Articles
DETECTION AND IDENTIFICATION OF CUCUMBER MOSAIC VIRUS ISOLATE FROM RED CURRANT ‘ROSETTA’
Article number
780_7
Pages
55 – 60
Language
English
Abstract
Cucumber mosaic virus was detected using enzyme-linked immunosorbent assay (ELISA) in a symptomless red currant plant Rosetta, during routine testing in the nursery in central Poland.
Mild leaf mosaic and vein yellowing symptoms were observed on the same plant a year later.
Serological properties of the virus (isolate PORZ) were determined using ELISA with four polyclonal antibodies: commercially available CMV-I and CMV-II specific for subgroup I and II, respectively (Agdia), and antisera locally prepared against CMV isolates M and Wic belonging to subgroup I and II, respectively.
The red currant isolate (PORZ) reacted positively with antibodies M, Wic and CMV-II, but did not react with CMV-I antibodies.
The coat protein (CP) of isolate PORZ co-migrated in SDS-PAGE with the CP of isolates belonging to CMV subgroup II, faster than the CP of CMV-I isolates.
The CP was detected in western blots with polyclonal antiserum against CMV. Fragments of the PORZ RNA3 were reverse transcribed and amplified using immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) and sequenced.
The PORZ isolate had greater than 95% nucleotide sequence identity in the CP and 3a genes with the members of CMV subgroup II, and less than 80% sequence identity with CMV-I isolates.
The identity of isolate PORZ in deduced amino acid sequence of the coat protein with CMV II isolates was 96% or higher, and less than 87% with CMV-I isolates.
On the basis of these results, isolate PORZ was classified as a member of subgroup II of CMV.
Mild leaf mosaic and vein yellowing symptoms were observed on the same plant a year later.
Serological properties of the virus (isolate PORZ) were determined using ELISA with four polyclonal antibodies: commercially available CMV-I and CMV-II specific for subgroup I and II, respectively (Agdia), and antisera locally prepared against CMV isolates M and Wic belonging to subgroup I and II, respectively.
The red currant isolate (PORZ) reacted positively with antibodies M, Wic and CMV-II, but did not react with CMV-I antibodies.
The coat protein (CP) of isolate PORZ co-migrated in SDS-PAGE with the CP of isolates belonging to CMV subgroup II, faster than the CP of CMV-I isolates.
The CP was detected in western blots with polyclonal antiserum against CMV. Fragments of the PORZ RNA3 were reverse transcribed and amplified using immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) and sequenced.
The PORZ isolate had greater than 95% nucleotide sequence identity in the CP and 3a genes with the members of CMV subgroup II, and less than 80% sequence identity with CMV-I isolates.
The identity of isolate PORZ in deduced amino acid sequence of the coat protein with CMV II isolates was 96% or higher, and less than 87% with CMV-I isolates.
On the basis of these results, isolate PORZ was classified as a member of subgroup II of CMV.
Authors
H. Śliwa, M. Kamińska, T. Malinowski
Keywords
CMV, serology, IC-RT-PCR
Online Articles (10)