DOI: 10.5061/dryad.t9140
title: Data from: A novel, enigmatic basal leafflower moth lineage pollinating a derived leafflower host illustrates the dynamics of host shifts, partner replacement, and apparent coadaptation in intimate mutualisms

description: Leafflower plant/leafflower moth brood pollination mutualisms are
 widespread in the Paleotropics. Leafflower moths pollinate leafflower
 plants, but their larvae consume a subset of the hosts’ seeds. These
 interactions are highly phylogenetically constrained: six clades of
 leafflower plants are each associated with a unique clade of leafflower
 moths (Epicephala). Here, we report a previously unrecognized basal
 seventh pollinating Epicephala lineage—associated with the highly derived
 leafflower clade Glochidion—in Asia. Epicephala lanceolaria is a
 pollinator and seed predator of Glochidion lanceolarium. Phylogenetic
 inference indicates that the ancestor of E. lanceolaria most likely
 shifted onto the ancestor of G. lanceolarium and displaced the ancestral
 allospecific Epicephala pollinator in at least some host populations. The
 unusual and apparently coadapted aspects of the G. lanceolarium/E.
 lanceolaria reproductive cycles suggest that plant-pollinator coevolution
 may have played a role in this displacement and provide insights into the
 dynamics of host shifts and trait coevolution in this specialized
 mutualism.
description: Sequence_data.fastaNovel DNA sequence data generated for this study from the species Glochidion lanceolarium (Phyllanthaceae) and Epicephala lanceolaria (Insecta: Lepidoptera: Gracillariidae). These data represent five loci: for Epicephala, mitochondrial COI, and nuclear EF1-α and ArgK; for Glochidion, nuclear ribosomal ITS and ETS. Sequences are the same as GenBank numbers KY078233-KY078291; however, GenBank numbers are not included in this file. For correspondences between sample numbers and GenBank sequences for Epicephala, please see Table 2 in the manuscript. Glochidion lanceolarium ITS sequence corresponds to GenBank number KY078253 and ETS sequence corresponds to GenBank number KY078252.Figures_3_4Data used in generating Figures 3 and 4 in the manuscript (see Excel spreadsheet tab labels). Figure 3 data represent the number of adult Epicephala lanceolaria moths found in individual Glochidion lanceolarium fruit at three sites in Guangdong Province, China. Figure 4 data represent the number of Epicephala lanceolaria pupae with and without cocoons found when dissecting Glochidion lanceolarium fruit at two sites in Guangdong Province, China.MrBayes_Glochidion.nex.conPhylogenetic hypothesis recovered using Bayesian inference implemented in MrBayes for Glochidion. Tip labels represent either specimen numbers or specific epithets for Glochidion species and are from the present study as well as Kawakita et al. (2004) Evolution and Hembry et al. (2013) Proc. Roy. Soc. B. Phyllanthus roseus is a very close relative to Glochidion, but is not pollinated by Epicephala (Kawakita and Kato 2009 Proc. R. Soc. B). Many tip labels follow the Supplementary Appendix in Hembry et al. (2013) Proc. R. Soc. B. This tree corresponds to Figure 5 and half of Figure 7 in the present manuscript.RAxML_GlochidionPhylogenetic hypothesis for Glochidion recovered using maximum likelihood inference implemented in RAxML. Tip labels represent either specimen numbers or specific epithets for Glochidion species and are from the present study as well as Kawakita et al. (2004) Evolution and Hembry et al. (2013) Proc. Roy. Soc. B. Phyllanthus roseus is a very close relative to Glochidion, but is not pollinated by Epicephala (Kawakita and Kato 2009 Proc. R. Soc. B). Many tip labels follow the Supplementary Appendix in Hembry et al. (2013) Proc. R. Soc. B. This tree corresponds to Supplementary Figure 1 in the present manuscript.MrBayes_Epicephala.nex.conPhylogenetic hypothesis for Epicephala, recovered using Bayesian inference implemented in MrBayes. Tip labels represent either specimen numbers and/or host species (in the genera Flueggea, Phyllanthus, Breynia, and Glochidion) from which specimens were reared, and are from the present study as well as Kawakita et al. (2004) Evolution, Kawakita et al. (2009) Proc. R. Soc. B, and Hembry et al. (2013) Proc. Roy. Soc. B. Many tip labels follow the Supplementary Appendix in Hembry et al. (2013) Proc. R. Soc. B. For E. lanceolaria OTUs, numbers refer to particular specimens; two-letter codes indicate collection localities (GB: Gaobangshan, LD: South China Botanical Garden, QA: Qiao Island, WT: Wutongshan); C = pupa with cocoon, NC = pupa with no cocoon. This tree corresponds to Figure 6 in the present manuscript.RAxML_EpicephalaPhylogenetic hypothesis for Epicephala, recovered using maximum likelihood implemented in RAxML. Tip labels represent either specimen numbers and/or host species (in the genera Flueggea, Phyllanthus, Breynia, and Glochidion) from which specimens were reared, and are from the present study as well as Kawakita et al. (2004) Evolution, Kawakita et al. (2009) Proc. R. Soc. B, and Hembry et al. (2013) Proc. Roy. Soc. B. Many tip labels follow the Supplementary Appendix in Hembry et al. (2013) Proc. R. Soc. B. For E. lanceolaria OTUs, numbers refer to particular specimens; two-letter codes indicate collection localities (GB: Gaobangshan, LD: South China Botanical Garden, QA: Qiao Island, WT: Wutongshan); C = pupa with cocoon, NC = pupa with no cocoon. This tree corresponds to Supplementary Figure 2 in the present manuscript.Epicephala_constrained.nex.conPhylogenetic hypothesis for Epicephala, recovered using Bayesian inference implemented in MrBayes constrained to force monophyly of all Glochidion-associated Epicephala. Tip labels represent either specimen numbers and/or host species (in the genera Flueggea, Phyllanthus, Breynia, and Glochidion) from which specimens were reared, and are from the present study as well as Kawakita et al. (2004) Evolution, Kawakita et al. (2009) Proc. R. Soc. B, and Hembry et al. (2013) Proc. Roy. Soc. B. Many tip labels follow the Supplementary Appendix in Hembry et al. (2013) Proc. R. Soc. B. For E. lanceolaria OTUs, numbers refer to particular specimens; two-letter codes indicate collection localities (GB: Gaobangshan, LD: South China Botanical Garden, QA: Qiao Island, WT: Wutongshan); C = pupa with cocoon, NC = pupa with no cocoon. This tree was used in Shimodaira-Hasegawa tests as described in the present manuscript.Epicephala_negconstrained.nex.conPhylogenetic hypothesis for Epicephala, recovered using Bayesian inference implemented in MrBayes constrained to force non-monophyly of Glochidion-associated Epicephala. Tip labels represent either specimen numbers and/or host species (in the genera Flueggea, Phyllanthus, Breynia, and Glochidion) from which specimens were reared, and are from the present study as well as Kawakita et al. (2004) Evolution, Kawakita et al. (2009) Proc. R. Soc. B, and Hembry et al. (2013) Proc. Roy. Soc. B. Many tip labels follow the Supplementary Appendix in Hembry et al. (2013) Proc. R. Soc. B. For E. lanceolaria OTUs, numbers refer to particular specimens; two-letter codes indicate collection localities (GB: Gaobangshan, LD: South China Botanical Garden, QA: Qiao Island, WT: Wutongshan); C = pupa with cocoon, NC = pupa with no cocoon. This tree was used in Shimodaira-Hasegawa tests as described in the present manuscript.Pollination_and_ovipositionVideo depicting pollination of and oviposition into flowers of Glochidion lanceolarium by a female Epicephala lanceolaria moth.