Samples collection and DNA extraction
Locations were selected to cover a large range of climatic conditions across E. coccineum natural distribution; with three locations (Chillán, Nahuelbuta and Curacautín) representing the northern part of the distribution, four locations (Puerto Montt, Chiloé Norte, Chiloé Sur and Pumalín) the center part of the distribution, and three locations (Coyhaique, Chile Chico and Torres del Paine) the southern part of the distribution (Fig. 1a and Supplementary table 1). These three regions are subsequently named North, Center and South. Three to six individuals were sampled within each location and five mature leaves were collected per tree. A total of 42 trees were sampled. To calibrate divergence time among E. coccineum genetic groups, two individuals ofLomatia hirsuta Diels a Proteaceae species from Embothrieae tribe (Sauquet et al., 2009) were included as outgroup.
Whole genomic DNA extraction was carried out using the Qiagen DNeasy Plant kit (Qiagen Inc., USA) following manufacturer’s instructions. Foliar tissue (50 mg) was kept in Precellys24 homogenizer (Precellys, USA) with two 1/4” ceramic spheres (MP BIOMEDICALS, USA) and AP1 buffer until processing in the laboratory. The genomic DNA integrity was evaluated using direct visualization in 1% agarose gels, and the DNA was quantified using a Qubit fluorometer (Invitrogen, USA).