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Journal of Virological Methods
Pasquini, G., CRA-Istituto Sperimentale per la Patologia Vegetale, Via C.G. Bertero 22, 00156 Rome, Italy
Barba, M., CRA-Istituto Sperimentale per la Patologia Vegetale, Via C.G. Bertero 22, 00156 Rome, Italy
Hadidi, A., Agricultural Research Service, United States Department of Agriculture, Beltsville, MD 20705, United States
Faggioli, F., CRA-Istituto Sperimentale per la Patologia Vegetale, Via C.G. Bertero 22, 00156 Rome, Italy
Negri, R., Laboratorio di Genomica Funzionale e Proteomica dei Sistemi Modello, Dipartimento di Biologia cellulare e dello Sviluppo, Università degli Studi La Sapienza, 00185 Rome, Italy
Sobol, I., Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agriculture, The Hebrew University of Jerusalem, Rehovot, 76100, Israel
Tiberini, A., CRA-Istituto Sperimentale per la Patologia Vegetale, Via C.G. Bertero 22, 00156 Rome, Italy
Caglayan, K., Department of Plant Protection, Faculty of Agriculture, Mustafa Kemal University, 31034 Antakya, Hatay, Turkey
Mazyad, H., Plant Pathology Research Institute, Agricultural Research Center, Ministry of Agriculture and Land Reclamation, Giza, 12619, Egypt
Anfoka, G., Department of Technology, Faculty of Agricultural Technology, Al-Balqa' Applied University, Al- Salt, 19117, Jordan
Ghanim, M., Department of Entomology, Agricultural Research Organization, The Volcani Center, Bet Dagan, 50250, Israel
Zeidan, M., Plant Protection and Inspection Services, Ministry of Agriculture, Bet Dagan, 50250, Israel
Czosnek, H., Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agriculture, The Hebrew University of Jerusalem, Rehovot, 76100, Israel
Plum pox virus (PPV) is the most damaging viral pathogen of stone fruits. The detection and identification of its strains are therefore of critical importance to plant quarantine and certification programs. Existing methods to screen strains of PPV suffer from significant limitations such as the simultaneous detection and genotyping of several strains of PPV in samples infected with different isolates of the virus. A genomic strategy for PPV screening based on the viral nucleotide sequence was developed to enable the detection and genotyping of the virus from infected plant tissue or biological samples. The basis of this approach is a long 70-mer oligonucleotide DNA microarray capable of simultaneously detecting and genotyping PPV strains. Several 70-mer oligonucleotide probes were specific for the detection and genotyping of individual PPV isolates to their strains. Other probes were specific for the detection and identification of two or three PPV strains. One probe (universal), derived from the genome highly conserved 3′ non-translated region, detected all individual strains of PPV. This universal PPV probe, combined with probes specific for each known strain, could be used for new PPV strain discovery. Finally, indirect fluorescent labeling of cDNA with cyanine after cDNA synthesis enhanced the sensitivity of the virus detection without the use of the PCR amplification step. The PPV microarray detected and identified efficiently the PPV strains in PPV-infected peach, apricot and Nicotiana benthamiana leaves. This PPV detection method is versatile, and enables the simultaneous detection of plant pathogens. © 2007 Elsevier B.V. All rights reserved.
פותח על ידי קלירמאש פתרונות בע"מ -
הספר "אוצר וולקני"
אודות
תנאי שימוש
Oligonucleotide microarray-based detection and genotyping of Plum pox virus
147
Pasquini, G., CRA-Istituto Sperimentale per la Patologia Vegetale, Via C.G. Bertero 22, 00156 Rome, Italy
Barba, M., CRA-Istituto Sperimentale per la Patologia Vegetale, Via C.G. Bertero 22, 00156 Rome, Italy
Hadidi, A., Agricultural Research Service, United States Department of Agriculture, Beltsville, MD 20705, United States
Faggioli, F., CRA-Istituto Sperimentale per la Patologia Vegetale, Via C.G. Bertero 22, 00156 Rome, Italy
Negri, R., Laboratorio di Genomica Funzionale e Proteomica dei Sistemi Modello, Dipartimento di Biologia cellulare e dello Sviluppo, Università degli Studi La Sapienza, 00185 Rome, Italy
Sobol, I., Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agriculture, The Hebrew University of Jerusalem, Rehovot, 76100, Israel
Tiberini, A., CRA-Istituto Sperimentale per la Patologia Vegetale, Via C.G. Bertero 22, 00156 Rome, Italy
Caglayan, K., Department of Plant Protection, Faculty of Agriculture, Mustafa Kemal University, 31034 Antakya, Hatay, Turkey
Mazyad, H., Plant Pathology Research Institute, Agricultural Research Center, Ministry of Agriculture and Land Reclamation, Giza, 12619, Egypt
Anfoka, G., Department of Technology, Faculty of Agricultural Technology, Al-Balqa' Applied University, Al- Salt, 19117, Jordan
Ghanim, M., Department of Entomology, Agricultural Research Organization, The Volcani Center, Bet Dagan, 50250, Israel
Zeidan, M., Plant Protection and Inspection Services, Ministry of Agriculture, Bet Dagan, 50250, Israel
Czosnek, H., Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agriculture, The Hebrew University of Jerusalem, Rehovot, 76100, Israel
Oligonucleotide microarray-based detection and genotyping of Plum pox virus
Plum pox virus (PPV) is the most damaging viral pathogen of stone fruits. The detection and identification of its strains are therefore of critical importance to plant quarantine and certification programs. Existing methods to screen strains of PPV suffer from significant limitations such as the simultaneous detection and genotyping of several strains of PPV in samples infected with different isolates of the virus. A genomic strategy for PPV screening based on the viral nucleotide sequence was developed to enable the detection and genotyping of the virus from infected plant tissue or biological samples. The basis of this approach is a long 70-mer oligonucleotide DNA microarray capable of simultaneously detecting and genotyping PPV strains. Several 70-mer oligonucleotide probes were specific for the detection and genotyping of individual PPV isolates to their strains. Other probes were specific for the detection and identification of two or three PPV strains. One probe (universal), derived from the genome highly conserved 3′ non-translated region, detected all individual strains of PPV. This universal PPV probe, combined with probes specific for each known strain, could be used for new PPV strain discovery. Finally, indirect fluorescent labeling of cDNA with cyanine after cDNA synthesis enhanced the sensitivity of the virus detection without the use of the PCR amplification step. The PPV microarray detected and identified efficiently the PPV strains in PPV-infected peach, apricot and Nicotiana benthamiana leaves. This PPV detection method is versatile, and enables the simultaneous detection of plant pathogens. © 2007 Elsevier B.V. All rights reserved.
Scientific Publication
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