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Optimalizace izolace RNA v kontextu plastového znečištění u akvatických mikroorganismů
Summary
Researchers examined microplastic contamination in edible fish species sold at markets across a coastal city, detecting particles in muscle tissue and gastrointestinal contents of all species tested. The study estimated human dietary intake of microplastics via fish consumption and found that smaller, lower-trophic-level species carried higher particle burdens per unit mass.
Microplastic are globaly widespread and perceived as a significant environmental contaminant. Little is yet known about their impact on aquatic microorganisms. Cyanobacteria are an important part of the ecosystem with great biotechnological potential. Due to their morphological diversity, RNA isolation is problematic and must be optimised for each species. In this study, RNA isolation was optimized for Synechococcus elongatus PCC 7942, Nostoc sp. PCC 7120, Chlorella sorokiniana, and Photobacterium phosphoreum. Then, the effect of PET -microplastic particles at concentrations of 0 mg/l, 150 mg/l, 300 mg/l was studied on their growth and vitality. It was found out that double homogenization in Synechococcus elongatus PCC 7942, Nostoc sp. PCC 7120, and Chlorella sorokiniana has a positive effect on RNA yield. Washing of biomass has an effect on RNA purity, as measured by A260/230 ratios. The organism Photobacterium phosphoreum always achieved the highest results. PET- microplastic particles decelerated growth in all used organisms except the genus Synechococcus, where was observed no significant difference in growth in medium with and without microplastics. RNA was isolated from the microplastic experiment from all organisms using the optimized method. For future analyses, the isolated RNA could be sent for sequencing and the effect of microplastics could be studied at the transcriptome level.