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.

Sugarcane ethanol is produced at large scale generating wastes that could be used for microalgae biomass production in a biorefinery strategy. In this study, forty microalgae strains were screened for growth in sugarcane vinasse at different concentrations. Two microalgae strains, Micractinium sp. Embrapa|LBA32 and C. biconvexa Embrapa|LBA40, presented vigorous growth in a light-dependent manner even in undiluted vinasse under non-axenic conditions. Microalgae strains presented higher biomass productivity in vinasse-based media compared to standard Bold's Basal Medium in cultures performed using 15L airlift flat plate photobioreactors. Chemical composition analyses showed that proteins and carbohydrates comprise the major fractions of algal biomass. Glucose was the main monosaccharide detected, ranging from 46% to 76% of the total carbohydrates content according to the strain and culture media used. This research highlights the potential of using residues derived from ethanol plants to cultivate microalgae for the production of energy and bioproducts.