Supporting Methods: A detailed description of the protocol used to construct a
Libellulidae phylogeny and subsequent tests for the detection of phylogenetic signal on various traits
Blomberg’s K (Blomberg & Garland 2002, Blomberg et al.
2003) analysis was performed in order to assess if phylogeny plays a role in determining range area, dispersal behavior, and niche width. This test for phylogenetic signal requires the construction of an ultrametric tree, which is outlined below.
Sequence data from several genes (Table S2) were downloaded from GenBank
(Benson et al.
2005) and managed within Geneious v6.1.6 (Drummond et al.
2013).
Geneious was then used to align and curate each gene alignment. The 28S sequence data was aligned via SINA (Pruesse et al.
2012) before being imported into Geneious and its alignment refined. All sequence alignments were concatenated into a single master alignment.
The master alignment and gene partition information was input into
PartitionFinder (Lanfear et al.
2012) in order to select appropriate models of evolution for each gene partition while simultaneously avoiding over-parameterization by joining partitions that are not significantly different. The initial gene partitions were as follows:
5.8S rDNA, 12S rDNA, 16S rDNA, 28S rDNA, tRNA-Val, cox1 (cytochrome oxidase subunit 1; split by codon position), & EF-1

(elongation factor 1

; split by codon position). A PHYML (Guindon et al.
2010) tree constructed from the non-partitioned master alignment using a GTR model was used as a starting tree for PartitionFinder.
PartitionFinder was used to construct appropriate partitions for MrBayes v3.2.2
(Ronquist et al.
2012). A phylogenetic tree was produced via MrBayes using the seven partitions selected by PartitionFinder: 28S + 5.8S + EF-1

(codon position 1); 12S + 16S
+ tRNA-Val; cox1 (codon position 1); cox1 (codon position 2); cox1 (codon position 3);
EF-1

(codon position 2); EF-1

(codon position 3). MrBayes was run for 10 million generations in which the data where sampled every 100 generations to ensure that the combined parameters from two simultaneous runs had Effective Sample Size (ESS) values above 200 within Tracer v1.5 (
). Alignment, partition information, run parameters, and resulting phylogeny are contained within
Supporting Information File 1 and Figure S1. Note that this phylogeny was constructed with limited available data for the taxa included in this study (Table S2). Further investigation into the phylogenetic structure of Libellulidae needed.
BEAST v1.7.5 (Drummond, et al.
2012) was used to construct an ultrametric tree.
Within BEAST, the substitution models and partitions were set as per PartitionFinder and unlinked. The resulting six partitions were: 28S + 5.8S + EF-1

(codon position 1); 12S
+ 16S + tRNA-Val; cox1 (codon position 1); cox1 (codon position 2); cox1 (codon position 3) + EF-1

(codon position 2); EF-1

(codon position 3). Exponential relaxed clock models were set for each partition and unlinked, and the tree model was linked and set to a “Birth-Death Incomplete Sampling” model). The phylogeny produced by
MrBayes in the previous step was used as a starting tree with a fixed topology constraint for BEAST analysis. See Supporting Information File 2 for the XML formatted BEAST input and run settings. In brief, BEAST sampled every 250 generations over 50 million generations, to ensure all parameters had ESS values above 200 within Tracer v1.5

(
). A final tree was summarized by BEASTs
TreeAnnotator program by burning away the first 10% of sampled trees.
The resulting ultrametric tree output from BEAST and trait data was input into R
(R Development Core Team, 2013). Blomberg’s K test for phylogenetic signal was carried out via the ‘phylosignal’ command within the ‘picante’ R package (Kembel et al.
2010). Range area, dispersal behavior, and niche breadth all lacked phylogenetic signal
(P = 0.34, 0.745, and 0.507 respectively). See Supporting Information File 3 for input data and R commands.
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