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Honors Theses

5-2017

Dating the Terrestrial Invasion of the (:) Using the Fossilized Birth-Death Model

Timothy Gervascio

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Recommended Citation Gervascio, Timothy, "Dating the Terrestrial Invasion of the Cyclophoroidea (Mollusca:Gastropoda) Using the Fossilized Birth-Death Model" (2017). Honors Theses. 115. https://digitalcommons.esf.edu/honors/115

This Thesis is brought to you for free and open access by Digital Commons @ ESF. It has been accepted for inclusion in Honors Theses by an authorized administrator of Digital Commons @ ESF. For more information, please contact [email protected], [email protected]. Dating the terrestrial invasion of the Cyclophoroidea (Mollusca:Gastropoda) using the Fossilized Birth-Death model.

By

Timothy Gervascio

Bachelor of Science with Honors

Department of Environmental and Forest Biology

May 2017

APPROVED

Thesis Project Advisor: /'l-- v / / ~

Second Reader: ~ ----,L------,<'---,,L--,,P.--~;.._,...~ ----

Honors Director: lJ~~ ~J} William M. Shields, Ph.D.

Date: 5 /u> / ,7 Abstract

Despite their vast diversity, few metazoan lineages have made the transition from marine to terrestrial . These transitions must have been accompanied by dramatic shifts in the physiology and ecology of the lineages, marking major evolutionary events that have shaped the diversity we see today. Among the Metazoa, gastropod molluscs have been among the most successful at making the transition, comprising at least 12 separate transitions. This unusual history makes them particularly suitable for studying the evolutionary context of these major events. The development of new relaxed molecular clock methods for seamlessly integrating molecular and data provides new possibilities for determining the timing of evolutionary events. Here we use the Fossilized Birth-Birth Death process implemented in BEAST v 2.4 to estimate the divergence time of the uniformly terrestrial Cyclophoroidea (Mollusca: ) from the rest of the Caenogastropoda. Our analysis shows that the terrestrialization of the Cyclophoroidea occurred around the end of the , and may have been associated with the End mass . This places the Cyclophoroidea as the first caenogastropod to make the transition to terrestriality. Table of Contents

Acknowledgements ...... i

Introduction ...... 1

Methods ...... 4

Results ...... 7

Discussion ...... 7

Conclusion ...... 10

References ...... 12

Appendix ...... 15

Acknowledgements

I thank Dr. Rundell for her endless generosity and support as my advisor and teacher; and Jesse Czekanski-Moir for his help and patience as I worked on this project.

I would also like to thank David Bullis, Teresa Rose Osborne, and Alyssa Lau for their role in developing the larger questions that this research addresses.

And I would like to thank the Rundell Lab team as a whole for their enthusiasm for science and for their extraordinary of humor. I could not have imagined such a phenomenal group before I joined the lab. Introduction

Transitions from marine to terrestrial habitats have been exceedingly rare among the Metazoa. These transitions must have necessitated novel physiological strategies, as requirements for osmoregulation, gas exchange, and thermoregulation are vastly different on land (Selden & Edwards, 1989; Selden, 2005). Such shifts in the biology of the colonizers have undoubtedly left a signature in the we see today.

Additionally, the first introductions of new lineages and functions to terrestrial ecosystems fundamentally altered the geochemical characteristics of the earth's surface and augmented the possibilities for future biotic interaction (Selden & Edwards, 1989;

Kenrick et al. , 2012). However rare, these transitions clearly mark major events in the history of the Metazoa that have shaped the geographic and biological template for evolution to act on, resulting in the particular range of terrestrial biodiversity we see today.

The earliest evidence of terrestrial metazoans comes from paleosols with Skolithos trace attributed to an ancestor of modern myriopods (Selden, 2005).

Millipedes were followed by arachnid mites and the first by the early

(Selden, 2005). A major molecular clock analysis of the has proposed somewhat older ages for these transitions than the fossil record alone, but supports the sequence of the fossil record (Rota-Stabelli et al., 2013). Among the Metazoa, gastropod molluscs have been the most numerous colonizers of land, comprising at least 12 separate transitions (Vermeij & Dudley, 2000; Kameda & Kato, 2011; Romero et al., 2016). This makes them a key group for studying the evolutionary context and repercussions of terrestrial izati on. Terrestrialization has taken place throughout the Gastropoda, with the

Stylommatophora containing the most rich terrestrial . Some of the earliest terrestrial gastropod fossils from the were originally attributed to the

Stylommatophora, but their placement has been in doubted (Bandel, 1993). Additionally, the age of the Stylommatophora has been estimated using molecular data to be somewhere in the late , making their terrestrial constituents substantially younger than previously thought (Dinapoli & Klussmann-Kolb, 20 I 0).

The Caenogastropoda is the most species rich group of gastropods and its species employ diverse ecological strategies and inhabit many different habitats, including

multiple independent transitions to terrestriality (Colgan et al., 2007). In contrast to the

Stylommatophora, the history of the Caenogastropoda reaches back at least to the

Carboniferous, where type gastropods with convincing caenogastropod larval characteristics are known from the Mississippian (358-323 mya) Tamworth Belt rocks

(Cook et al., 2008).

The Cyclophoroidea are a rich and highly endemic superfamily of terrestrial caenogastropods distributed throughout Southeast Asia, the Indo-Pacific, and South

America. Fossil cyclophoroideans are known mainly from the -

Purbeck Group of Europe, but some less certain Paleozoic specimens have been assigned

as well (Bandel, 1993). Convincing specimens have also come from (J.

Czekanski-Moir, personal communication, Feb. 2017). This, combined with their basal placement in the Caenogastropoda and age of closely related groups (such as the

Carboniferous fauna), suggests their history is substantially deeper than the

late Mesozoic (Bandel, 1993; Colgan et al., 2007).

2 Studying evolutionary events in the context of the depends upon

being able to reliably estimate their timing. This is often done in a phylogenetic context, using assumptions based on the clock-like accumulation of in all genomes. By applying a substitution rate (substitutions/time) model to the (the

branch lengths), it is theoretically possible to determine the timing of events on the tree.

However, the nucleotide sequence alone does not contain any information about the timing of substitutions, so external information about the timing of the evolutionary

process is required to calibrate the branch lengths to absolute time. Because it is

impractical to experimentally determine the rate for every loci and species on a given phylogenetic tree, other external information is required. Fossils are the most common type of external information because they contain phylogenetic information

about the evolutionary process and can be dated to absolute time. By placing a fossil with

a known age in the tree, the substitution rate model can be calibrated to absolute time and

the timing of evolutionary events can be estimated. The development of new relaxed

molecular clock methods allow for substitution rates to vary realistically across the tree,

and new methods of integrating the fossil data into the tree provide the most realistic

divergence time estimates available.

The Fossilized Birth-Death process (FBD) is a recently developed Bayesian

method for inferring time-calibrated phylogenies (Stadler 2010; Heath et al., 2014). In

contrast to node-dating methods, FBD treats the fossils and occurrence times as tips that

were produced by the same evolutionary process as the extant taxa. The model describes

the probability of a time tree and fossil placements based on the diversification rate,

3 turnover, and probability of sampling a fossil. This allows for the possibility that the fossils are extinct sister-taxa on branches of the extant tree, or that fossils are sampled ancestors of the extant taxa rather than an arbitrary minimum node age.

The most complete higher taxanomic level phylogenetic hypothesis for the

Caenogastropoda to date places the Cyclophoroidea among completely marine and freshwater groups (, ) (Colgan et al., 2007). Additionally, recent work has proposed the of the Cyclophoroidea, suggesting a single ancestral terrestrialization event some time in their history (Webster et al., 2012). Here, we date the divergence of the Cyclophoroidea from the rest of the Caenogastropoda using seven fossils for calibration, including two new Cyclophoroidea taxa from Burmese amber.

Methods

Taxon Sampling

Twenty-six taxa were selected to cover the Cyclophoroidea, ,

Campaniloidea, Cerithioidea, , and (Table 1) . Taxa, as well as publicly available sequence data for taxa within the Cyclophoroidea and for miles were drawn primarily from Webster and coworkers' study of cyclophoroidean phylogenetics and shell evolution (2012). Taxa from additional caenogastropod families,

Ampullariidae and , were included to facilitate the placement of additional fossils. Two ampullariid species, polita and bridgesii, were selected to cover the maximum distance within the (Hayes et al., 2009). Two species were selected from the Viviparidae by the same criteria (Richter, 2015).

4 symbolicum was selected as a potential sister taxa to the Cyclophoroidea based on previous phylogenetic work covering the whole of Caenogastropoda (Colgan et al.,

2006). sulcatus was selected for the same purpose based on BLAS TN searches of the NCBI nucleotide database.

Fossils

Five fossils within the Cyclophoroidea and two fossils from other groups were used to calibrate branch lengths to absolute time (Table 1) . Two fossils from Burmese amber (1 00mya) had more certain taxonomic placement, and were constrained to the

Cyclophoridae and the respectively (for discussion of constraints, see below) (J. Czekanski-Moir, personal communication, Feb. 2017). Three additional fossils from the European (late Jurassic, early Cretaceous), while certainly members of the Cyclophoroidea, had a less explicit position within the Cyclophoroidea

(Bandel, 1993). These three fo ssils were positioned within the Cyclophoroidea by the

FBD model.

One fossil, langtonensis (170 mya) was assigned to the Viviparidae

(Richter, 2015; Tracey et al., 1993). One fossil, nipponica (160 mya) was assigned to the Ampulariidae (Tracey et al., 1993).

Molecular Sequence Matrix

Publicly available sequence data for five loci was collected from GenBank (Table

2). Eukaryotic SSU and LSU rRNA sequences were aligned using the SILVA rRNA database, which maintains highly curated and quality controlled rRNA alignments (Quast et al., 2013). All other alignments were carried out using ClustalW as implemented in

5 MEGA v 7.0 (Kumar et al., 2016). 16s rRNA sequences were aligned as non-coding nucleotides, cytochrome oxidase 1 (COl) sequences were aligned as coding nucleotides using the mitochondrial code, and histone H3 (H3) was aligned as coding nucleotides with the standard genetic code. We identified the best fit substitution model using AIC calculated by the "Find Best Models" tool in MEGA 7. 16s, 28s, CO 1, and H3 were best fit by GTR + rand 18s was best fit by SYM.

The Fossilized Birth-Death Process and Phylogenetic Inference

Here, we use the Fossilized Birth-Death model with Sampled Ancestors implemented in BEAST v 2.4 (Drummond et al., 2012). In this method, fossils are assigned to a user defined clade based on prior information rather than to a node, and their placement in the topology of the clade is subject to the model parameters. Table 2 describes our user defined fossil placements. Substitution models used for phylogenetic inference are described above. Each gene was assigned an independent relaxed log normal molecular clock, meaning the substitution rate of a gene was drawn from a log normal distribution independently for each branch. A linked tree topology was inferred by BEAST using information from all five loci. We assigned 350mya as the origin time

(xo) of the branching process based on the earliest definitively caenogastropod fossils

(Cook et al. , 2008). The "Belau clade" of the Diplommatinidae, including Hungerfordia and , was constrained to be no older than 30mya by assigning a bounded uniform prior to the node to reflect the geological age of the archipelago they are endemic to

(Crombie & Pregill, 1999; Rundell, 2008).

Posterior probabilities were estimated from three MCMC chains run for 2 x I 08 generations, sampling every 1000 generations on the CIPRES Science Gateway (Miller et

6 al., 20 I 0). Convergence was assessed by viewing marginal probability distributions, and mixing was assessed by viewing time series plots in Tracer v 1.6 (Rambaut et al., 2014).

The MCMC chain was searched for the maximum clade credibility tree using

LogCombiner and TreeAnnotator with a burn-in of 200 trees (Drummond et al., 20 I 2).

Results

The maximum clade credibility tree from these FBD analyses produced a mid­

Carboniferous to mid-Permian age range for the divergence of the Cyclophoroidea from the other caenogastropods included in the analyses (Figure I). All other Caenogastropoda included were more closely related to each other than to the Cyclophoroidea. The analyses revealed a Permian - age range for the crown Cyclophoroidea and a

Jurassic age for the two largest clades within Cyclophoroidea, the Diplommatininae

(Diplommatinidae minus Cochlostomatinae) and the (Figure 1).

Discussion

The time calibrated phylogenetic hypothesis based on 26 extant taxa and 7 fossil calibration points revealed a mid-Carboniferous to mid-Permian age for the divergence of the Cyclophoroidea from the rest of the Caenogastropoda and a Permian-Triassic age for the uniformly terrestrial crown Cyclophoroidea, suggesting the transition to terrestriality may have been associated with the End-Permian mass extinction.

The End-Permian mass extinction was the most extreme of earth's five mass , resulting in the sudden loss of 90% of marine species and a significant turnover of terrestrial life. The timing of the Permian-Triassic (PT) boundary coincides

7 with a massive introduction of CO2 and methane, a global drop in 02, and a spike in atmospheric sulfates in the geologic record attributed to volcanism, declining sea level, and a general breakdown of the marine carbon cycle (Jin et al., 2000; Shen et al., 2011 ).

In addition to the sudden turnover of marine species, terrestrial environments were marked by a collapse of tropical Gigantopteris flora and global wildfires as a result of climate change (Shen et al., 2011 ). Marine conditions were characterized by regional sea level declines, a corresponding increase in salinity, and anoxia in deep waters beyond the reach of mixing (Erwin et al., 2002; Knoll et al., 2007).

Changes in marine conditions coinciding with the proposed timing of

Cyclophoroidea terrestrialization offer important information about selective pressures that may have factored into terrestrialization. Knoll and coworkers (2007) suggest that shallow waters could have acted as biological refugia during the PT event as deeper waters became anoxic due to a breakdown in mixing or to climatic factors. The survival of the ancestral Cyclophoroidea in those conditions could have been contingent upon making the transition to shallower waters (i.e. closer to land) or even to terrestrialization itself.

While the described analyses can provide context for the evolutionary shift from marine to terrestrial habitats in the Cyclophoroidea, some questions still remain. The divergence of the ancestrally marine Cyclophoroidea from the rest of the

Caenogastropoda happened earlier than the events surrounding the PT extinctions. If the divergence of the Cyclophoroidea was a result of terrestrial ization (before the PT events) the uniform terrestriality of extant Cyclophoroidea could be understood as contingent upon the terrestrial Cyclophoroidea avoiding the worst of the PT events.

8 Another interpretation is to view the crown age of extant Cyclophoroidea as a result of a radiation following terrestrialization. This hypothesis is more in line with the immediate PT mechanisms discussed above as the 95% CI is centered almost exactly on the PT boundary (256.8 mya). This could also indicate that terrestrialization occurred sometime through the Triassic as ecosystems recovered and previously occupied niches were opened. Unfortunately, we lack a full understanding of how recovery occurred following the PT extinction due to the depauperate fossil record (Chen &

Benton, 2012).

In comparison to other independent gastropod terrestrialization events, the

Cyclophoroidea were likely one of the first to make the transition. Members of the

Truncatellidae, , and made the transition in the , while the , made the transition in the Mesozoic (Kameda &

Kato, 2011; Kano et al., 2002). Carboniferous fossils previously thought to be terrestrial stylommatophorans are in doubt, and recent molecular dating of the suggests a much later Mesozoic diversification and terrestrialization (Bandel, 1993;

Dinapoli & Klussmann-Kolb, 2010). Only the extinct Dawsonellidae is known from the

Paleozoic (Kano et al. , 2002). This leaves the possibility that the Cyclophoroidea have the longest history on land among the Gastropoda, and confirms that they have the longest history on land of any caenogastropod clade.

The ecology of these extant lineages suggests the timing and context of terrestrialization matter greatly in determining the trajectory of the lineage. The modern

Cyclophoroidea mainly feed on lichens and , while stylommatophorans are more often specialized . The Stylommatophra also appear to have transitioned to

9 land during the late Mesozoic as angiosperms radiated, while the Cyclophoroidea predate the radiation of flowering plants. In the case of the Cyclophoroidea, this is likely the ancestral dietary state, suggesting significant conservation in their ecology despite comprising substantial species diversity. This may be characteristic of terrestrial gastropods as a whole, where modern examples of are common and low vagility makes geography a potent barrier to gene flow (Rundell & Price, 2009;

Rundell, 2011). Our data extends this observation into the realm of macroevolution.

Estimating of the timing of major evolutionary events is an important part of understanding the history of life on earth. Despite the limited communication between and paleontology, synthesis is a critical goal for modern evolutionary biology

(Grantham, 2004 ). Time calibrated phylogenies are a common source of interaction between the fields, and are only improved by the recent explosion of publicly available molecular data and the development of new methods for applying paleontological data to molecular based trees.

Conclusion

The transition from marine to terrestrial habitats marks a major event in the evolution of a lineage, necessitating novel evolutionary and ecological trajectories that shape the diversity of life we see today. Gastropod molluscs have been among the most successful metazoans crossing this seemingly impassable physical boundary, making gastropods important systems for studying the evolutionary context of terrestrialization events. Our analyses place the divergence of the completely terrestrial Cyclophoroidea from the other Caenogastropoda in the mid-Carboniferous to mid-Permian preceding the

PT mass extinction. The crown age of the Cyclophoroidea falls directly over the PT

10 boundary, suggesting the largest mass extinction in Earth' s history may have played a role in the transition. The terrestrialization of the Cyclophoroidea is likely the oldest among the caenogastropods and one of the earliest in all of Gastropoda. As the amount of available molecular and fossil data continues to swell, time calibrated phylogenies like the presented here are an important source of interaction between the historically isolated fields of neontology and paleontology.

11 References

Bandel, K. (1993). Caenogastropoda during Mesozoic times. Scripta Geologica, Special Issue, 2, 7-56.

Chen, Z. Q., & Benton, M. J. (2012). The timing and pattern of biotic recovery following the end-Permian mass extinction. Nature Geoscience, 5(6), 375-383.

Colgan, D. J., Ponder, W. F., Beacham, E., & Macaranas, J. (2007). Molecular phylogenetics of Caenogastropoda (Gastropoda: Mollusca). Molecular Phylogenetics and Evolution, 42(3), 717-737.

Cook, A., NUtzel, A. , & Fryda, J. (2008). Two Mississippian caenogastropod from Australia and their meaning for the ancestry of the Caenogastropoda. Journal ofPaleontology , 82(1 ), 183-187.

Crombie, R. I., & Pregill, G. K. (1999). A checklist of the herpetofauna of the Palau Islands (Republic of Belau), Oceania. Herpetological Monographs, 13, 29-80.

Dinapoli, A., & Klussmann-Kolb, A. (2010). The long way to diversity-phylogeny and evolution of the Heterobranchia (Mollusca: Gastropoda). Molecular Phylogenetics and Evolution, 55(1), 60-76.

Drummond, A. J., Suchard, M.A., Xie, D., & Rambaut, A. (2012). Bayesian phylogenetics with BEA Uti and the BEAST 1. 7. Molecular Biology and Evolution, 29(8), 1969-1973.

Erwin, D. H., Bowring, S. A ., & Yugan, J. (2002). End-Permian mass extinctions: a review. Special Papers-Geological Society ofAmerica , 356, 363-384.

Grantham, T. (2004). The role of fossils in phylogeny reconstruction: Why is it so difficult to integrate paleobiological and neontological evolutionary biology?. Biology and Philosophy, 19(5), 687-720.

Hayes, K. A., Cowie, R. H., & Thi en go, S. C. (2009). A global phylogeny of apple : Gondwanan origin, generic relationships, and the influence of outgroup choice (Caenogastropoda: Ampullariidae). Biological Journal of the Linnean Society, 98(1), 61-76.

Heath, T. A., Huelsenbeck, J. P., & Stadler, T. (2014). The fossilized birth-death process for coherent calibration of divergence-time estimates. Proceedings ofthe National Academy ofSciences , 111 (29), E2957-E2966.

12 Jin, Y. G., Wang, Y. , Wang, W., Shang, Q. H ., Cao, C. Q., & Erwin, D. H. (2000). Pattern of marine mass extinction near the Permian-Triassic boundary in South China. Science, 289(5478), 432-436.

Kameda, Y. , & Kato, M. (2011 ). Terrestrial invasion of pomatiopsid gastropods in the heavy-snow region of the Japanese Archipelago. BMC Evolutionary Biology, 11(1), 118.

Kenrick, P. , Wellman, C. H., Schneider, H. , & Edgecombe, G. D. (2012). A timeline for terrestrialization: consequences for the carbon cycle in the Palaeozoic Philosophical Transactions ofth e Royal Society of London B: Biological Sciences, 367(1588), 519-536.

Knoll, A.H., Bambach, R. K., Payne, J. L. , Pruss, S. , & Fischer, W.W. (2007). Paleophysiology and end-Permian mass extinction. Earth and Planetary Science Letters, 256(3), 295-313.

Kumar, S., Stecher, G. , & Tamura, K. (2016). MEGA?: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33, 1870-1874.

Larkin, M.A., Blackshields, G. , Brown, N. P. , Chenna, R., McGettigan, P.A., Mc William, H., Valentin, F., Wallace, I. M., Wilm, A. , Lopez, R., Thompson, J. D., Gibson T. J., & Higgins, D. G. (2007). Clustal Wand Clustal X version 2.0. Bioinformatics, 23(21), 2947-2948.

Quast, C., Pruesse, E. , Yilmaz, P., Gerken, J. , Schweer, T., Yarza, P., Peplies, J. , & Glockner, F. 0. (2012). The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Research, 41 (D 1) , D590-D596.

Rambaut, A. , Suchard, M.A., Xie, D., & Drummond, A.J. (2014) Tracer (Version 1.6) [Software]. Available from http://beast.bio.ed.ac.uk/Tracer

Richter, R. (2015). Die Evolution und Biogeographie der sudostasiatischen Sumpfdeckelschnecken (Viviparidae): ein molekularer und morphologischer Ansatz (Doctoral dissertation, Berlin, Humboldt Universitat zu Berlin, Diss., 2015).

Romero, P. E. , Pfenninger, M., Kano, Y. , & Klussmann-Kolb, A. (2016). Molecular phylogeny of the (Gastropoda: ) supports independent terrestrial invasions. Molecular Phylogenetics and Evolution, 97, 43-54.

Rota-Stabelli, 0., Daley, A. C., & Pisani, D. (2013). Molecular timetrees reveal a colonization of land and a new scenario for ecdysozoan evolution. Current Biology, 23(5), 392-398.

13 Rundell, R. J. (2008). Cryptic diversity, molecular phylogeny and biogeography of the rock-and leaf litter-dwelling land snails of Belau (Republic of Palau, Oceania). Philosophical Transactions ofthe Royal Society of London B: Biological Sciences, 363( 1508), 3401-3412.

Rundell, R. J. (2011 ). Snails on an evolutionary tree: Gulick, speciation, and isolation. American Malacological Bulletin, 29( 1/ 2), 145-157.

Rundell, R. J., & Price, T. D. (2009). Adaptive radiation, nonadaptive radiation, ecological speciation and nonecological speciation. Trends in Ecology & Evolution, 24(7), 394-399.

Selden, P.A. (2005). Terrestrialization (-Devonian). In Encyclopedia of Life Sciences, 174-177. John Wiley & Sons.

Selden, P.A., & Edwards, D. (1989). Colonisation of the land. In K.C. Allen & D.E.G. Briggs ( eds.) Evolution and the fossil record, 122-152. London: Belhaven.

Shen, S. Z., Crowley, J. L., Wang, Y., Bowring, S. A., Erwin, D. H., Sadler, P. M., Cao, C., Rotham, D. H., Henderson, C. M., Ramezani, J. , Zhang, H., Shen, Y., Wang, X., Wang, W., Mu, L., Li, W., Tang, Y., Liu, X., Liu, L. , Zeng, Y. , Jian, Y., & Jin, Y.(2011). Calibrating the end-Permian mass extinction. Science, 334(6061 ), 1367-1372.

Stadler, T. (2010). Sampling-through-time in birth- death trees. Journal of Theoretical Biology, 267(3), 396-404.

Tracey, S., J. A. Todd & D. H. Erwin (1993). Mollusca: Gastropoda. In M. J. Benton (ed.), The Fossil Record, S.131-167. London: Chapman and Hall.

Vermeij, G. J., & Dudley, R. (2000). Why are there so few evolutionary transitions between aquatic and terrestrial ecosystems?. Biological Journal of the Linnean Society, 70(4), 541-554.

Webster, N. B., Van Dooren, T. J. , & Schilthuizen, M. (2012). Phylogenetic reconstruction and shell evolution of the Diplommatinidae (Gastropoda: Caenogastropoda). Molecular Phylogenetics and Evolution, 63 (3), 625-638.

14

Table 1. Seven fossils used to calibrate the phylogeny of extant Cyclophoroidea. In contrast to node-dating methods, the Fossilized Birth-Death model assigns fossils to a monophyletic clade and allows them to be positioned as a stem taxa to the clade or as a sampled ancestor of an extant taxa. The placement of fossils is referred to in the "FBD Clade Assignment" column.

S~ecies FBD Clade Assignment Age (m~a) Source Viviparus langtonensis Yiviparidae 170 Tracey et al., 1993 Ampullaria nipponica Ampullariidae 160 Tracey et al., 1993 Maillardinus sp. Cyclophoroidea 140 Bandel, 1993 Loriolina sp. Cyclophoroidea 140 Bandel, 1993 Diplommaloptychia sp. Cyclophoroidea 140 Bandel, 1993 Undescribed Diplommatinidae 100 J. Czekanski-Moir, personal communication, Feb. 2017 Undescribed C~clophoridae 100 J. Czekanski-Moir, personal communication, Feb. 2017

16 Table 2. Extant taxa and associated Gen Bank accession numbers used to generate the phylogeny of the Cyclophoroidea.

Classification Species 16s 18s 28s COI H3 Cyclophoroidea Megalostomatidae Acroptychia bathiei HM753491 HM753437 HM753380 HM753335 Megalostomatidae Acroptychia mi/Ioli HM753492 HM75 3438 HM7533 8I HM753336 Cyclophoridae cf kelantanensis HM753481 HM753427 HM75337I HM753273 Cyclophoridae everetl i HM753480 HM75 3426 HM7533 70 HM753329 HM753272 Cyclophoridae Opisthoporus birostris HM753488 HM753434 HM753377 HM753333 HM753279 Cyclophoridae sp. nov. A MS 20 I 0 HM753486 HM753432 HM7533 76 HM753332 HM753277 Cyclophoridae latus HM753484 HM75 3430 HM753374 HM75333 l HM753275 Cochlostomatinae elegans HM753489 HM753435 HM7533 78 HM753334 HM753280 Cochlostomatinae Cochlosloma roseoli HM753490 HM753436 HM753281 Coch lostomati nae Cochlostoma septemspirale HM753497 HM753423 HM753367 HM753326 HM753269 Diplomatininae Palaina albata HM753531 HM753469 HM753409 HM753354 HM753312 l-funge1fordia sp. nov. A MS Diplomatininae 2010 HM753527 HM753463 HM753405 HM75 3352 HM753306 Diplomatininae .fraternum HM753496 HM753422 HM753366 HM753325 HM753268 Diplomatininae rubicunda HM753520 HM75 3477 HM753417 HM753363 HM753320 Diplomatininae Dip/ommatina suratensis HM753499 HM753425 HM753369 HM753328 HM753271 Diplomatininae DipLommatina demorgani HM753507 HM753448 HM75339I HM753340 HM753291 Aperostoma palmeri DQ093479.1 DQ093435 DQ093458 DQ093523. I DQ093514. I Pupinidae hosei HM753493. I HM75 3439 HM753382 HM753282. I Pupinidae Pupine//a swinhoei HM753494. I HM753440 HM753383 HM753283. I Ampullarioidea Ampullariidae KC 109970.1 DQ916521 HM003648 DQ916496 DQ916445 Ampullariidae Pila po/ita FJ710220.l FJ710256 EU27453 1 FJ7 I 0304. I EU274513 Campanilidae Campanile symbolicum AYOIOS07 DQ916524 DQ916548 AY296828 AF033683 Cerithioidea HQ833974. 1 KC904730 FJ606988 JF693416. I HQ834125. I Conoidea FJ868145. I DQ916538.1 DQ916564 AY588202. I AF033684.2 Yiviparoidea Yiviparidae Bel Iamya jeffreysi FJ405690.1 FJ405649 FJ405582 FJ405865. I FJ405763. l Viviparidae Angulyagra sp. / AJ 2009 FJ405723. 1 FJ405675 FJ405628 FJ405879. I FJ405769.I

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Cycklphoruslatus ~ Os,11thopo1u s bH0SfnS A.l ye,eu, ci kelanllrwnlil MSZOIO Cyclophoroidea I I I I -·- Chllmatrcaeus,..,.etti JoPo'N ,p rKW A MS 2010

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aleozoic 1- ":Mesozoic j Cenozoic

Figure 1. The maximum ciade credibility tree of the Cyclophoroidea based on five loci (16s, 18s, 28s, COi, H3). Time calibration was carried out using the Fossilized Birth-Death process and seven fossil calibration points. Node labels indicate the age of the node in billions of years and node bars indicate 95% Cl. The Cyclophoroidea diverged from the rest of the Caenogastropoda in the mid Carboniferous to mid Permian. The crown Cyclophoroidea are Permain to Triassic in age, centered close to the End Permian mass extinction.

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