The Taxonomy and Population Structure of the Buckeye Butterflies (GenusJunonia, Nymphalidae: Nymphalini) of Florida, USA

How old can we go? Evaluating the age limit for effective DNA recovery from historical insect specimens

“…To evaluate the maximum insect specimen age that still permits mitochondrial genotyping, we selected 48 New World Junonia butterfly specimens collected prior to 1910 (ranging in age from pre‐1813 to 1909) (Table S1) for comparison with specimens from reference populations that we had previously genotyped for other projects (Table S2) (Lalonde et al ., ; Lalonde & Marcus, , b). To reduce possible biases due to differences in storage techniques, we sampled specimens from five natural history museum collections [American Museum of Natural History (AMNH), Illinois Natural History Survey (INHS), Natural History Museum London (NHMUK), Smithsonian National Museum of Natural History (USMNH), Yale Peabody Museum of Natural History (YPM)] and two private collections (Thomas W. Turner and specimens maintained as part of the research collection within the Marcus laboratory) for inclusion in our dataset.…”

“…In the New World Junonia , due to retained ancestral polymorphism and/or lateral transfer between species, mitochondrial haplotypes are not species‐specific (Borchers & Marcus, ; Gemmell et al ., ; Gemmell & Marcus, ). However, regardless of species, all of the Junonia from any one locality typically share mitochondrial haplotype frequencies for the two most common haplotype groups (A and B) found in most of the western hemisphere (Pfeiler et al ., ; Gemmell & Marcus, ; Peters & Marcus, ; Lalonde et al ., ; Lalonde & Marcus, ). It is noteworthy that haplotype group A is virtually absent from all species of North American Junonia except in south Florida (Gemmell & Marcus, ; Lalonde et al ., ; Lalonde & Marcus, , b).…”

“…However, regardless of species, all of the Junonia from any one locality typically share mitochondrial haplotype frequencies for the two most common haplotype groups (A and B) found in most of the western hemisphere (Pfeiler et al ., ; Gemmell & Marcus, ; Peters & Marcus, ; Lalonde et al ., ; Lalonde & Marcus, ). It is noteworthy that haplotype group A is virtually absent from all species of North American Junonia except in south Florida (Gemmell & Marcus, ; Lalonde et al ., ; Lalonde & Marcus, , b). Two additional, rarer Junonia haplotypes are geographically restricted, with haplotype C known only from the Peruvian Andes, and a distinct variant of haplotype group B, haplotype B CA , is commonly found in the American southwest and Mexico (Gemmell & Marcus, ; Lalonde & Marcus, ).…”

“…10% of the cost of fluorescent dye‐terminated Sanger DNA sequencing (U.S.$0.83 vs 9.12 per sample at July 2019 prices and currency exchange rates). It should be noted that this new genotyping method does not involve sequencing DNA, but instead assigns mitochondrial genotypes based on the presence or absence of sequence‐ and haplotype‐specific restriction enzyme cut sites in the DNA fragments being evaluated [which were already known for each haplotype based on prior exploratory work that included conventional DNA sequencing (Borchers & Marcus, ; Gemmell et al ., ; Peters & Marcus, ; Lalonde et al ., )]. In our previous work, the fragment‐based genotyping technique worked with 100% efficiency for > 1800 specimens collected in the 20th century and contemporary specimens (1910–2016) from the butterfly genus Junonia (Lepidoptera: Nymphalidae)(Table ), as well as for a handful of 19th‐century specimens, with the oldest genotyped museum specimen dating from 1866 (Lalonde & Marcus, , b).…”

Taxonomy as a hypothesis: testing the status of the Bermuda buckeye butterfly Junonia coenia bergi (Lepidoptera: Nymphalidae)

“…Sequences from the wg gene of J. coenia bergi specimens (GenBank vouchers KX228459–KX228470) were compared with other New World Junonia wg sequences from the Caribbean, Florida, and California (GenBank vouchers KF292254–KF292275 and KR094173–KR094437) (Borchers & Marcus, ; Lalonde et al , ). Individuals heterozygous for single nucleotide polymorphisms (SNPs) in the coding sequence of wg were identified and analyzed as previously described (Borchers & Marcus, ; Gemmell et al , ).…”