Application of High-Resolution Melting and DNA Barcoding for Discrimination and Taxonomy Definition of Rocket Salad (Diplotaxis spp.) Species

Nuclear and cytoplasmic DNA barcoding regions are useful for plant identification, breeding, phylogenesis. In this study, the genetic diversity of 17 Diplotaxis species, was investigated with 5 barcode markers. The allelic variation based on sequences chloroplast markers including spacer between trnL trnF tRNA-Phe gene (trnL-F), rubisco (rbcl), maturase K (matk), as well internal transcribed (ITS) region nuclear ribosomal DNA. A highly polymorphic marker (HRM500) derived from a comparison genome in Brassicaceae, also included. Subsequently, real-time PCR method coupled HRM analysis implemented to better resolve taxonomic relationships identify assays suitable species identification. Integration five revealed grouping according common chromosomal set number. Clusters n = 11 (D. duveryrieriana or cretacea, D. tenuifolia, simplex acris), 8 ibicensis, brevisiliqua ilorcitana), 9 brachycarpa, virgata, assurgens, berthautii) chromosomes were identified. Both phylogenetic structure collection identified siifolia most distant species. Previous studies emphasized species’ extremely high glucosinolate content, particularly glucobrassicin. High-resolution melting showed specific curve patterns discrimination thus determining ITS1 best fingerprinting. Findings demonstrate that approach used study is effective taxa investigations studies.