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We exposed outbred SP8 strain to the 30 generations of experimental evolution in one of five regime: Control 20°C, in which the temperature was constant 20°C throughout 30 generations, Warm 25°C in which the temperature was 25°C for 30 generations, Increased warming in which the temperature gradually raised from 20°C to 25°C so that every 2.13 days temperature increased by 0.1°C, Slow temperature cycles where the temperature fluctuated every 10 generations between 20°C and 25°C, and Fast temperature cycles were the temperature fluctuated between 20°C and 25°C every two generations. Each regime was replicated six times. The experimental evolution procedure has been described in detail in Lind, et al. (2020). Phenotypic assays Worms were maintained for 2 generations in a common garden in 20°C. The generations were then tested for development time, size at maturity and growth rate in both 20°C and 25°C. Ten female worms (day 2 of adulthood) per population were left to lay eggs for 1 h of synchronized egg-laying and development time of the offspring females was recorded. Once 61 h (in 20°C) or 41 h (in 25°C) has passed, the plates were checked hourly and all mature females were collected and photographed using Lumenera Infinity2-5C digital microscope camera. The photographs were used to estimate size at maturity and a development time which was scored as number of hours between egg laying and the sexual maturity. Growth rate was calculated as log(size at maturity)/development time (mm2 h-1). To avoid using negative numbers, size at maturity was multiplied by a constant of 100. References: Lind, M. I., Zwoinska, M. K., Andersson, J., Carlsson, H., Krieg, T., Larva, T., & Maklakov, A. A. (2020). Environmental variation mediates the evolution of anticipatory parental effects. Evolution Letters, 4(4), 371–381. https://doi.org/doi:10.1002/evl3.177 |
