Continuous Silicate Utilization Over Multiple 14L:10D Day:Night Cycles Confirms Night Metabolism in Lake Michigan Diatom Enrichments Using Either Nitrate or Ammonium as a Nitrogen Source

Michelle Soderling, Carmen Aguilar and Russell Lee Cuhel, University of Wisconsin Milwaukee, Milwaukee, WI, United States
Diatoms are single-celled organelle containing eukaryotes living in “glass houses”. As diatoms only take up silica when they replicate, measuring the amounts of dissolved and particulate silicate were an important aspect of this study. Silica was used as a proxy of the diatom reproduction. Depending on growth conditions, some algal species divide throughout the day and night; this suggests that protein synthesis can be an important component of algal night metabolism and hence nitrogen utilization. The goal of this experiment was to measure the amount of night protein synthesis occurring in a culture of diatoms from Lake Michigan. Diatoms were enriched with light for energy and excess nutrients—including phosphate, silicate, nitrate and limited ammonium for some—along with use of physical separation methods. Growing conditions were prepared in a way which anticipated the diatoms would synchronize to a 14:10 day/night cycle and store energy, during their day phase, to use for night protein synthesis and replication. Their growth was monitored by taking samples before and after the transitions of light to dark along with midday and midnight samples. Assays of dissolved and particulate silicate were used to measure utilization, which confirmed their nighttime growth. As hypothesized, the diatoms had significant growth during their night phase. There were decreases in the nighttime dissolved silicate and increases in the nighttime particulate silicate. When available, the diatoms preferred to use ammonium instead of nitrate. Cell division during the night phase indicated sufficient daytime energy storage to fuel night protein synthesis and cell replication. Uptake of nutrients occurred at night almost as if the "sun" did not set. There was continuous growth of this photosynthetic community.