Volume 19 Part 1 Article 15
Title: A sensitive and quantitative test for >31 viral RNAs associated with the Mushroom Virus X diseases in both compost and mushrooms.
Author: Edward Dobbs, Greg Deakin, Julie Bennett, Caoimhe Fleming-Archibald, Helen Grogan, Ian Jones and Kerry Burton
Mushroom Virus X (MVX) disease is a syndrome affecting the commercial fungal crop Agaricus bisporus (the common white mushroom). The disease has been widespread across Europe for the last 25 years causing a number of symptoms: (1) brown discolouration of the mushroom cap, (2) delayed fruit body development and (3) misshapen fruit bodies. The symptoms have been found to coincide with an increased number of double stranded (ds)RNA elements which vary in size between ~20kb and ~600bp
Double-stranded (ds)RNA was purified from the fruit bodies of one symptomless sample and nine diseased samples and sequenced by Illumina MiSeq. The resulting sequences were compared to known sequence information for the MVX dsRNA banding patterns. Primers were designed and validated for all of the viral RNAs for qPCR detection.
RNA extraction from mushroom compost combined with a quantitative RT-qPCR technique were used to develop a test to measure relative levels of each of the novel viral RNAs, to be used by the growers and to aid epidemiological studies of the disease.