Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas-Universidad Politécnica de Valencia, Spain.
Instituto Valenciano de Investigaciones Agrarias, Moncada, Valencia, Spain.
Sci Rep. 2017 Feb 17;7:42825. doi: 10.1038/srep42825.
Natural populations of peach latent mosaic viroid (PLMVd) are complex mixtures of variants. During routine testing, TaqMan rtRT-PCR and RNA gel-blot hybridization produced discordant results with some PLMVd isolates. Analysis of the corresponding populations showed that they were exclusively composed of variants (of class II) with a structural domain different from that of the reference and many other variants (of class I) targeted by the TaqMan rtRT-PCR probe. Bioassays in peach revealed that a representative PLMVd variant of class II replicated without symptoms, generated a progeny with low nucleotide diversity, and, intriguingly, outcompeted a representative symptomatic variant of class I when co-inoculated in equimolecular amounts. A number of informative positions associated with the higher fitness of variants of class II have been identified, and novel sets of primers and probes for universal or specific TaqMan rtRT-PCR detection of PLMVd variants have been designed and tested.
桃潜隐花叶病毒(PLMVd)的自然种群是变异体的复杂混合物。在常规检测中,TaqMan rtRT-PCR 和 RNA 凝胶印迹杂交技术与一些 PLMVd 分离株产生了不一致的结果。对相应种群的分析表明,它们仅由与参考序列和 TaqMan rtRT-PCR 探针靶向的许多其他变体(I 类)不同的结构域的变体(II 类)组成。在桃树上的生物测定表明,II 类代表 PLMVd 变体在没有症状的情况下复制,产生的后代核苷酸多样性较低,而且,当等量共接种时,它出人意料地与 I 类有症状的代表变体竞争。已经鉴定出与 II 类变体更高适应性相关的多个信息性位置,并设计和测试了用于通用或特定 TaqMan rtRT-PCR 检测 PLMVd 变体的新的引物和探针。