Additional file 1: of Molecular genetic and biochemical evidence for adaptive evolution of leaf abaxial epicuticular wax crystals in the genus Lithocarpus (Fagaceae)

Figure S1. Detail of adaxial layer of leaf epidermis of the genus Lithocarpus using Scanning Electron Microscope (SEM). (A) L. amygdalifolius; (B) L. brevicaudatus; (C) L. cornea; (D) L. dodonaeifolius; (E) L. formosanus; (F) L. glaber; (G) L. hanceii; (H) L. harlandii; (I) L. kawakamii; (J) L. konishii; (K) L. lepidocarpus; (L) L. nantoensis; (M) L. shinsuiensis; (N) L. taitoensisx. The SEM shows that no epicuticular wax crystals covered the leaf adaxial leaf surface in Lithocarpus. The scale bar represents 200Â Îźm. Figure S2. The spatial distribution reconstructed according to the current sampling records from GBIF and predicted using a machine-learning algorithm, maximum entropy algorithms, implemented in Maxent [79]. Figure S3. Ancestral state inference of discrete characters using the Maximum likelihood framework assuming one-parameter equal rates (ER) of character transition model and summarizing the 300 simulated character reconstructions. Figure S4. Pairwise comparison of substitution rates (K) of LAEWC related genes and reference genes using simple linear regression (SLR) and dependent two-group Wilcoxon Signed Rank Test (WSRT). Table S1. Primer list and annealing temperatures used in this study. Table S2. Results of likelihood ratio test of hypotheses of positive selection on retaining the gain or loss of LAEWC trait. Table S3. Ecophysiological measurements and altitudinal distribution of Lithocarpus species tested in this study. (PDF 1094 kb)