Title In search of phylogenetic congruence between molecular and morphological data in bryozoans with extreme adult skeletal heteromorphy

Authors Taylor, PD; Waeschenbach, A; Smith, AM; Gordon, DP

Description peerreview_statement: The publishing and review policy for this title is described in its Aims & Scope. aims_and_scope_url: http://www.tandfonline.com/action/journalInformation? show=aimsScope&journalCode=tsab20

Date Submitted 2016-03-24 Systematics and Biodiversity

In search of phylogenetic congruence between molecular and morphological data in bryozoans with extreme adult skeletal heteromorphy

Journal: Systematics and Biodiversity

Manuscript ID: TSAB-2015-0002.R1

Manuscript Type: Original Research Article

Bryozoa, Cyclostomata, early astogeny, molecular phylogeny, morphology, Keywords: New Zealand, taxonomy

URL: http://mc.manuscriptcentral.com/tsab Page 1 of 48 Systematics and Biodiversity

1 2 3 4 5 6 In search of phylogenetic congruence between molecular and 7 8 morphological data in bryozoans with extreme adult skeletal 9 10 heteromorphy 11 12 1 2 3 13 PAUL D. TAYLOR , ANDREA WAESCHENBACH , ABIGAIL M. SMITH & 14 DENNIS P. GORDON4 15 16 17 1Department of Earth Sciences, Natural History Museum, London SW7 5BD 18 2 19 Department of Life Sciences, Natural History Museum, London SW7 5BD 20 3Department of Marine Science, University of Otago, Dunedin 9054, New Zealand 21 4 22 NIWA, Greta Point, P.O. Box 14901, Wellington, New Zealand 23 24 25 The taxonomy of cyclostome bryozoans, both Recent and fossil, is founded almost 26 entirely on characters of the mineralized skeleton. However, the adequacy of these 27 28 characters is now being questioned by molecular sequence data. In this study we 29 construct a molecular tree using ssrDNA and lsrDNA sequences and identify a clade 30 of New Zealand cyclostomes containing species exhibiting widely different 31 32 morphologies. In particular, Diaperoecia purpurascens (Hutton), a species assigned 33 to the suborder Tubuliporina on the basis of adult skeletal morphology, is shown to be 34 35 closely related to New Zealand species assigned to Heteropora, including H. 36 neozelanica Busk, which has a very different adult skeleton and is traditionally placed 37 38 in the suborder Cerioporina. A new species resembling the Antarctic genus 39 Hastingsia, ‘H’. whitteni sp. nov., from North Island, New Zealand, is found to 40 41 belong to the same clade, despite being placed conventionally in a different family 42 (Hastingsiidae) from both Diaperoecia (Diaperoeciidae) and Heteropora 43 44 (Cerioporidae). These results challenge the utility of adult skeletal morphology in 45 cyclostome taxonomy. In contrast to the striking dissimilarity between the adult 46 47 skeletons of D. purpurascens and ‘Hastingsia’ whitteni compared to New Zealand 48 species of Heteropora, morphological similarities in early colony development, as 49 50 well possibly as the presence of a gizzard, corroborate the molecular interpretation of 51 their close relationships. Greater attention should be paid in the future to early 52 53 astogenetic characters in cyclostome taxonomy. 54 55 56 57 58 1 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 2 of 48

1 2 3 4 5 6 Key words: Bryozoa, Cyclostomata, early astogeny, molecular phylogeny, 7 morphology, New Zealand, taxonomy 8 9 10 11 Introduction 12 A recurrent topic of discussion in taxonomy is the reliability of morphology in 13 14 interpreting phylogenetic relationships and hence in devising ‘natural’ classifications. 15 Molecular sequence data provide a powerful means of accessing the efficacy of 16 17 morphology for this purpose. In the ideal scenario there is a high degree of 18 congruence between molecules and morphology, corroborating the taxonomic value 19 20 of morphology. However, there are numerous instances where molecular sequence 21 data have undermined the taxonomic value of traditional morphological characters by 22 23 revealing close genetic relationships between morphologically distinct taxa (e.g. 24 Blackwell et al., 2003; Sotiaux et al., 2009; Pérez, 2011; Bourret et al., 2012). 25 26 Bryozoa is a phylum of lophotrochozoan invertebrates, distributed globally 27 across many aquatic habitats, and moderately diverse, comprising some 5689 living 28 29 species (Bock & Gordon, 2013). The two dominant orders of modern bryozoans – 30 Cheilostomata and Cyclostomata – are both marine and have calcareous skeletons that 31 furnish nearly all of the characters used in their taxonomy. Not only are these skeletal 32 33 characters immediately obvious in preserved material, but exactly the same characters 34 are available to palaeontologists studying fossil cheilostomes and cyclostomes. 35 36 Indeed, a common procedure in taxonomic studies of cheilostomes and cyclostomes is 37 to soak specimens in dilute bleach to remove all soft tissues, leaving only the 38 39 calcareous skeleton for identification. 40 A reasonably large number of skeletal morphological characters are available 41 42 for cheilostome taxonomy and molecular studies at low taxonomic levels generally 43 show good congruence with morphology (e.g., Hughes et al., 2008; HerreraCubilla 44 45 & Jackson, 2014), even though morphological convergence is evident between some 46 higher taxonomic groups (Waeschenbach et al., 2012). Cyclostomes, however, have a 47 48 simpler skeletal morphology and there can be significant incongruence between 49 molecules and morphology, as is apparent from mismatches when traditional 50 51 suborders founded on skeletal morphology are mapped onto molecular trees 52 (Waeschenbach et al., 2009; Taylor et al., 2011). 53 54 55 56 57 58 2 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 3 of 48 Systematics and Biodiversity

1 2 3 4 5 6 The purpose of this study is to use molecular sequence data to construct a 7 phylogenetic tree for cyclostome bryozoans and to focus particularly on the 8 9 unexpected relationships found between some species from New Zealand. These 10 species are revised taxonomically, based on traditional skeletal characters, and 11 12 morphological apomorphies supporting the molecular phylogeny are sought. One of 13 the commonest cyclostome bryozoans found in New Zealand waters is Diaperoecia 14 15 purpurascens (Hutton, 1877) (Fig. 1). Although the generic attribution of this species 16 has varied through time, with others placing it in Entalophora or Pustulopora, it has 17 18 always been considered as belonging to suborder Tubuliporina based on skeletal 19 morphology. We here generate ssrDNA and lsrDNA sequences for Diaperoecia 20 21 purpurascens for the first time, showing that the species groups within a clade 22 containing New Zealand material attributed to Heteropora (Fig. 2). Traditional 23 24 cyclostome taxonomy places Heteropora in a different suborder – Cerioporina – 25 reflecting the striking differences in the morphology of the zooidal skeletons of 26 27 Diaperoecia and Heteropora. However, our molecular findings are corroborated by a 28 study of skeletal characters from early colony development, notably the shape and 29 30 pattern of the pseudopores on the protoecium, which support a close relationship 31 among D. purpurascens, the New Zealand species of Heteropora and a previously un 32 33 named New Zealand species of ‘Hastingsia’ which molecular sequence data group 34 with these other two cyclostomes. Our findings point to the potential taxonomic value 35 36 of morphological characters seen in the early colony development of cyclostome 37 bryozoans. 38 39 40 Material and methods 41 42 43 Morphology 44 45 The material used for this study comprises historical specimens preserved in the 46 collections of the Natural History Museum, London (NHMUK), as well as some more 47 48 recently collected material from New Zealand which is now divided between the 49 collections of the NHMUK and the National Institute for Water and Atmospheric 50 51 Research (NIWA) in Wellington, New Zealand. 52 SEM of skeletal morphology was undertaken using LEO 1455VP and FEI 53 54 Quanta 650 ESEM scanning electron microscopes at the NHMUK. These instruments 55 56 57 58 3 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 4 of 48

1 2 3 4 5 6 allowed the study of uncoated, dried and bleached specimens imaged with back 7 scattered electrons. 8 9 10 Molecular study 11 12 Total genomic DNA was extracted from ethanolpreserved specimens using the 13 DNeasy tissue kit (QIAGEN) following the manufacturer’s instructions. lsrDNA was 14 15 amplified in two fragments using Steno250F + Steno1800R and Steno1800F + 16 Steno3490R (Waeschenbach et al., 2009). ssrDNA was amplified in two fragments 17 18 using 18e (Hillis & Dixon, 1991) + Stenolae600R (Waeschenbach et al. 2009) and 19 Stenolae600F (Waeschenbach et al., 2009) + 18p (Hillis & Dixon, 1991) (for PCR 20 21 cycling conditions, primer sequences and additional sequencing primer details, see 22 Waeschenbach et al., 2009). PCR products were purified using QIAquick PCR 23 24 purification Kit (QIAGEN). Sequencing was carried out on an Applied Biosystems 25 3730 DNA Analyser, using Big Dye version 1.1. Sequence identity was checked 26 27 using the Basic Local Alignment Search Tool (BLAST) (www.ncbi.nih.gov/BLAST/). 28 Contigs were assembled using SEQUENCHER 4.8 (GeneCodes Corporation). 29 30 Newly generated lsrDNA and ssrDNA sequences were aligned by eye to an 31 existing alignment, as published in the supplementary data for Waeschenbach et al. 32 33 (2009; see this reference for GenBank accession numbers of published sequences 34 used in the present study). GenBank accession numbers for data generated in this 35 36 study are as follows: lsrDNA ‘Hastingsia’ whitteni sp. nov. (KP331437), Diaperoecia 37 purpurascens (KP331438); ssrDNA ‘H.’ whitteni sp. nov. (KP331439), D. 38 39 purpurascens (KP331440),. The data were partitioned into three character sets: (1) 40 lsrDNA, (2) ssrDNA, and (3) lsr+ssrDNA. Modeltest (version 3.7macX, Posada and 41 42 Crandall, 1998) was used to select a model of evolution for the nucleotide data using 43 the Akaike Information Criterion. Phylogenetic trees were constructed using Bayesian 44 45 inference (BI) (MrBayes, version 3.2.1, Huelsenbeck & Ronquist, 2001). Likelihood 46 settings were set to nst=6, rates=invgamma, ngammacat=4 (equivalent to the 47 48 GTR+I+G model of evolution). In the combined lsr+ssrDNA analysis, parameters 49 were estimated separately for each gene. Two chains (temp = 0.2) were run for 50 51 5,000,000 generations and sampled every 1000 generations. Post‘burnin’ was 52 identified when the standard deviation of split frequency reached < 0.01. 2 million 53 and 4.5 million generations were discarded as burnin in the combined lsr+ssrDNA 54 55 analysis and single gene analyses, respectively. Nodes with < 0.95 posterior 56 57 58 4 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 5 of 48 Systematics and Biodiversity

1 2 3 4 5 6 probabilities (pp) were collapsed in all the figures. 7

8 9 Results 10 11 12 Taxonomy 13 14 15 Remarks. The three taxa forming the focus of this study are classified below 16 17 according to conventional morphologybased taxonomy and nomenclature. Changes 18 to this classification implied by the findings of the molecular phylogenetic analysis 19 20 can be found in the Discussion section at the end of the paper. 21 22 23 Suborder Tubuliporina Milne Edwards, 1838 24 Family Diaperoeciidae Canu, 1918 25 26 Genus Diaperoecia Canu, 1918 27 28 29 Type species. Pustulopora intricaria Busk, 1875; Recent, Australia. 30 Remarks. Several genera of tubuliporine cyclostome bryozoans, both living and 31 32 fossil, have ‘vinculariiform’ colonies comprising narrow bifurcating branches with 33 autozooids opening evenly around the entire circumference. They include 34 35 Entalophora Lamouroux, 1821, Pustulopora Blainville, 1830, Collapora Quenstedt, 36 1881, Diaperoecia Canu, 1918, Mecynoecia Canu, 1918, Entalophoroecia Harmelin, 37 38 1976 and Annectocyma Hayward & Ryland, 1985. With the exception of Entalophora, 39 which is characterized by having an axial lumen (Walter, 1970), characters of the 40 41 gonozooid provide the main means of distinction between these genera but are not 42 always clearcut and the taxonomy of these cyclostomes remains unclear. 43 44 Canu (1918, p. 329) diagnosed Diaperoecia thus: “L’ovicelle est elliptique ou 45 suborbiculaire; elle entoure les péristomes d’un assez grand nombre de tubes qui ne 46 47 sont pas dérangés dans leurs positions respectives. L’oeciostome isolé et subcentral.” 48 (The gonozooid is elliptical or suborbicular; it surrounds the peristomes [of the 49 50 autozooids] a fairly large number of which are not disturbed in their respective (?) 51 positions. The ooeciostome is isolated and subcentral.) 52 53 Brood (1976) redescribed Busk’s (1875) type specimens of Pustulopora 54 intricaria, noting that the material subsequently placed in this species by Harmer 55 56 57 58 5 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 6 of 48

1 2 3 4 5 6 (1915) from near Sorong in Indonesia was not conspecific. This is unfortunate 7 because Canu (1918) based his concept of Diaperoecia, and consequently on the 8 9 family Diaperoeciidae, on the gonozooids present in Harmer’s material, whereas 10 Busk’s types are infertile. Key skeletal characters visible in the Australian type 11 12 material of D. intricaria are illustrated in Figs 3–8. Flangelike kenozooidal 13 projections are developed on the sides of some branches (Figs 3, 4). The spacing of 14 15 apertures across the colony surface is irregular (Figs 3, 4), reflecting a high degree of 16 variability in the lengths of the autozooids. Peristomes are short and some apertures 17 18 are closed by terminal diaphragms (Fig. 5). Autozooidal frontal walls have closely 19 spaced, subcircular pseudopores (Fig. 6). Transversely fractured branches (Fig. 7) 20 21 show the presence of small buds near the branch axis and the triplelayered interior 22 walls illustrated in thin section by Brood (1976, fig. 1C). Mural spines are present, 23 24 some with a shaft and a barbed head (Fig. 8). A similar suite of skeletal characters is 25 evident in Pustulopora purpurascens (Hutton, 1877) and this species from New 26 27 Zealand can therefore be assigned on morphological grounds without reservation to 28 Diaperoecia. 29 30 31 Diaperoecia purpurascens (Hutton, 1877) 32 33 (Figs 1, 9–20, 47–49, 53–55) 34 35 36 Pustulipora [sic] purpurascens Hutton, 1877: 361. Pustulipora [sic] purpurascens. – 37 Hutton, 1880: 198. Entalophora purpurascens. – Hutton, 1891: 107. Diaperoecia 38 39 purpurascens. – Taylor et al., 2004: 57, fig. 2A, B. Diaperoecia purpurascens. – 40 Taylor et al., 2007: 220, fig. 1. Diaperoecia purpurascens. – Gordon et al., 2009: 293. 41 42 Diaperoecia purpurascens. – Smith & Gordon 2011: 36. 43 44 45 Material. NHMUK 2010.6.25.2 (Fig. 1), 75.1.5.33, 75.1.5.38, 99.7.1.4060–1 (Busk 46 Collection), all from unknown localities in New Zealand. NHMUK 2014.12.17.2 47 48 (Figs 9–12, 16), East Ulva Island, Paterson Inlet, Stewart Island, 8/6/1995, D. Foster 49 and A. M. Smith Collection. NHMUK 2017.12.17.3 (Figs 13, 14), 2017.12.17.4 (Fig. 50 51 15), craypot at Waitangi, Chatham Island, 4/11/1999, P. D. Taylor Collection. 52 NHMUK 2017.12.17.5 (Figs 17, 18), molecular voucher specimen AW578, 46.93°S; 53 168.16°E, 39 m, Otago Shelf, New Zealand, A. M. Smith Collection, 25/1/2010. 54 55 NHMUK 90.10.25.3, Wanganui. National Museum of New Zealand BS 862 (Figs 19, 56 57 58 6 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 7 of 48 Systematics and Biodiversity

1 2 3 4 5 6 47–49), outside Whangaroa Harbour, 25 m, 26/1/1981. NHMUK BZ 7733 (Fig. 20), 7 Pleistocene, Nukumaruan, Nukumaru Brown Sand, Nukumaru Beach, 01/2014, P. D. 8 9 Taylor Collection. 10

11 12 Description. Colony erect, bushlike (Fig. 1), up to 8 cm in diameter, consisting of 13 narrow, bifurcating branches; branch diameter 0.71–2.20 mm, typically 1.0–1.5 mm 14 15 (see Taylor et al., 2007, table 1). Colour in life creamybrown, purplishbrown post 16 mortem. All zooidal polymorphs fixedwalled, developing pseudoporous frontal walls 17 18 away from branch growth tips. Autozooids elongate, frontal walls ranging in length 19 from 0.65–2.24 mm (typically about 1 mm) by in width 0.19–0.48 mm (typically 0.25 20 21 mm), crossed by concave growth lines, zooidal boundaries slightly salient (Figs 9, 22 10); pseudopores teardropshaped, pointed distally (Fig. 11); apertures subcircular, 23 24 about 0.14–0.19 mm in diameter, disposed around entire branch circumference, 25 sometimes almost quincuncially arranged but in other cases unevenly spaced and 26 27 occasionally clustered into small groups; peristomes short; terminal diaphragms 28 closing older zooids (Fig. 18), sparsely to densely pseudoporous. Kenozooids 29 30 infrequent. Gonozooids longitudinally elongate (Figs 13–15), >4 mm in total length 31 by 0.6–0.8 mm wide, inconspicuous, the frontal wall only slightly convex, extending 32 33 distally of ooeciopore typically as lobes in daughter branches following bifurcation, 34 lateral edges indented by neighbouring autozooids but roof seldom pierced by 35 36 autozooids; pseudopore density higher than in autozooids; ooeciopore subcircular 37 (Fig. 14), slightly smaller than neighbouring autozooidal apertures, about 0.16 mm in 38 39 diameter, located nonterminally; ooeciostome very short. 40 Early astogeny observed in two colonies, one recent (Fig. 19), the other an 41 42 Early Pleistocene fossil (Fig. 20). Fanlike encrusting base giving rise to three erect 43 stems in both cases. Ancestrula (Figs 53–55) curved to the right, 0.83–1.09 mm long, 44 45 with longitudinally elliptical aperture 0.16–0.19 mm long by 0.14–0.17 mm wide, 46 occluded by a terminal diaphragm in the recent example but open in the fossil. 47 48 Protoecium large, 0.33–0.37 mm in transverse diameter; pseudopores confined to a 49 crescentic marginal band, closely but irregularly spaced, subcircular, countersunk, 50 51 external diameter c. 12 µm, internal diameter c. 6 µm. Nonpseudoporous distal part 52 of protoecium elevated and with chevronshaped transition to ancestrular tube which 53 has a rugose surface with noncountersunk pseudopores. 54 55 56 57 58 7 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 8 of 48

1 2 3 4 5 6 Mural spines developed on surfaces of interior walls and internal surfaces of 7 frontal exterior walls, with a moderately long shaft a head covered by sharp barbs, 8 9 either symmetrically or more often only on the side facing proximally. Distal edges of 10 interior walls exhibiting transversely fibrous ultrastructure. 11 12 13 Remarks. The whereabouts of Hutton’s (1877) material of this species is unknown. 14 15 Although it is known that he sent many of his samples to the then British Museum 16 (Natural History), none could be found appropriately labelled as having originated 17 18 from F. W. Hutton. Given that there is no controversy about the identity of this 19 species, there is currently no justification for erecting a neotype. 20 21 22 Distribution. Widespread in the seas around New Zealand at the presentday, this 23 24 species also occurs as a fossil back to at least the Late Pliocene and is especially 25 abundant in the Pleistocene Tainui Shell Bed of Wanganui (PDT, pers obs). Taylor et 26 27 al. (2004, fig. 4C) showed the distribution of D. purpurascens based on 137 NZOI 28 station samples, ranging in latitude from 33–54 °S, and in depth from 0–1156 m, 29 30 although material from the deeper stations may have been transported downslope and 31 90% of records are shallower than 250 m. 32 33 34 35 36 Family Hastingsiidae Borg, 1944 37 Genus Hastingsia Borg, 1944 38 39 40 Type species. Hastingsia irregularis Borg, 1944; Recent, Antarctica. 41 42 Remarks. Borg (1944) assigned three new species to his new genus Hastingsia which 43 he placed in the new monogeneric family Hastingsiidae. He remarked on how the 44 45 autozooids could open singly or in fascicles on the frontal sides of the branches, with 46 gonozooids located in branch axils. All three of the species assigned to Hastingsia 47 48 were recorded by Borg from Antarctica, although one – H. gracilis (MacGillivray, 49 1883) – was originally described from Victoria, Australia. 50 51 52 ‘Hastingsia’ whitteni sp. nov. 53 (Figs 21–26, 56–58) 54 55 56 57 58 8 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 9 of 48 Systematics and Biodiversity

1 2 3 4 5 6 Idmonea giebeliana Stoliczka – Hutton, 1873: 102 [non Idmonea giebeliana 7 Stoliczka, 1865]. Fasciculipora ramosa d’Orbigny – Gordon, 1967: 63, fig. 45 [non 8 9 d’Orbigny, 1842]. Fasciculipora ramosa – Morton & Miller, 1968: 227, 575, figs 76, 10 214 [non d’Orbigny, 1842]. Hastingsia maoriana Whitten MS 1979: 379, pl. 22, figs 11 12 4, 5. Hastingsia n. sp. 2 – Gordon et al. 2009: 293. Hastingsia cf. ‘giebeliana’ 13 Stolickza – Gordon et al. 2009: 293 [non Idmonea giebeliana Stoliczka, 1865]. 14 15 16 Etymology. Named for Ronald F. Whitten who first recognised this as a new species 17 18 and gave description of it in his unpublished PhD thesis (Whitten, 1979). 19 20 21 Material. Holotype: NIWA 98120 (Fig. 23), Auckland, on Sargassum sinclairii, 22 3/1/1996, D. P. Gordon Collection. Paratypes: NIWA 98121 (Figs 24, 56–58), details 23 24 as for holotype. NHMUK 2017.12.17.6 (Fig. 21), 2017.12.17.7 (Figs 22, 25), 25 2017.12.17.8 (Fig. 26), NZOI Station KAH1206/69, S39.9857°, E174.1988°, 44 m, 26 27 20/4/2012, molecular voucher specimen AW757. 28 29 30 Description. Colony encrusting (Fig. 23), becoming semierect (Fig. 26), small, 31 adnate branches bifurcating; pale brownishcream when alive, purplishbrown post 32 33 mortem. All zooidal polymorphs fixedwalled, developing pseudoporous frontal walls 34 away from branch growth tips. Autozooids elongate, frontal walls rugose with 35 36 moderately developed growth lines (Fig. 25), zooidal boundaries grooved slightly, 37 pseudopores subcircular, density decreasing into peristomes; apertures subcircular to 38 39 rounded quadrate when connate, about 0.18 mm in diameter, usually clustered into 40 small groups (Figs 21, 23, 25, 26) but sometimes isolated; peristomes long, up to at 41 42 least 1 mm (Fig. 25); terminal diaphragms not observed. Kenozooids not observed. 43 Gonozooids appearing in early astogeny, longitudinally elongate, at least 1.5 mm in 44 45 total length by 1.2 mm wide, inconspicuous, the frontal wall only slightly convex, 46 boundaries illdefined (Fig. 21), enclosing some autozooidal peristomes; pseudopore 47 48 density slightly higher than in the autozooids; ooeciopore not identified but a small 49 aperture, 0.10 by 0.13 mm, in the roof of one gonozooid may be an ooeciopore. 50 51 Ancestrula large (Figs 21, 23, 25, 56–58), only a little smaller than later 52 budded autozooids, 0.79–0.82 mm long, with subcircular aperture 0.16–0.18 mm in 53 diameter. Protoecium large, 0.39–0.44 mm in transverse diameter; pseudopores 54 55 confined to a crescentic marginal band, irregularly spaced, subcircular, countersunk, 56 57 58 9 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 10 of 48

1 2 3 4 5 6 external diameter c. 12 µm, internal diameter c. 6 µm. Ushaped transition to 7 ancestrular tube which has a rugose surface and noncountersunk pseudopores. 8 9 Mural spines present in autozooids, including the ancestrula, and gonozooids 10 (Fig. 24); branched (in brood chambers) or unbranched with barbed heads (Fig. 22). 11 12 Transversely fibrous ultrastructure visible at wall growing edges. 13

14 15 Remarks. This new species differs from the type species of Hastingsia, H. irregularis 16 Borg, 1944), in having larger autozooids and gonozooids, and in not developing 17 18 extensive erect growth. The linear fascicles depicted by Bock 19 (http://www.bryozoa.net/cyclostomata/hastingsiidae/hastgra.html) in H. gracilis 20 21 (MacGillivray, 1884) have no equivalent in ‘H’. whitteni sp. nov., and the gonozooid 22 in this Australian species is ovoidal with a strongly compressed ooeciopore, 23 24 contrasting with the new species. The tiny gonozooid of the erect H. pygmaea Borg, 25 1944, the only other species assigned to Hastingsia, contrasts with the voluminous 26 27 gonozooid of H. whitteni sp. nov. 28 This is the first formally published record of Hastingsia in New Zealand, 29 30 although Whitten (1979) referred an undescribed species to this genus in his 31 unpublished PhD thesis. In addition, three fragments from New Zealand donated to 32 33 the NHMUK collection by F.W. Hutton (NHMUK 75.1.5.35) and labelled by Hutton 34 Idmonea giebeliana Stoliczka, 1865 can be assigned to ‘H’. whitteni. In addition, 35 36 ‘Fasciculate sp. 2’ of Boardman (1998, p. 29, fig. 75) from Leigh, and the unnamed 37 fasciculate cyclostome also from New Zealand figured by Taylor (2000, fig. 6), are 38 39 both apparently this species. 40 Material in the NIWA collections identified tentatively as the type species of 41 42 Hastingsia, H. irregularis, includes a broken protoecium which seemingly lacks 43 pseudopores and therefore differs significantly from that of ‘H’. whitteni, 44 45 underscoring the problem of assigning the New Zealand species to a genus. 46 47 48 Distribution. Northeastern North Island, New Zealand, Auckland to Whangarei, 49 including Hauraki Gulf from where Whitten (1979) obtained his material. 50 51 52 Suborder Cerioporina von Hagenow, 1851 53 Family Cerioporidae Busk, 1859 54 55 Genus Heteropora de Blainville, 1830 56 57 58 10 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 11 of 48 Systematics and Biodiversity

1 2 3 4 5 6 7 Type species. Ceriopora cryptopora Goldfuss, 1826; Cretaceous, Maastrichtian, The 8 9 Netherlands. 10

11 12 Remarks. There are unresolved problems over the concept of the genus Heteropora 13 owing to the fact that the Cretaceous type species (see Nye, 1976) differs substantially 14 15 from the extant species, including H. neozelanica Busk, 1879 described below, which 16 have been assigned to the genus. One solution (e.g., Gordon et al., 2009) has been to 17 18 refer these recent species to another cerioporid genus Tetrocycloecia Canu, 1917, with 19 a Miocene type species (see Taylor & McKinney, 2006, p. 51) but this too may not be 20 21 congeneric with the species described below. 22 23 24 ‘Heteropora’ neozelanica Busk, 1879 25 (Figs 2, 27–45, 50–52, 59–61) 26 27 28 Heteropora neozelanica Busk, 1879: 724, pl. 15, figs 1–4. Heteropora neozelanica. – 29 30 Nicholson, 1880: 329, figs 1A–C, 2. Heteropora pelliculata Waters. – Hutton, 1891: 31 107. Heteropora neozelanica. – Borg, 1933: 306, textfigs 12–14, pl. 5, figs 1–5, pl. 32 33 6, figs 3, 4, pl. 7, figs 1–3, pl. 10, figs 1, 2. Heteropora neozelanica. – Borg, 1944: 34 210, pl. 16, fig. 1. Tetrocycloecia neozelanica. – Gordon et al., 2009: 293. 35 36 Tetrocycloecia spp. – Smith & Gordon 2011: 31. 37 38 39 Material. Lectotype (chosen here): NHMUK 99.7.1.4281 (Figs 27, 28, 30, 31), New 40 Zealand. It is evident that Busk (1879) had more than one specimen at his disposal 41 42 when he erected Heteropora neozelanica as he remarked that Nicholson had furnished 43 him with some very fine specimens of this species. However, his figures (Busk, 1879, 44 45 pl. 15, figs 1–4) could conceivably be from a single specimen. Busk’s figure 1, which 46 is reproduced here as Fig. 27, depicts an intact ramose colony. Unfortunately, it is not 47 48 possible to match this or the any of the higher magnifications views (pl. 15, figs 2–4) 49 with material labelled as ‘type’ in the Busk Collection at the NHMUK and it seems 50 51 possible that the intact colony is either lost or has been brokenup into the fragments 52 that are now mounted on slides. One of these (Fig. 28, 30, 31) is herein chosen as the 53 lectotype of ‘H’. neozelanica. Neither Busk (1879) nor Nicholson (1880) gave any 54 55 more detailed information than simply New Zealand. 56 57 58 11 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 12 of 48

1 2 3 4 5 6 Paralectotype: NHMUK 99.7.1.4280 (Figs 29, 32), Nicholson Collection. 7 Other material: NHMUK 2014.12.17.1 (Figs 2, 33, 36, 40–45, 50–52), Otago Shelf, 8 9 Stn Mu8829, 87–89 m, 11/5/1988, P. D. Taylor Collection. NHMUK 2014.12.17.9 10 (Figs 34, 37), molecular voucher specimen AW086, 46.42°S, 167.58°E, 54 m, The 11 12 Snares, New Zealand, A. M. Smith & J. Porter Collection, February 2008. NIWA 13 98122 (Figs 59–61), NZOI Stn Z9684, 34°23.55’S, 172°51.72’E, 40 m, 26/1/1999. 14 15 Comparative material (Heteropora sp.): NHMUK 2014.12.17.10 (Figs 35, 38), 16 molecular voucher specimen NZ064, 47.08°S, 168.12°E, 94 m, The Snares, New 17 18 Zealand, A. M. Smith & J. Porter Collection, 29/1/2008. 19

20 21 Description. Colony erect, bushlike (Figs 2, 27), 7 cm or more in diameter, 22 consisting of bifurcating branches. Pale yellow to pale brownishyellow in life, pale 23 24 creamy brown to purplishbrown postmortem. Branch diameter variable between 25 colonies, ranging from 0.7–2.2 mm, most often 2.0–6.0 mm, increasing modestly 26 27 from distal branch tips to branch bases, mean value variable between colonies. 28 Zooidal polymorphs predominantly freewalled, apart from gonozooids, which have 29 30 fixedwalled brood chamber roofs (Figs 42, 43), zooids from early astogeny, and old 31 branches and occasional patches of autozooids with short peristomes of exterior wall 32 33 and kenozooids closed by terminal diaphragms (Figs 39, 40, 43–45). Distinction 34 between autozooids and kenozooids sometimes obscure (Fig. 36). Autozooidal 35 36 apertures subcircular, about 0.19–0.24 mm in diameter, rim often slightly raised; 37 exteriorwalled peristomes rarely developed, sparsely pseudoporous, short, <1 mm 38 39 high (Figs 40, 44, 45). Basal diaphragms present close to endozone/exozone boundary 40 in some autozooids (Fig. 32). Kenozooids more numerous than the autozooids which 41 42 they surround entirely, apertures rounded polygonal, typically smaller and more 43 variable in diameter than autozooidal apertures, equidimensional or elongate, about 44 45 0.08–0.28 mm wide, thickwalled with sharp zooidal boundaries; some kenozooids 46 closed by terminal diaphragms accreted centripetally, with closelyspaced 47 48 pseudopores about 10 µm in size, often teardropshaped, located slightly to 49 appreciably proximally of apertural rims. Gonozooids subcircular (Fig. 42), variable 50 51 in size, 1.4–3 mm in diameter, roof bulging very slightly, formed of densely 52 pseudoporous exterior wall accreted centrifugally from the peristomes of autozooids 53 passing through brood chamber, margins becoming overgrown by surrounding 54 55 zooids; ooeciopore located about midway between centre and distal edge, 56 57 58 12 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 13 of 48 Systematics and Biodiversity

1 2 3 4 5 6 approximately the same size as an autozooidal aperture, compressed, 0.19 mm long 7 by 0.12 mm wide in the single example studied (Fig. 42); ooeciostome short, slightly 8 9 flared. 10 Early astogeny observed in two colonies, both from NZOI Station Z9684 in 11 12 Spirits Bay, northern New Zealand. Fanlike encrusting base giving rise to a single 13 erect stem. Ancestrula (Figs 59–61) 0.77–1.05 mm long, with subcircular aperture 14 15 0.17–0.18 mm long by 0.13–0.18 mm wide, occluded by a terminal diaphragm in one 16 example. Protoecium large, 0.38–0.46 mm in transverse diameter; pseudopores 17 18 confined to a crescentic marginal band, closely but irregularly spaced, subcircular, 19 countersunk, external diameter c. 10 µm, internal diameter c. 5 µm. Shallow U 20 21 shaped transition to ancestrular tube which has a rugose surface with noncountersunk 22 pseudopores. 23 24 Mural spines present in both autozooids and kenozooids, tending to be more 25 numerous and longer in the latter; varies in morphology, many with a smooth shaft 26 27 bearing a head covered by sharp barbs, longest around circumference of head and 28 downwardly curved; shaft sometimes dividing distally into two or three branches, 29 30 each with a barbed head; other mural spines with a single long barb directed 31 proximally and resembling the beak on a bird’s head. Distal edges of walls exhibit 32 33 transversely fibrous ultrastructure. 34 35 36 Remarks. Material provisionally assigned to this species varies widely in branch 37 diameter. Busk’s syntypes have branches 5–6 mm in diameter, whereas the branches 38 39 in colonies from the Otago Shelf vary from the same size (see colour photo in Smith 40 & Gordon, p. 31) to about half this size (e.g. Fig. 2). Other aspects of the skeletal 41 42 morphology, however, seem indistinguishable. In view of the depthrelated variability 43 in branch diameters between colonies of Heteropora pacifica Borg, 1933 from 44 45 Washington State described by Schopf et al. (1980), the possibility that branch 46 diameter in ‘H’. neozelanica also varies ecophenotypically cannot be excluded. 47 48 Therefore, ‘H’. neozelanica is here interpreted in a broad sense to encompass most 49 species of ‘Heteropora’ from New Zealand waters with widely ranging branch 50 51 diameters. 52 The molecular phylogenetic analysis below revealed more than one clade of 53 ‘Heteropora’ in New Zealand. Comprehensive research combining morphological 54 55 and molecular analyses of material from multiple localities around New Zealand will 56 57 58 13 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 14 of 48

1 2 3 4 5 6 be needed to unravel the complexity of this group of species. This is beyond the scope 7 of the current study. 8 9 Neither Busk (1879) nor Nicholson (1880) described the gonozooids of this 10 species. Borg (1933) too was unable to identify them with certainty in the material he 11 12 had available for study but he believed that some enlarged apertures visible on colony 13 surfaces were possibly ooeciopores of overgrown gonozooids. However, it is clear 14 15 from his figure (pl. 5, fig. 5) that these supposed ooeciopores are tubes formed by the 16 bryozoan around a symbiont such as a spionid polychaete (cf. Ernst et al., 2014). 17 18 Unequivocal examples of gonozooids are present in material of ‘H’. neozelanica from 19 the Otago Shelf. These show the exteriorwalled roof supported by autozooids passing 20 21 through the brood chamber (Figs 42, 43) and an ooeciopore of about the same size as 22 an autozooidal aperture but laterally compressed. 23 24 25 Distribution. Widely distributed in waters around New Zealand at the presentday, at 26 27 least from Spirits Bay in the north to Foveaux Strait in the south, as well as The 28 Snares. 29 30 31 Molecular phylogenetics 32 33 Figure 46 depicts the molecular phylogeny based on the concatenated dataset of 34 lsr+ssrDNA, in which the newly sequenced taxa ‘Hastingsia’ whitteni sp. nov. and 35 36 Diaperoecia purpurascens were analysed in the context of the previously published 37 cyclostome phylogenies of Waeschenbach et al. (2009) and Taylor et al. (2011). The 38 39 tubuliporine Annectocyma tubulosa forms the sister group to a clade composed of the 40 cerioporines ‘Heteropora’ sp. (molecular specimen NZ064), ‘Heteropora’ 41 42 neozelanica (molecular specimen AW086) and the tubuliporines ‘Hastingsia’ whitteni 43 sp. nov. (molecular specimen AW757) and Diaperoecia purpurascens (molecular 44 45 specimen AW578), where ‘Heteropora’ sp. forms the sister group to an unresolved 46 assemblage of the latter three taxa (Fig. 46). The same topology for this grouping was 47 48 obtained in analyses using the single gene partitions (Online Supplementary 49 Material), but the node that places ‘Heteropora’ sp. as the sister group to ‘Hastingsia’ 50 51 whitteni sp. nov. + ‘Heteropora’ neozelanica + Diaperoecia purpurascens is only 52 weakly supported in the lsrDNA only analysis (0.72 pp). 53

54 55 56 57 58 14 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 15 of 48 Systematics and Biodiversity

1 2 3 4 5 6 Discussion 7 8 9 The results of the molecular phylogenetic analysis, which is the most comprehensive 10 11 and uptodate for cyclostomes, are unexpected because they show a close 12 relationship between cyclostome bryozoans previously assigned to different suborders 13 14 based on morphological criteria. Whereas Heteropora is traditionally assigned to the 15 suborder Cerioporina, Diaperoecia and Hastingsia are placed in the suborder 16 17 Tubuliporina, although some classifications (e.g., Walter, 1970; Ryland, 1982) would 18 assign the latter genus to a third suborder – Fasciculina – not universally recognised. 19 20 There have been no previous suggestions in the literature that Heteropora might be 21 closely related to Diaperoecia and Hastingsia, and the only superficial similarity is 22 23 their pale yellowish to yellowishbrown colour in life and generally purplishbrown 24 postmortem pigmentation. But can this relationship be supported using 25 26 morphological characters? 27 The taxonomy of cyclostome bryozoans has been founded entirely on 28 29 characters of the mineralized skeleton. Skeletal characters are easily visible without 30 histological preparation, and are conserved in dried material as well as fossils. In 31 32 contrast, very little is known about the soft part anatomy of cyclostome bryozoans and 33 available data concerns only a small proportion of known species (see Boardman, 34 35 1998). Cyclostome suborders have been recognized mainly according to the types of 36 skeletal walls employed by the autozooids and gonozooids. 37 38 There are two basic types of skeletal walls in cyclostomes (and other 39 bryozoans): interior and exterior (see Taylor et al., 2014). Interior skeletal walls 40 41 partition body cavities and are secreted by an epithelium that is present on both sides 42 and wraps around the distal end of the wall. They lack a cuticular outer layer. Exterior 43 44 skeletal walls are located at the interface between the bryozoan and the environment, 45 and are secreted by an epithelium present on the body cavity side only, the opposite 46 47 side being covered by a layer of cuticle. When no frontal exterior walls are developed 48 a hypostegal cavity (coelom or pseudocoel) invests the colony, forming a continuous 49 50 layer that connects the body cavities of the zooids over the outer ends of their interior 51 walls. This skeletal organization was termed ‘doublewalled’ by Borg (1926), later 52 53 renamed ‘freewalled’. In contrast, cyclostomes with frontal exterior walls lack a 54 55 56 57 58 15 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 16 of 48

1 2 3 4 5 6 hypostegal cavity investing the colony, an organization termed ‘singlewalled’ by 7 Borg but now known as ‘fixedwalled’. 8 9 Two cyclostome suborders (Tubuliporina and Articulata) possess fixedwalled 10 autozooids and gonozooids (i.e., with exterior frontal walls), one order (Cerioporina) 11 12 has freewalled autozooids but fixedwalled gonozooids, while the other two 13 suborders (Rectangulata and Cancellata) have freewalled autozooids and gonozooids. 14 15 To Borg (1926) and many later authors, free vs. fixedwalled skeletal organizations 16 represented fundamental differences of high taxonomic value. Indeed, it has been 17 18 argued that modern freewalled cyclostomes are more closely related to various orders 19 of freewalled Palaeozoic bryozoans than they are to the fixedwalled cyclostomes 20 21 (e.g., Boardman, 1984; Viskova, 1992). 22 The value of skeletal organization in highlevel cyclostome taxonomy has 23 24 been challenged in recent years, firstly by the recognition of taxa showing mixtures of 25 the two organizational types, and subsequently by molecular phylogenetic evidence. 26 27 Taylor (2000) pointed to examples of mixing free and fixedwalled organizations in 28 seemingly monophyletic cyclostome taxa (e.g., Eleidae, Cinctiporidae), and even in 29 30 single colonies (e.g., Cinctipora elegans: see Boardman et al., 1992). A particularly 31 clear example is Spiritopora perplexa Taylor & Gordon, 2003, in which the zooids in 32 33 the extensive lamellar base of the colony are normally fixedwalled, whereas those 34 forming the erect fronds are predominantly freewalled. 35 36 Molecular phylogenetic studies of cyclostomes have recovered trees showing 37 no clear separation between free and fixedwalled species. Waeschenbach et al. 38 39 (2009) assembled a tree using sequence data from 22 species of cyclostomes, showing 40 poor congruence with traditional classifications based on skeletal organization; 41 42 transitions between freeand fixedwalled conditions, or viceversa, have apparently 43 occurred several times during cyclostome evolution. A subsequent molecular study 44 45 (Taylor et al., 2011) showed the dominantly freewalled cyclostome Tennysonia to 46 group with the fixedwalled tubuliporine Idmidronea, despite the additional difference 47 48 in the morphology of Tennysonia, which has kenozooids as spacers between the 49 autozooidal apertures. 50 51 In the light of this accumulating evidence for plasticity in skeletal 52 organization, it is perhaps unsurprising to find the fixedwalled tubuliporines 53 Diaperoecia purpurascens and ‘Hastingsia’ whitteni in the same molecular clade as 54 55 the freewalled cerioporine ‘Heteropora’ neozelanica. As with the clade containing 56 57 58 16 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 17 of 48 Systematics and Biodiversity

1 2 3 4 5 6 Idmidronea and Tennysonia, a further difference is apparent in the manner in which 7 the apertures of the autozooids – and hence the lophophores in living colonies – are 8 9 spaced apart. Areas of the colony surface between the autozooidal apertures of D. 10 purpurascens and ‘Hastingsia’ whitteni are occupied by the calcified exterior frontal 11 12 walls of the autozooids (e.g., Fig. 9), whereas in ‘Heteropora’ they are filled by 13 kenozooids with open apertures (e.g., Figs 36–38). These kenozooids are best 14 15 interpreted as ‘normal’ buds that failed to develop a polypide with a functional 16 lophophore. 17 18 It is also worth remarking on the various forms of frontal exterior wall 19 calcification that can be found in the predominantly freewalled ‘Heteropora’ 20 21 neozelanica. Aside from the fixedwalled gonozooids, which are found in other 22 cerioporine cyclostomes too, patches of exterior wall calcification are developed 23 24 elsewhere on colony surfaces. They are particularly common in the proximal parts of 25 branches (Figs 39, 40) but can also be seen in distal branches, for example adjacent to 26 27 gonozooids (Figs 42–44). The kenozooids in these patches are usually closed by 28 terminal diaphragms, either accreted distally almost as a lamina across the ends of 29 30 their walls, or more proximally, leaving the distal ends of the vertical kenozooidal 31 walls exposed above the diaphragms (Fig. 40). Autozooids in these patches often 32 33 develop short exteriorwalled peristomes with scattered pseudopores (Fig. 45). 34 Peristomes are also present in the autozooids passing through brood chambers and 35 36 supporting the roofs of the gonozooids (Figs 42, 43). 37 Three nominal species of Heteropora have been recorded from the Pacific 38 39 coast of North America. Ross (1973) described the skeletal morphology of 40 Heteropora colonies from Washington State, USA, noting the presence of what she 41 42 termed ‘collars’ around the aperture of autozooids proximal of the branch growing 43 tips. These collars are clearly exteriorwalled peristomes very similar to those seen in 44 45 New Zealand ‘Heteropora’. Fixedwalled growth, therefore, is more widely 46 distributed taxonomically in Recent species assigned to Heteropora than is generally 47 48 acknowledged. 49 50 51 Gizzard 52 Despite the paucity of information on cyclostome softpart anatomy, the molecular 53 tree recovered here (Fig. 46) is corroborated by at least one anatomical feature, the 54 55 presence of a gizzard. Gizzards are modified sections of stomach cardia with 56 57 58 17 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 18 of 48

1 2 3 4 5 6 microvillous plates or teeth (Gordon, 1975) and apparently function to allow 7 bryozoans to open diatom frustules (Markham & Ryland, 1987). They have been 8 9 reported in a relatively small number of bryozoans, including some ctenostome and 10 cheilostome gymnolaemates as well as four species of cyclostomes (Schäfer, 1986). 11 12 The cyclostome species having gizzards were identified as Diaperoecia major 13 (Johnston, 1847) and D. tubulosa (Busk, 1875) by Schäfer (1986), and as Pustulopora 14 15 purpurascens Hutton and ‘Fasciculate sp. 2’ by Boardman (1998). In all of these 16 cyclostomes the gizzard comprises two dental plates that act like jaws. The species 17 18 identified as Diaperoecia tubulosa is now Annectocyma tubulosa, a northern 19 hemisphere species recovered as the sistertaxon to the clade of New Zealand 20 21 cyclostomes described in the current paper. Boardman’s (1998) Pustulopora 22 purpurascens is Diaperoecia purpurascens, and his ‘Fasciculate sp. 2’, to judge from 23 24 fig. 75, is probably ‘Hastingsia’ whitteni sp. nov. Therefore, the clade including 25 Annectocyma tubulosa and all crownward taxa possess a gizzard as an apomorphic 26 27 character, although no histological data is yet available for New Zealand species of 28 Heteropora. The finding of a gizzard in ‘Heteropora’ would corroborate this 29 30 character as a cladedefining apomorphy. 31 32 33 Mural spines 34 Minute spines of uncertain function grow from the skeletal walls into the zooidal 35 36 chambers in many cyclostome taxa (Farmer, 1979; Taylor & Weedon, 2000 and 37 references therein). These zooecial or mural spines vary in morphology among 38 39 species, and also within taxa, both between polymorphs and within the same 40 polymorph. 41 42 Mural spines with long shafts supporting barbed heads covered by a few to a 43 moderate number of spinelets occur in the three related species described above, as 44 45 well as in the type species of Diaperoecia (Fig. 8). Most are unbranched (Figs 22, 47– 46 48, 52) but some bifurcate (Fig. 51) and have a barbed head on each branch. The long 47 48 spines of Heteropora neozelanica with globose densely barbed heads are also seen in 49 species of ‘Heteropora’ from the Pacific Coast of North America (Farmer, 1979, text 50 51 fig. 4; Weedon & Taylor, 1996, fig. 7e) and could be taxonomically diagnostic. 52 However, spines with flattened heads and a small number of spinelets (Figs 8, 47–49) 53 seem to be more taxonomically widespread. They occur, for example, in 54 55 lichenoporids (Farmer, 1979, pl. 1; Dick et al., 2006, fig. 16D) which are distant in 56 57 58 18 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 19 of 48 Systematics and Biodiversity

1 2 3 4 5 6 molecular trees (Waeschenbach et al., 2009, 2012) from the New Zealand 7 cyclostomes described here. 8 9 Unfortunately, too little is known about the morphological variants and 10 taxonomic distribution of mural spines for their potential in cyclostome taxonomy to 11 12 be evaluated. Additionally, they have little utility in fossil cyclostomes as they are too 13 fragile for routine preservation. 14 15 16 Early astogeny 17 18 Few taxonomic studies include information on the early astogeny of cyclostome 19 bryozoans. Nevertheless, Jenkins & Taylor (2014) considered early astogeny to be a 20 21 potentially important source of higherlevel taxonomic characters, given that early 22 development may be relatively conserved in evolution and can therefore provide 23 24 evidence of deep phylogenetic relationships among cyclostomes. Studying early 25 astogenetic stages in species with large erect colonies, including Diaperoecia 26 27 purpurascens and ‘Heteropora’, is difficult as the oldest parts of colonies may be lost 28 or obscured by subsequent growth of the colony or fouling organisms. However, it 29 30 has been possible to find examples of colonies preserving the ancestrula and other 31 early zooids in both of these species, as well as in the smaller colonies of ‘Hastingsia’ 32 33 whitteni. Contrary to the striking differences seen in the skeletal morphology of later 34 astogenetic stages in these three closely related species, their early astogeny is 35 36 remarkably similar. 37 Juvenile colonies of all three species are fanshaped encrustations comprising 38 39 fixedwalled autozooids with calcified exterior frontal walls (e.g., Figs 19, 20). One or 40 more erect stems grow upwards from the substrate from this encrusting base. In the 41 42 case of ‘Heteropora’, the zooids in the erect stems differ from those of the encrusting 43 base in being freewalled, but in Diaperoecia purpurascens and ‘Hastingsia’ whitteni 44 45 they have the same fixedwalled skeletal organization. 46 The greatest similarities are apparent in the ancestrula (Figs 53, 56, 59), and 47 48 particularly the protoecium, which is large in all three species: 0.33–0.37 mm wide in 49 D. purpurascens, 0.39–0.44 mm in ‘Hastingsia’ whitteni, and 0.38–0.46 mm in 50 51 ‘Heteropora’. These sizes compare with typical protoecial widths of 0.1–0.2 mm in 52 cyclostomes (e.g., Jenkins & Taylor, 2014). 53 Cyclostome protoecia exhibit interspecific variability in the number and 54 55 distribution of pseudopores (Weedon, 1998). All three species from New Zealand are 56 57 58 19 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 20 of 48

1 2 3 4 5 6 characterized by protoecia with numerous, somewhat irregularly arranged 7 pseudopores distributed in a crescentic band around the proximal and lateral edges of 8 9 the protoecium (Figs 54, 57, 60). Pseudopores are lacking from more distal parts of 10 the protoecium but reappear on the ancestrular tube. 11 12 Not only is the distribution of protoecial pseudopores very distinctive in 13 Diaperoecia purpurascens, ‘Hastingsia’ whitteni and ‘Heteropora’, but they are very 14 15 similar in size and shape. In all three species the pseudopores are circular to elliptical 16 and have a characteristic countersunk morphology (Figs 55, 58, 61), with an external 17 18 diameter of about 10–12 µm and an internal diameter of 5–6 µm. In contrast, 19 pseudopores on the ancestrular tube and later zooids are not countersunk. Weedon 20 21 (1998) described such pseudopores in a specimen from the Otago Shelf (New 22 Zealand) which he identified as Diaperoecia cf. purpurascens, noting that the 23 24 pseudopores are located not only at the boundaries between the strips of calcification 25 forming the protoecium but also, and unusually, within strips. 26 27 The ancestrula has not been described in Annectocyma tubulosa, the sister 28 taxon to the New Zealand clade forming the focus of the current study, and attempts 29 30 to find an example in the collections of the NHMUK have been unsuccessful. A 31 colony of the related A. major (Johnston, 1847), however, has a protoecium with a 32 33 nearmarginal band of countersunk pseudopores (Figs 62, 63). The corollary is that 34 this character may be apomorphic for a clade that includes not only the three New 35 36 Zealand species under study, but also some taxa such as Annectocyma from the 37 immediate outgroup. 38 39 Figure 642 summarises diagrammatically how the very similar skeletal 40 organization in the early astogenetic stages of Diaperoecia purpurascens and 41 42 ‘Heteropora’ changes in later astogeny. In D. purpurascens, the zooids in late 43 astogeny retain the same fixedwalled organization as those from early astogeny. 44 45 Only the growing tips of the erect branches are covered by hypostegal coelom, and 46 calcified exterior frontal walls of the autozooids form the branch surface between the 47 48 everted tentacle crowns. In contrast, in ‘Heteropora’, the zooids in late astogeny have 49 a freewalled organization, hypostegal coelom extends proximally from the branch 50 51 growth tips to cover the entire branch surface, and kenozooids function as spacers 52 between the everted tentacle crowns in the absence of calcified exterior frontal walls. 53 Transitions from one state to the other can be construed as heterochronous (see 54 55 Taylor, 2000). 56 57 58 20 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 21 of 48 Systematics and Biodiversity

1 2 3 4 5 6 7 Implications for cyclostome taxonomy 8 9 Our molecular phylogenetic analysis recovers, with strong support, a clade of 10 cyclostomes containing the heteromorphic assemblage of ‘Heteropora spp., 11 12 ‘Hastingsia’ whitteni and Diaperoecia purpurascens. While this clade at first seems 13 to have a scant morphological basis, it is apparently corroborated by at least one soft 14 15 part character – the presence of a gizzard – and by the skeletal morphology of the 16 ancestrula, with distinctive countersunk pseudopores arranged in a crescent on the 17 18 protoecium. These are not morphological characters previously applied in cyclostome 19 taxonomy. Therefore, it is unsurprising that the supraspecific classification of the taxa 20 21 studied here does not mirror their phylogenetic relationships. How then can these taxa 22 be reclassified and named to reflect phylogeny better? 23 24 Cyclostome families have been defined largely on characters of the 25 gonozooids since the work on F. Canu and R. S. Bassler in the early 20th century. 26 27 Indeed, Canu (1918) created Diaperoeciidae on the basis of the supposed morphology 28 of the gonozooid of Diaperoecia intricaria (Busk) (but see above), which he 29 30 described as elliptical or suborbicular, surrounding autozooidal apertures and with a 31 subcentral ooeciopore. While the gonozooids of Diaperoecia purpurascens, 32 33 ‘Hastingsia’ whitteni and ‘Heteropora’ could just about all be accommodated within 34 this diagnosis, there are major differences in structure of the gonozooids between the 35 36 first two species and ‘Heteropora’. Gonozooids of D. purpurascens and ‘H’. whitteni 37 have roofs that calcify in a distal direction, parallel to the frontal walls of the adjacent 38 39 autozooids. The roofs in ‘Heteropora’, however, calcify radially outwards from the 40 autozooids that penetrate the brood chamber. 41 42 Notwithstanding the problems posed by the lack of knowledge of gonozooid 43 morphology in the type species of Diaperoecia, and the substantial contrast in the 44 45 growth pattern of the gonozooids described above, the family Diaperoeciidae can be 46 used to accommodate the three species from New Zealand studied here if it is 47 48 redefined using other morphological characters, viz. the presence of a gizzard, and a 49 protoecium with a marginal band of countersunk pseudopores. As both of these 50 51 characters are also found in species of Annectocyma, this genus can also be included 52 in Diaperoeciidae. The family Annectocymidae Hayward & Ryland, 1985 then 53 becomes a junior synonym of Diaperoeciidae Canu, 1918. 54 55 56 57 58 21 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 22 of 48

1 2 3 4 5 6 More problematical are the most appropriate generic names for the species 7 here rererred to as ‘Hastingsia’ whitteni and ‘Heteropora’ spp. While it would be 8 9 justifiable to place all of these species in Diaperoecia given their close phylogenetic 10 affinities to D. purpurascens, this would conceal the strong morphological differences 11 12 between these taxa. On the other hand, the identities of both ‘Hastingsia’ whitteni and 13 ‘Heteropora’ require clarification through detailed studies of their generic type 14 15 species, including early astogeny and, in the case of the extant type species of 16 Hastingsia, softpart anatomy to ascertain whether a gizzard is present and sequence 17 18 data to fix its position on the molecular tree. Pending this research, the generic names 19 Hastingsia and Heteropora are retained as ‘formgenera’ for the species described 20 21 here from New Zealand. 22 Finally, a serious difficulty is posed by the nonmonophyletic status of the 23 24 species identified here as ‘Heteropora’. Sequenced specimens categorized 25 morphologically as belonging to this genus – taking into account the high degree of 26 27 variation in skeletal characters evident within colonies (see above) – appear as two 28 branches on the molecular tree (Fig. 46) One branch is part of an unresolved 29 30 assemblage including Diaperoecia purpurascens and ‘Hastingsia’ whitteni, while the 31 other is sister to the clade containing all these three taxa. There are two possible 32 33 evolutionary interpretations: (1) the ‘Heteropora’ morphotype with its predominantly 34 freewalled skeletal organization evolved twice, or (2) the fixedwalled ‘Hastingsia’ 35 36 whitteni and Diaperoecia purpurascens evolved from a heteroporid ancestor. The 37 unresolved nature of the node in question means that no inferences can be made about 38 39 the independent acquisition of the types of skeletal organization, although there is 40 moderate support for a sistergroup relationship between ‘Heteropora’ neozelanica 41 42 and Diaperoecia purpurascens in two of the analyses [lsr+ssrDNA: 0.93 pp (result 43 not shown); ssrDNA: 0.92 pp (Supplementary Fig. 2)], which points to independent 44 45 acquisitions of either fixedwalled skeletal organization from a freewalled ancestor 46 or vice versa. Sequence data from additional specimens of ‘Heteropora’ from New 47 48 Zealand and elsewhere should help in testing these alternatives. 49 50 51 Conclusions 52 53 1. The skeletal morphology is described in three species of cyclostome bryozoans 54 from New Zealand – Diaperoecia purpurascens (Hutton, 1877), ‘Hastingsia’ 55 56 57 58 22 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 23 of 48 Systematics and Biodiversity

1 2 3 4 5 6 whitteni sp. nov. and ‘Heteropora’ neozelanica Busk, 1879 – based on SEM of type 7 and other material. 8 9 2. Traditional taxonomy places these species in three separate families 10 (Diaperoeciidae, Hastingsiidae and Cerioporidae (=Heteroporidae)) and two or three 11 12 suborders, reflecting major differences in their skeletal morphology. 13 3. A molecular phylogenetic analysis showed, however, that the three heteromorphic 14 15 species are closely related and form a clade that is sister to Annectocyma. 16 4. Morphological support for this ‘diaperoeciid’ clade + Annectocyma comes from the 17 18 presence of a gizzard (yet to be proven as present in ‘Heteropora’ neozelanica) and 19 the skeletal morphology of the earliest part of the colony, the protoecium. In all of 20 21 these taxa, the protoecium is large and has a peripheral band of countersunk 22 pseudopores. 23 24 5. The occurrence of fixed and freewalled species in the diaperoeciid clade further 25 underscores the plasticity in skeletal organization exhibited by cyclostome bryozoans. 26 27 6. Until more is known about the type species of both Hastingia and Heteropora, 28 these genus names are best employed as formgenera for ‘Hastingsia’ whitteni, 29 30 ‘Heteropora’ neozelanica and ‘Heteropora’ sp. 31 7. Much work remains to be done on assembling a more complete molecular tree of 32 33 cyclostomes and in identifying morphological characters supportive of the clades that 34 can be identified on molecular grounds. Only then will it be possible to overhaul the 35 36 morphological classification of cyclostomes currently in use and replace it with one 37 that is more reflective of phylogeny. 38 39 40 Acknowledgements 41 42 Research on cyclostome bryozoans from New Zealand was commenced during the 43 tenure of a British Council Link Programme Grant in the mid1990s, and continued 44 45 during NERC Grant (NE/E015298/1; October 2007–October 2010). DPG's research 46 was supported by NIWA through Coasts and Oceans Research Programme 2 (Marine 47 48 Biological Resources) taxonomy project COBR1502. PDT is grateful to Dr Lee 49 Hsiang Liow (University of Oslo) for the opportunity to visit New Zealand for 50 51 geological fieldwork in January 2014 when the specimen shown in Fig. 20 was 52 collected. We thank Joanne Porter (HeriotWatt University, Edinburgh) for her help 53 54 with the collection of specimens, and the NHM sequencing facility for their 55 56 57 58 23 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 24 of 48

1 2 3 4 5 6 sequencing expertise. The sequenced material of 'Hastingsia' whitteni sp. nov. was 7 collected as part of a commercial project for Trans Tasman Resources (TTR) Ltd 8 9 (TTR11301 Benthos and TTR11301 Deepwater); TTR is thanked for permission to 10 use the specimen for research purposes. 11 12 13 14 References 15 16 17 Blackwell, M., Henk, D. A. & Jones, K. G. (2003). Extreme morphological 18 divergence: phylogenetic position of a tremite ectoparasite. Mycologia, 95, 987– 19 20 992. 21 Blainville, H. M. D. de (1830). Zoophytes. Dictionnaire des Sciences Naturelles, 60, 22 23 1–631. 24 Boardman, R. S. (1984). Origin of the PostTriassic Stenolaemata (Bryozoa): a 25 26 taxonomic oversight. Journal of Paleontology, 58, 19–39. 27 Boardman, R. S. (1998). Reflections on the morphology, anatomy, evolution, and 28 29 classification of the Class Stenolaemata (Bryozoa). Smithsonian Contributions 30 to Paleobiology, 86, 1–59. 31 Boardman, R. S., McKinney, F. K. & Taylor, P. D. (1992). Morphology, anatomy, 32 33 and systematics of the Cinctiporidae, new family (Bryozoa: Stenolaemata). 34 Smithsonian Contributions to Paleobiology, 70, 1–81. 35 36 Bock, P. E. & Gordon, D. P. (2013). Phylum Bryozoa Ehrenberg, 1831. Zootaxa, 37 3703, 67–74. 38 39 Borg, F. (1926). Studies on recent cyclostomatous Bryozoa. Zoologiska Bidrag fran 40 Uppsala, 10, 181–507. 41 42 Borg, F. (1933). A revision of the Recent Heteroporidae (Bryozoa). Zoologiska Bidrag 43 fran Uppsala, 14, 253–394. 44 45 Borg, F. (1944) The Stenolaematous Bryozoa. In S. Bock (Ed.), Further Zoological 46 Results of The Swedish Antarctic Expedition 1901-1903 (pp. 1–276). 47 48 Stockholm: Norstedt and Söner. 49 Bourret, A., McPeek, M. A. & Turgeon, J. (2011). Regional divergence and mosaic 50 51 spatial distribution of two closely related damselfly species (Enallagma hageni 52 and Enallagma ebrium). Journal of Evolutionary Biology, 25, 196–209. 53 54 55 56 57 58 24 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 25 of 48 Systematics and Biodiversity

1 2 3 4 5 6 Brood, K. (1976). Note on the bryozoan Pustulopora (Diaperoecia) intricaria Busk, 7 1875. Zoologica Scripta, 5, 231–233. 8 9 Busk, G. (1859). A monograph of the fossil Polyzoa of the Crag. London: 10 Palaeontographical Society, 136 pp. 11 12 Busk, G. (1875). Catalogue of the Marine Polyzoa in the Collection of the British 13 Museum. Part III. Cyclostomata. London: Trustees of the British Museum, 39 14 15 pp. 16 Busk, G. (1879). On Recent species of Heteropora. Journal of the Linnean Society 17 18 (Zoology), 14, 724–726. 19 Canu, F. (1917). Etudes sur les ovicelles des Bryozoaires Cyclostomes. Bulletin de la 20 21 Société Géologique de France, (4), 17, 345–347. 22 Canu, F. (1918). Les ovicelles des Bryozoaires Cyclostomes. Etudes sur quelques 23 24 familles nouvelles et anciennes. Bulletin de la Société Géologique de France, 25 (4), 16, 324–335. 26 27 Dick, M. H., Tilbrook, K. J. & Mawatari, S. F. (2006). Diversity and taxonomy of 28 rockyintertidal Bryozoa on the Island of Hawaii. Journal of Natural History, 29 30 40, 2197–2257. 31 Ernst, A., Taylor, P. D. & Bohatý, J. (2014). A new Middle Devonian cystoporate 32 33 bryozoan from Germany containing a new symbiont bioclaustration. Acta 34 Palaeontologica Polonica, 59, 173–183. 35 36 Farmer, J. D. (1979). Morphology and function of zooecial spines in cyclostome 37 Bryozoa: implications for paleobiology. In G. P. Larwood & M. B. Abbott (Eds) 38 39 Advances in bryozoology (pp. 219–246). London: Academic Press. 40 Goldfuss, G. A. (1826). Petrefacta Germaniae, Abbildung und Beschreibungen der 41 42 Petrefacten Deutschlands und der angreuzenden Länder. Teil 1. Dusseldorf: 43 Arnz & Co., 76 pp. 44 45 Gordon, D. P. (1967). A report on the ectoproct Polyzoa of some Auckland shores. 46 Tane (Journal of the Auckland University Field Club), 13, 43–76. 47 48 Gordon, D. P. (1975). The resemblance of bryozoan gizzard teeth to “annelidlike” 49 setae. Acta Zoologica, 56, 283–289. 50 51 Gordon, D. P., Taylor, P. D. & Bigey, F. P. (2009). Phylum Bryozoa moss animals, 52 sea mats, lace corals. In D. P. Gordon (Ed.), The New Zealand Inventory of 53 Biodiversity.Volume 1. Kingdom Animalia - Radiata, Lophotrochozoa, and 54 55 Deuterostomia, pp. 271–297. Christchurch: Canterbury University Press. 56 57 58 25 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 26 of 48

1 2 3 4 5 6 Hagenow, F. von. (1851). Die Bryozoen der Maastrichter Kreidebildung. Cassel: 7 Fischer, 106 pp. 8 9 Harmelin, J.G. (1976). Le sousordre des Tubuliporina (Bryozoaires Cyclostomes) en 10 Méditerranée. Memoires de l'Institut Oceanographique, Monaco, 10, 1–326. 11 12 Harmer, S. F. (1915). The Polyzoa of the Siboga Expedition. Part 1. Entoprocta, 13 Ctenostomata and Cyclostomata. Siboga-Expeditie, 28a, 1–180. 14 15 Hayward, P. J. & Ryland, J. S. (1985). Systematic notes on some British 16 Cyclostomata (Bryozoa). Journal of Natural History, 19, 1073–1078. 17 18 HerreraCubilla, A. & Jackson, J. B. C. (2014). Phylogeny of genus Cupuladria 19 (Bryozoa, Cheilostomata) in the Neogene of tropical America. Journal of 20 21 Paleontology, 88, 851–894. 22 Hillis, D. M. & Dixon, M. T. (1991). Ribosomal DNAMolecular Evolution and 23 24 Phylogenetic Inference. Quarterly Review of Biology, 66, 411–453. 25 Huelsenbeck, J. P. & Ronquist, F. (2001). MRBAYES: Bayesian inference of 26 27 phylogenetic trees. Bioinformatics, 17, 754–755. 28 Hughes, R.N., Gómez, A., Wright, P. J., Moyano, H. I., Cancinao, J. M., Cravalho, G. 29 30 R. & Lunt, D. H. (2008). Molecular phylogeny supports division of the 31 ‘cosmopolitan’ taxon Celleporella (Bryozoa; Cheilostomata) into four major 32 33 clades. Molecular Phylogenetics and Evolution, 46, 369–374. 34 Hutton, F. W. (1877). Corrections and additions to the list of Polyzoa in the Catalogue 35 36 of Marine Mollusca of New Zealand (1873). Transactions and Proceedings of 37 the New Zealand Institute, 9, 358–361. 38 39 Hutton, F. W. (1880). Manual of the New Zealand Mollusca. Wellington: James 40 Hughes, 224 pp. 41 42 Hutton, F. W. (1891). Revised list of the marine Bryozoa of New Zealand. 43 Transactions and Proceedings of the New Zealand Institute, 23, 102–107. 44 45 Jenkins, H. L. & Taylor, P. D. (2014). New or poorly known skeletal features in the 46 early astogeny of some cyclostome bryozoans. Studi Trentini di Scienze 47 48 Naturali, 94, 125–130. 49 Johnston, G. (1847). A history of the British Zoophytes. 2nd Edition. London: John van 50 51 Voorst, London, 488 pp. 52 Lamouroux, J. (1821). Exposition méthodique des genres de l'ordre des Polypiers. 53 Paris: Agasse, 115 pp. 54 55 56 57 58 26 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 27 of 48 Systematics and Biodiversity

1 2 3 4 5 6 MacGillivray, P. H. (1883). Descriptions of new or littleknown Polyzoa, Part 4. 7 Transactions and Proceedings of the Royal Society of Victoria, 19, 287–293. 8 9 Markham, J. B., & Ryland, J. S. (1987). Function of the gizzard in Bryozoa. Journal 10 of Experimental Marine Biology and Ecology, 107, 21–37. 11 12 Milne Edwards, H. (1838). Mémoire sur les Crisies, les Hornères et plusieurs autres 13 Polypes vivants ou fossils don’t l’organisation est analogue à celle des 14 15 Tubulipores. Annales des Sciences naturelles, Zoologie et Biologie animale, 16 Série 2, 9, 193238. 17 18 Morton, J. & Miller, M. (1968) The New Zealand Sea Shore. Auckland: Collins. 638 19 pp. 20 21 Nicholson, H. A. (1880). On the minute structure of the Recent Heteropora 22 neozelanica, Busk, and on the relations of the genus Heteropora to 23 24 Monticulipora. Annals and Magazine of Natural History, Series 5, 6, 329–339, 25 414–423. 26 27 Nye, O. B. Jr. (1976). Generic revision and skeletal morphology of some cerioporid 28 cyclostomes (Bryozoa). Bulletins of American Paleontology, 69, 1–222. 29 30 Orbigny, A. D. d’ (1835–47). Voyage dans l’Amérique méridionale … execute 31 pendant … 1826–33 par A. D. d’Orbigny, &c. Paris: P. Bertrand, Vol. + Atlas, 32 33 2 Vols. [Zoophytes in Vol. 5 (4), pp. 7–28, pls 1–13. Text, pp. 7–28, Livr. 89 34 (1847); pls 1, 3, 5, Livr. 56 (1841); pls 2, 6–8, Livr. 57 (1842); pls 4, 9, 10, 12, 35 36 13, Livr. 58 (1842); pl. 11, Livr. 59 (1842).] 37 Pérez, F. (2011). Discordant patterns of morphological and genetic divergence in the 38 39 closely related species Schizanthus hookeri and S. grahamii (Solanaceae). Plant 40 Systematics and Evolution, 293, 197–205. 41 42 Posada, D. & Crandall, K.A. (1998). Modeltest: testing the model of DNA 43 substitution. Bioinformatics, 14 (9), 817818. 44 45 Quenstedt, F. A. (1881). Atlas zu den Röhren- und Sternkorallen. Lepizig: Fues’s 46 Verlag, pls 155–184. 47 48 Ross, J. R. P. (1973). Calcification in the colonial stenolaemate, Heteropora. In G. P. 49 Larwood (Ed.), Living and Fossil Bryozoa. London: Academic Press, pp. 475– 50 51 483. 52 Ryland, J. S. (1982). Bryozoa. In S. P. Parker (Ed.), Synopsis and classification of 53 living organisms. New York: McGrawHill, pp. 743769. 54 55 56 57 58 27 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 28 of 48

1 2 3 4 5 6 Schäfer, P. (1986). On the gizzard in the bryozoan genus Diaperoecia Canu (Order 7 Tubuliporata). Senckenbergiana Maritima, 17, 253–277. 8 9 Schäfer, P. (1991). Brutkammern der Stenolaemata (Bryozoa); 10 Konstruktionsmorphologie und phylogenetische Bedeutung. Courier 11 12 Forschungsinstitut Senckenberg, 136, 1–263. 13 Schopf, T. J. M., Collier, K. O. & Bach, B. O. (1980). Relation of the morphology of 14 15 sticklike bryozoans to bottom currents and suspended matter and depth at 16 Friday Harbor, Washington, Paleobiology, 6, 466–476. 17 18 Smith, A. M. & Gordon, D. P. (2011). Bryozoans of southern New Zealand: a field 19 identification guide. New Zealand Aquatic Environment and Biodiversity Report, 20 21 75, 1–64. 22 Sotiaux, A., Enroth, J., Olsson, S., Quandt, D. & Vanderpoorten, A. (2009). When 23 24 morphology and molecules tell us different stories: a casein point with 25 Leptodon corsicus, a new and unique endemic moss species from Corsica. 26 27 Journal of Bryology, 31, 186–196. 28 Stoliczka, F. (1865). Fossile Bryozoen aus dem Tertiären Grünsandsteine der Orakei 29 30 Bay bei Auckland. Reise der Osterreichischen Fregatte “Novara” um die Erde in 31 der Jahren 1857, 1858, 1859. Geologischer Theil, 1(2), 89–158. 32 33 Taylor, P. D. (2000). Cyclostome systematics: phylogeny, suborders and the problem 34 of skeletal organization. In A. Herrera Cubilla & J. B. C. Jackson (Eds), 35 36 Proceedings of the 11th International Bryozoology Association Conference. 37 Balboa Republic of Panama: Smithsonian Tropical Research Institute, pp. 87– 38 39 103. 40 Taylor, P. D. & Gordon, D. P. (2003). Endemic new cyclostome bryozoans from 41 42 Spirits Bay, a New Zealand marinebiodiversity “hotspot”. New Zealand 43 Journal of Marine and Freshwater Research, 37, 653–669. 44 45 Taylor, P. D. & McKinney, F. K. (2006). Cretaceous Bryozoa from the Campanian 46 and Maastrichtian of the Atlantic and Gulf Coastal Plains, United States. Scripta 47 48 Geologica, 132, 1–346. 49 Taylor, P. D., Gordon, D. P. & Batson, P. B. (2004). Bathymetric distributions of 50 51 modern populations of some common Cenozoic Bryozoa from New Zealand, 52 and paleodepth estimation. New Zealand Journal of Geology and Geophysics, 53 47, 57–69. 54 55 56 57 58 28 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 29 of 48 Systematics and Biodiversity

1 2 3 4 5 6 Taylor, P. D., Kuklinski, P. & Gordon, D. P. (2007). Branch diameter and 7 depositional depth in cyclostome bryozoans: testing a potential 8 9 paleobathymetric tool. Palaios, 22, 220–224. 10 Taylor, P. D. & Weedon, M. J. (2000). Skeletal ultrastructure and phylogeny of 11 12 cyclostome bryozoans. Zoological Journal of the Linnean Society, 128, 337– 13 399. 14 15 Taylor, P. D., Waeschenbach, A. & Florence, W. (2011). Phylogenetic position and 16 systematics of the bryozoan Tennysonia: further evidence for convergence and 17 18 plasticity in skeletal morphology among cyclostome bryozoans. Zootaxa, 3010, 19 58–68. 20 21 Taylor, P. D., Lombardi, C. & Cocito, S. (2014). Biomineralization in bryozoans: 22 present, past and future. Biological Reviews, DOI 10.1111/brv.12148. 23 24 Viskova, L. V. (1992). Morskie Postpaleozoyskie Mshanki. Trudy 25 Paleontologicheskogo Instituta. Akademiya Nauk SSSR, Moscow, 250, 1–187. 26 27 Waeschenbach, A., Cox, C. C., Littlewood, D. T. J., Porter, J. S. & Taylor, P. D. 28 (2009). First molecular estimate of cyclostome bryozoan phylogeny confirms 29 30 extensive homoplasy among skeletal characters used in traditional taxonomy. 31 Molecular Phylogenetics and Evolution, 52, 241–251. 32 33 Waeschenbach, A., Taylor, P. D. & Littlewood, D. T. J. (2012). A molecular 34 phylogeny of bryozoans. Molecular Phylogenetics and Evolution, 62, 718–735. 35 36 Walter, B. (1970). Les Bryozoaires Jurassiques en France. Documents des 37 Laboratoires de Géologie de la Faculté des Sciences de Lyon, 35, 1–328. 38 39 Weedon, M. J. (1998). Skeletal ultrastructure of the early astogenetic stages of some 40 cyclostome bryozoans. Acta Zoologica, 79, 163–174. 41 42 Weedon, M. J. & Taylor, P. D. (1996). Skeletal ultrastructures in some cerioporine 43 cyclostome bryozoans. Acta Zoologica, 77, 249–265. 44 45 Whitten, R. F. (1979). Systematics and ecology of northern Hauraki Gulf Bryozoa 46 (Unpublished doctoral thesis). University of Auckland, Auckland, New 47 48 Zealand. 49 50 51 52 53 54 55 56 57 58 29 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 30 of 48

1 2 3 4 5 6 Figure captions 7

8 9 Figures 1, 2. Dried colonies of two cyclostome bryozoans from New Zealand 10 described here. 1. Diaperoecia purpurascens (Hutton), unknown locality in New 11 12 Zealand, NHMUK 2010.6.25.2. 2. ‘Heteropora’ neozelanica Busk, Otago Shelf, 13 NHMUK 2014.12.17.1. Scale bars: 10 mm. 14 15 16 Figures 3–8. Diaperoecia intricaria (Busk); backscattered scanning electron 17 18 micrographs of syntypes of the type species of the genus Diaperoecia; NHMUK 19 75.5.29.35, syntypes, SW Australia, Busk ex Gould Collection. 3. Bifurcating 20 21 branches and kenozooidal flange (arrow). 4. Detail showing irregular distribution of 22 autozooid apertures and a kenozooidal outgrowth (lower right). 5. Terminal 23 24 diaphragm. 6. Autozooids with short peristomes and pseudoporous frontal walls. 7. 25 Oblique transversely fractured branch. 8. Mural spine. Scale bars: 3, 4 = 1 mm; 5–7 = 26 27 100 µm; 8 = 10 µm. 28 29 30 Figures 9–18. Diaperoecia purpurascens (Hutton); backscattered scanning electron 31 micrographs. 9–12, 16. East Ulva Island, Paterson Inlet, Stewart Island, New Zealand, 32 33 D. Foster and A. M. Smith Collection, 8/6/1995, NHMUK 2014.12.17.2; 9. 34 Bifurcating branch; 10. Detail showing autozooids; 11. Pseudopores across a zooidal 35 36 boundary; 12. Transversely fractured branch; 16. Longitudinally sectioned branch 37 showing aperture (upper left), distantly spaced interzooidal pores in interior walls, 38 39 closely spaced pseudopores in exterior frontal wall (left), and mural spines growing 40 from frontal wall. 13–15. From craypot at Waitangi, Chatham Island, P. D. Taylor 41 42 Collection, 4/11/1999; 13, 14. NHMUK 2017.12.17.3; 13. Fertile branch showing 43 longitudinally elongate gonozooid. 14. Detail of gonozooid and ooeciopore. 15. 44 45 NHMUK 2017.12.17.4, another fertile branch with gonozooid scarcely visible 46 (possible ooeciopore arrowed). 17, 18. Molecular sequence voucher specimen, Otago 47 48 Shelf, A. M. Smith Collection, 25/1/2012, NHMUK 2017.12.17.5; 17. Broken branch; 49 18. Incomplete terminal diaphragm. Scale bars: 9, 13, 15 = 1 mm; 10 = 200 µm; 11 = 50 51 50 µm; 12, 17 = 500 µm; 14, 16, 18 = 100 µm. 52 53 Figures 19, 20. Diaperoecia purpurascens (Hutton); backscattered scanning electron 54 55 micrographs of encrusting colony bases with broken ends of erect stems; arrows point 56 57 58 30 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 31 of 48 Systematics and Biodiversity

1 2 3 4 5 6 to ancestrular protoecia. 19. Outside Whangaroa Harbour, 26/1/1981, National 7 Museum of New Zealand BS 862. 20. Pleistocene fossil, Nukumaru Brown Sand, 8 9 Nukumaru Beach, 01/2014, NHMUK BZ 7733, P. D. Taylor Collection. Scale bars = 10 1 mm. 11 12 13 Figures 21–26. ‘Hastingsia’ whitteni sp. nov.; backscattered scanning electron 14 15 micrographs. 21, 22, 25, 26. Paratypes (molecular voucher specimens AW757), NZOI 16 Station KAH1206/69, S39.9857°, E174.1988°, 44 m, 20/4/2012. 21. Fertile colony 17 18 (possible ooeciopore arrowed) with ancestrula (bottom), paratype, NHMUK 19 2017.12.17.6. 22, 25. Paratype, NHMUK 2017.12.17.7; 22. Mural spine; 25. Entire 20 21 colony showing ancestrula (left) with long peristome. 26. Paratype, semierect colony, 22 NHMUK 2017.12.17.8. 23, 24. Auckland, on Sargassum sinclairii, 3/1/1996, D. P. 23 24 Gordon Collection; 23. Holotype, NIWA 98120; 24. Incomplete gonozooid from 25 paratype, NIWA 98121. Scale bars: 21, 23, 25, 26 = 1 mm; 22 = 10 µm; 24 = 500 µm. 26 27 28 Figures 27–32. ‘Heteropora’ neozelanica Busk. 27. Reproduction of colony 29 30 illustrated by Busk (1879, fig. 1). 28–32. Backscattered scanning electron 31 micrographs of Busk’s syntypes. 28, 30, 31. Lectotype, NHMUK 99.7.1.4281; 28. 32 33 Branch fragment; 30. Autozooid surrounded by slightly smaller and more polygonal 34 kenozooids. 31. Mural spines visible in kenozooid aperture. 29, 32. Paralectotype, 35 36 NHMUK 99.7.1.4280; 29. Transversely sectioned branch showing axial endozone 37 surrounded by exozone with thickwalled zooids; 32. Aborally convex terminal basal 38 39 diaphragm. Scale bars: 27 = 10 mm; 28, 29 = 1 mm; 30 = 200 mm; 31 = 100 µm; 32 40 = 50 µm. 41 42 43 Figures 33–45. ‘Heteropora’ spp. from New Zealand. 33–34, 36–37, 39–45, ‘H’. 44 45 neozelanica Busk; backscattered scanning electron micrographs. 33, 36, 39–45. 46 NHM 2014.12.17.1, Otago Shelf, Stn Mu8829, 87–89 m, 11/5/1988, P. D. Taylor 47 48 Collection; 33. Bifurcating branch;. 36. Detail of apertures of varying size, lacking a 49 clear distinction between autozooids and kenozooids; 39. Branch with freewalled 50 51 zooids distally and fixedwalled proximally; 40. Detail of fixedwalled part of branch 52 showing autozooids with peristomes (some containing terminal diaphragms) and 53 kenozooids with terminal diaphragms well proximal of wall edges. 41. Transversely 54 55 fractured branch showing endozone and exozone; 42. Gonozooid with ooeciopore 56 57 58 31 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 32 of 48

1 2 3 4 5 6 arrowed (distal is to the left); 43. Broken gonozooid surrounded by band of fixed 7 walled zooids; 44. Closeup of fixedwalled zooids showing autozooids with 8 9 peristomes, kenozooids with terminal diaphragms almost flush with the ends of the 10 vertical walls (cf. Fig. 40), and a zooid of a fouling cheilostome (top); 45. Detail of 11 12 exteriorwalled peristome with pseudopore (arrowed) and base of interior wall 13 calcification. 34, 35, 37, 38. Molecular voucher specimens; 35, 38. Heteropora sp., Formatted: Font: Not Bold, Italic 14 15 NHMUK 2017.12.17.10, voucher NZ064, The Snares, 94 m, A. M. Smith & J. Porter Formatted: Font: Not Bold 16 Collection, February 2008; this species was distinguished using molecular evidence 17 18 but is morphologically almost identical to specimens shown here as ‘H.’ neozelanica; 19 35. Branch surface; 38. Raised autozooidal apertures surrounded by kenozooids. 34, 20 21 37. ‘ H’. neozelanica Busk‘Heteropora’ sp., NHMUK 2017.12.17.9, voucher AW086, 22 The Snares, 54 m; The Snares, A. M. Smith & J. Porter Collection, February 2008; 23 24 this species was distinguished using molecular evidence but is morphologically 25 almost identical to specimens shown here as ‘H.’ neozelanica; 34. Branch surface 26 27 showing freewalled zooids; 37. Autozooidal and kenozooidal apertures moderately 28 well differentiated; Scale bars: 33–35, 39, 41, 43 = 1 mm; 36–38, 40, 44 = 200 µm; 29 30 42 = 500 µm; 45 = 100 µm. 31 32 33 Figure 46. Bayesian analysis of the concatenated lsrDNA and ssrDNA dataset 34 constructed using MrBayes version 3.2.1 under the GTR+I+G model; 5,000,000 35 36 generations, 2,000,000 generations burnin. All nodes with <0.95 posterior probability 37 have been collapsed. The branch length scale bar indicates number of substitutions 38 39 per site. Box indicates the clade forming the focus of the current study. Cyclostome 40 suborders based on conventional morphological classifications are given on the right. 41 42 43 Figures 47–52. Mural spines; highresolution backscattered electron micrographs. 44 45 47–49. Diaperoecia purpurascens (Hutton), National Museum of New Zealand BS 46 862, outside Whangaroa Harbour; 47. Group of mural spines on inside of frontal wall; 47 48 48. Detail of barbed mural spine with flat head; 49. Mural spine on interior wall with 49 spinelets directed proximally. 50–52. ‘Heteropora’ neozelanica Busk, NHMUK 50 51 2014.12.17.1, Otago Shelf, Stn Mu8829; 50. Kenozooid containing numerous mural 52 spines; 51. Bifid mural spine; 52. Mural spine with long shaft. Scale bars: 47, 50 = 50 53 µm; 48, 49, 51, 52 = 5 µm. 54 55 56 57 58 32 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 33 of 48 Systematics and Biodiversity

1 2 3 4 5 6 Figures 53–61. Comparative morphology of ancestrulae; backscattered scanning 7 electron micrographs. 53–55. Diaperoecia purpurascens (Hutton), outside 8 9 Whangaroa Harbour, 26/1/1981, National Museum of New Zealand BS 862; 53. 10 Ancestrula; 54. Protoecium; 55. Protoecial pseudopores. 56–58. ‘Hastingsia’ whitteni 11 12 sp. nov., Auckland, on Sargassum sinclairii, 3/1/1996, D. P. Gordon Collection, 13 paratype NIWA 98121; 56. Ancestrula; 57. Protoecium; 58. Protoecial pseudopores. 14 15 59–61. ‘Heteropora’ neozelanica Busk, NZOI Stn Z9684, NIWA 98122; 59. 16 Ancestrula; 60. Protoecium; 61. Protoecial pseudopores. Scale bars: 53, 56, 59 = 200 17 18 µm; 54, 57, 60 = 100 µm; 55, 58, 61 = 20 µm. 19 20 21 Figure 62–63. Annectocyma major (Johnston, 1847), English Channel, 49°19.9,N Formatted: Font: Bold 22 3°22.4’W, 77 m, 3/2/1962. 62, ancestrula with aperture closed by a terminal Formatted: Font: Italic 23 Formatted: Font: Times New Roman, 12 24 diaphragm, and early budded zooids. 63, damaged protoecium showing countersunk pt 25 pseudopores. Scale bars: 62 = 500 µm; 63 = 200 µm. Formatted: Font: Bold 26 Formatted: Font: Bold 27 28 Figure 642. Simplified vertical sections through young colonies of Diaperoecia 29 30 purpurascens and ‘Heteropora’ showing skeletal organization similarity during early 31 astogenetic stages compared with its dissimilarity in later stages when erect growth 32 33 develops (see text for full explanation). Solid black lines indicate calcified skeletal 34 walls; dashed lines uncalcified outer body walls enclosing hypostegal coelom. 35 36 Everted tentacle crowns of autozooids are shown diagrammatically; open apertures 37 between the autozooids in ‘Heteropora’ are kenozooids. 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 33 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 34 of 48

1 2 3 4 5 6 Supplementary Material 7

8 9 Supplementary Figure 1. Bayesian analysis of the lsrDNA dataset constructed using 10 MrBayes version 3.2.1 under the GTR+I+G model; 5,000,000 generations, 4,500,000 11 12 generations burnin. The branch length scale bar indicates number of substitutions per 13 site. 14 15 16 Supplementary Figure 2. Bayesian analysis of the ssrDNA dataset constructed using 17 18 MrBayes version 3.2.1 under the GTR+I+G model; 5,000,000 generations, 4,500,000 19 generations burnin. The branch length scale bar indicates number of substitutions per 20 21 site. 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 34 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 35 of 48 Systematics and Biodiversity

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 Figures 1, 2. Dried colonies of two cyclostome bryozoans from New Zealand described here. 1. Diaperoecia purpurascens (Hutton), unknown locality in New Zealand, NHMUK 2010.6.25.2. 2. ‘Heteropora’ neozelanica 48 Busk, Otago Shelf, NHMUK 2014.12.17.1. Scale bars: 10 mm. 49 99x160mm (300 x 300 DPI) 50 51 52 53 54 55 56 57 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 36 of 48

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 Figures 3–8. Diaperoecia intricaria (Busk); backscattered scanning electron micrographs of syntypes of the 33 type species of the genus Diaperoecia; NHMUK 75.5.29.35, syntypes, SW Australia, Busk ex Gould Collection. 3. Bifurcating branches and kenozooidal flange (arrow). 4. Detail showing irregular distribution of 34 autozooid apertures and a kenozooidal outgrowth (lower right). 5. Terminal diaphragm. 6. Autozooids with 35 short peristomes and pseudoporous frontal walls. 7. Oblique transversely fractured branch. 8. Mural spine. 36 Scale bars: 3, 4 = 1 mm; 5–7 = 100 µm; 8 = 10 µm. 37 189x140mm (300 x 300 DPI) 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 37 of 48 Systematics and Biodiversity

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 189x189mm (300 x 300 DPI) 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 38 of 48

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 Figures 19, 20. Diaperoecia purpurascens (Hutton); back-scattered scanning electron micrographs of encrusting colony bases with broken ends of erect stems; arrows point to ancestrular protoecia. 19. Outside 48 Whangaroa Harbour, 26/1/1981, National Museum of New Zealand BS 862. 20. Pleistocene fossil, Nukumaru 49 Brown Sand, Nukumaru Beach, 01/2014, NHMUK BZ 7733, P. D. Taylor Collection. Scale bars = 1 mm. 50 99x180mm (300 x 300 DPI) 51 52 53 54 55 56 57 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 39 of 48 Systematics and Biodiversity

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 189x129mm (300 x 300 DPI) 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 40 of 48

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 Figures 27–32. ‘Heteropora’ neozelanica Busk. 27. Reproduction of colony illustrated by Busk (1879, fig. 1). 39 28–32. Backscattered scanning electron micrographs of Busk’s syntypes. 28, 30, 31. Lectotype, NHMUK 40 99.7.1.4281; 28. Branch fragment; 30. Autozooid surrounded by slightly smaller and more polygonal 41 kenozooids. 31. Mural spines visible in kenozooid aperture. 29, 32. Paralectotype, NHMUK 99.7.1.4280; 29. 42 Transversely sectioned branch showing axial endozone surrounded by exozone with thickwalled zooids; 32. 43 Aborally convex terminal diaphragm. Scale bars: 27 = 10 mm; 28, 29 = 1 mm; 30 = 200 mm; 31 = 100 44 µm; 32 = 50 µm. 45 189x180mm (300 x 300 DPI) 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 41 of 48 Systematics and Biodiversity

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 Figures 33–45. ‘Heteropora’ spp. from New Zealand. 33–34, 36–37, 39–45, ‘H’. neozelanica Busk; back scattered scanning electron micrographs. 33, 36, 39–45. NHM 2014.12.17.1, Otago Shelf, Stn Mu8829, 87– 48 89 m, 11/5/1988, P. D. Taylor Collection; 33. Bifurcating branch. 36. Detail of apertures of varying size, 49 lacking a clear distinction between autozooids and kenozooids; 39. Branch with freewalled zooids distally 50 and fixedwalled proximally; 40. Detail of fixedwalled part of branch showing autozooids with peristomes 51 (some containing terminal diaphragms) and kenozooids with terminal diaphragms well proximal of wall 52 edges. 41. Transversely fractured branch showing endozone and exozone; 42. Gonozooid with ooeciopore 53 arrowed (distal is to the left); 43. Broken gonozooid surrounded by band of fixedwalled zooids; 44. Close 54 up of fixedwalled zooids showing autozooids with peristomes, kenozooids with terminal diaphragms almost 55 flush with the ends of the vertical walls (cf. Fig. 40), and a zooid of a fouling cheilostome (top); 45. Detail of exteriorwalled peristome with pseudopore (arrowed) and base of interior wall calcification. 34, 35, 37, 38. 56 Molecular voucher specimens; 35, 38. NHMUK 2017.12.17.10, voucher NZ064, The Snares, 94 m, A. M. 57 Smith & J. Porter Collection, February 2008; 35. Branch surface; 38. Raised autozooidal apertures 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 42 of 48

1 2 3 surrounded by kenozooids. 34, 37. ‘Heteropora’ sp., NHMUK 2017.12.17.9, voucher AW086, The Snares, 54 4 m; The Snares, A. M. Smith & J. Porter Collection, February 2008; this species was distinguished using 5 molecular evidence but is morphologically almost identical to specimens shown here as ‘H.’ neozelanica; 34. 6 Branch surface showing freewalled zooids; 37. Autozooidal and kenozooidal apertures moderately well 7 differentiated; Scale bars: 33–35, 39, 41, 43 = 1 mm; 36–38, 40, 44 = 200 µm; 42 = 500 µm; 45 = 100 8 µm. 189x230mm (300 x 300 DPI) 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 43 of 48 Systematics and Biodiversity

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 Figure 46. Bayesian analysis of the concatenated lsrDNA and ssrDNA dataset constructed using MrBayes 39 version 3.2.1 under the GTR+I+G model; 5,000,000 generations, 2,000,000 generations burn-in. All nodes 40 with <0.95 posterior probability have been collapsed. The branch length scale bar indicates number of 41 substitutions per site. Box indicates the clade forming the focus of the current study. Cyclostome suborders 42 based on conventional morphological classifications are given on the right. 43 450x425mm (300 x 300 DPI) 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 44 of 48

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 Figures 47–52. Mural spines; highresolution backscattered electron micrographs. 47–49. Diaperoecia 39 purpurascens (Hutton), National Museum of New Zealand BS 862, outside Whangaroa Harbour; 47. Group of 40 mural spines on inside of frontal wall; 48. Detail of barbed mural spine with flat head; 49. Mural spine on 41 interior wall with spinelets directed proximally. 50–52. ‘Heteropora’ neozelanica Busk, NHMUK 2014.12.17.1, 42 Otago Shelf, Stn Mu8829; 50. Kenozooid containing numerous mural spines; 51. Bifid mural spine; 52. 43 Mural spine with long shaft. Scale bars: 47, 50 = 50 µm; 48, 49, 51, 52 = 5 µm. 44 189x180mm (300 x 300 DPI) 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 45 of 48 Systematics and Biodiversity

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 Figures 53–61. Comparative morphology of ancestrulae; backscattered scanning electron micrographs. 53– 41 55. Diaperoecia purpurascens (Hutton), outside Whangaroa Harbour, 26/1/1981, National Museum of New 42 Zealand BS 862; 53. Ancestrula; 54. Protoecium; 55. Protoecial pseudopores. 56–58. ‘Hastingsia’ whitteni 43 sp. nov., Auckland, on Sargassum sinclairii, 3/1/1996, D. P. Gordon Collection, paratype NIWA 98121; 56. 44 Ancestrula; 57. Protoecium; 58. Protoecial pseudopores. 59–61. ‘Heteropora’ neozelanica Busk, NZOI Stn 45 Z9684, NIWA 98122; 59. Ancestrula; 60. Protoecium; 61. Protoecial pseudopores. Scale bars: 53, 56, 59 = 200 µm; 54, 57, 60 = 100 µm; 55, 58, 61 = 20 µm. 46 189x189mm (300 x 300 DPI) 47 48 49 50 51 52 53 54 55 56 57 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Page 46 of 48

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 99x180mm (300 x 300 DPI) 48 49 50 51 52 53 54 55 56 57 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Page 47 of 48 Systematics and Biodiversity

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 180x324mm (300 x 300 DPI) 48 49 50 51 52 53 54 55 56 57 58 59 60 URL: http://mc.manuscriptcentral.com/tsab Systematics and Biodiversity Pectinatella magnifica Page 48 of 48

1 2 Entalophoroecia cf. robusta 3 4 5 6 Diplosolen obelia 7 0.6 8 0.9 9 10 Cardioecia watersi 11 12 13 0.75 14 Frondipora verrucosa 15 16 17 18 0.46 Hornera foliacea 19 20 21 1 22 Hornera cf. caespitosa 23 24 0.43 25 1 26 27 Hornera robusta 28 29 30 31 Cinctipora_elegans 32 33 34 35 Favosipora_rosea 36 37 38 0.53 39 Plagioecia patina 40 1 41 1 42 43 Plagioecia patina 44 45 46 0.93 47 Disporella pristis 48 49 50 1 51 Disporella hispida fimbriata 52 53 1 54 55 Disporella cf. neapolitana 56 57 0.65 58 59 60 Crisia sigmoidea

0.97 Crisia denticulata 0.67 Crisia fistulosa 0.06 Tubulipora liliacea 0.95

1 Idmidronea atlantica

0.92 Tennysonia stellata 1 1 Tennysonia stellata

Annectocyma tubulosa

1 ‘Heteropora’ sp.

0.72 ‘Hastingsia’ whitteni sp. nov.

1 ‘Heteropora’ neozelanica URL: http://mc.manuscriptcentral.com/tsab 0.43 Diaperoecia purpurascens Page 49 of 48 Systematics and Biodiversity Pectinatella magnifica

1 2 Cinctipora elegans 3 4 5 6 Disporella pristis 7 8 0.52 9 0.99 10 Disporella hispida fimbriata 11 12 1 13 14 Disporella cf. neapolitana 15 1 16 17 18 Favosipora rosea 19 20 21 0.9 22 Plagioecia patina 23 24 1 25 26 Plagioecia patina 27 28 29 30 Entalophoroecia cf. robusta 31 1 32 33 34 Diplosolen obelia 35 0.61 36 1 37 38 Cardioecia watersi 39 40 41 0.6 42 Frondipora verrucosa 43 44 45 0.64 46 Hornera robusta 47 48 49 0.7 50 Hornera foliacea 51 52 0.87 53 0.49 54 Hornera cf. caespitosa 55 56 57 58 Crisia sigmoidea 59 60 1 Crisia denticulata 0.96 Crisia fistulosa

0.05 Tubulipora liliacea 0.96 0.91 Idmidronea atlantica

0.97 Tennysonia stellata 1 0.98 Tennysonia stellata

Annectocyma tubulosa

1 ‘Heteropora’ sp.

1 ‘Hastingsia’ whitteni sp. nov.

1 ‘Heteropora’ neozelanica URL: http://mc.manuscriptcentral.com/tsab 0.92 Diaperoecia purpurascens