A qPCR method for distinguishing biomass from non-axenic terrestrial cyanobacteria cultures in hetero- or mixotrophic cultivations

Abstract

AbstractThe cultivation of cyanobacteria with the addition of an organic carbon source (meaning as heterotrophic or mixotrophic cultivation) is a promising technique to increase their slow growth rate. However, most cyanobacteria cultures are infected by non-separable heterotrophic bacteria. While their contribution to the biomass is rather insignificant in a phototrophic cultivation, problems may arise in heterotrophic and mixotrophic mode. Heterotrophic bacteria can potentially utilize carbohydrates quickly, thus preventing any benefit for the cyanobacteria. In order to estimate the advantage of the supplementation of a carbon source, it is essential to quantify the proportion of cyanobacteria and heterotrophic bacteria in the resulting biomass. In this work, the use of quantitative polymerase chain reaction (qPCR) is proposed. To prepare the samples, a DNA extraction method for cyanobacteria was improved to provide reproducible and robust results for the group of terrestrial cyanobacteria. Two pairs of primers were used, which bind either to the 16S rRNA gene of all cyanobacteria or all bacteria including cyanobacteria. This allows a determination of the proportion of cyanobacteria in the biomass. The method was established with the two terrestrial cyanobacteria Trichocoleus sociatus SAG 26.92 and Nostoc muscorum SAG B-1453-12a. As proof of concept, a heterotrophic cultivation with T. sociatus with glucose was performed. After 2 days of cultivation, a reduction of the biomass partition of the cyanobacterium to 90% was detected. Afterwards, the proportion increased again.