Borrero-Pérez, Giomar H.; Solís-Marín, Francisco A.; Lessios, Harilaos
Understanding the color variability and resolving taxonomic confusion in the sea cucumber Isostichopus badionotus (Echinodermata, Holothuroidea): a revision of the genus Isostichopus
Fig. 1 (continued on next page). Color patterns of species and subspecies of Isostichopus Deichmann, 1958. A–J. Isostichopus badionotus (Selenka, 1867). A. Pink background and dark brown spots (Chips pattern – CH) (BT99, Panamá). B. Close-up of A, note large dark brown spots and small dots. C. Light brown background and darker ocellar spots (CH pattern) (INV TEJ1125-Ma10, Colombia). D. Close-up of C, note large ocellar spots and small black dots. E. Uniform black background (Uniform pattern – U) (BT59, Panamá). F. Beige background color with a reticulum of darker brown and yellow papillae (Reticulated pattern – R) (USNM 1659460-BT20, Panamá). G. Close-up of F, reticulated pattern in detail. H. Reticulated pattern with brown papillae (R pattern) (INV TEJ1167-SM4R, Colombia). I. Close-up of H, reticulated pattern in detail. J. Black background and yellow papillae (Black and yellow pattern-BY) (INV TEJ1176-SM13, Colombia). K–Y. Isostichopus maculatus phoenius (Clark, (BI), Maximum Likelihood (ML), and Neighbour-Joining (NJ). BI and ML were performed, using the GTR evolutionary model and NJ using K2P distances. BI was performed with MrBayes ver. 3.2.6 (Ronquist & Huelsenbeck 2003); the data set was run twice, using four Markov chains for ten million generations; trees were sampled every 500th generation, the first 2.5 million generations were discarded, and a 50% majority tree was obtained. ML analysis was performed in MEGA X (Kumar et al. 2018); support was assessed with 1000 bootstrap reiterations.
Fig. 3A. Bayesian inference tree of concatenated sequences of the mitochondrial COI-Fr1 (Barcoding region), COI-Fr2 and 16S gene fragments. The numbers on the nodes indicate Bayesian posterior probability/Maximum Likelihood (bootstrap %)/Neighbor Joining (bootstrap %). A hyphen (-) indicates that a node was absent in the tree estimated by a particular method. Individuals are indicated by the field number, which includes the initial letter of the locality and the color pattern when available (see Table 1, Fig. 1). Text color meaning: green = Isosostichopus maculatus maculatus (Greeff, 1882); blue = I. maculatus phoenius (Clark, 1922); pink = I. badionotus (Selenka, 1867) Chips (CH) pattern; purple = Uniform (U) pattern; red = reticulated (R) pattern; orange = Black and Yellow (BY) pattern. Sequences from GenBank are indicated with their accession number and species ID as included in GenBank (see Table 1). Outgroup species: Apostichopus japonicus (Selenka, 1867) and Stichopus horrens Selenka, 1867.
Fig. 8 (continued on next page). Color patterns and morphological variation of Isostichopus badionotus (Selenka, 1867) (DNA sequences and detailed information of some specimens are indicated in Table 1, see Photo ID column for correspondence). A–U, I’. Specimens with “Chips” color pattern from Bocas del Toro, Panamá (A–I), Neguanje, Colombia (J–L), La Guajira, Colombia (M–Q); juveniles from Neguanje, Colombia (R–U). V–B’. Specimens with “Uniform” pattern from Bocas del Toro (Panama) (V–X), La Guajira, Colombia (Y), Curaçao (Z), Magdalena, Colombia (A’–B’). C’–K’. Specimens with “Reticulated” pattern from Bocas del Toro, Panama (C’–E’), Juvenile from Neguanje, Colombia (F’), Punta Betin, Magdalena, Colombia (G’–I’), La Guajira, Colombia (J’–K’). L’–O’. Specimens with “Black and yellow” pattern from Bocas del Toro, Panamá (L’), Magdalena, Colombia (M’–O’). Photos: A–I, Q, V–X, C’–E’, L’, O’ by G. Borrero; R–U by G. Ospina; J–L by E. Acosta; M–P, Y, J’–K’ by E. Ortiz; Z, M’ by M. González-Wangüemert; G’–I’ by J. Gómez; and A’–B’, N’ by S. Zea. Scale bars = 10 mm.
Fig. 7. Isostichopus badionotus (Selenka, 1867) ossicles (specimen USNM 1659454-BT111, 190 mm long). A. Thin C-shaped rods, tables, perforated plates and large, curved rods from dorsal body wall and papillae. B. Thin C- and S-shaped rods, tables, perforated plates, large, curved rods and end plate (not complete, ¼ of the plate) from ventral body wall and tube feet. C. Rods and tables from tentacles. D. Large tables from mouth membrane. E. Thin C-shaped rods and simple rods from longitudinal muscles. F. Thin C-shaped rods, simple, cross-shaped and irregular plate-like branched rods in the cloaca. G. Table from respiratory trees. H. Cross-shaped ossicles from intestine. I. Simple rod from gonads. Photos by G. Borrero.
Fig. 6. Preserved lectotype of Isostichopus badionotus (Selenka, 1867) from Florida, USA (MCZ HOL- 509, 190 mm long). A. Dorsal view. B. Ventral view with original labels. Scale bar = 35 mm. Photos by G. Borrero.
Fig. 4. Comparison of the calcareous rings of species and subspecies of Isostichopus Deichmann, 1958. A. I. badionotus (Selenka, 1867). B. I. maculatus phoenius (Clark, 1922) (A–B showing calcareous rings from specimens of different sizes). C. I. maculatus maculatus (Greeff, 1882). D. I. fuscus (Ludwig, 1875). E. I. macroparentheses (Clark, 1922). Dimensions of the individuals from which each ring was obtained are shown next to their museum catalog number. Abbreviations: ip = interradial plate; rp = radial plate. Scale bar = 4 mm.
Fig. 11. Light microscope photographs showing ossicles from the mouth membrane, anterior cloaca, posterior cloaca, and gonads of the species and subspecies of Isostichopus Deichmann, 1958. A. I. badionotus (Selenka, 1867) (mouth membrane and gonads from the lectotype MCZ HOL-509; anterior and posterior cloaca from USNM 1659457-IbBT116). B. I. maculatus phoenius (Clark, 1922) (all ossicles from the holotype MCZ HOL-1182). C. I. maculatus maculatus (Greeff, 1882) (all ossicles from the neotype USNM E16150; male gonads at the bottom from the neoparatype USNM E16151). D. I. fuscus (Ludwig, 1875) (mouth membrane from MCZ HOL-742; anterior cloaca from MBMLP- If212; posterior cloaca and gonads from MCZ HOL-743). E. I. macroparentheses (Clark, 1922) (mouth membrane, anterior and posterior cloaca from USNM E47524; gonads from the holotype MCZ HOL- 921). Photos by G. Borrero. Scale bars = 100 µm.
Fig. 10. Light microscope photographs of the same specimens as in Fig. 9 (see the length (L) for correspondence), comparing ossicle size and shape from dorsal papillae and dorsal body wall from small to large specimens from each of the species and subspecies of Isostichopus Deichmann, 1958. A. I. badionotus (Selenka, 1867). B. I. maculatus phoenius (Clark, 1922). C. I. maculatus maculatus (Greeff, 1882). D. I. fuscus (Ludwig, 1875). E. I. macroparentheses (Clark, 1922). Photos by G. Borrero. Scale bars = 100 µm.
Fig. 9. Box-plots comparing ossicle size from dorsal papillae from small to large specimens(along x axis) from each of the species and subspecies of Isostichopus Deichmann, 1958. A. Dorsal papillae: C-shaped ossicle length. B. Dorsal papillae: table height. C. Dorsal papillae: table disc diameter. Photos of the ossicles of the same specimens are presented in Fig. 10. Specimens IDs: I. badionotus (Selenka, 1867) L = 40 mm, USNM E11783; L = 130 mm, USNM 1659462-BT48; L = 195 mm, USNM 1659454- BT111; L = 230 mm, USNM 1659447–BT116; I. maculatus phoenius (Clark, 1922) L = 25 mm, INV EQU4565; L = 90 mm, USNM 1659481-Ga224; L = 125, USNM 1659472-BT192; L = 185, USNM 1659479-BT53; I. maculatus maculatus (Greeff, 1882) L = 55 mm, ASI1; L = 110 mm, ASI4; L = 150 mm, CV1; I. fuscus (Ludwig, 1875) L = 40 mm, INV EQU4229; L = 115 mm, INV EQU4324; L = 55 mm, MBMLP-If212; L = 180 mm, INV EQU4323; I. macroparentheses (Clark, 1922) L = 20 mm, USNM 1659484-Be99; L = 40 mm, USNM 1659485-Be107; L = 65 mm, MCZ HOL-1214; L = 135 mm, USNM E47524.
Fig. 5. Geographic distribution of species and subspecies of Isostichopus Deichmann, 1958. A. I. badionotus (Selenka, 1867) (gray circles), I. maculatus phoenius (Clark, 1922) (red squares) and I. maculatus maculatus (Greeff, 1882) (blue circles). B. I. fuscus (Ludwig, 1875) (green circles) and I. macroparentheses (Clark, 1922) (orange circles). Locations for I. badionotus and I. fuscus were constructed based on observation records in Global Biodiversity Information Facility data and from museum specimens. Locations forI. maculatus phoenius, I. maculatusmaculatus andI. macroparentheses were constructed based only on confirmed localities of collected or preserved museum specimens.
Georgia Marine Resources for Minerals Management Service
Continental Shelf Associes for BLM / MMS
N. Cruz-Macrofauna- Corpoguajira Project
N. Cruz- Macrofauna-Corpoguajira Project
C. C. Nutting & State University of Iowa