Freshwater fungi play an important role in the decomposition of organic matter of leaf-litter in rivers and streams. They also possess the necessary mechanisms to endure lower temperatures caused by habitat and weather variations. This includes the production of cold-active enzymes and antifreeze proteins. To better understand the physiological activities of freshwater fungi in their natural environment, different methods are being applied, and genome sequencing is one in the spotlight. In our study, we sequenced the first genome of the freshwater fungus Filosporella fistucella (45.7 Mbp) and compared the genome with the evolutionary close-related species Tricladium varicosporioides (48.2 Mpb). The genomes were annotated using the carbohydrate-active enzymes database where we then filtered for leaf-litter degradation-related enzymes (cellulase, hemicellulase, laccase, pectinase, cutinase, amylase, xylanase, and xyloglucanase). Those enzymes were analyzed for antifreeze properties using a machine-learning approach. We discovered that Filosporella fistucella has more enzymes to participate in the breakdown of sugar, leaf, and wood than Tricladium varicosporioides (855 and 719, respectively). F. fistucella shows a larger set of enzymes capable to resist cold temperatures than T. varicosporioides (75 and 66 respectively). Our findings indicate that in comparison with T. varicosporioides, F. fistucella has a greater capacity for aquatic growth, adaptability to freshwater environments, and resistance to low temperatures.