Abstract

ABSTRACTViral interactions during multiple viral infections were examined in Agaricus bisporus cultures harboring 9 viruses (comprising 18 distinct viral RNAs) by statistically analyzing their relative abundance in fruitbodies. Four clusters of viral RNA were identified that suggested synergism and coreplication. Pairwise correlations revealed negative and positive correlations between clusters, indicating further synergisms and an antagonism involving a group containing a putative hypovirus and four nonhost ORFan RNAs (RNAs with no similarity to known sequences) possibly acting as defective interfering RNAs. The disease phenotype was observed in 10 to 15% of the fruitbodies apparently randomly located among asymptomatic fruitbodies. The degree of symptom expression consistently correlated with the levels of the multipartite virus AbV16. Diseased fruitbodies contained very high levels of AbV16 and AbV6 RNA2; these levels were orders of magnitude higher than those in asymptomatic tissues and were shown statistically to be discretely higher populations of abundance, indicating an exponential shift in the replicative capacity of the virus. High levels of AbV16 replication were specific to the fruitbody and not found in the underlying mycelium. There appeared to be a stochastic element occurring in these viral interactions, as observed in the distribution of diseased symptoms across a culture, differences in variance between experiments, and a number of additional viruses undergoing the step-jump in levels between experiments. Possible mechanisms for these multiple and simultaneous viral interactions in single culture are discussed in relation to known virus-host regulatory mechanisms for viral replication and whether additional factors could be considered to account for the 1,000-fold increase in AbV16 and AbV6 RNA2 levels.

Highlights

  • Viral interactions during multiple viral infections were examined in Agaricus bisporus cultures harboring 9 viruses by statistically analyzing their relative abundance in fruitbodies

  • The complexity of interactions is massively increased in multiple viral infections where a host harbors large numbers of viruses, a condition that appears to be widespread in some perennial plants and fungi

  • A preliminary experiment was carried out to determine how many of the 30 viral RNAs identified by Deakin et al [10] were present in mushroom virus X (MVX)-4569-infected mushrooms using quantitative PCR of a single brown mushroom and a single white mushroom from each experiment

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Summary

Introduction

Viral interactions during multiple viral infections were examined in Agaricus bisporus cultures harboring 9 viruses (comprising 18 distinct viral RNAs) by statistically analyzing their relative abundance in fruitbodies. There appeared to be a stochastic element occurring in these viral interactions, as observed in the distribution of diseased symptoms across a culture, differences in variance between experiments, and a number of additional viruses undergoing the step-jump in levels between experiments Possible mechanisms for these multiple and simultaneous viral interactions in single culture are discussed in relation to known virus-host regulatory mechanisms for viral replication and whether additional factors could be considered to account for the 1,000-fold increase in AbV16 and AbV6 RNA2 levels. The name MVX was originally used to describe both of these emerging diseases, but they are considered distinct Fruitbodies of both diseases contain multiple viral infections, and 30 RNAs have been sequenced, consisting of 18 viruses and 8 nonhost ORFans (RNAs with no similarity to known sequences) [10]. Mushroom bacilliform virus (MBV) has been characterized and identified in the deep-sequenced RNAs of mushrooms with BCMD symptoms and occurs as a coinfection with AbV1 in La France disease [10, 25]

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