Temperature treatments across the experimental period for the entire temperature assays. from Temperature- versus precipitation-limitation shape local temperature tolerance in a Holarctic freshwater crustacean
2019-07-08T08:48:30Z (GMT) by
Supplementary figure 1. Temperature treatments across the experimental period for the entire temperature assays. At the beginning, animals were taken from the stock cultures (at 20 °C) and cultured in 380-mL jars filled with 350 mL Daphnia medium at 23 °C. Every two weeks, the temperature was raised (23, 25, 26, 27 and 28 °C), and population survival and the presence of juveniles (as an indicator of successful reproduction) were recorded. All replicate populations that were able to reproduce were then kept at 28 °C (high temperature stock). We then took animals from these 28 °C cultures and tested for reproduction and survival at nine higher temperatures (29 to 37 °C). For this, we collected three juvenile females (1 – 3 days old) from each of the 28 °C replicate populations and placed them in a 100-mL jar with 80 mL medium. These jars were kept in an incubator at the higher temperature (assay temperature). Animals were checked for survival and reproduction. If at least one animal of a replicate survived the 14-day assay period, the same replicate was tested at the next higher temperature, but starting with juveniles taken from the 28 °C cultures. If none of the animals survived 14 days at a certain temperature, this replicate was still tested at the next higher temperature. If all animals died again, we assumed that we had exceeded its upper temperature tolerance and discontinued with this replicate.