The heterokont benthic multicellular algae Schizocladia ischiensis EC Henry, K. Okuda et H. Kawai (Schizocladiophyceae), Hecatonema terminale (Kützing) Kylin and Striaria attenuata (Greville) Greville (Phaeophyceae) are reported for the first time from the waters around the Maltese islands in the central Mediterranean. They were identified through algal isolation from incubated natural substrata, coupled with DNA barcoding targeting the biomarkers COI and rbcL plus the RuBisCO spacer. For three additional brown algae, Colpomenia sinuosa (Mertens ex Roth) Derbès et Solier, Asperococcus bullosus JV Lamouroux and Sphacelaria sp., DNA sequences confirmed previous morphology-based records from Malta. This paper also provides an updated literature-based species list of the marine macroalgae present in Malta.