Correlation between the green-island phenotype and Wolbachia infections during the evolutionary diversification of Gracillariidae leaf-mining moths

Abstract

Internally feeding herbivorous insects such as leaf miners have developed the ability to manipulate the physiology of their host plants in a way to best meet their metabolic needs and compensate for variation in food nutritional composition. For instance, some leaf miners can induce green-islands on yellow leaves in autumn, which are characterized by photosynthetically active green patches in otherwise senescing leaves. It has been shown that endosymbionts, and most likely bacteria of the genus Wolbachia, play an important role in green-island induction in the apple leaf-mining moth Phyllonorycter blancardella. However, it is currently not known how widespread is this moth-Wolbachia-plant interaction. Here, we studied the co-occurrence between Wolbachia and the green-island phenotype in 133 moth specimens belonging to 74 species of Lepidoptera including 60 Gracillariidae leaf miners. Using a combination of molecular phylogenies and ecological data (occurrence of green-islands), we show that the acquisitions of the green-island phenotype and Wolbachia infections have been associated through the evolutionary diversification of Gracillariidae. We also found intraspecific variability in both green-island formation and Wolbachia infection, with some species being able to form green-islands without being infected by Wolbachia. In addition, Wolbachia variants belonging to both A and B supergroups were found to be associated with green-island phenotype suggesting several independent origins of green-island induction. This study opens new prospects and raises new questions about the ecology and evolution of the tripartite association between Wolbachia, leaf miners, and their host plants.

References Andrew, R., L. Bernatchez, A. Bonin, C. A. Buerkle, B. C. Carstens, B. C. Emerson, et al. 2013. A road map for molecular ecology. Mol. Ecol. 22:2605–2626. Arthofer, W., M. Riegler, D. N. Avtzis, and C. Stauffer. 2009. Evidence for low-titre infections in insect symbiosis: Wolbachia in the bark beetle Pityogenes chalcographus (Coleoptera, Scolytinae). Environ. Microbiol. 11:1923–1933. Baldo, L., N. Lo, and J. H. Werren. 2005. Mosaic nature of the Wolbachia surface protein. J. Bacteriol. 187:5406–5418. Baldo, L., J. C. Dunning Hotopp, K. A. Jolley, S. R. Bordenstein, S. A. Biber, R. R. Choudhury, et al. 2006. Multilocus sequence typing system for the endosymbiont Wolbachia pipientis. Appl. Environ. Microbiol. 72:7098–7110. Barker, D., A. Meade, and M. Pagel. 2007. Constrained models of evolution lead to improved prediction of functional linkage from correlated gain and loss of genes. Bioinformatics 23:14–20. Barr, K. L., L. B. Hearne, S. Briesacher, T. L. Clark, and G. E. Davis. 2010. Microbial symbionts in insects influence down-regulation of defense genes in maize. PLoS ONE 5:e11339. Bazzocchi, C., W. Jamnongluk, S. L. O'Neill, T. J. Anderson, C. Genchi, and C. Bandi. 2000. Wsp gene sequences from the Wolbachia of _filarial nematodes. Curr. Microbiol. 41:96–100. Biere, A., and A. E. Bennett. 2013. Three-way interactions between plants, microbes and insects. Funct. Ecol. 27:567–573. Body, M., W. Kaiser, G. Dubreuil, J. Casas, and D. Giron. 2013. Leaf-Miners Co-opt microorganisms to enhance their nutritional environment. J. Chem. Ecol. 39:969–977. Boto, L. 2010. Horizontal gene transfer in evolution: facts and challenges. Proc. Biol. Sci. 277:819–827. Braig, H. R., W. Zhou, S. L. Dobson, and S. L. O'Neill. 1998. Cloning and characterization of a gene encoding the major surface protein of the bacterial endosymbiont Wolbachia pipientis. J. Bacteriol. 180:2373–2378. Brower, A. V., and R. DeSalle. 1998. Patterns of mitochondrial versus nuclear DNA sequence divergence among nymphalid butterflies: the utility of wingless as a source of characters for phylogenetic inference. Insect Mol. Biol. 7:73–82. Colgan, D. J., A. McLauchlan, G. D. F. Wilson, S. P. Livingston, G. D. Edgecombe, J. Macaranas, et al. 1998. Histone H3 and U2 snRNA DNA sequences and arthropod molecular evolution. Aust. J. Zool. 46:419–437. Dedeine, F., F. Vavre, F. Fleury, B. Loppin, M. E. Hochberg, and M. Boulétreau. 2001. Removing symbiotic Wolbachia bacteria specifically inhibit oogenesis in a parasitic wasp. Proc. Natl Acad. Sci. USA 98:6247–6252. Dedeine, F., M. E. Ahrens, L. Calcaterra, and D. D. Shoemaker. 2005. Social parasitism in fire ants (Solenopsis spp): a potential mechanism for interspecies transfer of Wolbachia. Mol. Ecol. 14:1543–1548. Dinca, V., E. V. Zakharov, P. D. N. Hebert, and R. Vila. 2011. Complete DNA barcode reference library for a country's butterfly fauna reveals high performance for temperate Europe. Proc. R. Soc. Lond. B Biol. Sci., 278:347–355. Fernandes, G. W., H. Duarte, and U. Lüttge. 2003. Hypersensitivity of Fagus sylvatica L. against leaf galling insects. Trees 17:407–411. Frago, E., M. Dicke, and H. C. J. Godfray. 2012. Insect symbionts as hidden players in insect-plant interactions. Trends Ecol. Evol. 27:705–711. Frost, C. L., H. Fernandez-Marin, J. E. Smith, and W. O. H. Hughes. 2010. Multiple gains and losses of Wolbachia symbionts across a tribe of fungus-growing ants. Mol. Ecol. 19:4077–4085. Giron, D., and G. Glevarec. 2014. Cytokinin-induced phenotypes in plant-insect interactions: learning from the bacterial world. J. Chem. Ecol. 40:826–835. Giron, D., E. Frago, G. Glevarec, C. M. J. Pieterse, and M. Dicke. 2013. Cytokinins as key regulators in plant–microbe–insect interactions: connecting plant growth and defence. Funct. Ecol. 27:599–609. Gouy, M., S. Guindon, and O. Gascuel. 2010. SeaView Version 4: a multiplatform graphical user interface for sequence alignment and phylogenetic tree building. Mol. Biol. Evol. 27:221–224. Harris, M., T. Friesen, S. Xu, M.-S. Chen, D. Giron, and J.Stuart. 2015. Pivoting from Arabidopsis to wheat to understand how agricultural plants integrate responses to biotic stress. J. Exp. Bot., 66:513–531. Hebert, P. D. N., A. Cywinska, S. L. Ball, and J. R. de Waard. 2003. Biological identifications through DNA barcodes. Proc. R. Soc. Lond. B Biol. Sci., 270:313–321. Hebert, P. D. N., E. H. Penton, J. M. Burns, D. H. Janzen, and W. Hallwachs. 2004. Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator. Proc. Natl Acad. Sci. USA 101:14812–14817. Hosokawa, T., Y. Kikuchi, M. Shimada, and T. Fukatsu. 2007. Obligate symbiont involved in pest status of host insect. Proc. Biol. Sci. 274:1979–1984. Hosokawa, T., R. Koga, Y. Kikuchi, X. Y. Meng, and T. Fukatsu. 2010. Wolbachia as a bacteriocyte-associated nutritional mutualist. Proc. Natl. Acad. Sci. USA 107:769–774. Ivanova, N. V., J. R. Dewaard, and P. D. N. Hebert. 2006. An inexpensive, automation-friendly protocol for recovering high-quality DNA. Mol. Ecol. Notes 6:998–1002. Kaiser, W., E. Huguet, J. Casas, C. Commin, and D. Giron. 2010. Plant green-island phenotype induced by leaf-miners is mediated by bacterial symbionts. Proc. Biol. Sci. 277:2311–2319. Kawahara, A. Y., I. Ohshima, A. Kawakita, J. C. Regier, C. Mitter, M. P. Cummings, et al. 2011. Increased gene sampling provides stronger support for higher-level groups within gracillariid leaf mining moths and relatives (Lepidoptera: Gracillariidae). BMC Evol. Biol. 11:182. Kent, B. N., L. J. Funkhouser, S. Setia, and S. R. Bordenstein. 2011. Evolutionary genomics of a temperate bacteriophage in an obligate intracellular bacteria (Wolbachia). PLoS ONE 6:e24984. Kirichenko, N., H. Deutsch, P. Triverti, R. Rougerie, P. Huemer, and C. Lopez-Vaamonde. 2015. Integrative taxonomy reveals a new species of Callisto (Lepidoptera: Gracillariidae) in the Alps. ZooKeys 473:157–176. Kodandaramaiah, U., T. J. Simonsen, S. Bromilow, N. Wahlberg, and F. Sperling. 2013. Deceptive single-locus taxonomy and phylogeography: Wolbachia-associated divergence in mitochondrial DNA is not reflected in morphology and nuclear markers in a butterfly species. Ecol. Evol. 3:5167–5176. Lees, D. C., A. Y. Kawahara, O. Bouteleux, I. Ohshima, A. Kawakita, R. Rougerie, et al. 2013. DNA barcoding reveals a largely unknown fauna of Gracillariidae leaf-mining moths in the Neotropics. Mol. Ecol. Resour. 14:286–296. Lopez-Vaamonde, C., H. C. J. Godfray, and J. M. Cook. 2003. Evolutionary dynamics of host plant use in a genus of leaf-mining moths. Evolution 57:1804–1821. Lopez-Vaamonde, C., N. Wikström, C. Labandeira, H. C. J. Godfray, S. J. Goodman, and J. M. Cook. 2006. Fossil-calibrated molecular phylogenies reveal that leaf-mining moths radiated several million years after their host plants. J. Evol. Biol. 19:1314–1326. Meier, R., K. Shiyang, G. Vaidya, and P. K. Ng. 2006. DNA barcoding and taxonomy in Diptera: a tale of high intraspecific variability and low identification success. Syst. Biol. 55:715–728. Mutanen, M., L. Kaila, and J. Tabell. 2013. Wide-ranging barcoding aids discovery of one-third increase of species richness in presumably well-investigated moths. Sci. Rep. 3:2901. Nikoh, N., T. Hosokawa, M. Moriyama, K. Oshima, M. Hattori, and T. Fukatsu. 2014. Evolutionary origin of insect-Wolbachia nutritional mutualism. Proc. Natl Acad. Sci. USA 111:10257–10262. Pagel, M. 1994. Detecting correlated evolution on phylogenies: a general method for the comparative analysis of discrete characters. Proc. Biol. Sci. 255:37–45. Posada, D. 2008. jModelTest: phylogenetic model averaging. Mol. Biol. Evol. 25:1253–1256. Ratnasingham, S., and Hebert P. D. N. 2013. BOLD: the barcode of life data system (http://www.barcodinglife.org). Mol. Ecol. Resour., 7:355–364. Robert, C. A. M., D. L. Frank, K. A. Leach, T. C. J. Turlings, B. E. Hibbard, and M. Erb. 2013. Direct and indirect plant defences are not suppressed by endosymbionts of a specialist root herbivore. J. Chem. Ecol. 39:507–515. Ronquist, F., M. Teslenko, P. Van der Mark, D. L. Ayres, A. Darling, S. Höhna, et al. 2012. MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Syst. Biol. 61:539–542. Rougerie, R., I. J. Kitching, J. Haxaire, S. E. Miller, A. Hausmann, and P. D. N. Hebert. 2014. Australian Sphingidae – DNA barcodes challenge current species boundaries and distributions. PLoS ONE 9:e101108. Schäfer, M., I. D. Meza-Canales, A. Navarro-Quezada, C. Brütting, R. Vanková, I. T. Baldwin, et al. 2014. Cytokinin levels and signaling respond to wounding and the perception of herbivore elicitors in Nicotiana attenuata. J. Integr. Plant Biol., 57:198–212. Schneider, D. I., M. Riegler, W. Arthofer, H. Merçot, C. Stauffer, and W. J. Miller. 2013. Uncovering Wolbachia diversity upon artificial host transfer. PLoS ONE 8:e82402. Serbielle, C., S. Moreau, F. Veillard, E. Voldoire, A. Bézier, M.-A. Mannucci, et al. 2009. Identification of parasite-responsive cysteine proteases in Manduca sexta. Biol. Chem. 390:493–502. Simões, P. M., G. Mialdea, D. Reiss, M.-F. Sagot, and S. Charlat. 2011. Wolbachia detection: an assessment of standard PCR protocols. Mol. Ecol. Resour. 11:567–572. Smith, M. A., C. Bertrand, K. Crosby, E. S. Eveleigh, J. Fernandez-Triana, B. L. Fisher, et al. 2012. Wolbachia and DNA barcoding insects: patterns, potential, and problems. PLoS ONE 7:e36514. Song, H., J. E. Buhay, M. F. Whiting, and K. A. Crandall. 2008. Many species in one: DNA barcoding overestimates the number of species when nuclear mitochondrial pseudogenes are coamplified. Proc. Natl Acad. Sci. USA 105:13486–13491. Stone, G., and K. Schönrogge. 2003. The adaptive significance of insect gall morphology. Trends Ecol. Evol. 18:512–522. Sugio, A., G. Dubreuil, D. Giron, and J.-C. Simon. 2015. Plant-insect interactions under bacterial influence: ecological implications and underlying mechanisms. J. Exp. Bot., 66:467–478. Wahlberg, N., and C. W. W. Wheat. 2008. Genomic outposts serve the phylogenomic pioneers: designing novel nuclear markers for genomic DNA extractions of Lepidoptera. Syst. Biol. 57:231–242. Walters, D. R., N. McRoberts, and B. D. Fitt. 2008. Are green islands red herrings? Significance of green islands in plant interactions with pathogens and pests. Biol. Rev. Camb. Philos. Soc. 83:79–102. Werren, J. H., and D. M. Windsor. 2000. Wolbachia infection frequencies in insects: evidence of a global equilibrium? Proc. Biol. Sci. 267:1277–1285. West, S. A., J. M. Cook, J. H. Werren, and H. C. J. Godfray. 1998. Wolbachia in two insect host-parasitoid communities. Mol. Ecol. 7:1457–1465. Wiemers, M., and K. Fiedler. 2007. Does the DNA barcoding gap exist? - a case study in blue butterflies (Lepidoptera: Lycaenidae). Front. Zool. 4:8. Zhu, F., E. H. Poelman, and M. Dicke. 2014. Insect herbivore-associated organisms affect plant responses to herbivory. New Phytol. 204:315–321.