Skip to main content
Log in

New smut-specific primers for the ITS barcoding of Ustilaginomycotina

  • Original Article
  • Published:
Mycological Progress Aims and scope Submit manuscript

Abstract

The smut fungi (Ustilaginomycotina) are a highly diverse group, containing about 115 genera and 1700 species, most of which are biotrophic plant pathogens. As for other fungal groups, the ITS rDNA region is widely used to determine smut fungi at species level due to its high discriminatory power and for phylogenetic reconstructions within genera. So far, two primer sets, ITS1/ITS4 and M-ITS1/ITS4, were generally used to amplify smut fungi, but these often co-amplify host plants or contaminant fungi and do not yield satisfactory amplification for a variety of smut fungi. In the present study, based on a selection of genera that include more than 90% of the species of smut fungi (more than half of the genera of smut fungi), three new primers, smITS-F, smITS-R1 and smITS-R2, situated in the SSU or LSU region, were designed to avoid the amplification of host plants and to extend the coverage of PCR amplification for as many smut genera as possible.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1

Similar content being viewed by others

References

  • Albul S, Toome M, Aime MC (2015) Violaceomyces palustris gen. et sp. nov. and a new monotypic lineage, Violaceomycetales ord. nov. in Ustilaginomycetes. Mycologia 107:1193–1204

    Article  Google Scholar 

  • Bauer R, Oberwinkler F, Vánky K (1997) Ultrastructural markers and systematics in smut fungi and allied taxa. Can J Bot 75:1273–1314

    Article  Google Scholar 

  • Bauer R, Begerow D, Oberwinkler F, Piepenbring M, Berbee ML (2001) Ustilaginomycetes. In: McLaughlin DJ, Spatafora JW (eds) The Mycota, vol. VII, systematics and evolution part B, 2nd edn. Springer, Berlin, pp 57–83

    Google Scholar 

  • Bauer R, Garnica S, Oberwinkler F, Riess K, Weiß M, Begerow D (2015) Entorrhizomycota: a new fungal phylum reveals new perspectives on the evolution of fungi. PLoS ONE 10:e0128183

    Article  PubMed  PubMed Central  Google Scholar 

  • Begerow D, Bauer R, Oberwinkler F (1997) Phylogenetic studies on the nuclear large subunit ribosomal DNA of smut fungi and related taxa. Can J Bot 75:2045–2056

    Article  CAS  Google Scholar 

  • Begerow D, Bauer R, Boekhout T (2000) Phylogenetic placements of ustilaginomycetous anamorphs as deduced from nuclear LSU rDNA sequences. Mycol Res 104:53–60

    Article  CAS  Google Scholar 

  • Begerow D, Stoll M, Bauer R (2006) A phylogenetic hypothesis of Ustilaginomycotina based on multiple gene analyses and morphological data. Mycologia 98:906–916

    Article  PubMed  Google Scholar 

  • Begerow D, Schäfer AM, Kellner R, Yurkov A, Kemler M, Oberwinkler F, Bauer R (2014) Ustilaginomycotina. In: McLaughlin DJ, Spatafora JW (eds) The Mycota, Vol. VII, systematics and evolution part a, 2nd edn. Springer, Berlin, pp 295–329

    Google Scholar 

  • Boekhout T, Theelen B, Houbraken J, Robert V, Scorzetti G, Gafni A, Gerson U, Sztejnberg A (2003) Novel anamorphic mite-associated fungi belonging to the Ustilaginomycetes: Meira geulakonigii gen. nov., sp. nov., Meira argovae sp. nov. and Acaromyces ingoldii gen. nov., sp. nov. Int J Syst Evol Microbiol 53:1655–1664

    Article  CAS  PubMed  Google Scholar 

  • Castlebury LA, Carris LM, Vánky K (2005) Phylogenetic analysis of Tilletia and allied genera in order Tilletiales (Ustilaginomycetes; Exobasidiomycetidae) based on large submit nuclear RDNA sequences. Mycologia 97:888–900

    Article  CAS  PubMed  Google Scholar 

  • Cunnington JH, Vánky K, Shivas RG (2005) Lundquistia is a synonym of Sporisorium (Ustilaginomycetes). Mycol Balc 2:95–99

    Google Scholar 

  • Gardes M, Bruns TD (1993) ITS primers with enhanced specificity for basidiomycetes--application to the identification of mycorrhizae and rusts. Mol Ecol 2:113–118

    Article  CAS  PubMed  Google Scholar 

  • Kämper J, Kahmann R, Bölker M, Ma LJ, Brefort T, Saville BJ, Banuett F, Kronstad JW, Gold SE, Müller O, Perlin MH, Wösten HA, de Vries R, Ruiz-Herrera J, Reynaga-Peña CG, Snetselaar K, McCann M, Pérez-Martín J, Feldbrügge M, Basse CW, Steinberg G, Ibeas JI, Holloman W, Guzman P, Farman M, Stajich JE, Sentandreu R, González-Prieto JM, Kennell JC, Molina L, Schirawski J, Mendoza-Mendoza A, Greilinger D, Münch K, Rössel N, Scherer M, Vranes M, Ladendorf O, Vincon V, Fuchs U, Sandrock B, Meng S, Ho EC, Cahill MJ, Boyce KJ, Klose J, Klosterman SJ, Deelstra HJ, Ortiz-Castellanos L, Li W, Sanchez-Alonso P, Schreier PH, Häuser-Hahn I, Vaupel M, Baggins B, Koopmann E, Friedrich G, Voss H, Schlüter T, Margolis J, Platt D, Swimmer C, Gnirke A, Chen F, Vysotskaia V, Mannhaupt G, Güldener U, Münsterkötter M, Haase D, Oesterheld M, Mewes HW, Mauceli EW, DeCaprio D, Wade CM, Butler J, Young S, Jaffe DB, Calvo S, Nusbaum C, Galagan J, Birren BW (2006) Insights from the genome of the biotrophic fungal plant pathogen Ustilago maydis. Nature 444:97–101

    Article  PubMed  Google Scholar 

  • Katoh K, Standley DM (2013) MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Biol Evol 30:772–780

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mahmoud AGY, Zaher EHF (2015) Why nuclear ribosomal internal transcribed spacer (ITS) has been selected as the DNA barcode for fungi? Adv Genet Eng 4:119

    Google Scholar 

  • McTaggart AR, Shivas RG, Geering AD, Callaghan B, Vánky K, Scharaschkin T (2012) Soral synapomorphies are significant for the systematics of the Ustilago-Sporisorium-Macalpinomyces complex (Ustilaginaceae). Persoonia 29:63–77

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nasr S, Soudi MR, Shahzadeh Fazeli SA, Nguyen HDT, Lutz M, Piątek M (2014) Expanding evolutionary diversity in the Ustilaginomycotina: Fereydouniaceae fam. nov. and Fereydounia gen. nov., the first urocystidalean yeast lineage. Mycol Prog 13:1012

    Article  Google Scholar 

  • O’Donnell K (1993) Fusarium and its near relatives. In: Reynolds DR, Taylor JW (eds) The fungal holomorph: mitotic, meiotic and pleomorphic speciation in fungal systematics. CAB International, Wallingford, pp 225–233

    Google Scholar 

  • Piątek M, Riess K, Karasiński D, Yorou NS, Lutz M (2016) Integrative analysis of the West African Ceraceosorus africanus sp. nov. provides insights into the diversity, biogeography, and evolution of the enigmatic Ceraceosorales (Fungi: Ustilaginomycotina). Org Divers Evol. doi:10.1007/s13127-016-0285-3, in press

    Google Scholar 

  • Redchenko O, Vondrakand J, Kosnar J (2012) The oldest sequenced fungal herbarium sample. Short communications. Lichenologist 44:715–718

    Article  Google Scholar 

  • Savchenko KG, Carris LM, Castlebury LA, Heluta VP, Wasser SP, Nevo E (2014) Stripe smuts of grasses: one lineage or high levels of polyphyly? Persoonia 33:169–181

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schoch CL, Seifert KA, Huhndorf S, Robert V, Spouge JL, Levesque CA, Chen W, Fungal Barcoding Consortium (2012) Nuclear ribosomal internal transcribed spacer (ITS)region as a universal DNA barcode marker for fungi. Proc Natl Acad Sci U S A 109:6241–6246

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sharma R, Gassel S, Steiger S, Xia X, Bauer R, Sandmann G, Thines M (2015) The genome of the basal agaricomycete Xanthophyllomyces dendrorhous provides insights into the organization of its acetyl-CoA derived pathways and the evolution of Agaricomycotina. BMC Genomics 16:233

    Article  PubMed  PubMed Central  Google Scholar 

  • Stielow JB, Lévesque CA, Seifert KA, Meyer W, Irinyi L, Smits D, Renfurm R, Verkley GJM, Groenewald M, Chaduli D, Lomascolo A, Welti S, Lesage-Meessen L, Favel A, Al-Hatmi AMS, Damm U, Yilmaz N, Houbraken J, Lombard L, Quaedvlieg W, Binder M, Vaas LAI, Vu D, Yurkov A, Begerow D, Roehl O, Guerreiro M, Fonseca A, Samerpitak K, van Diepeningen AD, Dolatabadi S, Moreno LF, Casaregola S, Mallet S, Jacques N, Roscini L, Egidi E, Bizet C, Garcia-Hermoso D, Martín MP, Deng S, Groenewald JZ, Boekhout T, de Beer ZW, Barnes I, Duong TA, Wingfield MJ, de Hoog GS, Crous PW, Lewis CT, Hambleton S, Moussa TAA, Al-Zahrani HS, Almaghrabi OA, Louis-Seize G, Assabgui R, McCormick W, Omer G, Dukik K, Cardinali G, Eberhardt U, de Vries M, Robert V (2015) One fungus, which genes? Development and assessment of universal primers for potential secondary fungal DNA barcodes. Persoonia 35:242–263

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Stoll M, Piepenbring M, Begerow D, Oberwinkler F (2003) Molecular phylogeny of Ustilago and Sporisorium species (Basidiomycota, Ustilaginales) based on internal transcribed spacer (ITS) sequences. Can J Bot 81:976–984

    Article  CAS  Google Scholar 

  • Stoll M, Begerow D, Oberwinkler F (2005) Molecular phylogeny of Ustilago, Sporisorium, and related taxa based on combined analyses of rDNA sequences. Mycol Res 109:342–356

    Article  CAS  PubMed  Google Scholar 

  • Telle S, Thines M (2008) Amplification of cox2 (620 bp) from 2 mg of Up to 129 years Old herbarium specimens, comparing 19 extraction methods and 15 Polymerases. PLoS ONE 3:e3584

    Article  PubMed  PubMed Central  Google Scholar 

  • Vánky K (2012) Smut fungi of the world. APS, St. Paul

    Google Scholar 

  • Vilgalys R, Hester M (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. J Bacteriol 172:4238–4246

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang Q-M, Theelen B, Groenewald M, Bai F-Y, Boekhout T (2014) Moniliellomycetes and Malasseziomycetes, two new classes in Ustilaginomycotina. Persoonia 33:41–47

    Article  PubMed  PubMed Central  Google Scholar 

  • Wang Q-M, Begerow D, Groenewald M, Liu X-Z, Theelen B, Bai F-Y, Boekhout T (2015) Multigene phylogeny and taxonomic revision of yeasts and related fungi in the Ustilaginomycotina. Stud Mycol 81:55–83

    Article  PubMed  PubMed Central  Google Scholar 

  • White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA sequences for phylogenetics. In: Innis MA et al (eds) PCR protocols: a guide to methods and applications. Academic, San Diego, pp 315–322

    Google Scholar 

Download references

Acknowledgements

This study was funded by the LOEWE research funding programme in the framework of the Cluster for Integrative Fungal Research (IPF). We are grateful to the curators of PDD, BPI, VPRI, HBG, WSP, M, GLM and TUB (mostly HUV samples) for allowing the investigation of specimens in their keeping. We thank the private collectors, Udo and Heidrun Richter and Volker Kummer, for allowing us to include some of their collections in this study.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Marco Thines.

Additional information

Section Editor: Dominik Begerow

Electronic supplementary material

Below is the link to the electronic supplementary material.

Table S1

(DOCX 45 kb)

Table S2

(DOCX 46 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kruse, J., Choi, YJ. & Thines, M. New smut-specific primers for the ITS barcoding of Ustilaginomycotina. Mycol Progress 16, 213–221 (2017). https://doi.org/10.1007/s11557-016-1265-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11557-016-1265-x

Keywords

Navigation