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The role of ecology as a modulator of sexual conflict in the wild: an experimental approach with Drosophila melanogaster

Periodic Reporting for period 1 - SexWarEcol (The role of ecology as a modulator of sexual conflict in the wild: an experimental approach with Drosophila melanogaster)

Reporting period: 2022-09-01 to 2024-08-31

The overall aim of the project was to improve the general understanding of how sexual conflict unfolds in nature and how ecological factors such as temperature may affect evolutionary processes (i.e. sexual selection/conflict) that impact a much wider range of phenotypic traits than those directly shaped by temperature. Answering the above questions could also contribute to ascertain how directional temperature changes (e.g. global warming) impact population viability. We identified genes linked to variation in sexual conflict phenotypes in an experimental evolution study. Furthermore, we identified major changes in SFPs production and shed light on the underlying gene expression patterns responsible for the divergent reaction norms described under different temperatures regimes.
First, We extracted DNA from pools of flies from each of the four replicates of the 3 experimental evolution lines (EELs) and did Illumina genome sequencing. Moreover, we also sequenced wild flies from 2023 and the Requena lab stock to have a “time zero” control. This enabled us to shed light on the underlying genomic basis of thermally selected sexual traits, with a special emphasis on sexual conflict. My work identified potential causal genes underlying divergent sexual conflict traits such as behaviour (learning, memory and female mating behaviour), metabolism and reproduction (oogenesis and spermatogenesis). Surprisingly we did not find any SFP genes under selection during experimental evolution. Second, we designed an experiment to investigate the effect of temperature on the head, accessory gland and testis gene expression. Thus, our goal is two-fold: examine the role of phenotypic plasticity in sexual conflict responses to temperature (including SFP production) and, simultaneously, understand the physiological and genetic pathways underlying the observed variation in sexual conflict mechanisms. To this aim, we used 15 isolines derived from our wild population which show clear thermal GxE for male reproductive success (as we’ve found to be typical of this population); whereby some genotypes show a higher male reproductive success at high and low temperatures (28ºC and 20ºC, respectively), having low reproductive success at intermediate temperatures (24ºC), whereas others show maximal reproductive success at intermediate (24ºC) vs lower at high and low temperatures (28ºC and 20ºC, respectively). In this experiment we raised all the isolines in a common garden at 24ªC and exposed adult males of the 15 isolines to females for 2 hours under low, mean and high temperatures (20ºC, 24ªC and 28ºC, respectively). Finally, we extracted RNA in pools of 12 testis, accessory glands and heads; and did RNA seq for each tissue. Moreover, we sequenced the genomes of 29 isolines, including the ones used in this experiment.

I presented 3 posters to disseminate my results in the 54th Course of the International School of Ethology “Danilo Mainardi” on Sexual selection in a changing world (Erice, Italy) and the IX Congress of the Spanish Society of Evolutionary Biology (SESBE, Málaga, Spain). The final publications all will be available as open access, while files (e.g. databases and R scripts) will be stored and made accessible through public repositories such as the Dryad Digital Repository and GenBank. Afterwards we will use both my personal and Carazo´s lab social media accounts to post a comprehensive thread detailing the major achievements of our research. In addition, the University of Valencia will do a press-release to disseminate our main findings. Eventually, the publications resulting from the project SexWarEcol will include a reference to EU funding.
The direct results from our SexWarEcol project will be a total of 2 scientific publications. One that identifies genes linked to variation in sexual conflict phenotypes (behaviour, metabolism and reproduction) and temperature variation. Our second publication, will show the major changes in SFPs production and shed light on the underlying gene expression patterns responsible for the divergent reaction norms described under different temperatures regimes. Moreover, and thanks to other ongoing experiments that build upon the results presented here sequencing data generated (either RNA, DNA or protein sequences), we expect more than 3 upcoming publications that will be partially derived from our results. Overall, our results will be highly valuable to the scientific community, from evolutionary ecologists studying adaptation in short temporal scales and conservation biologists focused on the impact of global change to evolutionary biologists interested in major evolutionary trends. In addition, the society will benefit from a greater understanding of evolutionary processes that produce either short-term or long-term adaptation and its implication in challenges such as global change.
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