Cuticle function can be pivotal to plant success in different environments. Yet, the occurrence of intraspecific adjustments in cuticle traits resulting from acclimation or adaptation to different habitats remains poorly understood. Here, we used genetically well-characterised populations of Arabidopsis arenosa to investigate whether cuticle traits were adjusted as part of the parallel evolution from a foothill to an alpine ecotype. Six alpine and six foothill populations, representing at least three independent evolutionary origins of an alpine ecotype, were used in reciprocal transplantation experiments, to investigate cuticle traits at the eco-physiological, biochemical and structural levels. The genetic basis behind these traits was assessed by combining selection scans and differential gene expression analysis. Overall, alpine populations showed reduced cuticular transpiration in conjunction with consistently altered cuticular wax composition, with higher accumulation of two fatty alcohols and two iso-alkanes. Genomic analysis unravelled nine genes associated with cuticular wax metabolism showing allelic differentiation in alpine compared to lowland populations. In silico gene expression analysis revealed differences between ecotypes for several genes related to cuticle metabolism. Repeated ecotypic differentiation in cuticle traits together with the genetic architecture of the alpine ecotype points at an adaptive value of cuticle adjustments for the colonisation of alpine habitats.

Department of Botany and Biodiversity Research University of Vienna Rennweg 14 Vienna 1030 Austria

Department of Botany Charles University of Prague Benátská 2 Prague 128 01 Czech Republic

Department of Botany University of Innsbruck Sternwartestraße 15 Innsbruck 6020 Austria

Institute of Ecology and Evolution University of Bern Baltzerstraße 6 Bern 3012 Switzerland

Polish Academy of Sciences Institute of Nature Conservation al Adama Mickiewicza 33 Krakow 31 120 Poland

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