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Abstract Background The soil-borne fungi, Rhizoctonia solani and Sclerotium rolfsii, are major pathogens of Brassicae crops. This study was performed to clarify the relationship between the accumulation pattern of the genus Trichoderma and disease suppression in frequently inoculated soils with binucleate Rhizoctonia (BNR), Rhizoctonia solani and Sclerotium rolfsii. Results As compared to the control group, five Trichoderma virens strains isolated from soil inoculated with R. solani or BNR significantly reduced the severity of S. rolfsii (85.6–100% covering percentage) and R. solani (95.7–100% covering percentage). Similarly, five T. hamatum strains obtained from soil inoculated with R. solani were shown to be highly suppressive against S. rolfsii (83.9–97.1% covering percentages) and R. solani (60.2–96.2% covering percentages). Four out of five T. hamatum strains obtained from soil infected with S. rolfsii exhibited considerable suppression against S. rolfsii (63.7–91.2% covering percentages), while the SM5 strain did not. The phylogenetic analysis of the TEF and ITS regions of Trichoderma hamatum revealed that most isolates were classified into the same cluster with homology of 99–100%. Five strains of each T. virens and T. hamatum were isolated from the suppressive soil with high antagonistic potentials against R. solani and S. rolfsii. Suppression and antagonistic activity of T. hamatum isolated from soil frequently inoculated with sterile barley grains were negligible, whereas T. hamatum isolated from frequently inoculated soil with BNR and R. solani demonstrated considerable suppression of the pathogens and antagonistic activity. Accumulation and quantification of T. virens and T. hamatum were confirmed using loop-mediated isothermal amplification (LAMP). Conclusion In conclusion, disease suppression in frequently inoculated soil with BNR, R. solani and S. rolfsii was due to Trichoderma spp. accumulated selectively in each replicate of soil inoculation.

Abstract Background The phylogenetic relationships and phylogeny of twenty-six Trichoderma species collected from various Egyptian locations were investigated. The genetic diversity among the examined isolates was tested using the rep-PCR marker. Trichoderma species were screened for their cellulase activities. Results Three isolates demonstrated highly significant FPase activities, namely MNF-MAS-Tricho 1, MNF-MAS-Tricho 2, and MNF-MAS-Tricho 3 (0.50, 0.39, and 0.49 IU ml−1, respectively). MNF-MAS-Tricho1 showed the highest significant CMCase activity (0.80 IU ml−1). Concerning β-glucosidase, MNF-MAS-Tricho 1 was the highest (0.78 IU ml−1), while MNF-MSH-Trich 11 and MNF-MAS-Tricho 15 were the lowest (0.36 IU mL−1). The percentage of polymorphism ranged from 46.15 to 83.33%. (GTG)5 marker produced the greatest number of polymorphic loci (13 loci out of 18 loci) with about 83.33% polymorphism, followed by rep-10 with 69.2% polymorphism. Furthermore, the polymorphism information content (PIC) estimates ranged between 0.285 for Rep-10 and 0.340 for (GTG) 5 with an average of 0.306. The tested primers exhibited high discriminating and resolving powers. Conclusion The findings of this investigation were used to classify Trichoderma species, evaluate their genetic variability using ITS sequencing, rep-PCR, and measure their cellulase activities. These markers can facilitate more rapid and less complicated studies of Trichoderma population dynamics and evaluate their establishment after release into agricultural environments. The results will help to evaluate the genetic diversity of Trichoderma in future research.

Abstract Background Postharvest diseases cause a wide loss to tomato fruits during handling and storing from harvest to consumers. Fungicides are mainly used to control postharvest diseases. Biological control is the eco-friendly substitute strategy used for postharvest diseases management as which becoming promise worldwide. Six bacterial bioagent (i.e., Bacillus subtilis, B. amyloliquefaciens, Pseudomonas resinovorans, P. alcaligenes, P. putida and P. stutzeri) were tested to suppress both Geotrichum candidum and Alternaria alternata causal agents of tomato fruit rots during storage. Results In vitro, most of bioagents significantly reduced mycelial growth rate of G. candidum. Both of B. subtilis and P. stutzeri were the most superior bacterial bioagents with values 67.03 and 72.2%, respectively. In addition, B. subtilis and B. amyloliquefaciens resulted in the most superior over all antagonists against A. alternata. The lowest percent of disease incidence and severity of G. candidum were obtained by B. subtilis and P. stutzeri. The maximum reduction percent of it on tomato fruits was recorded by applying B. subtilis and P. stutzeri with values (90 and 87%) and (91, 89%) in both seasons, respectively. Also, the highest reduction of A. alternata was obtained by using B. subtilis, which resulted in 85 and 84% in both seasons, respectively. The application of bioagents against both pathogens was significantly improved fruit quality aspects (weight loss, vitamin C, TSS and acidity %) during storage period compared to infected control fruits. Conclusion The findings revealed that both of B. subtilis and P. stutzeri could be potential biological control agents against most postharvest pathogens of tomato fruits. This might be an alternative control strategy instead of fungicides which service the sustainable and organic farming.

Abstract Background Yellow (stripe) rust, caused by Puccinia striiformis f. sp. tritici (Pst), is an economic disease of wheat. Growth-promoting fungi (GPF) such as Trichoderma asperellum and Penicillium simplicissimum have been investigated for their potential to control yellow rust and their involvement in gene expression of four PR proteins for all-stage resistance. Results Wheat plants (cv. Sids-12) treated individually with each of the biocontrol agents, P. simplicissimum and T. asperellum, at 24 and 48 hpi showed a resistance response (infection type = 2) to yellow rust, compared to the non-treated plants, which showed highly susceptible response (infection type = 9). Both biocontrol agents induced resistance against yellow rust on wheat plants, exhibiting a moderate resistance (10 MR) and reduced the colony size of Pst (0.6 mm2). Moreover, P. simplicissimum and T. asperellum increased (P ≤ 0.05) the grain yields of wheat plants infected with Pst. Scanning electron microscope (SEM) of yellow rust infected wheat leaves treated with P. simplicissimum and T. asperellum at 24 and 48 hpi showed hyperparasitism on Pst urediniospores and inhibition of the spore germination. Expressions of pathogenesis-related (PR) protein genes, PR1, PR2, PR3 and PR4 were higher in wheat plants treated with both biocontrol agents than the non-treated checks. Conclusion P. simplicissimum and T. asperellum exhibited biocontrol potential against yellow rust disease caused by P. striiformis f. sp. tritici (Pst) on wheat plants. It was found that wheat defence mechanism against Pst was activated by a high expression of PR protein genes induced by both biocontrol agents.

Abstract Background Bhendi or okra (Abelmoschus esculentus) is an important crop widely cultivated in the Indian subcontinent. The production of okra in tropical regions is constrained by several abiotic and biotic factors. Among biotic stresses, yellow vein mosaic disease (YVMD), transmitted by the whitefly (Bemisia tabaci Genn.), causes significant production losses. Results Leaves showing symptoms of YVMD were collected from okra plants in the district of Mardan, Pakistan, from which the total nucleic acid was extracted. It was found that the viral genome was 2739 bp in the length and had seven conserved open reading frames, comparable to those of monopartite begomovirus species from the Old World. Nucleotide sequence comparison revealed that the genome has 97.7% identity with bhendi yellow vein mosaic virus (BYVMV). Additionally, alpha- and betasatellite DNA components were amplified and sequenced. The alpha- and betasatellite DNA sequences were 1367 and 1346 nt in length, respectively. Sequence analysis revealed that the alpha- and betasatellite sequences shared 97.9 and 98.7% similarity with cotton leaf curl Multan alphasatellite DNA (CLCuMuA) and croton yellow vein mosaic betasatellite DNA (CroYVMB), respectively. Conclusions Based on the prevailing classification system, the isolate was identified as a variant of BYVMV, CLCuMuA, and CroYVB. Alphasatellite presence in the begomovirus betasatellite complex detected in the present study indicated a recent mobilization into the viral complex infecting okra in this region. The study findings may facilitate the design of new management strategies to protect this valuable crop against begomovirus infection.

Background The carcinogenic, mutagenic, and teratogenic chemicals such as aflatoxin are a worldwide health problem. Aspergillus spp., responsible for most cases of aflatoxin contamination, are common in the environment and spread easily to many different types of food. The objectives of this study were to conduct a survey of fungi associated with three soil invertebrates in Taif, Saudi Arabia, identify these isolates and explore mycotoxins formation. Methods In total, 114 fungal isolates were collected from various soil invertebrates (millipedes, Armadillidium vulgare and Porcellio laevis) in Taif, Saudi Arabia, among them, 22 isolates were identified as Aspergillus spp. based on morphological and molecular characteristics followed by both Fusarium and Penicillium. Results The sequences of ITS 1 and ITS 4 were utilized. Using bootstrap analysis, phylogenetic tree was split into two distinct clusters. Five sub clusters were included inside the first major cluster, and their bootstrap value was 99%. While, there were two small clusters in the second major cluster. All the tested Aspergillus strains were able to have a single PCR fragment amplified using the primer AspTef. TEF-1 DNA sequence bootstrap analysis with 1,000 replicates revealed two distinct groups. Additionally, the Aspergillus isolates were grouped into two different clusters with about 65% genetic similarity using ISSR-PCR analysis. The standard polymerase chain reaction was used to effectively amplify the Aopks, afl-A and omt-A genes in aflatoxigenic Aspergillus strains. Four Aspergillus strains used in this investigation were shown to generate aflatoxin B1. While, three Aspergillus stains showed ochratoxin genes. Conclusions In conclusion, the results indicate significant differences in the fungal community between ecoregions and soil invertebrates. Moreover, mycotoxin detection and identification among Aspergillus isolates were elucidated. This study could shed light on the risk of mycotoxin contamination along the supply chain.

Abstract Background Tomato mosaic virus (ToMV) is a dangerous disease of tomato (Lycopersicon esculentum) that reduces dramatically the yield. To reduce ToMV infection, microalgal isolates were utilized. Microalgal species (Chlorella vulgaris, Anabaena oryzae, Spirulina platensis, Nostoc linckia and Dunaliella salina) were shown to be responsible for the stimulation of tomato resistance against ToMV. Results Initial signs of discoloration and mosaic in ToMV-inoculated plants were detected and identified on inoculated leaves at 6 and 12 dpi in control and treated plants, respectively, suggesting that microalgae may inhibit ToMV growth. Treatment with microalgae resulted in a significant decrease in symptoms (up to 63% reduction in disease severity) and negative ELISA readings, indicating that the microalgae induced resistance in tomato against ToMV infection. The isolates also enhanced the activity of pathogenesis-related enzymes (PPO and POX reaching to 0.033 and 0.054 in D. salina, respectively), as well as tomato growth characters in comparison with the control. Microalgal treatments demonstrated that the salicylic acid (SA) and jasmonic acid (JA) pathways were involved in tomato plant defense responses. The relative gene expressions of PR1 and phenylalanine ammonia lyase (PAL), which are involved in the SA and JA pathways, respectively, were improved in treated plants compared to the control. Conclusion The findings indicated that algal-induced ToMV resistance was mediated via several defense pathways in tomato. The antiviral mechanism was described, which provides a light on the potential of algae in plant viral disease management.

Abstract Background Powdery mildew was found in most of the sunflower fields in Egypt, causing severe yellowing of the blade, petiole, stem and calyx, as well as a considerable defoliation during the summer season of 2018. Out of the fungal mycelium from infected leaves, collected from sunflower fields in the four Egyptian Governorates (Kafr El-Sheikh, Gharbia, Giza and El-Beheira), five isolates of powdery mildew pathogen were obtained and identified using morphological and molecular identification methods. Results In 2019 and 2020 seasons, five biocontrol agents (Bacillus subtilis, B. pumilus, Trichoderma harzianum, T. viride and T. koningii) were used to control powdery mildew disease on sunflower plants under field conditions compared with the fungicide (Vectra 10% SC). Treatments were significantly effective for controlling the powdery mildew disease relative to the control. The best treatment for reducing disease parameters (final disease severity (FDS%), area under disease progress curve (AUDPC) and efficacy) than the control was T. koningii. Foliar application of all the tested treatments improved plant height, head and stem diameters and seed yield in relation to untreated plants (control). The components (FDS%, AUDPC and efficacy) were extracted and described approximately 95.251% of the pooled data of seasons 2019 and 2020. In such pooled data, the principal components (PC1, PC2 and PC3) of all disease parameters, plant development parameters and yield components were recorded 76.305, 86.635 and 96.265% of the total variance, respectively. Conclusion A biological control agent, such as T. koningii, can be suggested for disease control based on the experimental findings.

This research was performed to evaluate the potential of carvone, cuminaldehyde, cineole, and linalool for the control of root-knot of tomato. The tested control agents were evaluated for their ability to stimulate systemic resistance to Meloidogyne incognita in tomato by monitoring the transcription levels of defense-related genes. Moreover, the ability of the tested agents to induce nematicidal activity concerning second-stage juveniles (J2) hatching and mortality was evaluated. Furthermore, the effect of the tested agents on certain tomato growth and yield parameters was assessed. The tested monoterpenes showed high nematicidal activity against M. incognita concerning J2 hatching inhibition and mortality. Carvone, cuminaldhyde, linalool, and cineole had LC50 values of 123.5, 172.2, 354.9, 466.4, and 952.3 μg/mL, respectively. Carvone was found to be the most efficient hatching inhibitor. The tested monoterpenes showed a high potential against root-knot under greenhouse and field conditions with respect to root-galling, egg masses, and the number of J2. Carvone was the most effective treatment. The growth and yield characters of treated tomato were significantly increased in monoterpenes treatments compared to untreated control. Treated tomato plants showed expression of defense-related genes (PR1 and PAL) 5-8 folds higher than the control. The results also showed that cuminaldhyde, followed by carvone, linalool, and cineole, had the greatest levels of expression in tomato plants. Taken together, the selected monoterpenes could be used as alternatives to control the root-knot of tomato.

Extensive use of chemical control agents and fungicides typically leads to numerous risks to human health and the environment. Using plant extracts as natural substances represents a dual key for the environment and sustainable food production, as it reduces the input of synthetic pesticides into the environment and/or controls plant pathogens. For the first time, a Plantago lagopus ethanolic extract has been characterized and evaluated for its protective and curative effects against Rhizoctonia solani in tomato plants. The results showed that P. lagopus extract (10 μg/ml) completely inhibited R. solani mycelial growth in vitro. At 20 days of post fungal inoculation, the results demonstrated that using P. lagopus extract (100 μg/ml) in vivo enhanced tomato plant growth by significantly increasing shoot and root parameters in protective and curative treatments. Furthermore, the protective and curative treatments significantly reduced the disease index by 18.66 and 38.66%, respectively. Induction of systemic resistance with upregulation of PR-1 and PR-2 and a significant increase in the transcriptional levels of PR-3 and CHS in all P. lagopus extract-treated tomato plants were reported compared to untreated plants. HPLC analysis showed that the most common polyphenolic components detected in P. lagopus extract were rutin (74206.3 mg/kg), naringenin (2388.74 mg/kg), quercetin (1249.13 mg/kg), and p-hydroxybenzoic acid (1035.87 mg/kg). In addition, the ellagic acid (798.47 mg/kg), vanillic acid (752.55 mg/kg), catechol (648.89 mg/kg), cinnamic acid (332.51 mg/kg), ferulic acid (296.32 mg/kg), benzoic acid (295.95 mg/kg), and chlorogenic acid (116.63 mg/kg) were also reported. Our study is the first to show that P. lagopus extract can help plants fight off R. solani fungal infection. Furthermore, the findings imply that using the P. lagopus extract as a natural biocontrol agent could be a sustainable strategy to manage plant fungal diseases.

Soil contamination with toxic heavy metals [such as lead (Pb)] is becoming a serious global problem due to the rapid development of the social economy. However, accumulation of Pb in plant parts is very toxic for plant growth and decreases crop yield and productivity. In the present study, we have investigated the different concentrations of Pb in the soil i.e., [0 (no Pb), 50, and 100 mg kg–1] to study plant growth and biomass, photosynthetic pigments and gas exchange characteristics, oxidative stress indicators and the response of various antioxidants (enzymatic and non-enzymatic), nutritional status of the plant, organic acid exudation pattern and also Pb accumulation in the roots and shoots of the plants of two varieties of tomato (Solanum lycopersicum L.) i.e., Roma and Cchuas, grown under different levels of synergic acid [no spray (NS), water spray (WS), 0.3-0.5°μM]. Results from the present study showed that the increasing levels of Pb in the soil decreased non-significantly (P < 0.05) shoot length, root length, shoot fresh weight, root fresh weight, shoot dry weight, root dry weight, chlorophyll-a, chlorophyll-b, total chlorophyll, carotenoid content, net photosynthesis, stomatal conductance, transpiration rate, soluble sugar, reducing sugar, non-reducing sugar contents, calcium (Ca2+), magnesium (Mg2+), iron (Fe2+), and phosphorus (P) contents in the roots and shoots of the plants. However, Pb toxicity also induced oxidative stress in the roots and shoots of the plants by increasing malondialdehyde (MDA), hydrogen peroxide (H2O2), and electrolyte leakage (EL) which also induced increased the compounds of various enzymatic and non-enzymatic antioxidants and also organic acids exudation pattern in the roots such as fumaric acid, acetic acid, citric acid, formic acid, malic acid, oxalic acid contents and increased the concentration of Pb in different parts of the plants. Results also show that the Cchuas showed better growth and development compared to Roma, under the same levels of Pb in the soil. The alleviation of Pb toxicity was induced by the application of synergic acid, and results showed that the application of synergic acid increased plant growth and biomass and also increased the gas exchange characteristics and antioxidant capacity in the roots and shoots of the plants. Research findings, therefore, suggested that synergic acid application can ameliorate Pb toxicity in S. lycopersicum varieties and result in improved plant growth and composition under metal stress as depicted by balanced exudation of organic acids.

Klebsiella is a common dangerous pathogen for humans and animals and is widely present in the digestive system. The genus Klebsiella is ubiquitous, as it is endemic to surface water, soil, and sewage. In this study, 70 samples were obtained from soil-dwelling invertebrates from September 2021 to March 2022 from Taif and Shafa in different altitudinal regions of Saudi Arabia. Fifteen of these samples were identified as Klebsiella spp. The Klebsiella isolates were genetically identified as Klebsiella pneumoniae using rDNA sequencing. The antimicrobial susceptibility of the Klebsiella isolates was determined. Amplification of virulence genes was performed using PCR. In this study, 16S rDNA sequencing showed a similarity from 98% to 100% with related K. pneumonia from the NCBI database, and the sequences were deposited in the NCBI GenBank under accession numbers ON077036 to ON077050. The growth inhibition properties of ethanolic and methanolic extracts of the medicinal plant Rhazya stricta’s leaves against K. pneumoniae strains using the minimum inhibitory concentration (MIC) method and disc diffusion were evaluated. In addition, the biofilm inhibitory potential of these extracts was investigated using crystal violet. HPLC analysis identified 19 components divided into 6 flavonoids, 11 phenolic acids, stilbene (resveratrol), and quinol, and revealed variations in the number of components and their quantities between extracts. Both extracts demonstrated interesting antibacterial properties against K. pneumoniae isolates. The 2 extracts also showed strong biofilm inhibitory activities, with percentages of inhibition extending from 81.5% to 98.7% and from 35.1% to 85.8% for the ethanolic and methanolic extracts, respectively. Rhazya stricta leaf extract revealed powerful antibacterial and antibiofilm activities against K. pneumoniae isolates and could be a good candidate for the treatment or prevention of K. pneumonia-related infections.

Abstract The influence of the inoculation with plant growth-promoting fungi (PGPF) isolates on growth enhancement of onion plants and progress of white rot disease in onions, caused by Sclerotium cepivorum, was evaluated. The tested PGPF isolates were Phoma sp. GS8-1, Phoma sp. GS 8-3, Trichoderma asperellum SKT-1, Fusarium equiseti GF18-3 and Penicillium simplicissmum GP17-2. Abnormal shape and lysis in the mycelia of the pathogen were reported in dual cultures of PGPF and S. cepivorum using scanning electron microscopy. The germination of sclerotia was reduced after soaking in culture filtrates of PGPF. The highest reduction was recorded in P. simplicissmum GP17-2 treatment (70.85%). The results demonstrated that the treatments with PGPF isolates significantly enhanced the plant height, root length, bulb perimeter and plant dry weight. Additionally, PGPF clearly reduced disease severity of white rot disease of onions. Among PGPF isolates, T. asperellum SKT-1 and P. simplicissmum GP17-2 showed the best effects in reducing the incidence and severity of white rot and enhancing onion growth. On the other hand, the levels of peroxidase and polyphenol oxidase were improved in the treated onion plants than in the untreated plants at 7 days after induction treatments. Similarly, early and strong expressions of defence genes, PR1 and PR2, were reported in plants treated with PGPF. Overall, this research suggested that PGPF treatments generated favourable effects on the suppression of white rot disease of onions.

Wastewater irrigation is a substitute for surface water scarcity, but traces of heavy metals (HMs) result in deleterious implications for soil, crop productivity, and in humans. Crops presenting HMs tolerance in genetic behavior are important for producing tolerant genotypes cultivated under wastewater irrigation. In the first part of this experiment, the results obtained previously are re-assessed in a hydroponic system and similar patterns and concentrations of HMs are found in different tomato organs. Following this trial, the tomato’s (Solanum lycopersicum L.) genetic basis of traits conferring HMs tolerance and yield are assessed when irrigated with waste or canal water. The North Carolina Mating II analysis illustrate the amount of gene action, nature, and inheritance pattern. Genetic components depict the involvement of non-additive, additive, and maternal genetic effects in HMs tolerance inheritance and yield. A noticeable increase in cumulative additive variance for the number of flowers (11,907.2) and the number of fruits (10,557.9) is recorded for tomato plants irrigated with wastewater, illustrating additive gene action. However, female and male (MSf/MSm) square ratios also show an association with cytoplasmic inheritance. For HMs tolerance, both additive and dominant variances appeared to be significant; cumulative dominance variance (4.83, 16.1, 4.69, 76.95, and 249.37) is higher compared to additive variance (0.18, 2.36, 0.19, −0.27, and 14.14) for nickel (Ni), chromium (Cr), lead (Pb), manganese (Mn), and zinc (Zn), respectively, indicating dominance gene action. The genotype RIOGRANDI accumulated and translocated fewer HMs to the aerial part of the plant compared to CLN-2418A and PB-017906, thus presenting a tolerant tomato genotype according to the hydroponic experiment. This also exhibited a differential pattern of gene action for HMs tolerance, suggesting that genotypes possess significant differences for HMs tolerance.

The Red Palm Weevil (Rhynchophorus ferrugineus (Oliv.) (Coleoptera, Dryophthoridae) is a well-known palm tree pest that has caused enormous economic damage all over the globe. Insecticides are still the primary method of controlling this pest at this period. However, field populations of RPW have been shown to be resistant to pesticides. Using Bacillus spp. might be one of the options for controlling R. ferruginous. In this study, 23 species of Bacillus spp. were isolated from the rhizosphere of date palm trees in Al Ahsa Oasis, Saudi Arabia. The isolates were identified using 16S rRNA gene sequencing. R. ferrugineus larvae and adults were tested on sugarcane pieces that were treated with the B. thuringiensis strain PDC-AHSAA1 and B. cereus strains (PDC-AHSAA2, PDC-AHSA3 and PDC-AHSA4). The LC50 values for larvae and adults were quite low when they were compared with those of the other isolated strains. The B. thuringiensis strain PDC-AHSAA1 was more effective against both the larvae and adults. The determined LC50 values for B. thuringiensis ranged from 4.19 × 107–3.78 × 109. After 21 days, the data on larval mortality and body weight were evaluated. The surviving larvae that were treated with the bacterial isolates did not acquire a substantial weight. For the RPW larvae and adults, the mortality and corrected mortality death rates were increased by increasing the concentration of B. thuringiensis. In conclusion, Bacillus-treated diets negatively influenced the growth and development of the RPW. This research reported on the interaction between the RPW and the rhizosphere Bacillus spp. and highlighted the tremendous potential for the development of microbial resource-based control strategies for this pest.

Chemical pesticides and fungicides are used extensively, negatively affecting people’s health and the environment. Reducing synthetic pesticides and increasing the efficiency of sustainable food production using plant extracts as natural chemicals is a win–win. Here, we first describe and evaluate an ethanolic extract of Chorisia speciosa for its protective and curative activities against Rhizoctonia solani in greenhouse-grown tomato plants. The results showed that the mycelial growth of R. solani was completely suppressed in vitro by C. speciosa extract (10 µg/mL). Twenty days after fungal inoculation, the results demonstrated that using C. speciosa extract (10 µg/mL) in vivo significantly improved shoot and root growth parameters in protective and curative treatments. Further, the protective and curative treatments decreased the disease index by 26.67% and 53.33%, respectively. C. speciosa-treated tomato plants showed significantly increased antioxidant enzyme production (PPO, CAT, and SOD) and up-regulated PR-1, PR-2,PR-3, PAL, and CHS expression levels compared to untreated plants. According to HPLC examination, the most prevalent phenolic acids or flavonoid components quantities (µg/mL) noticed in C. speciosa extract were 7-OH-flavone (10.36), kaempferol (9.23), p-coumaric acid (8.65), ferulic acid (8.14), caffeic acid (7.59), gallic acid (6.33), and iso-ferulic (5.71). Our findings are the first to demonstrate that a C. speciosa extract can assist plants in combating fungal infestation. Therefore, the data imply that C. speciosa extract, as a natural and renewable product, could be adopted as a long-term approach for regulating plant fungus.

Potato Virus Y (PVY) is a serious potato disease that may significantly decrease potato production. To suppress potato virus infection, several measures have been undertaken. The utilization of plant growth-promoting rhizobacteria is one of these methods. Biochar soil treatment is believed to provide plants with a number of advantages, including increased plant growth and the development of systemic resistance to a variety of plant diseases. The goal of this research was to see whether adding biochar and Klebsiella oxytoca to the soil might cause PVY resistance and enhance the involved mechanisms in PVY resistance. Potato and tobacco seedlings treated with Klebsiella oxytoca and biochar exhibited the same impact of significant symptom reduction, with complete negative ELISA findings, supporting the antiviral activity of K. oxytoca and biochar. Furthermore, owing to the connection between the ISR implicated substrates, significant amounts of polyphenol oxidase, catalase, and superoxide dismutase were observed in treated plants, with the same behavior as defense genes expression levels. It may be a step forward in the development of biochar and K. oxytoca as potential environmentally friendly disease control strategies against PVY.

Metal oxide nanoparticles have recently garnered interest as potentially valuable substances for the management of plant diseases. Copper oxide nanoparticles (Cu2ONPs) were chemically fabricated to control root rot disease in cucumbers. A scanning electron microscope (SEM), X-ray diffraction (XRD) and photoluminescence (PL) were employed to characterize the produced nanoparticles. Moreover, the direct antifungal activity of Cu2ONPs against Fusarium solani under laboratory, greenhouse, and field conditions were also evaluated. In addition, the induction of host-plant resistance by Cu2ONPs was confirmed by the results of enzyme activities (catalase, peroxidase, and polyphenoloxidase) and gene expression (PR-1 and LOX-1). Finally, the effect of Cu2ONPs on the growth and productivity characteristics of the treated cucumber plants was investigated. The average particle size from all the peaks was found to be around 25.54 and 25.83 nm for 0.30 and 0.35 Cu2O, respectively. Under laboratory conditions, the study found that Cu2ONPs had a greater inhibitory effect on the growth of Fusarium solani than the untreated control. Cu2ONP treatment considerably reduced the disease incidence of the root rot pathogen in cucumber plants in both greenhouse and field environments. Defense enzyme activity and defense genes (PR1 and LOX1) transcription levels were higher in cucumber plants treated with Cu2ONPs and fungicide than in the untreated control. SEM analysis revealed irregularities, changes, twisting, and plasmolysis in the mycelia, as well as spore shrinking and collapsing in F. solani treated with Cu2ONPs, compared to the untreated control. The anatomical analysis revealed that cucumber plants treated with Cu2ONPs had thicker cell walls, root cortex, and mesophyll tissue (MT) than untreated plants. Cucumber growth and yield characteristics were greatly improved after treatment with Cu2ONPs and fungicide. To the best of our knowledge, employing Cu2ONPs to treat cucumber rot root disease is a novel strategy that has not yet been reported.

The current study focuses on the effects of Cucumber mosaic virus (CMV) infection on phytochemical changes and pathogenesis- and phenylpropanoid pathway-associated gene activities in squash (Cucurbita pepo L.) plants during a time course of 2 to 12 days post inoculation (dpi). The identity of the CMV isolate was confirmed by DAS-ELISA, TEM, and coat protein gene sequence. The CMV infection initially boosts and then suppresses transcript levels of the defense-related genes PR-1, PR-2, PAL, HQT, and CHS during the investigated time course compared to controls. The expression profile during the time-course study indicated that early, transient induction of PR-1 occurs during CMV infection, while CMV induced the expression of PR-2 in systemically infected squash tissues at all time points and suppressed the expression of PAL and HQT at 8-12 dpi. CHS transcript levels fluctuated between up- and down-regulation, but by 12 dpi, CHS expression reached its peak. The HPLC and GC–MS analyses of CMV-infected squash extracts revealed that different phenolic, flavonoid, and fatty acid compounds could be induced or suppressed upon CMV infection. In particular, CMV could suppress the synthesis of most phenolic compounds, specifically chlorogenic acid, possibly leading to the virus’s rapid spread.

Modified whey proteins with quercetin (WPI-QU) and onion extract (WPI-OE), as a control approach, could be applicable because it is available, safe and cheap. The modified whey protein isolate (WPI) with quercetin dihydrate and onion extract powder rich with quercetin were evaluated for induction of systemic resistance against Pepper mild mottle virus (PMMoV) in pepper plants. Data of mass spectrometry illustrated that one or more of Qu isomers covalently attached to WPI. Unmodified whey protein (UWPI), WPI-QU and WPI-OE significantly decreased PMMoV concentration and severity at two weeks after inoculation. Plant height, number of leaves, and shoot fresh and dry weights were substantially increased in WPI-QU- and WPI-OE-treated pepper plants compared to the control. Total antioxidant status (TAS) and vitamin C contents were highly increased in WPI-OE-treated plants compared with other treatments. The expression levels of defense related genes (PR4, PR9, TIN1 and PIN2) were enormously elevated in WPI-OE and WPI-QU treatments using qRT-PCR. In conclusion, the results give novel insights to possible applications of the WPI–quercetin bioconjugates in designing a wide range of functional products. Moreover, this study is the first to establish the effective control of PMMoV by modified whey proteins.

This study was conducted to evaluate eco-friendly control agents (carvone, cuminaldehyde, and linalool) against Rhizoctonia solani, which causes root rot disease either by induction of defense response or direct antifungal activity. The induction of resistance was examined by detecting the transcription of defense genes and the effect of the tested control agents on the growth and the yield of common bean plants. The growth of R. solani was significantly inhibited after treatment with the tested compounds compared to the untreated control under laboratory conditions. The disease severity of root rot was decreased in common bean plants treated with the tested compounds compared to untreated control plants under greenhouse conditions. Common bean plants treated with the tested control agents expressed defense genes (Phenylalanine ammonia lyase and β-1,3-Glucanase) involved in jasmonic acid (JA) and salicylic acid (SA) signaling pathways with 2–5 fold higher than the control. Treatment of common beans with the tested control agents and fungicide significantly improved the growth and yield characteristics of common bean. Therefore, the use of monoterpenes could be a novel strategy to control this pathogen and consider the first report.

Phage therapy is a promising biocontrol management on plant diseases caused by bacterial pathogens due to its specificity, efficiency and environmental friendliness. The emergence of natural phage-resistant bacteria hinders the application of phage therapy. Xanthomonas oryzae pv. oryzae (Xoo) is the causal agent of the devastating bacterial leaf blight disease of rice. Here, we obtained a spontaneous mutant C2R of an Xoo strain C2 showing strong resistance to the lytic phage X2. Analysis of the C2R genome found that the CDS2289 gene encoding glycosyltransferase acquired a frameshift mutation at the 180th nucleotide site, which also leads to a premature stop mutation at the 142nd amino acid. This mutation confers the inhibition of phage adsorption through the changes in lipopolysaccharide production and structure and bacterial surface morphology. Interestingly, glycosyltransferase-deficient C2R and an insertional mutant k2289 also showed reduced virulence, suggesting the trade-off costs of phage resistance. In summary, this study highlights the role of glycosyltransferase in interactions among pathogenic bacteria, phages and plant hosts, which provide insights into balanced coevolution from environmental perspectives.

Pepper is vulnerable to soil-borne fungal pathogens such as Rhizoctonia solani and Fusarium oxysporum. The potential of beneficial rhizosphere microorganisms to control R. solani and F. oxysporum f.sp. capsici was evaluated in pepper plants. Paenibacillus polymyxa and Trichoderma longibrachiatum were isolated from rhizospheric soil samples of healthy pepper plants. In vitro, both isolates caused clear reductions in the radial growth of root rot and wilt pathogens. Scanning electron microscopy displayed lysis and abnormal shape of the pathogens in dual cultures with P. polymyxa and T. longibrachiatum. The incidence and severity of root rot and wilt diseases were significantly reduced in pepper plants treated with the growth-promoting fungi (PGPF isolates; Fusarium equiseti GF19-1, Fusarium equiseti GF18-3, and Phoma sp. GS8-3), P. polymyxa, or T. longibrachiatum in comparison to the control. Moreover, the induction treatments led to increased pepper growth compared with their control. The defense related gene (CaPR4) expression was shown to be significantly higher in the treated plants than in the control plants. In conclusion, the antagonistic isolates (P. polymyxa and T. longibrachiatum) and PGPF isolates have a clear impact on the prevention of root rot and wilt diseases in pepper plants incited by R. solani and F. oxysporum f.sp. capsici. The expression of the CaPR4 gene added to the evidence that PGPF isolates generate systemic resistance to pathogen infections.

Wheat leaf rust is one of the world’s most widespread rusts. The progress of the disease was monitored using two treatments: chitosan nanoparticles and salicylic acid (SA), as well as three application methods; spraying before or after the inoculation by 24 h, and spraying both before and after the inoculation by 24 h. Urediniospore germination was significantly different between the two treatments. Wheat plants tested for latent and incubation periods, pustule size and receptivity and infection type showed significantly reduced leaf rust when compared to untreated plants. Pucciniatriticina urediniospores showed abnormalities, collapse, lysis, and shrinkage as a result of chitosan nanoparticles treatment. The enzymes, peroxidase and catalase, were increased in the activities. In both treatments, superoxide (O2−) and hydrogen peroxide (H2O2), were apparent as purple and brown discolorations. Chitosan nanoparticles and SA treatments resulted in much more discoloration and quantitative measurements than untreated plants. In anatomical examinations, chitosan nanoparticles enhanced thickness of blade (µ), thickness of mesophyll tissue, thickness of the lower and upper epidermis and bundle length and width in the midrib compared to the control. In the control treatment’s top epidermis, several sori and a large number of urediniospores were found. Most anatomical characters of flag leaves in control plants were reduced by biotic stress with P. triticina. Transcription levels of PR1-PR5 and PR10 genes were activated in chitosan nanoparticles treated plants at 0, 1 and 2 days after inoculation. In light of the data, we suggest that the prospective use of chitosan nanoparticles might be an eco-friendly strategy to improve growth and control of leaf rust disease.

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.

Sheath blight disease is a fungal pathogen that causes leaf blight in rice plants, resulting in significant yield losses throughout the growing season. Pseudomonas spp. have long been used as biocontrol agents for a variety of plant diseases. Four Pseudomonas isolates were tested for their ability to promote rice growth and generate systemic resistance to Rhizoctonia solani, the causal pathogen of sheath blight disease. In vitro, Pseudomonas isolates produced the growth hormone indole acetic acid (0.82–1.82 mg L−1). Additionally, seed treatment with Pseudomonas putida suspension outperformed P. brassicacearum, P. aeruginosa and P. resinovorans in terms of germination and vigor evaluation. The maximum seed germination of 89% was recorded after seed treatments with a fresh suspension of P. putida, followed by 87% germination in P. aeruginosa treatment, compared with only 74% germination in the untreated controls. When compared with the infected control plants, all Pseudomonas isolates were non-pathogenic to rice and their co-inoculation considerably enhanced plant growth and health by reducing the disease index to 37% and improving plant height (26%), fresh weight (140%) and dry weight (100%). All Pseudomonas isolates effectively reduced sheath blight disease incidence, as well as the fungicide carbendazim, which is recommended for field management of R. solani. In comparison to untreated control seedlings, treatment with Pseudomonas isolates enhanced the production of peroxidase and polyphenol oxidase enzymes and the expression of the phenylalanine ammonia lyase (PAL) and NPR1 genes, which could be involved in disease incidence reduction. In conclusion, the use of Pseudomonas spp. has been demonstrated to improve rice growth and resistance to R. solani while also providing an environmentally acceptable option to the agroecosystems.

The soil-borne pathogens Rhizoctonia solani and Sclerotium rolfsii have emerged as major pathogens of radish (Raphanus sativus) worldwide. The induction of soil suppressive of radish root rot disease was evaluated in soil repeatedly inoculated with R. solani, nonpathogenic binucleate Rhizoctonia sp. AG-A W1 (BNR) and S. rolfsii. The repeated inoculations of soil with R. solani and BNR significantly suppressed the disease severity of R. solani and S. rolfsii compared to the control. In contrast, the repeated inoculation of soil with S. rolfsii significantly suppressed only the pathogen, S. rolfsii. The community structure was examined using PCR-DGGE (polymerase chain reaction denaturing gradient gel electrophoresis) method. The bands of Trichoderma sp. were observed in the first, second and third inoculations of the soil with BNR. Similarly, bands of Trichoderma sp. were observed in the second and third inoculations of the soil with S. rolfsii and R. solani. Compared to the control, disease severity was significantly reduced in the soil repeatedly inoculated with S. rolfsii and R. solani . In conclusion, Trichoderma species were accumulated in specific patterns depending on the applied fungal inoculum in the suppressive soil.

Characterization of the genetic structure and the physiological races of Puccinia triticina is a growing necessity to apply host genetic resistance against wheat leaf rust as a successful control strategy. Herein, we collected and identified about 130 isolates of P. triticina from 16 Egyptian commercial wheat cultivars grown at different locations, over two seasons (2019/2020 and 2020/2021). The 130 isolates of P. triticina were segregated into 17 different physiological races. TTTST and TTTKS were the most common virulent races, whereas TTTST and MTTGT were the most frequent races. The races were classified into three groups, based on their distinct DNA band sizes (150 bp, 200 bp, and 300 bp) after RAPD analysis. The new wheat cultivars (Sakha-94, Sakha-95, and Shandweel-1) infected with the most virulent race (TTTST), Gemmeiza-12, and Misr-3 were resistant to all physiological races. The resistance of these cultivars was mostly due to the presence of Lr19- and Lr28-resistant genes. Our results serve as a warning about emerging aggressive races capable of supplanting resistance to leaf rust, and help in the understanding of the pathotype–cultivar–location association and its role in the susceptibility/resistance of new wheat cultivars to P. triticina.

Twenty-three wheat genotypes were evaluated for stripe and leaf rusts, caused by Puccinia striiformis f. sp. tritici and Puccinia triticina f. sp. tritici, respectively, at seedling and adult stages under greenhouses and field conditions during the 2019/2020 and 2020/2021 growing seasons. The race analysis revealed that 250E254 and TTTST races for stripe and leaf rusts, respectively were the most aggressive. Eight wheat genotypes (Misr-3, Misr-4, Giza-171, Gemmeiza-12, Lr34/Yr18, Lr37/Yr17, Lr46/Yr29, and Lr67/Yr46) were resistant to stripe and leaf rusts at seedling and adult stages. This result was confirmed by identifying the resistance genes: Lr34/Yr18, Lr37/Yr17, Lr46/Yr29, and Lr67/Yr46 in these genotypes showing their role in the resistance. Sids-14 and Shandweel-1 genotypes were susceptible to stripe and leaf rusts. Twelve crosses between the two new susceptible wheat genotypes and the three slow rusting genes (Lr34/Yr18, Lr37/Yr17, and Lr67/Yr46) were conducted. The frequency distribution of disease severity (%) in F2 plants of the twelve crosses was ranged from 0 to 80%. Resistant F2 plants were selected and the resistance genes were detected. This study is important for introducing new active resistance genes into the breeding programs and preserving diversity among recently released wheat genotypes.