Delimitation of cryptic species of the Scytosiphon lomentaria complex (Scytosiphonaceae, Phaeophyceae) in Japan, based on mitochondrial and nuclear molecular markers

Scytosiphon lomentaria (Scytosiphonaceae, Ectocarpales) is believed to include some cryptic species, particularly in the Pacific. We attempted to delimit these species in Japan using mitochondrial cox1 and cox3 and nuclear ITS2 and the second intron of the centrin gene (cetn-int2). Fifty-three cox1+cox3 mitotypes, 26 ITS2 ribotypes and 45 cetn-int2 haplotypes were found in 107 samples collected from 33 localities in Japan. Based on phylogenetic analyses, similar sequence types were grouped into ten mitogroups, eight ribogroups and six cetn-int2 haplogroups (sequence-type groups). From the molecular trees and combinations of the mito-, ribo- and haplogroups, three cryptic species were apparent (Groups I-III). Group I, widely distributed on Pacific coasts, was highly supported by all molecular trees, whereas Groups II (North Pacific) and III (Northwestern Pacific and Australasia) were more closely related to each other. However, sequence-type-group combinations that would be characteristic of hybrids between Groups II and III were not detected, suggesting no gene flow between the two Groups. Further investigations of an additional 127 sympatrically growing plants supported the absence of gene flow between Groups II and III. Four samples did not belong to any of the Groups I-III and possibly represent additional species.