Systematics of neotropical Psiguria (Cucurbitaceae) : identifying low-copy nuclear markers, molecular phylogenetics, and taxonomic revision

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2009-08

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Abstract

Psiguria Arn. is a small genus of Neotropical vines in the Cucurbitaceae that grows in both wet and dry tropical forests from southern Mexico to Paraguay, and on Caribbean islands. The genus is estimated to be very young with natural history characteristics that have contributed to confusing species circumscriptions. The unique relationship of plants in the group with their butterfly pollinators makes Psiguria an interesting and important genus in tropical ecosystems. Both molecular and morphological approaches were used to investigate the monophyly of Psiguria, to elucidate the number of species in the genus, to discover sister relationships, and to identify characteristics for delineating species. Toward that end, an intensive screening of 141 primer combinations in search of phylogenetically informative low-copy nuclear markers was conducted along with a molecular phylogenetic analysis and a complete taxonomic revision of Psiguria. From the screening study, three potentially phylogenetically informative low-copy nuclear markers were discovered for Psiguria, 11 were found to be potentially useful in rosids, and 32 in other angiosperms. DNA sequences for eight chloroplast intergenic spacers (ndhF-rpL32, ndhC-trnV, rps16-trnQ, trnS-trnG, psbZ-trnM, psbM-trnD, rpoB-trnC, and psbE-petL), ITS, and the nuclear serine/threonine phosphatase intron were obtained for 70 samples of Psiguria plus 14 outgroups. Phylogenetic analyses support the monophyly of Psiguria and a sister relationship between P. umbrosa and P. warscewiczii. In the final chapter, two reviews on the genus are presented – one encapsulating the nomenclatural history, and one summarizing 35 years of ecological and natural history studies. In addition, morphological characters were databased, descriptions were written, and maps of geographic distribution were produced for all species. Considering both molecular and morphological data, six species of Psiguria are defined. To distinguish those species missing identifiable morphological characters, a set of DNA barcodes was developed. At least four chloroplast regions are required to differentiate species (ndhC-trnV, rps16- trnQ, rpoB-trnC, and ndhF-rpL32). Because of the absence of many morphological characters, two taxonomic keys are presented – one using male flowers, and the other using the set of DNA barcodes along with consistent leaf characteristics and geographic distribution.

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