@ARTICLE{TreeBASE2Ref27473, author = {Tao Fujiwara and Ayumi Uehara and Tsukasa Iwasina and Sadamu Matsumoto and Yi Shan Chao and Yi Hang Chang and Yasuyuki Watano}, title = {Allotetraploid cryptic species in Asplenium normale in the Japanese Archipelago, detected by chemotaxonomical and multi-locus genotype approaches}, year = {2017}, keywords = {Allopolyploidy; Aspleniaceae; chemotaxonomy; cryptic species; fern; multi-locus genotype; speciation}, doi = {}, url = {http://}, pmid = {}, journal = {American Journal of Botany}, volume = {}, number = {}, pages = {}, abstract = {Premise of the study Delimitation of cryptic species provides an understanding of biodiversity and opportunities to elucidate speciation processes. Extensive flavonoid variation has been reported in the tetraploid cytotype of the fern, Asplenium normale, although related species have no intraspecific variations in flavonoid composition. We hypothesized that Japanese A. normale still harbors multiple cryptic species with different flavonoid compositions, and tested this hypothesis using chemotaxonomical and multi-locus genotyping approaches. Methods We determined the multi-locus genotypes (MLGs) of 230 samples from 37 populations for one chloroplast DNA region and three nuclear genes. MLGs were used to delimit reproductively isolated lineages by population-genetic approaches. We also tested the correspondence between genetically recognized groups and flavonoid compositions. To identify the origins of putative cryptic species, we conducted phylogenetic analysis of the DNA markers used in genotyping. Key results The genetic clusters and flavonoid compositions showed clear correspondence. We recognized three putative cryptic species in tetraploid A. normale in Japan. Phylogenetic analyses revealed that cryptic species I and III originated from allopolyploidization between diploid A. normale and unknown diploid of A. boreale, and cryptic species II originated from allopolyploidization between diploid A. normale and A. oligophlebium. Conclusions Our study demonstrated that intraspecific variation of secondary metabolites can be a good indicator of cryptic species in ferns. The presence of the two cryptic species having the same progenitor diploid pair suggests that speciation between allopolyploid lineages of independent origin may be more common than previously considered. } }

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