Small-RNA analysis of pre-basic mother plants and conserved accessions of plant genetic resources for the presence of viruses

PLoS One. 2019 Aug 7;14(8):e0220621. doi: 10.1371/journal.pone.0220621. eCollection 2019.

Abstract

Pathogen-free stocks of vegetatively propagated plants are crucial in certified plant production. They require regular monitoring of the plant germplasm for pathogens, especially of the stocks maintained in the field. Here we tested pre-basic mother plants of Fragaria, Rubus and Ribes spp., and conserved accessions of the plant genetic resources of Rubus spp. maintained at research stations in Finland, for the presence of viruses using small interfering RNA (siRNA) -based diagnostics (VirusDetect). The advance of the method is that unrelated viruses can be detected simultaneously without resumptions of the viruses present. While no virus was detected in pre-basic mother plants of Fragaria and Ribes species, rubus yellow net virus (RYNV) was detected in pre-basic mother plants of Rubus. Raspberry bushy dwarf virus (RBDV), black raspberry necrosis virus (BRNV), raspberry vein chlorosis virus (RVCV) and RYNV were detected in the Rubus genetic resource collection. The L polymerase encoding sequence characterized from seven RVCV isolates showed considerable genetic variation. The data provide the first molecular biological evidence for the presence of RYNV in Finland. RYNV was not revealed in virus indexing by indicator plants, which suggests that it may be endogenously present in some raspberry cultivars. In addition, a putative new RYNV-like badnavirus was detected in Rubus spp. Blackcurrant reversion virus (BRV) and gooseberry vein banding associated virus (GVBaV) were detected in symptomatic Ribes plants grown in the field. Results were consistent with those obtained using PCR or reverse transcription PCR and suggest that the current virus indexing methods of pre-basic mother plants work as expected. Furthermore, many new viruses were identified in the collections of plant genetic resources not previously tested for viruses. In the future, siRNA-based diagnostics could be a useful supplement for the currently used virus detection methods in certified plant production and thus rationalize and simplify the current testing system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Finland
  • Fragaria / virology
  • Methods
  • Plant Viruses / genetics
  • Plant Viruses / isolation & purification*
  • Polymerase Chain Reaction
  • RNA, Small Interfering*
  • Ribes / virology
  • Rubus / virology*

Substances

  • RNA, Small Interfering

Grants and funding

This study was supported by Ministry of Agriculture and Forestry, Finland (grants 1872/312/2012 and 2221/03.01.02/2015 to JPTV); https://mmm.fi/en/about-the-site. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.