was induced in response to many abiotic and hormonal stresses especially in leaves after treatment for 48 h. This is the first study exhibiting the expression profiles of the AvPR-1 gene under different stresses in oat

Identification of a Cucumber Mosaic Virus from <i>Cucurbita pepo</i> on New Reclamation Land in Egypt and the Changes Induced in Pumpkin Plants

In 2020, the leaves and fruit of 50 pumpkin plants with suspected cucumber mosaic virus (CMV) symptoms of leaf mosaic, vein yellowing, and mild leaf curling were collected from Sharq El-Owainat (Al Wadi El-Gaded governorate), a new reclamation land in Egypt. This study was aimed at deciphering and characterizing the causal agent of the leaf yellowing disease associated with pumpkin plants in Egypt. The causal agent was identified by serological, cytological, and molecular means. The serological identification by DAS–ELISA confirmed the presence of CMV in 20% of the plants. The cytological identification by electron microscopy revealed typical cucumovirus isometric particles of 28–30 nm diameter in the cytoplasm of the leaf parenchyma, epidermal cells, the integument, and the nucleus. Molecular characterization by one-step reverse transcriptase-PCR yielded the required size of amplicon (678 bp) for CMV. Additionally, mechanical sap inoculation was used to determine the host range and symptomatology of the isolated CMV in seventeen different plant species belonging to six different plant families. CMV replicated, moved systemically, and induced a range of symptoms in sixteen plant species. The isolated CMV was transmitted to pumpkin plants at a 16.4% rate by seeds. CMV-infected pumpkin plant leaves were characterized by a substantially low concentration of photosynthetic pigments, a high level of reducing sugars, relatively low protein levels, and a significant increase in total phenol contents, implying their potential role as antiviral agents. Ultrathin sections of infected cells revealed histological changes and cytological abnormalities in comparison to healthy plants. This is the first identification of CMV on new reclamation land in Egypt, pinpointing its swift spread, which could pose a major constraint to pumpkin production in Egypt

Host Resistance to <i>Uromyces appendiculatus</i> in Common Bean Genotypes

Rust, induced by the fungus Uromyces appendiculatus, is one of the most serious bean diseases. The involved mechanisms in rust resistance were evaluated in 10 common bean genotypes during the 2019/2020 and 2020/2021 growing seasons. The disease parameters such as final rust severity (FRS%), area under the disease progress curve (AUDPC) and disease increase rate (r-value) were lower in the resistant genotypes than in highly susceptible genotypes. Biochemical compounds such as total phenols and the activity of antioxidant enzymes such as catalase, peroxidase and polyphenol oxidase were increased in the resistant genotypes compared to susceptible genotypes. In the resistance genotypes, the levels of oxidative stress markers such as hydrogen peroxide (H2O2) and superoxide (O2•−) increased dramatically after infection. The electrolyte leakage percentage (EL%), was found to be much greater in susceptible genotypes than resistant genotypes. The resistant gene SA14, which was found in genotypes Nebraska and Calypso at 800 bp, had an adequate level of resistance to bean rust with high grain yield potential. After infection, the transcriptions levels of 1,3-D-glucanases and phenylalanine ammonia lyase) were higher in the resistant genotypes than susceptible genotypes. In conclusion, the resistant genotypes successfully displayed desirable agronomic traits and promising expectations in breeding programs for improving management strategies of common bean rust disease. The resistance was mediated by antioxidant enzymes, phenolic compounds, and defense gene expressions, as well as the resistant gene SA14

Exogenous Postharvest Application of Calcium Chloride and Salicylic Acid to Maintain the Quality of Broccoli Florets

The importance of broccoli (Brassica oleracea var. italica) consumption has increased in recent years due to its significant amount of anticarcinogenic and antioxidant compounds, as well as its many vitamins. However, broccoli florets are a highly perishable product which rapidly senesce and turn yellow after harvest, resulting in losses in nutritional and bioactive compounds. Thus, in this study, we evaluated the effect of postharvest exogenous of salicylic acid (SA) and calcium chloride (CaCl2) and their combination on the quality of broccoli florets stored at 5 &deg;C for 28 days to minimize the rapid senescence of broccoli florets. Samples treated with 2 mM SA alone or in combination with 2% CaCl2 showed lower weight loss and lower losses of chlorophyll content, vitamin C, phenolic compounds, carotenoids, flavonoids, and glucosinolates compared with the control samples. Additionally, antioxidant activity was maintained by either SA or SA + CaCl2 treatments while peroxidase activity was decreased. For higher quality and lower losses in antioxidant compounds of broccoli florets during refrigerated storage at 5 &deg;C, SA + CaCl2 treatment could be helpful for up to 21 days

Diversity among <i>Lasiodiplodia</i> Species Causing Dieback, Root Rot and Leaf Spot on Fruit Trees in Egypt, and a Description of <i>Lasiodiplodia newvalleyensis</i> sp. nov.

Lasiodiplodia (family Botryosphaeriaceae) is a widely distributed fungal genus that causes a variety of diseases in tropical and subtropical regions. During 2020–2021, a routine survey of fruit tree plants was conducted in five Egyptian Governorates, and fresh samples exhibiting dieback, decline, leaf spot and root rot symptoms were collected. Collection from eight different symptomatic leaves, twigs, branches and roots of fruit trees yielded 18 Lasiodiplodia-like isolates. The sequencing data from the internal transcribed spacer region (ITS), partial translation elongation factor 1-alpha (tef1-a) and β-tubulin (tub2) were used to infer phylogenetic relationships with known Lasiodiplodia species. Two isolates obtained from black necrotic lesions on Phoenix dactylifera leaves were identified as a putative novel species, L. newvalleyensis sp. nov., and were thus subjected to further morphological characterization. The results of isolation and molecular characterization revealed that L. theobromae (n = 9) was the most common species on Mangifera indica, Citrus reticulata, C. sinensis, Ficus carica, Prunus persica, Prunus armeniaca and Pyrus communis trees. Lasiodiplodia pseudotheobromae (n = 5) was isolated from M. indica, Prunus persica and C. sinensis. Lasiodiplodia laeliocattleyae (n = 2) was isolated from C. reticulata. Pathogenicity test results suggested that all Lasiodiplodia species were pathogenic to their hosts. The present study is considered the first to characterize and decipher the diversity of Lasiodiplodia species associated with fruit trees in Egypt, using the multi-locus ITS, tef1-a and tub2 sequence data, along with morphological and pathogenic trials. To our knowledge, this is the first report of L. newvalleyensis on Phoenix dactylifera and L. laeliocattleya on C. reticulata in Egypt and worldwide

Humic Acid-Coated Fe₃O₄ Nanoparticles Confer Resistance to Acremonium Wilt Disease and Improve Physiological and Morphological Attributes of Grain Sorghum

Acremonium wilt disease affects grain quality and reduces sorghum yield around the globe. The present study aimed to assess the efficacy of humic acid (HA)-coated Fe3O4 (Fe3O4/HA) nanoparticles (NPs) in controlling acremonium wilt disease and improving sorghum growth and yields. During the season 2019, twenty-one sorghum genotypes were screened to assess their response to Acremonium striticum via artificial infection under field conditions and each genotype was assigned to one of six groups, ranging from highly susceptible to highly resistant. Subsequently, over the two successive seasons 2020 and 2021, three different concentrations of 10, 40 and 80 mg L−1 of Fe3O4/HA NPs were tested against A. striticum. The concentrations of 40 and 80 mg L−1 were found to be highly effective in controlling acremonium wilt disease on different sorghum genotypes: LG1 (highly susceptible), Giza-3 (susceptible), and Local 119 (resistant) genotypes. After harvest, the physiological (growth and yield) and biochemical (peroxidase, catalase, and gibberellic acid) attributes of sorghum plants were determined, and the results demonstrated that concentrations of 40 and 80 mg L−1 increased peroxidase and catalase activities in healthy (uninoculated) sorghum genotypes compared to inoculated sorghum genotypes. Additionally, the toxicity of Fe3O4/HA NPs on male albino rats was investigated via hematological (CBC), chemical (ALT and AST) and histopathological analyses. The concentration 80 mg L−1 of Fe3O4/HA NPs caused a marked increase in ALT and creatinine level after 51 days of feeding. Severe pathological alterations were also observed in liver and kidney tissues of rats administered with grain sorghums treated with 80 mg L−1. In comparison with the untreated control plants, a concentration of 40 mg L−1 significantly increased the growth, yield and gibberellic acid levels (p ≤ 0.05) and was found to be safe in male albino rats. Conclusively, a concentration of 40 mg L−1 of Fe3O4/HA NPs showed promising results in curtailing A. striticum infections in sorghum, indicating its great potential to substitute harmful fertilizers and fungicides as a smart agriculture strategy