University of Helsinki, Department of Agricultural Sciences, Helsinki, Finland.
Natural Resources Institute Finland (Luke), Jokioinen, Finland.
PLoS One. 2019 Aug 7;14(8):e0220621. doi: 10.1371/journal.pone.0220621. eCollection 2019.
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.
无病原体的营养繁殖植物种系对于经认证的植物生产至关重要。它们需要定期监测植物种质中的病原体，特别是田间保存的种系。在这里，我们使用基于小干扰 RNA (siRNA) 的诊断方法 (VirusDetect) 测试了在芬兰研究站保存的草莓、悬钩子和醋栗属植物遗传资源的基础前母株和基础材料，以检测病毒的存在。该方法的优势在于，可以同时检测到不相关的病毒，而无需重新检测现有病毒。虽然在草莓和醋栗属植物的基础前母株中未检测到病毒，但在悬钩子属植物的基础前母株中检测到了覆盆子黄网病毒 (RYNV)。在覆盆子属植物遗传资源中还检测到了黑莓坏死病毒 (BRNV)、黑莓褪绿斑驳病毒 (RVCV) 和 RYNV。从七个 RVCV 分离株中鉴定出的 L 聚合酶编码序列表现出相当大的遗传变异。这些数据首次提供了 RYNV 在芬兰存在的分子生物学证据。在指示植物的病毒索引中未发现 RYNV，这表明它可能在某些覆盆子品种中内源性存在。此外，在悬钩子属植物中还检测到一种可能的新 RYNV 样香蕉束顶病毒。在田间生长的有症状的黑穗醋栗和醋栗叶脉带相关病毒 (GVBaV) 中检测到了黑加仑回复病毒 (BRV)。结果与使用 PCR 或反转录 PCR 获得的结果一致，表明目前用于基础前母株的病毒索引方法按预期运行。此外，在以前未测试过病毒的植物遗传资源收集物中还鉴定出了许多新病毒。在未来，siRNA 诊断可能成为经认证的植物生产中当前使用的病毒检测方法的有用补充，从而使当前的测试系统合理化和简化。
