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      Fifteen microsatellite markers for Herbertia zebrina (Iridaceae): An endangered species from South American grasslands 1

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          Abstract

          Premise of the study:

          Polymorphic microsatellite loci were developed and used to genotype individuals of Herbertia zebrina (Iridaceae) as a first step for assessment of intraspecific genetic diversity.

          Methods and Results:

          Primer pairs for 47 markers were developed: 20 from a microsatellite-enriched library and 27 from a next-generation sequencing run using the Illumina MiSeq platform. Of those, 15 loci were considered successful, of which 12 were polymorphic and three were monomorphic. The primers were tested in 50 individuals from three populations of H. zebrina. Two to 14 alleles per locus were identified, and observed and expected heterozygosity were 0.00–0.95 and 0.18–0.89, respectively. Tests of cross-amplification to evaluate the applicability of these markers showed positive results in one congeneric species, H. darwinii, and in a phylogenetically closely related species, Calydorea crocoides.

          Conclusions:

          These microsatellite markers can be used for studies of genetic variation and genetic population structure, as well as to support conservation efforts.

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          Most cited references9

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          Primer3Plus, an enhanced web interface to Primer3

          Here we present Primer3Plus, a new web interface to the popular Primer3 primer design program as an enhanced alternative for the CGI- scripts that come with Primer3. Primer3 consists of a command line program and a web interface. The web interface is one large form showing all of the possible options. This makes the interface powerful, but at the same time confusing for occasional users. Primer3Plus provides an intuitive user interface using present-day web technologies and has been developed in close collaboration with molecular biologists and technicians regularly designing primers. It focuses on the task at hand, and hides detailed settings from the user until these are needed. We also added functionality to automate specific tasks like designing primers for cloning or step-wise sequencing. Settings and designed primer sequences can be stored locally for later use. Primer3Plus supports a range of common sequence formats, such as FASTA. Finally, primers selected by Primer3Plus can be sent to an order form, allowing tight integration into laboratory ordering systems. Moreover, the open architecture of Primer3Plus allows easy expansion or integration of external software packages. The Primer3Plus Perl source code is available under GPL license from SourceForge. Primer3Plus is available at http://www.bioinformatics.nl/primer3plus.
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            Phytochemical Ecology, Phytochemical Society Symposia Series no. 8

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              Model-based conservation planning of the genetic diversity of Phellodendron amurense Rupr due to climate change

              Climate change affects both habitat suitability and the genetic diversity of wild plants. Therefore, predicting and establishing the most effective and coherent conservation areas is essential for the conservation of genetic diversity in response to climate change. This is because genetic variance is a product not only of habitat suitability in conservation areas but also of efficient protection and management. Phellodendron amurense Rupr. is a tree species (family Rutaceae) that is endangered due to excessive and illegal harvesting for use in Chinese medicine. Here, we test a general computational method for the prediction of priority conservation areas (PCAs) by measuring the genetic diversity of P. amurense across the entirety of northeast China using a single strand repeat analysis of twenty microsatellite markers. Using computational modeling, we evaluated the geographical distribution of the species, both now and in different future climate change scenarios. Different populations were analyzed according to genetic diversity, and PCAs were identified using a spatial conservation prioritization framework. These conservation areas were optimized to account for the geographical distribution of P. amurense both now and in the future, to effectively promote gene flow, and to have a long period of validity. In situ and ex situ conservation, strategies for vulnerable populations were proposed. Three populations with low genetic diversity are predicted to be negatively affected by climate change, making conservation of genetic diversity challenging due to decreasing habitat suitability. Habitat suitability was important for the assessment of genetic variability in existing nature reserves, which were found to be much smaller than the proposed PCAs. Finally, a simple set of conservation measures was established through modeling. This combined molecular and computational ecology approach provides a framework for planning the protection of species endangered by climate change.
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                Author and article information

                Journal
                Appl Plant Sci
                Appl Plant Sci
                apps
                Applications in Plant Sciences
                Botanical Society of America
                2168-0450
                May 2017
                12 May 2017
                : 5
                : 5
                : apps.1700035
                Affiliations
                [2 ]Department of Biosciences, Programa de Pós-Graduação em Botânica, Universidade Federal do Rio Grande do Sul, Brazil
                [3 ]Institute of Integrative Nature Conservation Research, University of Natural Resources and Life Science, Vienna, Austria
                [4 ]Department of Ecology and Ecosystem Management, Chair of Restoration Ecology, Technical University of Munich, Munich, Germany
                Author notes
                [1]

                The authors thank laboratory technician Holger Paetsch for assistance in the molecular laboratory. Financial support was received from Deutscher Akademischer Austauschdienst (DAAD) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) (to C.F.), the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; grant no. 304197/2012-2 to T.T.S.C.), the German Research Foundation (grant no. KO1741/3-1 to J.K.), and the Fundação Grupo Boticário de Proteção a Natureza (project 1018/20142).

                [5 ]Author for correspondence: crisforgiarini@ 123456gmail.com
                Article
                apps1700035
                10.3732/apps.1700035
                5435410
                28529837
                877e1ea6-fb98-4c60-b52b-cb4a35049b07
                © 2017 Forgiarini et al. Published by the Botanical Society of America

                This is an open access article distributed under the terms of the Creative Commons Attribution License ( CC-BY-NC-SA 4.0), which permits unrestricted noncommercial use and redistribution provided that the original author and source are credited and the new work is distributed under the same license as the original.

                History
                : 14 April 2017
                : 23 April 2017
                Categories
                Primer Note

                calydorea crocoides,herbertia darwinii,herbertia zebrina,illumina miseq,iridaceae,next-generation sequencing,simple sequence repeat (ssr) marker

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