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Open access

Beata Hasiów, Natasza Borodynko and Henryk Pospieszny

Development of a Real Time RT-PCR Assay for Detecting Genetically Different Pepino Mosaic Virus Isolates

Over a period of few years, Pepino mosaic virus (PepMV) has become one of the most important viral pathogen in tomato production worldwide. So far, five PepMV genotypes (EU, LP, CH2, US2 and US1) have been detected. A real time reverse transcription polymerase chain reaction (RT-PCR) procedure, using the fluorescence dye SYBR Green was developed for a rapid and reliable detection of genetically diverse Pepino mosaic virus isolates. This procedure was used for the detection and identification of PepMV in both Solanum lycopersicum and Nicotiana benthamiana species. The melting temperature (T m) for members of a particular strain was very similar, with a host effect that did not hinder strain identification. Under optimal reaction conditions, sensitivity of the detection was as low as 100 fg of viral RNA from infected plants. This level of sensitivity indicated that the real time RT-PCR developed in the present study could be used for routine plant health assays.

Open access

Beata Hasiów-Jaroszewska, Natasza Borodynko, Paulina Jackowiak, Marek Figlerowicz and Henryk Pospieszny

Pepino Mosaic Virus - A Pathogen of Tomato Crops in Poland: Biology, Evolution and Diagnostics

Pepino mosaic virus (PepMV) has the potential to cause serious tomato disease. Several isolates of PepMV were collected between 2003-2010 in the Department of Virology and Bacteriology, Institute of Plant Protection - National Research Institute, Poznań, Poland. The isolates were obtained from greenhouse tomato plants (Solanum lycopersicum) collected from different regions of Poland. Clear biological and genetic differences were detected among the Polish PepMV isolates. Since the latter induced a wide range of symptoms on tested plant species, five different pathotypes were distinguished. Sequence analysis showed that the Polish PepMV isolates share a very high sequence identity with those representing European and Chilean 2 genotype. The analysis of the genetic diversity of the Polish PepMV CH2 isolates suggested that each of them exists as a genetically diverse collection of variants called a quasispecies.

Open access

Julia Minicka, Beata Hasiów-Jaroszewska, Natasza Borodynko-Filas, Henryk Pospieszny and Inge Maria Hanssen

Abstract

Pepino mosaic virus (PepMV) has emerged as an important pathogen of greenhouse tomato crops and is currently distributed worldwide. Population genetic studies have revealed a shift in the dominant PepMV genotype from European (EU) to Chilean 2 (CH2) in North America and several European countries. New genetic variants are constantly being created by mutation and recombination events. Single nucleotide substitutions in different parts of the genome were found to affect on development of symptoms resulting in new pathotypes and accumulation of viral RNA. The variability of the PepMV population has a great impact on designing specific diagnostic tools and developing efficient and durable strategies of disease control. In this paper we review the current knowledge about the PepMV population, the evolutionary dynamics of this highly infective virus, methods for its detection and plant protection strategies.

Open access

Tadeusz Kobyłko, Piotr Dańda, Beata Hasiów, Henryk Pospieszny and Natasza Borodynko

Abstract

A virus was isolated from Lavandula angustifolia Mill. plants exhibiting yellow mottling and distortion of leaves. After mechanical inoculation it induced in the major part of used test plants symptoms characteristic for Cucumber mosaic virus (CMV). Its standard properties regarding the stability in crude plant sap were as follows: longevity in vitro 1-2 days, thermal inactivation point 55-60°C, dilution end point log10minus 3 - 4. The virus reacted positive with diagnostic antiserum against CMV in DAS-ELISA test. RT-PCR reaction revealed similarity between the investigated isolate and the isolate of CMV from the Netherlands belonging to subgroup II. In the light of the foregoing facts the isolated pathogen can be identified as the Cucumber mosaic virus and Lavandula angustifolia may be regarded as its natural host.

Open access

Tadeusz Kobyłko, Piotr Dańda, Beata Hasiów, Natasza Borodynko and Henryk Pospieszny