title: Diversity of Clonostachys species assessed by molecular phylogenetics and MALDI-TOF mass spectrometry.
abstract: We assessed the species diversity among 45 strains of Clonostachys from different substrates and localities in Brazil using molecular phylogenetics, and compared the results with the phenotypic classification of strains obtained from matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Phylogenetic analyses were based on beta tubulin (Tub), ITS-LSU rDNA, and a combined Tub-ITS DNA dataset. MALDI-TOF MS analyses were performed using intact conidia and conidiophores of strains cultivated on oatmeal agar and 4% malt extract agar. Six known species were identified: Clonostachys byssicola, Clonostachys candelabrum, Clonostachys pseudochroleuca, Clonostachys rhizophaga, Clonostachys rogersoniana, and Clonostachys rosea. Two clades and two singleton lineages did not correspond to known species represented in the reference DNA dataset and were identified as Clonostachys sp. 1-4. Multivariate cluster analyses of MALDI-TOF MS data classified the strains into eight clusters and three singletons, corresponding to the ten identified species plus one additional cluster containing two strains of C. rogersoniana that split from the other co-specific strains. The consistent results of MALDI-TOF MS supported the identification of strains assigned to C. byssicola and C. pseudochroleuca, which did not form well supported clades in all phylogenetic analyses, but formed distinct clusters in the MALDI-TOF dendrograms.
authors.affiliation: Departamento de Fitopatologia, Universidade Federal de Lavras, PO Box 3037, 37200-000 Lavras, MG, Brazil. Electronic address: lmabreu@gmail.com.
authors.forename: Lucas M
authors.initials: LM
authors.lastname: Abreu
authors.affiliation: Departamento de Fitopatologia, Universidade Federal de Lavras, PO Box 3037, 37200-000 Lavras, MG, Brazil.
authors.forename: Gláucia M
authors.initials: GM
authors.lastname: Moreira
authors.affiliation: Departamento de Química, Universidade Federal de São Carlos, PO Box 676, 13.565-905 São Carlos, SP, Brazil.
authors.forename: Douglas
authors.initials: D
authors.lastname: Ferreira
authors.affiliation: Departamento de Química, Universidade Federal de São Carlos, PO Box 676, 13.565-905 São Carlos, SP, Brazil.
authors.forename: Edson
authors.initials: E
authors.lastname: Rodrigues-Filho
authors.affiliation: Departamento de Fitopatologia, Universidade Federal de Lavras, PO Box 3037, 37200-000 Lavras, MG, Brazil.
authors.forename: Ludwig H
authors.initials: LH
authors.lastname: Pfenning
mesh_terms: D044822:Biodiversity; D001938:Brazil; D004271:DNA, Fungal / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D006999:Hypocreales / Q000145:classification* / Q000235:genetics; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013172:Spores, Fungal / Q000254:growth & development
chemical_list: D004271:DNA, Fungal; D004275:DNA, Ribosomal
keywords: Bionectriaceae; Mass fingerprinting; Neotropical fungi; Phylogenetic species; Species diagnosis