Detection and Molecular Characterization of Grapevine Virus A in Jordan

In a study on grapevines in Jordan conducted between 2002 and 2003, grapevine virus A (GVA) was detected in all areas where grapevines were planted. DAS-ELISA analysis of samples from symptomatic trees found that 16.1% of samples were infected with GVA. Using a GVA- specific primer pair (H587/C995), a portion of the coat protein gene of the virus was amplified by IC-RT-PCR and RT-PCR, using leaf extracts and RNA extracted from infected grapevines respectively. After cloning and sequencing the coat protein gene of the Jordanian isolate of GVA (GVA-Jo), the sequence of the amplified product was compared with sequences of other GVA isolates from different countries

Distribution of Fig Mosaic in Jordan

Fig mosaic (FM) is one of the most important diseases of figs in Jordan. A nationwide survey was conducted to determine the incidence and severity of this disease in trees and in seedlings propagated by cuttings in orchards and nurseries in 13 provinces and cities all over the country. Cultivars surveyed included Khdari, Mwazi, Zraki, Khartamani, Dafoori, Turki, Hamari, Esaili, Ajlouni, in addition to an Italian and a French cultivar. Disease severity varied from moderately severe to extremely severe with leaf malformation and fruit drop FM was found in all provinces. Incidence of FM, averaged over trees of all cultivars and all age categories, was 95.3%. Fig trees 3 years and older had the highest disease incidence, ranging from 93.3% to 100% in the different orchards. The Esaili cultivar had the lowest incidence ranging between 50% and100%, with an average of 76.5%. The highest FM incidence was on Dafoori. Of the most common cultivars, Khdari was the most susceptible. Jerash province had the highest percentage (12.5%) of fig seedlings and trees in the most severe disease category. The highest percentage (27.8%) of healthy fig seedlings and trees was in Irbid province. This paper reports the incidence of FM in various local and imported fig cultivars of different ages, and relates the spread of the disease to the method of fig propagation practiced in Jordan. Suggested solutions for the problem, which include the introduction of disease and pest free fig seedlings derived from tissue culture and the establishment of new rules and regulations to prevent the spread of the disease are discussed

Occurrence and Distribution of Citrus tristeza virus (CTV) in the Jordan Valley

In a survey conducted in 2002 and 2003, Citrus tristeza virus (CTV) was detected in the Jordan Valley. The direct tissue blot immunoassay (DTBIA) indicated that 12.7 and 15.2% of samples tested in the central and northern Jordan Valley respectively were infected with CTV. Similar results showed that all citrus species grown in the Jordan Valley were susceptible to CTV. DAS-ELISA analysis of samples from a citrus orchard in the Dir Alla area with severe CTV symptoms indicated that 49% of samples were infected with CTV. Using a CTV specific primer pair (CTV1/CTV10), the coat protein gene of the virus was successfully amplified from leaf extracts obtained from CTVinfected trees by IC-RT-PCR. After cloning and sequencing the coat protein gene, the sequence of the amplified product was deposited in the GenBank

Molecular characterization of Grapevine virus A isolates from Jordan

A total of 1141 samples of petioles and canes of grapevines from different locations in Jordan were tested by DAS-ELISA for Grapevine virus A (GVA). About 14.2% of samples were infected with GVA. Using Reverse Transcription- Polymerase Chain Reaction (RT-PCR) or Immunocapture (IC)-RT-PCR, a fragment of 430 bp of the coat protein gene was amplified using the primer pair H587/C995. Fifteen clones were sequenced and aligned with each other and with the Jordanian isolate (GVA-Jo) using the DNAMAN program. Alignment analysis showed that all Jordanian isolates shared 100% nucleotide identity with each other and with the Jordanian isolate GVA-Jo. To classify the collected GVA isolates in one of the GVA groups a pairwise nucleotide alignment between isolate GVA-5R and isolates from South Africa; 92/778, JP98 and P163-1 representing GVA group I, II and III respectively was done. Alignment results indicated that isolate GVA-5R shared 90, 83, and 76% nucleotide similarity with GVA groups I, II, and III, respectively

Antifungal activity of olive cake extracts

Powdered, dried olive (Olea europaea) cake was extracted with hexane, methanol and butanol. Six phenolic compounds, coumaric acid, ferulic acid, oleuropein, caffeic acid, protocatechuic acid and cinnamic acid, were isolated from these extracts after fractionation. The fractions were tested for their antifungal activity against Verticillium sp., Fusarium oxysporum, Rhizopus sp., Penicillium italicum, Rhizoctonia solani, Stemphylium solani, Cladosporium sp., Mucor sp., Colletotrichum sp. and Pythium sp. Strongest activity was reported against Fusarium oxysporum and Verticillium sp. No effect was observed against Alternaria sp

Validation of a microarrays protocol for detection and genotyping isolates of Plum pox virus

A genomic strategy for PPV identification has been recently developed (Pasquini et al., 2008). The method is based on using a 70-mer oligonucleotide DNA microarray chip capable of simultaneously detecting and genotyping PPV strains. Universal and specific probes have been identified and used with a sensitive protocol of hybridization using an indirect fluorescent labelling of cDNA product with cyanine able to enhance the sensitivity of the virus detection avoiding the use of the PCR amplification step. In order to evaluate the protocol fitness for diagnostic use, about 30 samples belonging to a PPV isolates collection, including M, D, EA and C strains, have been used for its validation, that was determined, estimating the performance criteria that include the following parameters: diagnostic sensitivity (D-SN), diagnostic specificity (D-SP) and diagnostic accuracy (D-AC). Keywords: oligonucleotides chip, PPV, sensitivity, specificity, accuracy, performance criteri

Hexanoic Acid Treatment Prevents Systemic MNSV Movement in Cucumis melo Plants by Priming Callose Deposition Correlating SA and OPDA Accumulation

[EN] Unlike fungal and bacterial diseases, no direct method is available to control viral diseases. The use of resistance-inducing compounds can be an alternative strategy for plant viruses. Here we studied the basal response of melon to Melon necrotic spot virus (MNSV) and demonstrated the efficacy of hexanoic acid (Hx) priming, which prevents the virus from systemically spreading. We analysed callose deposition and the hormonal profile and gene expression at the whole plant level. This allowed us to determine hormonal homeostasis in the melon roots, cotyledons, hypocotyls, stems and leaves involved in basal and hexanoic acid-induced resistance (Hx-IR) to MNSV. Our data indicate important roles of salicylic acid (SA), 12-oxo-phytodienoic acid (OPDA), jasmonic-isoleucine, and ferulic acid in both responses to MNSV. The hormonal and metabolites balance, depending on the time and location associated with basal and Hx-IR, demonstrated the reprogramming of plant metabolism in MNSV-inoculated plants. The treatment with both SA and OPDA prior to virus infection significantly reduced MNSV systemic movement by inducing callose deposition. This demonstrates their relevance in Hx-IR against MNSV and a high correlation with callose deposition. Our data also provide valuable evidence to unravel priming mechanisms by natural compounds.This work has been supported by grants from the Spanish Ministry of Science and Innovation (AGL2010-22300-C03-01-02, AGL2013-49023-C03-01-02-R and BIO2014-54862-R), co-funded by the European Regional Development Fund.Fernandez-Crespo, E.; Navarro Bohigues, JA.; Serra Soriano, M.; Finiti, I.; García Agustín, P.; Pallás Benet, V.; Gonzalez-Bosch, C. (2017). Hexanoic Acid Treatment Prevents Systemic MNSV Movement in Cucumis melo Plants by Priming Callose Deposition Correlating SA and OPDA Accumulation. Frontiers in Plant Science. 8:1-15. https://doi.org/10.3389/fpls.2017.01793S1158Alazem, M., & Lin, N. (2014). Roles of plant hormones in the regulation of host–virus interactions. Molecular Plant Pathology, 16(5), 529-540. doi:10.1111/mpp.12204Ando, S., Obinata, A., & Takahashi, H. (2014). WRKY70 interacting with RCY1 disease resistance protein is required for resistance to Cucumber mosaic virus in Arabidopsis thaliana. Physiological and Molecular Plant Pathology, 85, 8-14. doi:10.1016/j.pmpp.2013.11.001Anfoka, G. H. (2000). Benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester induces systemic resistance in tomato (Lycopersicon esculentum. Mill cv. Vollendung) to Cucumber mosaic virus. 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An untargeted global metabolomic analysis reveals the biochemical changes underlying basal resistance and priming in Solanum lycopersicum, and identifies 1-methyltryptophan as a metabolite involved in plant responses to Botrytis cinerea and Pseudomonas sy. The Plant Journal, 84(1), 125-139. doi:10.1111/tpj.12964Clarke, S. F., Guy, P. L., Burritt, D. J., & Jameson, P. E. (2002). Changes in the activities of antioxidant enzymes in response to virus infection and hormone treatment. Physiologia Plantarum, 114(2), 157-164. doi:10.1034/j.1399-3054.2002.1140201.xCollum, T. D., & Culver, J. N. (2016). The impact of phytohormones on virus infection and disease. Current Opinion in Virology, 17, 25-31. doi:10.1016/j.coviro.2015.11.003Conti, G., Rodriguez, M. C., Venturuzzi, A. L., & Asurmendi, S. (2016). Modulation of host plant immunity by Tobamovirus proteins. Annals of Botany, mcw216. doi:10.1093/aob/mcw216Culver, J. N., & Padmanabhan, M. S. (2007). 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Molecular Plant Pathology, 15(6), 550-562. doi:10.1111/mpp.12112Flors, V., Leyva, M. de la O., Vicedo, B., Finiti, I., Real, M. D., García-Agustín, P., … González-Bosch, C. (2007). Absence of the endo-β-1,4-glucanases Cel1 and Cel2 reduces susceptibility toBotrytis cinereain tomato. The Plant Journal, 52(6), 1027-1040. doi:10.1111/j.1365-313x.2007.03299.xFlors, V., Ton, J., Van Doorn, R., Jakab, G., García-Agustín, P., & Mauch-Mani, B. (2007). Interplay between JA, SA and ABA signalling during basal and induced resistance against Pseudomonas syringae and Alternaria brassicicola. The Plant Journal, 54(1), 81-92. doi:10.1111/j.1365-313x.2007.03397.xFriedrich, L., Lawton, K., Ruess, W., Masner, P., Specker, N., Rella, M. G., … Ryals, J. (1996). A benzothiadiazole derivative induces systemic acquired resistance in tobacco. The Plant Journal, 10(1), 61-70. doi:10.1046/j.1365-313x.1996.10010061.xFurch, A. C. U., Zimmermann, M. R., Kogel, K.-H., Reichelt, M., & Mithöfer, A. (2014). 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Molecular Plant Pathology, 9(4), 447-461. doi:10.1111/j.1364-3703.2008.00474.xHanley-Bowdoin, L., Bejarano, E. R., Robertson, D., & Mansoor, S. (2013). Geminiviruses: masters at redirecting and reprogramming plant processes. Nature Reviews Microbiology, 11(11), 777-788. doi:10.1038/nrmicro3117Hernandez, J. A., Diaz-Vivancos, P., Rubio, M., Olmos, E., Ros-Barcelo, A., & Martinez-Gomez, P. (2006). Long-term plum pox virus infection produces an oxidative stress in a susceptible apricot, Prunus armeniaca, cultivar but not in a resistant cultivar. Physiologia Plantarum, 126(1), 140-152. doi:10.1111/j.1399-3054.2005.00581.xHipper, C., Brault, V., Ziegler-Graff, V., & Revers, F. (2013). Viral and Cellular Factors Involved in Phloem Transport of Plant Viruses. Frontiers in Plant Science, 4. doi:10.3389/fpls.2013.00154Inaba, J., Kim, B. M., Shimura, H., & Masuta, C. (2011). Virus-Induced Necrosis Is a Consequence of Direct Protein-Protein Interaction between a Viral RNA-Silencing Suppressor and a Host Catalase. Plant Physiology, 156(4), 2026-2036. doi:10.1104/pp.111.180042Lange, L., & Insunza, V. (1977). Root-inhabiting Olpidium species: The O. radicale complex. Transactions of the British Mycological Society, 69(3), 377-384. doi:10.1016/s0007-1536(77)80074-0Lee, J.-Y., Wang, X., Cui, W., Sager, R., Modla, S., Czymmek, K., … Lakshmanan, V. (2011). A Plasmodesmata-Localized Protein Mediates Crosstalk between Cell-to-Cell Communication and Innate Immunity in Arabidopsis. The Plant Cell, 23(9), 3353-3373. doi:10.1105/tpc.111.087742Leyva, M. O., Vicedo, B., Finiti, I., Flors, V., Del Amo, G., Real, M. D., … González-Bosch, C. (2008). Preventive and post-infection control ofBotrytis cinereain tomato plants by hexanoic acid. Plant Pathology, 57(6), 1038-1046. doi:10.1111/j.1365-3059.2008.01891.xLi, J., Brader, G., Kariola, T., & Tapio Palva, E. (2006). WRKY70 modulates the selection of signaling pathways in plant defense. The Plant Journal, 46(3), 477-491. doi:10.1111/j.1365-313x.2006.02712.xManacorda, C. A., Mansilla, C., Debat, H. J., Zavallo, D., Sánchez, F., Ponz, F., & Asurmendi, S. (2013). Salicylic Acid Determines Differential Senescence Produced by Two Turnip mosaic virus Strains Involving Reactive Oxygen Species and Early Transcriptomic Changes. Molecular Plant-Microbe Interactions®, 26(12), 1486-1498. doi:10.1094/mpmi-07-13-0190-rMandadi, K. K., & Scholthof, K.-B. G. (2013). Plant Immune Responses Against Viruses: How Does a Virus Cause Disease? The Plant Cell, 25(5), 1489-1505. doi:10.1105/tpc.113.111658Mauch-Mani, B., & Mauch, F. (2005). The role of abscisic acid in plant–pathogen interactions. Current Opinion in Plant Biology, 8(4), 409-414. doi:10.1016/j.pbi.2005.05.015Mayers, C. N., Lee, K.-C., Moore, C. A., Wong, S.-M., & Carr, J. P. (2005). Salicylic Acid-Induced Resistance to Cucumber mosaic virus in Squash and Arabidopsis thaliana: Contrasting Mechanisms of Induction and Antiviral Action. Molecular Plant-Microbe Interactions®, 18(5), 428-434. doi:10.1094/mpmi-18-0428Mittler, R. (2017). ROS Are Good. Trends in Plant Science, 22(1), 11-19. doi:10.1016/j.tplants.2016.08.002Miura, K., & Tada, Y. (2014). Regulation of water, salinity, and cold stress responses by salicylic acid. Frontiers in Plant Science, 5. doi:10.3389/fpls.2014.00004Naumann, M., Somerville, S. C., & Voigt, C. A. (2013). Differences in early callose deposition during adapted and non-adapted powdery mildew infection of resistantArabidopsislines. Plant Signaling & Behavior, 8(6), e24408. doi:10.4161/psb.24408Navarro, J. A., Genovés, A., Climent, J., Saurí, A., Martínez-Gil, L., Mingarro, I., & Pallás, V. (2006). RNA-binding properties and membrane insertion of Melon necrotic spot virus (MNSV) double gene block movement proteins. Virology, 356(1-2), 57-67. doi:10.1016/j.virol.2006.07.040Nicaise, V. (2014). Crop immunity against viruses: outcomes and future challenges. Frontiers in Plant Science, 5. doi:10.3389/fpls.2014.00660Nieto, C., Morales, M., Orjeda, G., Clepet, C., Monfort, A., Sturbois, B., … Bendahmane, A. (2006). AneIF4Eallele confers resistance to an uncapped and non-polyadenylated RNA virus in melon. The Plant Journal, 48(3), 452-462. doi:10.1111/j.1365-313x.2006.02885.xNováková, S., Flores-Ramírez, G., Glasa, M., Danchenko, M., Fiala, R., & Skultety, L. (2015). Partially resistant Cucurbita pepo showed late onset of the Zucchini yellow mosaic virus infection due to rapid activation of defense mechanisms as compared to susceptible cultivar. Frontiers in Plant Science, 6. doi:10.3389/fpls.2015.00263Ohki, T., Akita, F., Mochizuki, T., Kanda, A., Sasaya, T., & Tsuda, S. (2010). The protruding domain of the coat protein of Melon necrotic spot virus is involved in compatibility with and transmission by the fungal vector Olpidium bornovanus. Virology, 402(1), 129-134. doi:10.1016/j.virol.2010.03.020Pacheco, R., García-Marcos, A., Manzano, A., de Lacoba, M. G., Camañes, G., García-Agustín, P., … Tenllado, F. (2012). Comparative Analysis of Transcriptomic and Hormonal Responses to Compatible and Incompatible Plant-Virus Interactions that Lead to Cell Death. Molecular Plant-Microbe Interactions®, 25(5), 709-723. doi:10.1094/mpmi-11-11-0305Padmanabhan, M. S., Shiferaw, H., & Culver, J. N. (2006). The Tobacco mosaic virus Replicase Protein Disrupts the Localization and Function of Interacting Aux/IAA Proteins. Molecular Plant-Microbe Interactions®, 19(8), 864-873. doi:10.1094/mpmi-19-0864Pallas, V., & García, J. A. (2011). How do plant viruses induce disease? Interactions and interference with host components. 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Actualización del estatus de las moscas blancas (Hemiptera: Aleyrodidae) en Jordania con énfasis en el complejo Bemisia tabaci

Whiteflies are economically important plant pests that cause damage to crops worldwide. This study aimed to update the status of whiteflies in Jordan by combining the classical morphological identification and the DNA markers using the mitochondrial cytochrome oxidase I (mtCOI) gene. Over the course of three consecutive years, 111 whiteflies were collected from different geographical regions and different plant hosts in Jordan. The results showed that, in addition to Bemisia tabaci, another nine different whitefly species were identified, including two species that were recorded for the first time in Jordan: Africaleurodes coffeacola, and Tetraleurodes neemani. A special focus has been given to economically important plant pests like the B. tabaci species complex. Three different diagnostic techniques were used to identify B. tabaci putative species based on mtCOI gene. All the collected samples of B. tabaci species complex were identified as Middle East–Asia Minor 1 (MEAM1) putative species.Las moscas blancas son plagas de plantas de importancia económica, que causan daños a cultivos en todo el mundo. Este estudio tuvo como objetivo actualizar el estado de conocimiento sobre las moscas blancas en Jordania, combinando la identificación morfológica clásica y la tecnica del gen de citocromo oxidasa I mitocondrial (mtCOI) como un marcador de ADN. En el transcurso de tres años consecutivos se recolectaron 111 moscas blancas de diferentes regiones geográficas, y de diferentes plantas hospederas. Los resultados mostraron que, además de Bemisia tabaci, existen nueve especies diferentes de mosca blanca; incluso se registraron por primera vez en Jordania dos especies: Africaleurodes coffeacola y Tetraleurodesneemani. Se hizo especial énfasis en el complejo de especies de B. tabaci por su importancia económica. Se utilizaron tres técnicas de diagnóstico diferentes para identificar especies cercanas a B. tabaci basadas en el gen mtCOI. Sin embargo, todas las muestras recolectadas del complejo de especies de B. tabaci se identificaron como especies del complejo de Oriente Medio-Asia Menor 1 (MEAM1)