A Multilocus Integrative Framework to Reassess Species Boundaries Within the Cystoseira Sensu Stricto Complex...

Cystoseira sensu lato (s.l.) (Fucales, Phaeophyceae) form structurally complex marine forests along the warm-temperate coasts of the Mediterranean Sea and the eastern Atlantic Ocean. They are experiencing widespread decline, yet their conservation and restoration are hindered by taxonomic uncertainty. Within Cystoseira sensu stricto (s.s.), species boundaries among taxa traditionally referred to as C. compressa, C. foeniculacea, C. humilis (including the C. canariensis morphotype), and C. pustulata have long been debated due to the high plasticity and partial overlap of diagnostic morphological traits. Here we reassess species boundaries within the Cystoseira s.s. complex using an integrative approach combining morphology, ecology, and multilocus genetic data. We collected specimens from 20 sites spanning the Mediterranean-Atlantic distribution. Genomic DNA was extracted from 220 specimens and Sanger sequencing generated 243 new sequences (ITS2: n = 99; cox1: n = 88; rbcL-rbcS: n = 56). We inferred single-locus and concatenated phylogenies, reconstructed haplotype networks, and applied multiple single-locus species delimitation approaches to test the robustness of inferred boundaries. We consistently recovered two primary lineages corresponding to C. foeniculacea and a C. compressa complex, the latter including specimens historically identified as C. pustulata and the C. humilis with the C. canariensis morphotype. Within an integrative framework that prioritises cross-locus concordance and diagnosability, we recognise two species-level lineages in Cystoseira s.s. and treat the morpho-ecologically coherent entities within C. compressa at the variety rank. Accordingly, we propose the new status and combination Cystoseira compressa var. pustulata (Ercegović) D'Ambros Burchio & Falace, comb. et stat. nov.