Invasive Species Compendium

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Abstract

Establishing a protocol for IC-RT-PCR detection of prune dwarf virus in almond.

Abstract

Most of the Portuguese almond trees, are strongly infected with viruses. Prune dwarf virus (PDV) is widespread in Portuguese almond orchards and is thought to be one of the main factors leading to their low productivity. To monitor the sanitary status of the material obtained from the sanitation programmes, a protocol using immunocapture/reverse transcriptional-polymerase chain reaction (IC-RT-PCR) was established for PDV detection in almond. Conditions like composition of the extraction buffer, age of the extract, incubation period and magnesium concentration were tested to optimize the IC-RT-PCR reactions. Using the optimized conditions, a cDNA fragment of 722 bp, containing the coat protein gene of PDV, was amplified from infected samples.