title: DNA Barcoding Green Microalgae Isolated from Neotropical Inland Waters.
abstract: This study evaluated the feasibility of using the Ribulose Bisphosphate Carboxylase Large subunit gene (rbcL) and the Internal Transcribed Spacers 1 and 2 of the nuclear rDNA (nuITS1 and nuITS2) markers for identifying a very diverse, albeit poorly known group, of green microalgae from neotropical inland waters. Fifty-one freshwater green microalgae strains isolated from Brazil, the largest biodiversity reservoir in the neotropics, were submitted to DNA barcoding. Currently available universal primers for ITS1-5.8S-ITS2 region amplification were sufficient to successfully amplify and sequence 47 (92%) of the samples. On the other hand, new sets of primers had to be designed for rbcL, which allowed 96% of the samples to be sequenced. Thirty-five percent of the strains could be unambiguously identified to the species level based either on nuITS1 or nuITS2 sequences' using barcode gap calculations. nuITS2 Compensatory Base Change (CBC) and ITS1-5.8S-ITS2 region phylogenetic analysis, together with morphological inspection, confirmed the identification accuracy. In contrast, only 6% of the strains could be assigned to the correct species based solely on rbcL sequences. In conclusion, the data presented here indicates that either nuITS1 or nuITS2 are useful markers for DNA barcoding of freshwater green microalgae, with advantage for nuITS2 due to the larger availability of analytical tools and reference barcodes deposited at databases for this marker.
authors.affiliation: Embrapa Agroenergy, Brasília, DF, Brazil.|Universidade Federal do Tocantins, Gurupi, TO, Brazil.|Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
authors.forename: Sámed I I A
authors.initials: SI
authors.lastname: Hadi
authors.affiliation: Embrapa Agroenergy, Brasília, DF, Brazil.|Universidade Federal da Bahia, Vitória da Conquista, BA, Brazil.
authors.forename: Hugo
authors.initials: H
authors.lastname: Santana
authors.affiliation: Embrapa Agroenergy, Brasília, DF, Brazil.
authors.forename: Patrícia P M
authors.initials: PP
authors.lastname: Brunale
authors.affiliation: Universidade Federal do Tocantins, Gurupi, TO, Brazil.
authors.forename: Taísa G
authors.initials: TG
authors.lastname: Gomes
authors.affiliation: Embrapa Pantanal, Corumbá, MS, Brazil.
authors.forename: Márcia D
authors.initials: MD
authors.lastname: Oliveira
authors.affiliation: Embrapa Swine and Poultry, Concórdia, SC, Brazil.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Matthiensen
authors.affiliation: Embrapa Amazônia Oriental, Belém, PA, Brazil.
authors.forename: Marcos E C
authors.initials: ME
authors.lastname: Oliveira
authors.affiliation: Embrapa Agroenergy, Brasília, DF, Brazil.
authors.forename: Flávia C P
authors.initials: FC
authors.lastname: Silva
authors.affiliation: Embrapa Agroenergy, Brasília, DF, Brazil.
authors.forename: Bruno S A F
authors.initials: BS
authors.lastname: Brasil
mesh_terms: D001938:Brazil; D000460:Chlorophyta / Q000145:classification* / Q000235:genetics*; D058893:DNA Barcoding, Taxonomic; D018744:DNA, Plant; D021903:DNA, Ribosomal Spacer; D005819:Genetic Markers; D058086:Microalgae / Q000145:classification* / Q000235:genetics*; D010802:Phylogeny; D017422:Sequence Analysis, DNA
chemical_list: D018744:DNA, Plant; D021903:DNA, Ribosomal Spacer; D005819:Genetic Markers