Abstract

BackgroundCitrus sudden death (CSD), a disease that rapidly kills orange trees, is an emerging threat to the Brazilian citrus industry. Although the causal agent of CSD has not been definitively determined, based on the disease's distribution and symptomatology it is suspected that the agent may be a new strain of Citrus tristeza virus (CTV). CTV genetic variation was therefore assessed in two Brazilian orange trees displaying CSD symptoms and a third with more conventional CTV symptoms.ResultsA total of 286 RNA-dependent-RNA polymerase (RdRp) and 284 heat shock protein 70 homolog (HSP70h) gene fragments were determined for CTV variants infecting the three trees. It was discovered that, despite differences in symptomatology, the trees were all apparently coinfected with similar populations of divergent CTV variants. While mixed CTV infections are common, the genetic distance between the most divergent population members observed (24.1% for RdRp and 11.0% for HSP70h) was far greater than that in previously described mixed infections. Recombinants of five distinct RdRp lineages and three distinct HSP70h lineages were easily detectable but respectively accounted for only 5.9 and 11.9% of the RdRp and HSP70h gene fragments analysed and there was no evidence of an association between particular recombinant mosaics and CSD. Also, comparisons of CTV population structures indicated that the two most similar CTV populations were those of one of the trees with CSD and the tree without CSD.ConclusionWe suggest that if CTV is the causal agent of CSD, it is most likely a subtle feature of population structures within mixed infections and not merely the presence (or absence) of a single CTV variant within these populations that triggers the disease.

Highlights

  • Citrus sudden death (CSD), a disease that rapidly kills orange trees, is an emerging threat to the Brazilian citrus industry

  • Previous analyses of Citrus tristeza virus (CTV) mixed infections have revealed within host CTV population structures that are expected for RNA virus quasispecies: One or two predominant genotypes (>30% of the population) and a number of closely related variants of these [2,6,9]

  • By contrast sequence analysis of the 286 RNA-dependent-RNA polymerase (RdRp) and 284 heat shock protein 70 homolog (HSP70h) gene fragments sampled from amongst the three Brazilian orange trees here revealed a total of 76 RdRp and 153 HSP70h haplotypes

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Summary

Introduction

Citrus sudden death (CSD), a disease that rapidly kills orange trees, is an emerging threat to the Brazilian citrus industry. The causal agent of CSD has not been definitively determined, based on the disease's distribution and symptomatology it is suspected that the agent may be a new strain of Citrus tristeza virus (CTV). Diseases caused by the aphid-borne Closterovirus, Citrus tristeza virus (CTV), are among the greatest threats to citrus production worldwide. Since 2001, an emergent disease, called citrus sudden death (CSD), has caused the death or eradication of four million orange trees throughout the Brazilian states of Minas Gerais and São Paulo [10]. One curiosity of the disease is that it has almost exclusively been reported in trees comprising a sweet orange graft onto a Rangpur lime (Citrus limonia Osb.) rootstock. Neither Rangpur lime nor sweet orange appear sensitive to the disease in isolation and diseased trees can sometimes be salvaged by removing the sweet orange graft

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