ISHS Acta Horticulturae 656: X International Symposium on Small Fruit Virus Diseases
PENTAPLEX RT-PCR FOR THE SIMULTANEOUS DETECTION OF FOUR APHID-BORNE VIRUSES IN COMBINATION WITH A PLANT MRNA SPECIFIC INTERNAL CONTROL IN FRAGARIA SPP. |
| Authors: | J.R. Thompson, S. Wetzel, W. Jelkmann |
| Abstract:
The principal aphid-borne viruses infecting strawberry (Fragaria spp.) Strawberry crinkle virus (SCV), Strawberry mild yellow edge virus (SMYEV), Strawberry mottle virus (SMoV) and Strawberry vein banding virus (SVBV) can cause serious crop losses. In this paper we describe a multiplex reverse transcriptase polymerase chain reaction (RT-PCR) method for the simultaneous detection of all four viruses in combination with a plant mRNA specific internal control which can be used as an indicator of the effectiveness of the extraction and RT-PCR. In total 18 naturally infected strawberry isolates were analysed by this method. Every combination of RNA virus was detected and a full complement of all four viruses was found together in three isolates, all from wild strawberry (Fragaria chiloensis (L., Duch.) in Chile. The upper detection limit for the four viruses was at an extract dilution of 1/200. The broad applicability of the RNA specific internal control primers which produced a PCR fragment of the expected size in 25 of 27 plant species tested, provides an internal standard for RT-PCR analyses in a wide variety of plant species. | |
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