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Double trouble for grasshopper molecular systematics: intra‐individual heterogeneity of both mitochondrial 12S‐valine‐16S and nuclear internal transcribed spacer ribosomal DNA sequences in Hesperotettix viridis (Orthoptera: Acrididae)
Author(s) -
SWORD GREGORY A.,
SENIOR LAURA B.,
GASKIN JOHN F.,
JOERN ANTHONY
Publication year - 2007
Publication title -
systematic entomology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 66
eISSN - 1365-3113
pISSN - 0307-6970
DOI - 10.1111/j.1365-3113.2007.00385.x
Subject(s) - biology , mitochondrial dna , nuclear dna , internal transcribed spacer , genetics , ribosomal dna , acrididae , phylogenetic tree , ribosomal rna , spacer dna , concerted evolution , evolutionary biology , dna , molecular phylogenetics , orthoptera , zoology , gene
Hesperotettix viridis grasshoppers (Orthoptera: Acrididae: Melanoplinae) exhibit intra‐individual variation in both mitochondrial 12S‐valine‐16S and nuclear internal transcribed spacer (ITS) ribosomal DNA sequences. These findings violate core assumptions underlying DNA sequence data obtained via polymerase chain reaction (PCR) amplification for use in molecular systematics investigations. Undetected intra‐individual variation of this sort can confound phylogenetic analyses at a range of taxonomic levels. The use of a DNA extraction protocol designed to enrich mitochondrial DNA as well as an initial long PCR of approximately 40% of the grasshopper mitochondrial genome failed to control for the presence of paralogous mitochondrial DNA‐like sequences within individuals. These findings constitute the first demonstration of intra‐individual heterogeneity in mitochondrial DNA‐like sequences in the grasshopper subfamily, Melanoplinae, and only the second report of intra‐individual variation in nuclear ITS ribosomal DNA sequences in grasshoppers. The fact that intra‐individual variation was detected in two independent DNA marker sets in the same organism strengthens the notion that the orthology of PCR‐derived DNA sequences should be examined thoroughly prior to their use in molecular phylogenetic analyses or as DNA barcodes.