52 research outputs found

    Detection and characterization of phytoplasmas infecting five plant species in Egypt

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    Samples of orange, hibiscus, peach, olive and pepper plants showing symptoms of dwarf branches with shortened internodes, leaf deformation and chlorosis, proliferation of axillary buds, yellowing, twisting and streaks together with samples from asymptomatic plants of the same age were collected from El-Behira and Alexandria Governorates, Egypt during 2015-2016. The samples were tested to verify phytoplasma presence by nested PCR assays using the ribosomal gene amplifying primers R16F2n/R2 on the P1/P7 1: 30 diluted amplicons as template. Amplification bands were obtained only from samples collected from symptomatic plants, and the RFLP analyses with informative restriction enzymes indicated that detected phytoplasmas belonged to 16SrII-D subgroup. These phytoplasmas are reported for the first time to infect olive trees, and for the first time in Egypt in peach, orange and hibiscus plants

    Effectiveness of root-bark extract from Salvadora persica against the growth of certain molecularly identified pathogenic bacteria

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    The acetone extract from root-bark of Salvadora persica L. (Salvadoraceae), is assayed for its antibacterial activity against some bacterial pathogens. By GC/MS analysis, the main chemical components of the acetone extract were found to be benzylisothiocyanate (39.4%), and benzyl nitrile (benzeneacetonitrile) (37.9%). According the extract concentrations used, the measured inhibition zones observed were between from 13.6 to 18.6 mm, 15.3–23 mm, 13.3–18.3 mm, 13.3–18.3 mm, and 12.3–19 mm, against the isolated plant bacterial pathogens namely Agrobacterium tumefaciens, Pectobacterium atrosepticum, Enterobacter cloacae, Dickeya solani and Ralstonia solanacearum, respectively, whilst it was between 8 and 12 mm, 8–9.6 mm, 8–11.6 mm, and 8–10.3 mm against Bacillus subtilis, Sarcina lutea, Escherichia coli and Staphylococcus aureus, respectively. The minimum inhibitory concentration values of the extract were between 16 and 32 μg/mL against the growth of plant bacterial, and from 1000 to 2000 μg/mL against the growth of the human bacteria. In conclusion, the acetone extract of rootbark of S. persica showed strong antibacterial activity against the plant pathogens and some activity against the human pathogens were reported. The results suggested that using the acetone extract from root-bark of S. persica as bioactive agent against the growth of the studied plant bacterial pathogens

    Bacillus velezensis PEA1 Inhibits Fusarium oxysporum Growth and Induces Systemic Resistance to Cucumber Mosaic Virus

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    Bacillus velezensis manifests robust biocontrol activity against fungal plant pathogens; however, its antiviral activity has rarely been investigated. Bacillus velezensis strain PEA1 was isolated, characterized, and evaluated for antifungal and antiviral activities against Fusarium oxysporum MT270445 and cucumber mosaic virus (CMV) MN594112. Our findings proved that strain PEA1 had intense antagonist activity against F.oxysporum. Under greenhouse conditions, the antiviral activities (protective, curative, and inactivation) of PEA1-culture filtrate (CF) on Datura stramonium plants were assayed, using a half-leaf method. The inactivation treatment exhibited the highest inhibition rate (97.56%) and the most considerable reduction of CMV-CP accumulation levels (2.1-fold) in PEA1-CF-treated plants when compared with untreated plants (26.9-fold). Furthermore, PEA1-CF induced systemic resistance with significantly elevated transcriptional levels of PAL, CHS, HQT, PR-1, and POD genes in D. stramonium leaves after all treatments. Gas chromatography‒mass spectrometry analysis showed that pyrrolo[1,2-a]pyrazine-1,4-dione is the main compound in the PEA1-CF ethyl acetate extract, which may act as an elicitor molecule that induces plant systemic resistance and inhibits both fungal growth and viral replication. Consequently, B. velezensis can be considered as a potential source for the production of bioactive compounds for the management of plant diseases. To our knowledge, this is the first report of the antiviral activity of B. velezensis against plant viral infection

    First Report of Protective Activity of <i>Paronychia argentea</i> Extract against Tobacco Mosaic Virus Infection

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    The widespread use of chemical control agents and pesticides for plant-pathogen control has caused many human health and environmental issues. Plant extracts and biocontrol agents have robust antimicrobial activity against different plant pathogens. However, their antiviral activities are still being investigated. In the present study, the methanol extract of Paronychia argentea was characterized and evaluated for its protective activity against the tobacco mosaic virus (TMV) infection in tomato plants under greenhouse conditions at 21 days post-inoculation. The results showed that the foliar application of P. argentea extract (10 µg/mL) enhanced tomato plant growth, resulting in significant increases in shoot and root parameters and total chlorophyll contents. Moreover, a significant reduction in TMV accumulation level in P. argentea-treated plants of 77.88% compared to non-treated plants was reported. Furthermore, induction of systemic resistance with significant elevation in production of antioxidant enzymes (PPO, CAT, and SOD) and transcriptional levels of the pathogenesis-related proteins (PR-1 and PR-7) and polyphenolic genes (CHS and HQT) were also observed. Out of 16 detected compounds, HPLC analysis revealed that the most abundant polyphenolic compounds found in P. argentea extract were gallic acid (5.36 µg/mL), kaempferol (7.39 µg/mL), quercetin (7.44 µg/mL), ellagic acid (7.89 µg/mL), myricetin (8.36 µg/mL), and ferulic acid (8.69 µg/mL). The findings suggest that the use of P. argentea extract as an effective and safe source for the production of bioactive compounds may offer a solution for a promising approach for the management of plant viral infections. To the best of our knowledge, this is the first report of the protective activity of P. argentea extract against plant viral diseases

    Toxicity effects of <i>Eriocephalus africanus</i> L. leaf essential oil against some molecularly identified phytopathogenic bacterial strains

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    Essential oil (EO) from Eriocephalus africanus L. leaves was evaluated against the growth of some phytopathogenic bacteria including Agrobacerium tumifaciens, Dickeya solani, Erwinia amylovora, Pseudomonas cichorii and Serratia pulmithica using the disc diffusion method and minimum inhibitory concentration (MIC) evaluation. Ten compounds in the EO with dominance of Artemisia ketone (2,5,5-trimethyl-2,6-heptadien-4-one) (77.92%) and ledol (19.92%) were revealed. The antibacterial activity indicated efficacy of essential oil against majority of strains isolated. The most effective action was recorded against D. solani, by 7.5 and 10 µL of oil, with 18.33 mm and 100 μg/mL as zone inhibition and MIC, respectively, whereas the lowest activity was exhibited against P. cichorii (diameter inhibition = 6.66 mm at 10 µL of oil, MIC = 100 μg/mL). The strain S. pulmithica appears to be resistant to the oil when the activity is measured by 10 µL of oil but its growth inhibition was reported with a MIC of 100 μg/mL.</p

    Rhizobium leguminosarum bv. viciae-Mediated Silver Nanoparticles for Controlling Bean Yellow Mosaic Virus (BYMV) Infection in Faba Bean Plants

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    The faba bean plant (Vicia faba L.) is one of the world’s most important legume crops and can be infected with various viral diseases that affect its production. One of the more significant viruses in terms of economic impact is bean yellow mosaic virus (BYMV). The current study used the molecularly identified Rhizobium leguminosarum bv. viciae strain 33504-Borg1, a nitrogen-fixing bacteria, to biosynthesize silver nanoparticles (AgNPs) to control BYMV disease in faba bean plants. Scanning electron microscopy (SEM), a particle size analyzer (PSA) with dynamic light scattering (DLS), transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDX), and Fourier transform infrared spectroscopy (FTIR) were used to characterize the prepared AgNPs. The DLS, SEM, and TEM analyses revealed that the AgNPs were spherical and rough, with sizes ranging from 13.7 to 40 nm. The FTIR analysis recognized various functional groups related to AgNP capping and stability. Under greenhouse conditions, spraying faba bean leaves with the AgNPs (100 µg/mL) 24 h before BYMV inoculation induced plant resistance and reduced plant disease severity and virus concentration levels. Contrarily, the AgNP treatment enhanced plant health by raising photosynthetic rates, increasing the fresh and dry weight of the faba bean plants, and increasing other measured metrics to levels comparable to healthy controls. Antioxidant enzymes (peroxidase and polyphenol oxidase) inhibited the development of BYMV in the faba bean plants treated with the AgNPs. The AgNPs decreased oxidative stress markers (H2O2 and MDA) in the faba bean plants. The plants treated with the AgNPs showed higher expression levels of PR-1 and HQT than the control plants. The study findings could be used to develop a simple, low-cost, and environmentally friendly method of protecting the faba bean plant from BYMV

    Antigenic and pathogenicity activities of Ralstonia solanacearum race 3 biovar 2 molecularly identified and detected by indirect ELISA using polyclonal antibodies generated in rabbits

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    The efficiency of the antiserum was compared among 42 isolates that cause potato brown rot disease; our polyclonal antiserum (14 days) reacted positively with all tested isolates at a dilution of 1:6.4×103. Data indicated the different reactions of eight R. solanacearum isolates at various dilutions (1:1.6×103 to 1:5.12×106) at 14 days against polyclonal antiserumat a concentration of approximately 1×108 CFU/mL and we found the lowest detection level by the indirect ELISA technique was 106 CFU/mL. Finally we recommended the reasonable sensitivity results of the ELISA technique to detect the bacterial pathogen given than the cost of this technique if much lower than that of other expensive molecular techniques.Eight molecular-characterized isolates of Ralstonia solanacearum from potato belonging to race 3 biovar 2, their virulence were evaluated on potato cv. Lady Rosette, tomato cv. Strain B, eggplant cv. Balady and pepper cv. Balady and showed high virulence on potato and tomato, and lower virulence on eggplant and pepper. A laboratory study conducted to produce polyclonal antibodies against the potato brown rot bacterium; R. solanacearum cells were generated in female New Zealand white rabbits. A modification were made on the technique of indirect enzyme-linked immunosorbent assay (ELISA) to improve the sensitivity of detection, including antigenic and sensitivity to R. solanacearum race 3 biovar 2 isolates. Determination of the optimum period to collect the antiserum (including, polyclonal antibodies) showed that the best collection dates were at 14, 3 and 7 days, in that order

    Comparative Analysis of the Expression Profiles of Pathogenesis-Related Genes in Tomato Systemically Infected with Tobacco Mosaic and Cucumber Mosaic Viruses

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    In this study, we used RT-qPCR to examine how PR genes were expressed in model tomato (Solanum lycopersicum L.) plants that had been infected with TMV or CMV. Under greenhouse conditions, the indirect ELISA data showed that both viruses were detected for the first time at 6 dpi. Then, the levels of accumulation increased very quickly, reaching a peak of 15 dpi. During the course of the study (1–15 dpi), the Delta CT, NormFinder, BestKeeper, and GeNorm software tools revealed that the β-actin gene was the most informative reference gene in the virally infected tomato tissues. For both the TMV- and CMV-infected tomato plants, the transcriptional expression levels of most tested genes changed between activation and repression, especially in the first 12 dpi. Compared to mock-inoculated plants, the expression levels of PR-1 were induced at all time intervals except at 8 dpi for CMV and at 6, 7, and 8 dpi for TMV infection. Conversely, the greater activation and accumulation of both viruses were associated with the greater up-regulation of PR-2 at 8 dpi, with relative expression levels of 7.28- and 5.84-fold for TMV and CMV, respectively. The up-regulated expression of PR-3, PR-4, and PR-7 was shown at 4 dpi. In contrast, the PR-5 gene was inhibited in TMV at 1 dpi until 9 dpi, and the induction of this gene at 10 dpi increased by 1.72-fold, but PR-5 was observed to up-regulate the expression of CMV at 1 dpi. This study provides the first valuable information on the comparative transcriptional levels of these tomato genes between TMV and CMV infections

    Protective and Curative Effects of Trichoderma asperelloides Ta41 on Tomato Root Rot Caused by Rhizoctonia solani Rs33

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    Two molecularly identified tomato isolates, Trichoderma asperelloides Ta41 and Rhizoctonia solani Rs33, were characterized and antagonistically evaluated. The dual culture technique showed that Ta41 had a high antagonistic activity of 83.33%, while a light microscope bioassay demonstrated that the Ta41 isolate over-parasitized the pathogen completely. Under greenhouse conditions, the application of Ta41 was able to promote tomato plant growth and had a significant increase in plant height, root length, and shoot fresh, shoot dry, root fresh, and root dry weight. It also improved chlorophyll content and total phenol content significantly, both in protective and in curative treatments. The protective treatment assay exhibited the lowest disease index (16.00%), while the curative treatment showed a disease index of 33.33%. At 20 days post-inoculation, significant increases in the relative expression levels of four defense-related genes (PR-1, PR-2, PR-3, and CHS) were observed in all Ta41-treated plants when compared with the non-treated plants. Interestingly, the plants treated with Ta41 alone showed the highest expression, with relative transcriptional levels of CHS, PR-3, PR-1, and PR-2 that were, compared with the control, 3.91-, 3.13-, 2.94-, and 2.69-fold higher, respectively, and the protective treatment showed relative transcriptional levels that were 3.50-, 3.63-, 2.39-, and 2.27-fold higher, respectively. Consequently, the ability of Ta41 to promote tomato growth, suppress Rs33 growth, and induce systemic resistance supports the incorporation of Ta41 as a potential bioagent for controlling root rot disease and increasing the productivity of crops, including tomatoes

    The Phytochemical, Antifungal, and First Report of the Antiviral Properties of Egyptian Haplophyllum tuberculatum Extract

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    In this study, ethanol whole plant extract (WPE) of Haplophyllum tuberculatum was characterized and tested for its antifungal and antiviral activities against Fusarium culmorum, Rhizoctonia solani and tobacco mosaic virus (TMV). High Performance Liquid Chromatography (HPLC) analysis showed that the main phytochemical constituents of H. tuberculatum WPE were resveratrol (5178.58 mg/kg), kaempferol (1735.23 mg/kg), myricetin (561.18 mg/kg), rutin (487.04 mg/kg), quercetin (401.04 mg/kg), and rosmarinic acid (387.33 mg/kg). By increasing H. tuberculatum WPE at concentrations of 1%, 2%, and 3%, all of the fungal isolates were suppressed compared to the two positive and negative controls. Under greenhouse conditions, WPE-treated Chenopodium amaranticolor plants strongly inhibited TMV infection and significantly reduced TMV accumulation levels when compared to non-treated plants. Moreover, the induction of systemic resistance with significant increases in the transcriptional levels of the pathogenesis-related protein-1 (PR-1), chalcone synthase (CHS), and hydroxycinnamoyl-CoA quinate transferase (HQT) genes for treated plants were noticed at 3 and 5 days post-inoculation (dpi) for both assays. To the best of our knowledge, this is the first reported observation of the antiviral activity of H. tuberculatum extract against plant viral infections. Finally, the results obtained suggest that H. tuberculatum WPE can be considered a promising source of both antifungal and antiviral substances for practical use and for developing plant-derived compounds for the effective management of plant diseases
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