Evolution and phylogenetic utility of CAD (rudimentary) among Mesozoic-aged Eremoneuran Diptera (Insecta).
We sequenced nearly the entire carbomoylphosphate synthase (CPS) domain of CAD, or rudimentary, (ca. 4 kb) from 29 species of flies representing all major clades within Eremoneura, or higher flies, and several orthorrhaphous brachyceran outgroups. We compared these sequences with orthologs from Anopheles gambiae and Drosophila melanogaster to assess structure, compositional bias, and phylogenetic utility. CAD is large (6.6+ kb), complex (comprised of three major and myriad minor functional domains) and relatively free of introns, extreme nucleotide bias (except third codon positions), and large hypervariable regions. The CPS domain possesses moderate levels of nonsynonymous divergence among taxa of intermediate evolutionary age and conveys considerable phylogenetic signal. Phylogenetic analysis of CPS sequences under varying methods and assumptions resulted in well-resolved, strongly supported trees concordant with many traditional ideas about higher dipteran phylogeny and with prior inferences from 28S rDNA. The most robustly supported major eremoneuran clades were Cyclorrhapha, Platypezoidea, Eumuscomorpha, Empidoidea, Atelestidae, Empidoidea exclusive of Atelestidae, Hybotidae s.l., Microphoridae+Dolichopodidae, and Empididae s. str. Because CAD is ubiquitous, apparently single copy (at least within holometabolous insects), readily obtained from several insect orders using primers described herein, and exhibits considerable phylogenetic utility, it should have wide applicability in insect molecular systematics.
Moulton
John K
JK
Department of Entomology and Plant Pathology, 2431 Center Drive, 205 Ellington Plant Sciences Building, The University of Tennessee, Knoxville, TN 37996-4560, USA. jmoulton@utk.edu
Wiegmann
Brian M
BM
D000818:Animals; D001482:Base Composition; D019731:Carbon-Nitrogen Ligases / Q000235:genetics*; D004275:DNA, Ribosomal; D004175:Diptera / Q000235:genetics / Q000502:physiology*; D019143:Evolution, Molecular; D014644:Genetic Variation; D006579:Heterozygote; D019476:Insect Proteins / Q000235:genetics*; D007438:Introns; D008969:Molecular Sequence Data; D010802:Phylogeny; D017434:Protein Structure, Tertiary
D004275:DNA, Ribosomal; D019476:Insect Proteins; D019731:Carbon-Nitrogen Ligases