Of fourCharacter coding and reconstruction of ancestral statesIn order to infer the evolutionary history of traits connected to niche use and defense inside Tenthredinidae, we collected data on diet program breadths, host-plant HLCL-61 (hydrochloride) cost associations, and host attributes, at the same time as on larval ecology, behavior, morphology, and chemically-based defensive approaches inside the species incorporated inside the phylogenetic trees. We then coded the information as unordered binary or multistate characters, and reconstructed ancestral states by singlerate (Mk1) maximum likelihood (ML) optimization in Mesquite v. two.75 [63]. Grey shaded bars show 95 highest posterior density intervals for relative node ages for nodes with posterior probabilities more than 50 .deleted from the matrix and connected trees before every single reconstruction. Host plants have been compiled from literature sources [55,64] and every single sawfly was coded for its diet regime breadth (Figure 3). Inside the case of species for which dependable hostplant records were missing, eating plan 
breadth was coded as unknown, unless revealed by additional laboratory testing with larvae from the similar population.To evaluate the toxicity of plants on which sawflies feed, each and every plant genus (and species, as far as you can) was associated together with the occurrence of toxins, by referring to typical operates around the chemistry of plants [65-70] and to smaller andor much more current works (e.g., [45]). A plant taxon was deemed toxic if the leaves contain secondary metabolites from one particular or more of your following chemical classes: alkaloids (like steroid alkaloids and steroidBoevet al. BMC Evolutionary Biology 2013, 13:198 http:www.biomedcentral.com1471-214813Page six ofFigure 3 (See legend on subsequent page.)Boevet al. BMC Evolutionary Biology 2013, 13:198 http:www.biomedcentral.com1471-214813Page 7 of(See figure on previous page.) Figure 3 Relaxed molecular-clock phylogeny from the Tenthredinidae, as well as the distribution of many larval ecological and defensive traits inside the group. The BEAST MCC tree is according to analysis of Dataset two, which incorporates all sequenced tenthredinids too as representatives from three non-blasticotomid families in Tenthredinoidea. Posterior probabilities ( ) resulting from analyses in BEAST and MrBayes are given above and below branches, respectively (clades not present within the MrBayes tree are indicated by hyphens). Grey shaded bars show the 95 highest posterior density intervals for relative node ages for nodes with posterior probabilities exceeding 50 . Branch colors denote host plant classes with the sawfly species (see legend) and ancestral reconstructions depending on maximum-likelihood optimization across 1,000 post-burnin trees (see Additional file 4A). In the table to the right of the tree, eating plan breadth, plant toxicity, and defensive traits (from left to suitable) are coded as shown in Table 1. () Unknown; (x) not applicable.saponins, that are closely associated with regards to biosynthesis and metabolism; [71]), glucosinolates, PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/21337810 cyanogenic glycosides, and non-protein amino acids. All these compounds, except steroid saponins, have in prevalent the inclusion of one particular or more nitrogen atoms. Nitrogencontaining secondary metabolites show acute toxicity andor strong feeding deterrence towards vertebrates andor invertebrates, and they may be essentially the most popular defensive chemical substances of plants [65,68-70,72]. A plant taxon was regarded as non-toxic if it only contains secondary metabolites that usually do not contain nitrogen, like phenolics (e.g., coumarins, phenolic glycosides, and the w.
