Department of Plant Pathology and Emerging Pathogens Institute, University of Florida, Gainesville, FL 32611 U.S.A.
Facultad de Ciencias de la Vida, Escuela Superior Politécnica del Litoral, Campus Gustavo Galindo, Km 30.5 Vía Perimetral. P.O. Box 09-01-5863, Guayaquil, Ecuador.
Plant Dis. 2019 Sep;103(9):2295-2304. doi: 10.1094/PDIS-12-18-2291-RE. Epub 2019 Jul 29.
species hybrids have been repeatedly reported as causing damaging diseases to cultivated and wild plants. Two known hybrids, and × , are pathogens of Solanaceae and ornamentals, respectively, although the extent of their host ranges are unknown. emerged from hybridization of and an unidentified related species, whereas × emerged from and . Considering that hybrids and parental species can coexist in the same regions and to distinguish them usually requires cloning or whole genome sequencing, we aimed to develop a rapid tool to distinguish them. Specifically, we used high-resolution melting (HRM) assays to differentiate genotypes based on their amplicon melting profiles. We designed primers for × and parental species based on available sequences of and nuclear genes containing polymorphisms between species. For , heterozygous sites from Illumina short reads were used for the same purpose. We identified multiple amplicons exhibiting differences in melting curves between parental species and hybrids. We propose HRM as a rapid method for differentiation of and × hybrids from parental species that could be employed to advance research on these pathogens.
种间杂交已被多次报道会导致栽培植物和野生植物的破坏性疾病。两个已知的杂种， 和 × ，分别是茄科和观赏植物的病原体，尽管它们的宿主范围未知。 是由 和一个未鉴定的相关物种杂交产生的，而 × 则是由 和 杂交产生的。考虑到杂种和亲代物种可能在同一地区共存，并且通常需要克隆或全基因组测序才能将它们区分开来，我们旨在开发一种快速区分它们的工具。具体来说，我们使用高分辨率熔解（HRM）分析来根据扩增子的熔解曲线区分基因型。我们根据 和 核基因中的多态性为 × 和亲代物种设计了引物，这些基因在物种之间存在。对于 ，则使用 Illumina 短读序列中的杂合位点来达到同样的目的。我们鉴定出了多个在亲本物种和杂种之间的熔解曲线存在差异的扩增子。我们提出 HRM 是一种快速区分 和 × 杂种与亲本物种的方法，可用于推进对这些病原体的研究。
