A phylogenetic survey based on ITS and trnT‐L sequences was conducted in the Campanula lusitanica complex, a group of annual species of Campanula sect. Rapunculus (Campanulaceae) distributed in the western Mediterranean region. Both nuclear and plastid data unambiguously demonstrated the existence of two non‐related lineages within this complex (C. lusitanica and C. dieckii). The C. lusitanica lineage is monophyletic and comprises five species (C. broussonetiana, C. cabezudoi, C. lusitanica s.str., C. specularioides, C. transtagana) that have all been referred to C. lusitanica s.l. by various authors; the C. dieckii lineage includes only one extant species, C. dieckii. Furthermore, our data strongly indicate that the affinities of C. dieckii are outside of the Campanula sect. Rapunculus core group. Divergence times indicate that the C. lusitanica lineage most likely originated in the Betic‐Rifian area in the late Miocene (Tortonian, approx. 9.4 My). In the C. lusitanica lineage a vicariant speciation pattern is observed in the recently evolved C. transtagana/C. broussonetiana clade (ca. 5.1 My) probably as a result of continental isolation caused by the opening of the Gibraltar Strait (ca. 5.3 My). Our evolutionary study strongly supports the taxonomic recognition of these otherwise neglected microspecies.

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