Reliable RT-PCR detection of Apple stem pitting virus in pome fruits and its association with quince fruit deformation disease
Abstract
In this study a spot nested RT-PCR assay was developed for the detection of Apple stem pitting virus (ASPV). A one step RT-PCR for the generic detection of foveaviruses using degenerate primers that target a conserved region of the RNA-dependent RNA polymerase (RdRp) gene was followed by a nested PCR that amplifies a 312 bp ASPV specific product. The method is rapid, simple and displays high sensitivity and broad detection range, overcoming the virus molecular variability. The optimum sampling conditions for reliable virus detection were also investigated. ASPV was detected throughout the year in different plant tissues of affected trees, thus the method could be used for routine screening and in certification schemes of pome fruits. ASPV was detected in quince orchards in Greece in all trees that were tested, showing a fruit deformation disorder. Sequencing and phylogenetic analysis of amplicons generated by RT-PCR from plant tissue affected with the deformation disease indicated that... the agent responsible was a variant of ASPV.
Keywords:
quince fruit deformation disease / Foveavirus / Cydonia oblonga / ASPVSource:
Plant Pathology, 2009, 58, 2, 228-236Publisher:
- Wiley, Hoboken
DOI: 10.1111/j.1365-3059.2008.01952.x
ISSN: 0032-0862
WoS: 000264084700004
Scopus: 2-s2.0-62349097432
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Institut za voćarstvoTY - JOUR AU - Mathioudakis, M. M. AU - Maliogka, V. I. AU - Dovas, C. I. AU - Paunović, Svetlana A. AU - Katis, N I. PY - 2009 UR - https://refri.institut-cacak.org/handle/123456789/148 AB - In this study a spot nested RT-PCR assay was developed for the detection of Apple stem pitting virus (ASPV). A one step RT-PCR for the generic detection of foveaviruses using degenerate primers that target a conserved region of the RNA-dependent RNA polymerase (RdRp) gene was followed by a nested PCR that amplifies a 312 bp ASPV specific product. The method is rapid, simple and displays high sensitivity and broad detection range, overcoming the virus molecular variability. The optimum sampling conditions for reliable virus detection were also investigated. ASPV was detected throughout the year in different plant tissues of affected trees, thus the method could be used for routine screening and in certification schemes of pome fruits. ASPV was detected in quince orchards in Greece in all trees that were tested, showing a fruit deformation disorder. Sequencing and phylogenetic analysis of amplicons generated by RT-PCR from plant tissue affected with the deformation disease indicated that the agent responsible was a variant of ASPV. PB - Wiley, Hoboken T2 - Plant Pathology T1 - Reliable RT-PCR detection of Apple stem pitting virus in pome fruits and its association with quince fruit deformation disease EP - 236 IS - 2 SP - 228 VL - 58 DO - 10.1111/j.1365-3059.2008.01952.x UR - conv_1375 ER -
@article{ author = "Mathioudakis, M. M. and Maliogka, V. I. and Dovas, C. I. and Paunović, Svetlana A. and Katis, N I.", year = "2009", abstract = "In this study a spot nested RT-PCR assay was developed for the detection of Apple stem pitting virus (ASPV). A one step RT-PCR for the generic detection of foveaviruses using degenerate primers that target a conserved region of the RNA-dependent RNA polymerase (RdRp) gene was followed by a nested PCR that amplifies a 312 bp ASPV specific product. The method is rapid, simple and displays high sensitivity and broad detection range, overcoming the virus molecular variability. The optimum sampling conditions for reliable virus detection were also investigated. ASPV was detected throughout the year in different plant tissues of affected trees, thus the method could be used for routine screening and in certification schemes of pome fruits. ASPV was detected in quince orchards in Greece in all trees that were tested, showing a fruit deformation disorder. Sequencing and phylogenetic analysis of amplicons generated by RT-PCR from plant tissue affected with the deformation disease indicated that the agent responsible was a variant of ASPV.", publisher = "Wiley, Hoboken", journal = "Plant Pathology", title = "Reliable RT-PCR detection of Apple stem pitting virus in pome fruits and its association with quince fruit deformation disease", pages = "236-228", number = "2", volume = "58", doi = "10.1111/j.1365-3059.2008.01952.x", url = "conv_1375" }
Mathioudakis, M. M., Maliogka, V. I., Dovas, C. I., Paunović, S. A.,& Katis, N. I.. (2009). Reliable RT-PCR detection of Apple stem pitting virus in pome fruits and its association with quince fruit deformation disease. in Plant Pathology Wiley, Hoboken., 58(2), 228-236. https://doi.org/10.1111/j.1365-3059.2008.01952.x conv_1375
Mathioudakis MM, Maliogka VI, Dovas CI, Paunović SA, Katis NI. Reliable RT-PCR detection of Apple stem pitting virus in pome fruits and its association with quince fruit deformation disease. in Plant Pathology. 2009;58(2):228-236. doi:10.1111/j.1365-3059.2008.01952.x conv_1375 .
Mathioudakis, M. M., Maliogka, V. I., Dovas, C. I., Paunović, Svetlana A., Katis, N I., "Reliable RT-PCR detection of Apple stem pitting virus in pome fruits and its association with quince fruit deformation disease" in Plant Pathology, 58, no. 2 (2009):228-236, https://doi.org/10.1111/j.1365-3059.2008.01952.x ., conv_1375 .