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      Fungal Identification Using Molecular Tools: A Primer for the Natural Products Research Community

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          Abstract

          Fungi are morphologically, ecologically, metabolically, and phylogenetically diverse. They are known to produce numerous bioactive molecules, which makes them very useful for natural products researchers in their pursuit of discovering new chemical diversity with agricultural, industrial, and pharmaceutical applications. Despite their importance in natural products chemistry, identification of fungi remains a daunting task for chemists, especially those who do not work with a trained mycologist. The purpose of this review is to update natural products researchers about the tools available for molecular identification of fungi. In particular, we discuss (1) problems of using morphology alone in the identification of fungi to the species level; (2) the three nuclear ribosomal genes most commonly used in fungal identification and the potential advantages and limitations of the ITS region, which is the official DNA barcoding marker for species-level identification of fungi; (3) how to use NCBI-BLAST search for DNA barcoding, with a cautionary note regarding its limitations; (4) the numerous curated molecular databases containing fungal sequences; (5) the various protein-coding genes used to augment or supplant ITS in species-level identification of certain fungal groups; and (6) methods used in the construction of phylogenetic trees from DNA sequences to facilitate fungal species identification. We recommend that, whenever possible, both morphology and molecular data be used for fungal identification. Our goal is that this review will provide a set of standardized procedures for the molecular identification of fungi that can be utilized by the natural products research community.

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          Most cited references141

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          Creating the CIPRES Science Gateway for inference of large phylogenetic trees

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            Two divergent intragenomic rDNA ITS2 types within a monophyletic lineage of the fungus Fusarium are nonorthologous.

            The evolutionary history of the phytopathogenic Gibberella fujikuroi complex of Fusarium and related species was investigated by cladistic analysis of DNA sequences obtained from multiple unlinked loci. Gene phylogenies inferred from the mitochondrial small subunit (mtSSU) rDNA, nuclear 28S rDNA, and beta-tubulin gene were generally concordant, providing strong support for a fully resolved phylogeny of all biological and most morphological species. Discordance of the nuclear rDNA internal transcribed spacer 2 (ITS2) gene tree is due to paralogous or xenologous ITS2 sequences. PCR and sequence analysis demonstrated that every strain of the ingroup species tested possesses two highly divergent nonorthologous ITS2 types designated type I and type II. Only the major ITS2 type, however, is discernable when PCR products are amplified and sequenced directly with conserved primers. The minor ITS2 type was recovered using ITS2 type-specific PCR primers. Distribution of the major ITS2 type within the species lineages exhibits a homoplastic pattern of evolution, thus obscuring true phylogenetic relationships. The results suggest that the ancestral ITS2 types may have arisen following an ancient interspecific hybridization or gene duplication which occurred prior to the evolutionary radiation of the Gibberella fujikuroi complex and related species of Fusarium. The results also indicate that current morphological-based taxonomic schemes for these fungi are unnatural and a new classification is required.
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              Phylogenetic species recognition and species concepts in fungi.

              The operational species concept, i.e., the one used to recognize species, is contrasted to the theoretical species concept. A phylogenetic approach to recognize fungal species based on concordance of multiple gene genealogies is compared to those based on morphology and reproductive behavior. Examples where Phylogenetic Species Recognition has been applied to fungi are reviewed and concerns regarding Phylogenetic Species Recognition are discussed.
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                Author and article information

                Journal
                J Nat Prod
                J. Nat. Prod
                np
                jnprdf
                Journal of Natural Products
                American Chemical Society and American Society of Pharmacognosy
                0163-3864
                1520-6025
                15 February 2017
                24 March 2017
                : 80
                : 3 , Special Issue in Honor of Phil Crews
                : 756-770
                Affiliations
                []Department of Chemistry and Biochemistry, University of North Carolina at Greensboro , Greensboro, North Carolina 27402, United States
                []Illinois Natural History Survey, University of Illinois , Champaign, Illinois 61820, United States
                [§ ]Mycosynthetix, Inc. , 505 Meadowland Drive, Suite 103, Hillsborough, North Carolina 27278, United States
                Author notes
                [* ]Phone: 336-334-5474. E-mail: Nicholas_Oberlies@ 123456uncg.edu .
                Article
                10.1021/acs.jnatprod.6b01085
                5368684
                28199101
                2769a64b-2056-4e34-9dbd-04d42bd3629c
                Copyright © 2017 American Chemical Society and American Society of Pharmacognosy

                This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes.

                History
                : 22 November 2016
                Categories
                Review
                Custom metadata
                np6b01085
                np-2016-01085v

                Organic & Biomolecular chemistry
                Organic & Biomolecular chemistry

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