Electronic Supplementary Material Note S1 Figures S1-S4 Tables S1, S2

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1 Electronic Supplementary Material Note S Figures S-S4 Tables S, S Note S: Description of glaucinid reproductive system Glaucinins have diaulic (i.e., having two pallial gonoducts, 30) reproductive systems [0]. After developing in a proximal hermaphroditic otestis, autosperm and eggs are sent distally to the respective male and female reproductive tracts via the muscular ampulla (Fig. ; 0,, ). The male system consists of a long duct (prostate) connected to a protrusible penis, which is housed within a penial sheath, opening at the gonopore (Fig. ). In Glaucus atlanticus the penis is armed with a cuticular spine, whereas it is unarmed in G. marginatus [0]. The female system consists of a proximal receptaculum seminis, which stores oriented allosperm, and female gland complex consisting of a proximal caule gland, followed by a membrane gland, and finally a distal mucous gland [3]. All of these structures open into the iduct. Fertilization occurs in the iduct, immediately distal to the receptaculum seminis and proximal to the female gland complex. The iduct extends distally to the vagina (here it may contain egg strings and/or unoriented sperm), and the vaginal opening is located at the gonopore [9]. In G. atlanticus and two of the G. marginatus lineages (A and C), the iduct connects to a bursa copulatrix, a thinwalled epithelial sac opening immediately proximal to the vagina (Figs., S, S). In G. marginatus lineages B and D, respectively the sister lineages of A and C, the bursa copulatrix is absent; the thin vaginal duct leads directly to the female gland complex and receptaculum seminis (Figs., S, S). Supplementary references: 30. Ghiselin, M. T. 966 The evolution of hermaphroditism among animals Quart. Rev. Biol. 44, Klussmann-Kolb, A. 00 The reproductive systems of the Nudibranchia (Gastropoda, Opisthobranchia): Comparative histology and ultrastructure of the nidamental glands with aspects of functional morphology Zool. Anz. 40, 9-36.

2 Supplementary Figure Legends (Figure S is on p. 4) Figure S. Two reproductive morphologies in Glaucus spp. viewed via dissections. The topology of the Glaucus clade is presented at left with color-coding corresponding to four G. marginatus lineages (A-D) and G. atlanticus (E) (Figs.,, S3). Images on the right are dorsal view photographs of glaucinid reproductive systems with most or all of the otestis remed. Dashed black circles outline the gonopore, black arrows indicate position of the vagina, and the bursa copulatrix (when present) is indicated with red letters. The penis is everted in all G. marginatus specimens (A-D) and inverted in G. atlanticus (E). A. G. marginatus (Indo-Pacific). B. G. marginatus (South Pacific). C. G. marginatus (Eastern North Pacific). D. G. marginatus (Central North Pacific). E. G. atlanticus. Abbreviations: am, ampulla;, bursa copulatrix;, female gland complex; ot, otestis;, penis;, prostate;, vagina (dashed red arrow). Scale bars =.0 mm. (Figure S is on p. 5) Figure S. Two reproductive morphologies in Glaucus spp. viewed via histological sections. The topology of the Glaucus clade is presented at left with color-coding corresponding to four G. marginatus lineages (A-D) and G. atlanticus (E) (Figs.,, S3). Images on the right are photographs of histological sections of distal glaucinid reproductive systems. The bursa copulatrix (when present) is indicated with red letters. A. G. marginatus (Indo-Pacific) dorsal view, penis inverted. The bursa copulatrix is located immediately proximal to the vagina. A. G. marginatus (Indo-Pacific) dorsal view. Detail of bursa copulatrix containing unoriented allosperm. B. G. marginatus (South Pacific) ventral view, outer body wall remed, penis everted. No bursa copulatrix is present; proximal to the vagina, the iduct leads to the female gland complex. B. G. marginatus (South Pacific). Detail of iduct. C. G. marginatus (Eastern North Pacific) dorsal view, penis everted. The bursa copulatrix is located immediately proximal to the vagina. C. G. marginatus (Eastern North Pacific). Detail of bursa copulatrix containing unoriented allosperm. D. G. marginatus (Central North Pacific) dorsal view, penis everted. No bursa copulatrix is present; proximal to the vagina, the iduct leads to the female gland complex. D. G. marginatus (Central North Pacific). Detail of iduct. E. G. atlanticus (ventral view), outer body wall remed, penis inverted. The vagina opens immediately to the bursa copulatrix and the iduct. E. G. atlanticus. Detail of the vagina. E3. G. atlanticus sagittal section, lateral view. Detail of the bursa copulatrix containing unoriented allosperm, which is separate from the iduct containing part of an egg string. Abbreviations:, bursa copulatrix;, female gland complex;, penis;, prostate; ot, otestis;, iduct; rs, receptaculum seminis;, vagina. Scale bars: a-a = 00 µm; b =.0 mm; b = 500 µm; c-c = 00 µm; d-d = 00 µm; e =.0 mm; e =.0 mm; e3 = 500 µm. (Figure S3 is on p. 6) Figure S3. Bayesian consensus phylogram of aeolid nudibranchs based on four loci (mt 6S rdna and COI; nuclear 8S rdna and Histone-H3). Statistical support percentages are shown abe internal branches in the form: Bayesian posterior probability/maximum likelihood bootstrap value. Asterisks indicate values of 00. Clades Aeolidida and Doridida (outgroup) are indicated to the far right of the topology. Superfamilies Aeolidioidea (Aeolidiidae+Glaucidae, blue), Fionoidea (green), and Flabellinoidea (orange) are indicated in color and labeled near

3 internal branches. Presence (+) or absence (-) of the bursa copulatrix is indicated in red on the topology. Glaucus lineage color coding appears as in Figs.,, S3. Four cryptic lineages of G. marginatus are denoted by bold letters: A, Indo-Pacific; B, South Pacific; C, Eastern North Pacific; D, Central North Pacific. (Figure S4 is on p. 7) Figure S4. Glaucus clade individual topologies for four loci. Bayesian phylograms depict within-glaucus relationshi for A, mt 6S rdna; B, mt COI; C, nuclear 8S rdna and D, Histone-H3. Glaucus atlanticus clades are collaed (see Fig. S3). Four cryptic lineages of G. marginatus were recered in mt 6S rdna, three of which were also recered in mt COI. Cryptic lineages of G. marginatus are denoted by bold letters: A, Indo-Pacific; B, South Pacific; C, Eastern North Pacific; D, Central North Pacific. All nodal support values (Bayesian and Maximum likelihood) = 00. 3

4 Fig. S A am - B ot C ot G. marginatus am - D ot am E ot G. atlanticus rs 4

5 Fig. S A ot - B rs - D C G. marginatus ot 3 G. atlanticus E 5

6 Fig. S3 Aeolidioidea 93/77 96/7 63/ 9/ 9/ */78 99/87 9/57 */97 */94 89/55 */83 56/ A8 A7 */93 A5 A3 A A */83 A4 A6 A9 A0 B3 B8 - B0 B7 B B B B4 B5 B6 B9 */98 C C C3 C5 C6 C7 C8 C9 C4 C0 - D6 D D D3 D4 D Glaucus atlanticus Favorinus elenalexarium Learchis poica Hermosita hakunamatata Aeolidiella alba Phidiana pugnax Aeolidiella chromosoma Facelina stearnsi Pteraeolidia ianthina Hermissenda crassicornis Nanuca sebastiani Dondice occidentalis Phyllodesmium parangatum Phyllodesmium longicirrum Aeolidia papillosa Spurilla neapolitana Spurilla sargassicola Caloria indica Protaeolidia juliae A C D Glaucus marginatus Aeolidida Fionoidea 98/66 99/63 Flabellinoidea */98 Calma glaucoides Cuthona abronia */7 Cuthona divae */90 Cuthona lagunae Cuthona fulgens Flabellina exoptata Flabellina fusca Flabellina trilineata.4 Cryptobranchia kuragiensis Doridida 0. 6

7 Fig. S4 A mt 6S rdna A B C D G. marginatus B mt COI A C D G. marginatus G. atlanticus G. atlanticus C D 8S rdna G. marginatus Histone H-3 G. marginatus A B C D A B C D G. atlanticus G. atlanticus 7

8 Table S. Locality data for Glaucus spp. used in the molecular and morphological analyses. Color coding corresponds to the four glaucinin molecular lineages (Figs., S3). Species Genus Gyre System Latitude ( Decimal) Longitude ( Decimal) atlanticus Glaucus North Atlantic atlanticus Glaucus North Atlantic atlanticus 3 Glaucus North Atlantic atlanticus 4 Glaucus North Atlantic atlanticus 5 Glaucus North Atlantic atlanticus 6 Glaucus North Atlantic atlanticus 7 Glaucus North Atlantic atlanticus 8 Glaucus North Atlantic atlanticus 9 Glaucus North Atlantic atlanticus 0 Glaucus North Atlantic atlanticus Glaucus South Pacific atlanticus Glaucus North Pacific atlanticus 3 Glaucus North Pacific atlanticus 4 Glaucus South Pacific atlanticus 5 Glaucus North Pacific atlanticus 6 Glaucus North Pacific atlanticus 7 Glaucus South Pacific atlanticus 8 Glaucus North Pacific atlanticus 9 Glaucus South Pacific atlanticus 0 Glaucus South Pacific atlanticus Glaucus Indian atlanticus Glaucus Indian atlanticus 3 Glaucus Indian atlanticus 4 Glaucus Indian atlanticus 5 Glaucus Indian atlanticus 6 Glaucus Indian atlanticus 7 Glaucus Indian atlanticus 8 Glaucus Indian atlanticus 9 Glaucus Indian atlanticus 30 Glaucus Indian marginatus A Glaucus Indian marginatus A Glaucus Indian marginatus A3 Glaucus Indian marginatus A4 Glaucus Indian marginatus A5 Glaucus North Pacific marginatus A6 Glaucus North Pacific marginatus A7 Glaucus North Pacific marginatus A8 Glaucus North Pacific marginatus A9 Glaucus North Pacific marginatus A0 Glaucus South Pacific marginatus B Glaucus South Pacific marginatus B Glaucus South Pacific marginatus B3 Glaucus South Pacific marginatus B4 Glaucus South Pacific marginatus B5 Glaucus South Pacific marginatus B6 Glaucus South Pacific marginatus B7 Glaucus South Pacific marginatus B8 Glaucus South Pacific marginatus B9 Glaucus South Pacific

9 marginatus B0 Glaucus South Pacific marginatus C Glaucus North Pacific marginatus C Glaucus North Pacific marginatus C3 Glaucus North Pacific marginatus C4 Glaucus North Pacific marginatus C5 Glaucus North Pacific marginatus C6 Glaucus North Pacific marginatus C7 Glaucus North Pacific marginatus C8 Glaucus North Pacific marginatus C9 Glaucus North Pacific marginatus C0 Glaucus North Pacific marginatus D Glaucus North Pacific marginatus D Glaucus North Pacific marginatus D3 Glaucus North Pacific marginatus D4 Glaucus North Pacific marginatus D5 Glaucus North Pacific marginatus D6 Glaucus North Pacific

10 Table S. Taxonomy, museum catalog number of voucher specimen, and GenBank accession numbers for sequences for four molecular markers (nuclear 8S rrna, Histone-H3, mitochondrial 6S rrna, COI) from the nudibranch species included in the molecular phylogeny (Figs. b, S3). Museums and universities holding voucher specimens are identified by prefixes before catalog numbers: AMS, Australian Museum, Sydney, Australia; CASIZ, California Academy of Sciences; CPIC, California Polytechnic State University, Pomona, USA; FLMNH, Florida Museum of Natural History, University of Florida, Gainesville, USA; LACM, Los Angeles County Museum of Natural History, Los Angeles, USA; NM, Natal Museum, Pietermaritzburg, South Africa; NUGB, Nagoya University, Nagoya, Japan (outgroup previously in GenBank); UMMZ, University of Michigan Museum of Zoology, Ann Arbor, USA; WAM, Western Australian Museum, Perth, Australia. For Glaucus spp. with no voucher, the specimen size was so small that the whole animal was used for DNA extraction. Whole genomic DNA template for these samples is available at UMMZ. All accession numbers not beginning with JQ come from other studies and were previously published in GenBank. Asterisks indicate no sequence data. Species Genus Family Voucher 8S H3 6S COI papillosa Aeolidia Aeolidiidae CPIC 0077 JQ69993 JQ JQ JQ alba Aeolidiella Aeolidiidae LACM 750 JQ69994 JQ * * chromosoma Aeolidiella Aeolidiidae LACM 7507 JQ69995 JQ JQ JQ glaucoides Calma Calmidae CPIC 0056 JQ69996 JQ JQ JQ indica Caloria Glaucidae FLMNH JQ69997 JQ DQ4773 DQ4735 kuragiensis Cryptobranchia Dorididae NUGB-L505 AB8779 AB AB38346 AB38457 abronia Cuthona Tergipedidae CASIZ JQ69998 JQ JQ JQ divae Cuthona Tergipedidae CASIZ JQ69999 JQ69939 JQ JQ fulgens Cuthona Tergipedidae CASIZ JQ JQ69939 JQ * lagunae Cuthona Tergipedidae no voucher JQ69930 JQ JQ69948 * occidentalis Dondice Glaucidae LACM JQ69930 JQ JQ69948 JQ stearnsi Facelina Glaucidae no voucher JQ JQ JQ JQ69957 elenalexiarum Favorinus Glaucidae LACM 5338 JQ JQ JQ HM6755 exoptata Flabellina Flabellinidae LACM JQ JQ JQ JQ69957 fusca Flabellina Flabellinidae LACM 7497 JQ JQ JQ JQ trilineata Flabellina Flabellinidae CPIC JQ JQ JQ GQ904 atlanticus Glaucus Glaucidae UMMZ JQ JQ JQ JQ atlanticus Glaucus Glaucidae UMMZ JQ JQ69940 JQ JQ atlanticus 3 Glaucus Glaucidae no voucher JQ69930 JQ69940 JQ JQ atlanticus 4 Glaucus Glaucidae UMMZ JQ6993 JQ JQ69949 JQ atlanticus 5 Glaucus Glaucidae UMMZ JQ6993 JQ JQ69949 JQ atlanticus 6 Glaucus Glaucidae no voucher JQ69933 JQ JQ JQ699579

11 atlanticus 7 Glaucus Glaucidae UMMZ JQ69934 JQ JQ JQ atlanticus 8 Glaucus Glaucidae UMMZ JQ69935 JQ JQ JQ69958 atlanticus 9 Glaucus Glaucidae UMMZ JQ69936 JQ JQ JQ69958 atlanticus 0 Glaucus Glaucidae UMMZ JQ69937 JQ JQ JQ atlanticus Glaucus Glaucidae UMMZ JQ69938 JQ69940 JQ JQ atlanticus Glaucus Glaucidae UMMZ 3098 JQ69939 JQ6994 JQ JQ atlanticus 3 Glaucus Glaucidae UMMZ 3098 JQ69930 JQ6994 JQ JQ atlanticus 4 Glaucus Glaucidae AMS C JQ6993 JQ69943 JQ69950 JQ atlanticus 5 Glaucus Glaucidae UMMZ JQ6993 JQ69944 JQ69950 JQ atlanticus 6 Glaucus Glaucidae UMMZ JQ69933 JQ69945 JQ JQ atlanticus 7 Glaucus Glaucidae AMS C JQ69934 JQ69946 JQ JQ atlanticus 8 Glaucus Glaucidae UMMZ JQ69935 JQ69947 JQ JQ69959 atlanticus 9 Glaucus Glaucidae AMS C JQ69936 JQ69948 JQ JQ69959 atlanticus 0 Glaucus Glaucidae AMS C JQ69937 JQ69949 JQ JQ atlanticus Glaucus Glaucidae WAM S59354 JQ69938 JQ69940 JQ JQ atlanticus Glaucus Glaucidae WAM S59354 JQ69939 JQ6994 JQ JQ atlanticus 3 Glaucus Glaucidae WAM S59354 JQ JQ6994 JQ69950 JQ atlanticus 4 Glaucus Glaucidae WAM S59354 JQ69933 JQ69943 JQ6995 JQ atlanticus 5 Glaucus Glaucidae WAM S59354 JQ69933 JQ69944 JQ6995 JQ atlanticus 6 Glaucus Glaucidae WAM S59354 JQ JQ69945 JQ69953 JQ atlanticus 7 Glaucus Glaucidae NM W7469 JQ JQ69946 JQ69954 JQ atlanticus 8 Glaucus Glaucidae NM W7469 JQ JQ69947 JQ69955 JQ69960 atlanticus 9 Glaucus Glaucidae NM W7469 JQ JQ69948 JQ69956 JQ69960 atlanticus 30 Glaucus Glaucidae NM W7469 JQ JQ69949 JQ69957 JQ marginatus A Glaucus Glaucidae CASIZ JQ JQ JQ69958 JQ marginatus A Glaucus Glaucidae CASIZ JQ JQ69943 JQ69959 JQ marginatus A3 Glaucus Glaucidae CASIZ JQ JQ69943 JQ69950 JQ marginatus A4 Glaucus Glaucidae CASIZ JQ69934 JQ JQ6995 JQ marginatus A5 Glaucus Glaucidae UMMZ JQ69934 JQ JQ6995 JQ marginatus A6 Glaucus Glaucidae UMMZ JQ JQ JQ69953 JQ marginatus A7 Glaucus Glaucidae UMMZ JQ JQ JQ69954 JQ69960 marginatus A8 Glaucus Glaucidae UMMZ JQ JQ JQ69955 JQ6996 marginatus A9 Glaucus Glaucidae UMMZ JQ JQ JQ69956 JQ6996 marginatus A0 Glaucus Glaucidae UMMZ JQ JQ JQ69957 JQ69963 marginatus B Glaucus Glaucidae AMS C JQ JQ JQ69958 * marginatus B Glaucus Glaucidae AMS C JQ JQ69944 JQ69959 *

12 marginatus B3 Glaucus Glaucidae AMS C JQ JQ69944 JQ * marginatus B4 Glaucus Glaucidae AMS C JQ69935 JQ JQ69953 * marginatus B5 Glaucus Glaucidae AMS C JQ69935 JQ JQ69953 * marginatus B6 Glaucus Glaucidae AMS C JQ JQ JQ * marginatus B7 Glaucus Glaucidae AMS C JQ JQ JQ * marginatus B8 Glaucus Glaucidae AMS C JQ JQ JQ * marginatus B9 Glaucus Glaucidae AMS C JQ JQ JQ * marginatus B0 Glaucus Glaucidae AMS C JQ JQ JQ * marginatus C Glaucus Glaucidae UMMZ JQ JQ JQ JQ69964 marginatus C Glaucus Glaucidae UMMZ JQ JQ69945 JQ JQ69965 marginatus C3 Glaucus Glaucidae UMMZ JQ JQ69945 JQ JQ69966 marginatus C4 Glaucus Glaucidae UMMZ JQ69936 JQ JQ69954 JQ69967 marginatus C5 Glaucus Glaucidae UMMZ JQ69936 JQ JQ69954 JQ69968 marginatus C6 Glaucus Glaucidae UMMZ JQ JQ JQ JQ69969 marginatus C7 Glaucus Glaucidae UMMZ JQ JQ JQ JQ69960 marginatus C8 Glaucus Glaucidae UMMZ JQ JQ JQ JQ6996 marginatus C9 Glaucus Glaucidae UMMZ JQ JQ JQ JQ6996 marginatus C0 Glaucus Glaucidae UMMZ JQ JQ JQ JQ69963 marginatus D Glaucus Glaucidae UMMZ JQ JQ JQ JQ69964 marginatus D Glaucus Glaucidae UMMZ JQ JQ69946 JQ JQ69965 marginatus D3 Glaucus Glaucidae UMMZ JQ JQ69946 JQ JQ69966 marginatus D4 Glaucus Glaucidae UMMZ JQ69937 JQ JQ69955 JQ69967 marginatus D5 Glaucus Glaucidae UMMZ JQ69937 JQ JQ69955 JQ69968 marginatus D6 Glaucus Glaucidae UMMZ JQ JQ JQ JQ69969 crassicornis Hermissenda Glaucidae LACM JQ JQ JQ JQ hakunamatata Hermosita Glaucidae LACM 7503 JQ JQ JQ JQ69963 poica Learchis Glaucidae CPIC 0037 JQ JQ JQ JQ69963 sebastiani Nanuca Glaucidae CPIC 005 JQ JQ JQ JQ pugnax Phidiana Glaucidae LACM JQ JQ JQ * longicirrum Phyllodesmium Glaucidae LACM 5383 JQ JQ69947 JQ JQ parangatum Phyllodesmium Glaucidae CASIZ JQ JQ69947 JQ JQ juliae Protaeolidia Aeolidiidae LACM 5385 JQ69938 * JQ69956 * ianthina Pteraeolidia Glaucidae LACM JQ69938 JQ JQ69956 * neapolitana Spurilla Aeolidiidae LACM JQ JQ JQ JQ sargassicola Spurilla Aeolidiidae CPIC 0078 JQ * JQ *

A New Species of Glossodoris (Mollusca: Nudibranchia), of the Glossodoris atromarginata Color Group, from Indonesia 1

A New Species of Glossodoris (Mollusca: Nudibranchia), of the Glossodoris atromarginata Color Group, from Indonesia 1 A ngel Valdés 2,3 and Mary Jane Adams 2 Abstract: A new species, Glossodoris tibboeli