Systematic Evaluation of Isoëtes asiatica Makino (Isoëtaceae) based on AFLP, nrITS, and Chloroplast DNA Sequences
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Abstract
On the basis of amplified fragment length polymorphism (AFLP) and nucleotide sequence data from nuclear ribosomal internal transcribed spacer (nrITS) and three chloroplast DNA regions (rbcL, cpITS, and trnS-psbC spacer), we investigated the species delimitation and the evolutionary lineage of Isoëtes asiatica from Hokkaido, Japan. The neighbor-joining (NJ) dendrogram based on AFLP markers revealed the well-defined clusters (bootstrap value = 100%) of I. asiatica. Results from the principal component analysis are largely congruent with those obtained in the NJ dendrogram. The maximum parsimony analysis, based on data from nrITS and three chloroplast DNA sequences, supported a monophyly of three species, I. asiatica, Isoëtes echinospora, and Isoëtes maritima from Hokkaido, Kamchatka, and Alaska regions. The distinct species status of I. asiatica was also well supported in the combined chloroplast DNA phylogeny. Therefore, I. asiatica appear to represent example of gradual speciation due to spatial isolation of ancestral populations followed by genetic divergence. Our results also suggest that I. asiatica is probably not the ancestral diploid of the polyploids occurring in East Asia.
Keywords
AFLP Chloroplast DNA Isoëtes I. asiatica Molecular phylogeny nrITSNotes
Acknowledgments
The authors thank Dr. Edith Kapinos (KEW Botanical Garden, UK) for I. asiatica DNA sample from Kamchatka and Y. Ito (University of Tokyo) for collecting the plant materials of I. asiatica in Hokkaido, Japan. We also thank two anonymous reviewers for comments on the manuscript. This work was supported by grants from the Ministry of Environment as parts of the Eco-Technopia 21 project (KIEST 052-071-049) and in part by the Korea Research Foundation Grant (KRF-2009-0073766) to C. K.
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