|Authors: ||K.I. Posthuma, Y.G. Hong, A.N. Adams|
|Keywords: ||Cytorhabdovirus, Plant rhabdovirus, Mononegavirales, Fragaria, RT-PCR, RACE.|
Strawberry crinkle virus (SCV) causes crinkle disease of strawberry, Fragaria x ananassa Duch., which can result in a decrease in fruit yield and quality.
SCV is routinely detected by leaf grafting to sensitive indicator plants, a method that is labour intensive.
The concentration of SCV in the plant is low, purification of the virus particles is difficult and attempts to develop a serological detection method for the virus have been unsuccessful.
In order to develop a PCR-based detection method for SCV, sequences of the large protein genes of seven rhabdoviruses were compared.
Degenerate primers were designed on highly conserved regions of the large protein gene.
Reverse-transcription polymerase chain reaction (RT-PCR) was performed on total RNA isolated from healthy Physalis pubescens L. and P. pubescens plants infected with SCV. Two SCV-specific products were cloned and sequenced.
The known sequence was extended towards the 3’ end of the large protein gene using rapid amplification of cDNA ends (RACE). Diagnostic primer pairs were designed and tested on SCV isolates in P. pubescens and strawberry.
It was found that SCV could not always be detected in strawberry whereas detection in P. pubescens was much more reliable.
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