Molecular Phylogenetics of Bat Flies of Cave-Roosting...
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Molecular Phylogenetics of Bat Flies of Cave-Roosting Bats Yisen Zheng, Solon Morse, Katharina Dittmar Department of Biological Sciences, University at Buffalo References 1) Dittmar, K., Porter, M.L., Murray, S., Whiting, M.F. (2006) Molecular phylogenetic analysis of nycterbiid and streblid bat flies (Diptera: Brachycera, Calyptrate):Implications for host associations and phylogeographic origins. Molecular Phylogenetics and Evolution, 38,155-170. 2) Hosokawa,T., Nikoh, N., Koga, R., Sato, M., Tanahashi, M., Meng, X.Y., Fukatsu, T. (2011) Reductive genome evolution, host-symbiont co-speciation and uterine transmission ofendosymbiotic bacteria in bat flies. The ISME Journal, 6, 577-587. 3) McAlpine, J. (1989) Phylogeny and classification of the Muscomorpha.Manual of Nearctic Diptera(ed. by J. F. McAlpine.), pp. 1397-1581. Canadian Government Publishing Centre. Research Branch Agriculture Canada, 3, Monograph No. 32: Hull. 4) Morse, S., Dick, C., Patterson, B., Dittmar, K. (2012) Some Like It Hot: Evolution and Ecology of Novel Endosymbionts in Bat Flies of Cave-Roosting Bats (Hippoboscoidea, Nycterophiliinae). Appl. Environ. Microbiol. December 2012 vol. 78 no. 24 8639-8649 Background Ectoparasitic, blood-sucking bat flies (Hippoboscidae; Nycteribiidae) are known to be ubiquitous on cave-roosting bats. While there is abundant data of their endosymbiont bacteria, there exists a lack of sampled host genes. To date, only one gene (Cytochrome Oxidase I) has been widely sampled among members of Nycteribiidae. Studies of this single gene have strongly suggested the presence of co-evolutionary trends between host flies and endosymbionts. However, a wider sampling of genetic data is necessary to provide the tools to unravel relations between host species that are currently understudied, to identify with confidence the presence of co-evolutionary trends between hosts and their parasites, and to better understand the role endosymbiotic parasites play in their host specimens. The results of this experiment were incorporated into a more encompassing study that analyzed the topology of both host flies and their endosymbionts, indicating phylogenetic concordance and highly similar rates of divergence 4 . Methods/Procedure Total genomic DNA extracted from 11 specimens was utilized in the conduction of Polymerase Chain Reactions for the purpose of amplifying genetic material. Primers specific for targeting a wide variety of genes (COII, 12S, 16S, 18S, 28S, H3, CAD, and cytB) were incorporated. PCR products then underwent gel electrophoresis, sequencing via HTSeq, and BLAST analysis via GenBank. Using GENEious, sequences were aligned, assembled, and concatenated for the purposes of determining evolutionary relationships. jModelTest was used to identify the best-fit models of nucleotide substitution and trees were constructed accordingly through Bayesian methods using BEAST. Specimen Country Location Host species BDP5030 Peru Arequipa, Atiquipa Platalina genovensium MEXA4.1 Mexico Tamaulipas Pteronotus parnellii MEXA4.2 Mexico Tamaulipas Pteronotus parnellii DR05203.1 Dominican Republic Baoruco Macrotus waterhousii DR05161.2 Dominican Republic Pedernales Macrotus waterhousii FE20 Puerto Rico Arecibo Glue trap KD17070809 Puerto Rico Arecibo Monophyllus redmani MEXG3 Mexico Tamaulipas Pteronotus davyi MEXB2 Mexico Tamaulipas Pteronotus davyi DR05256.1 Dominican Republic Baoruco Pteronotus parnellii DR05258.2 Dominican Republic Baoruco Pteronotus parnellii A member of the Pteronotus sp. from which various fly specimens were obtained. Image courtesy of Solon Morse. Results/Discussion A concatenated super-alignment of nine total genes (including COI) yielded a phylogenic distribution as presented in Figure 1. jModelTest identified GTR+I+G and TIM3+G as the best fit substitution models for these various genes. The Hippoboscidae family incorporates members of two genera utilized in this study: Nycterophilia and Phalconomus. Among the 11 bat fly genomes analyzed, only one was a member of the relatively obscure and little known genus, Phalconomus (BDP5030). As indicated in Figure 1, BDP5030 falls out as a closely related sister taxa to specimens belonging to Nycterophilia. As suspected, a strong correlation exists between the origin and host of a specimen (see Table 1) and their phylogenetic placement. For instance, two specimens that share a locational origin and host species, MEXA4.1 and MEXA4.2, have been classified as sister taxa on the basis of genetic analysis. Data was utilized towards conducting a more encompassing study which has indicated a pattern of co-evolution of bat flies with endosymbiotic bacterial associates. Character mapping has shown that a mutualistic relationship has evolved between host flies and their bacterial symbionts, allowing for survival in abnormally hot cave environments 4 . Figure 1 Table 1
Transcript of Molecular Phylogenetics of Bat Flies of Cave-Roosting...
Molecular Phylogenetics of Bat Flies of Cave-Roosting Bats Yisen Zheng, Solon Morse, Katharina Dittmar
Department of Biological Sciences, University at Buffalo
References 1) Dittmar, K., Porter, M.L., Murray, S., Whiting, M.F. (2006) Molecular
phylogenetic analysis of nycterbiid and streblid bat flies (Diptera: Brachycera, Calyptrate):Implications for host associations and phylogeographic origins. Molecular Phylogenetics and Evolution, 38,155-170.
2) Hosokawa,T., Nikoh, N., Koga, R., Sato, M., Tanahashi, M., Meng, X.Y., Fukatsu, T. (2011) Reductive genome evolution, host-symbiont co-speciation and uterine transmission ofendosymbiotic bacteria in bat flies. The ISME Journal, 6, 577-587.
3) McAlpine, J. (1989) Phylogeny and classification of the Muscomorpha.Manual of Nearctic Diptera(ed. by J. F. McAlpine.), pp. 1397-1581. Canadian Government Publishing Centre. Research Branch Agriculture Canada, 3, Monograph No. 32: Hull.
4) Morse, S., Dick, C., Patterson, B., Dittmar, K. (2012) Some Like It Hot: Evolution and Ecology of Novel Endosymbionts in Bat Flies of Cave-Roosting Bats (Hippoboscoidea, Nycterophiliinae). Appl. Environ. Microbiol. December 2012 vol. 78 no. 24 8639-8649
Background Ectoparasitic, blood-sucking bat flies (Hippoboscidae; Nycteribiidae) are known to be ubiquitous on cave-roosting bats. While there is abundant data of their endosymbiont bacteria, there exists a lack of sampled host genes. To date, only one gene (Cytochrome Oxidase I) has been widely sampled among members of Nycteribiidae. Studies of this single gene have strongly suggested the presence of co-evolutionary trends between host flies and endosymbionts. However, a wider sampling of genetic data is necessary to provide the tools to unravel relations between host species that are currently understudied, to identify with confidence the presence of co-evolutionary trends between hosts and their parasites, and to better understand the role endosymbiotic parasites play in their host specimens. The results of this experiment were incorporated into a more encompassing study that analyzed the topology of both host flies and their endosymbionts, indicating phylogenetic concordance and highly similar rates of divergence4.
Methods/Procedure Total genomic DNA extracted from 11 specimens was utilized in the conduction of Polymerase Chain Reactions for the purpose of amplifying genetic material. Primers specific for targeting a wide variety of genes (COII, 12S, 16S, 18S, 28S, H3, CAD, and cytB) were incorporated. PCR products then underwent gel electrophoresis, sequencing via HTSeq, and BLAST analysis via GenBank. Using GENEious, sequences were aligned, assembled, and concatenated for the purposes of determining evolutionary relationships. jModelTest was used to identify the best-fit models of nucleotide substitution and trees were constructed accordingly through Bayesian methods using BEAST.
Specimen Country Location Host species BDP5030 Peru Arequipa, Atiquipa Platalina genovensium
MEXA4.1 Mexico Tamaulipas Pteronotus parnellii
MEXA4.2 Mexico Tamaulipas Pteronotus parnellii
DR05203.1 Dominican Republic Baoruco Macrotus waterhousii
DR05161.2 Dominican Republic Pedernales Macrotus waterhousii
FE20 Puerto Rico Arecibo Glue trap
KD17070809 Puerto Rico Arecibo Monophyllus redmani
MEXG3 Mexico Tamaulipas Pteronotus davyi
MEXB2 Mexico Tamaulipas Pteronotus davyi
DR05256.1 Dominican Republic Baoruco Pteronotus parnellii
DR05258.2 Dominican Republic Baoruco Pteronotus parnellii
A member of the Pteronotus sp. from which various fly specimens were obtained. Image courtesy of Solon Morse.
Results/Discussion A concatenated super-alignment of nine total genes (including COI) yielded a phylogenic distribution as presented in Figure 1. jModelTest identified GTR+I+G and TIM3+G as the best fit substitution models for these various genes. The Hippoboscidae family incorporates members of two genera utilized in this study: Nycterophilia and Phalconomus. Among the 11 bat fly genomes analyzed, only one was a member of the relatively obscure and little known genus, Phalconomus (BDP5030). As indicated in Figure 1, BDP5030 falls out as a closely related sister taxa to specimens belonging to Nycterophilia. As suspected, a strong correlation exists between the origin and host of a specimen (see Table 1) and their phylogenetic placement. For instance, two specimens that share a locational origin and host species, MEXA4.1 and MEXA4.2, have been classified as sister taxa on the basis of genetic analysis. Data was utilized towards conducting a more encompassing study which has indicated a pattern of co-evolution of bat flies with endosymbiotic bacterial associates. Character mapping has shown that a mutualistic relationship has evolved between host flies and their bacterial symbionts, allowing for survival in abnormally hot cave environments4.
Figure 1
Table 1
