Conolophus marthae C. pallidus C. subcristatus C. subcristatus Amblyrhynchus cristatus Bell, 1825 C. subcristatus Conolophus marthae C. marthae Conolophus marthae Conolophus marthae sp. nov. (Squamata, Iguanidae), a new species of land iguana from the Galápagos archipelago Gentile, Gabriele Snell, Howard Zootaxa 2009 2201 1 10 [151,406,1648,1674] Reptilia Iguanidae Conolophus Animalia Squamata 1 2 Chordata species marthae sp. nov.    Holotype.A free-ranging adult male permanently branded with the number 117. A Passive Integrated Transponder (PIT) with the number 091-601-303 was hypodermically inserted in one of the posterior legs. The individual was captured and released approximately four km north of the Equator on the top of Volcan Wolf, IslaIsabela, Galápagos National Park, Ecuador( 0.03792° N; 91.36324°W, datum WGS84, as recorded by a Garmin 12CX handheld GPS). The individual was captured by A. Jaramillo on June 8th 2006, blood was drawn by G. Gentile. Photos were shot by G. Gentile. Blood in lysis buffer voucher n. MCZRR450 (as reported in the Genbank records FJ716129and FJ716130) is hosted in the reptile collection (as specimen n. R450) of the Civic Museum of Zoology ( MCZR, Rome, Italy,). Original photo files, named as “Morphobank_m27772.jpg” ( Figure 2), “Morphobank_m27773.jpg, Morphobank_m27774.jpg, M o r p h o b a n k_m 2 7 7 7 5. j p g, M o r p h o b a n k _m2 7 7 7 6. j p g, M o r p h o b a n k _m2 7 7 7 7. j p g, a n d Morphobank_m27778.jpg” ( Figures 3A, 3B, 3C, 3D, 3E, and 3F, respectively), and the movie “Morphobank_m27779.wmv” are included in a project titled as the present paper, hosted in Morphobank (http://www.morphobank.org). Such photos and video form a basis of the description and should be considered also as illustrating the typespecimen, for purposes of Article 73.1.4 of the Code ( ICZN, 1999), but see also the paragraph “Notes added in proofs”. All material refers to the same individual (free ranging, with PIT number 091-601-303), elected as Holotype.   Diagnosis.  Conolophus marthae  sp. nov.is distinguished from  C. pallidusand  C. subcristatusby the following color pattern: pinkish head, pinkish and black (dark) body and legs, with a typical black-striped pattern on the mid to posterior dorsal body; stripes are along the dorsal-ventral axis, may be irregular and their number variable; stripes may join to form a more complex pattern; stripes occur on the ventral body, but are less evident; dark tail. Other distinctive, but slightly variable morphological traits co-occur in males: i) adipose nuchal crest with small or reduced conic scales, ii) poorly elevated (pyramid-shaped) or almost flat dorsal head scales.   Conolophus marthae  sp. nov.is also distinguished from the other two congeneric species by a distinctive pattern of head-bob behavior ( Fig. 6, see Morphobank accession code: p241). The new species is unequivocally distinguished from  C. pallidusand  C. subcristatusby the several diagnostic sites in the sequence of the control region and cytochrome bgene of the mtDNA, reported in Table 1, and by a completely different, non overlapping, size-range of alleles at the microsatellite locus CS7 ( Tzika et al., 2008; Gentile et al.2009). Alleles at locus CS7 range between 245 and 333 bases (as defined in Gentile et al.2009).   Description of Holotypein life.  Sex:Male   Age:Adult  Weight:5.0 Kg.  Morphological measurements:SVL: 47.0 cm; VTL: 61.4 cm; head length: 78.22 mm; head width: 63.76 mm; internostril distance: 17.89 mm; eye-eye distance: 35.19 mm.   TABLE 1.Diagnostic sites in the sequence of the control region and cytochrome bgene in the mtDNA. Position are based on the Genbank records FJ716129(Control region) and FJ716130(cytochrome b). The sites 471 and 474 of the cytochrome bsequence are polymorphic in  C. subcristatus, but still diagnostic.  Control region  786 465 12 24  795 471 51 25  796 474 69 63   C. subcristatusand C.   pallidusG 807T T A C T C T 492T A T C T G C 75A G A C G G T C 85T  C. marthae  sp. nov.C 814C A T T G T C 498A G C T A A A 117G A C T A A C T 92C  831 525 135 93  847 528 147 167  Cytochrome b  867 536 171 168  868 547 207 179  873 550 249 198  C. subcristatusC 889C C C C C C C 553C C A T T T C 255A A A G C C T 204C G  C. pallidus. 890....... 561....... 267....... 205..  C. marthae  sp. nov.T 891T T T T T T T 573A T C C C C T 291G T T A T T C 247T A  914 600 295 318  948 633 315 335  967 666 321 338  1014 683 363 508  C. subcristatusC Y R T C C C T G A C C T C T C T A A G T T G A  1053 693 369 509  C. pallidus. T G.....................  1059 700 372 512  C. marthae  sp. nov.T G C C T T T C A G A T C T C A C G G C C C A C  1062 702 408 651  1068 706 411 696  1071 721 417 834  1081 723 426 850  1087 747 463 1098  C. subcristatusA T T C C C T C C T C T C C T G C G C A T A G C  C. pallidus.... A...................  C. marthae  sp. nov.G C C T T T C T T C T C T G C A T A T G C G T A Meristic characteristics:N. supralabial scales: 7 (left side) and 9 (right side); n. infralabial scales: 10 (left side) and 9 (right side); n. scales around the parietal scale: 8; n. scales around the mental scale: 9; n. scales around the rostral scale: 8; n. scales along the middle-dorsal line: 17; n. scales around the inguinal scar: 46. Number of femoral pores: 19 (left leg) and 18 (right leg).  Morphological characteristics:Snout elongated, not shortened. Tympanum taller than wide. Scales flat or almost flat above the tympanum, in the post-orbital region. Slightly more elevated pyramid-shaped scales occur in the dorsal head. Nuchal crest pronounced, adipose, with small conic scales which are reduced or almost flat along the ridge of the anterior half of the crest. Conic scales are more prominent, but not spinose, along the ridge of the posterior half. Dorsal crest less developed, with small conic scales along the ridge. Caudal crest poorly developed. Round-cross-section tail, not laterally compressed. Fingers of fore and hind legs with short claws, not recurved.   FIGURE 3.  Conolophus marthae  sp. nov.Holotype. Adult male with brand number 117 and PIT number 091-601-303. A–D) views of the head: right, frontal, left, and from above, respectively; E) dorsal view; F) ventral view. The individual is changing skin.   FIGURE 4.  Conolophus marthae  sp. nov.Adult male with brand number 118 and PIT number 091-062-369, held by A. Jaramillo, field assistant of the Charles Darwin Research Station. This individual is not the Holotype.   FIGURE 5.  Conolophus marthae  sp. nov.Adult male with brand number 36 and PIT number 091-336-546. Details of the head. This individual is not the Holotype.   FIGURE 6.Head-bob display of  Conolophus marthae  sp. nov.Multiple series (a, b, c, and d) of head-bob performed by the Holotype (June 2006). The series were performed with the following time intervals between each other: a–b, 27 seconds; b–c, 34 seconds; c–d, 11 seconds.  Coloration:pinkish head, pinkish and black (dark) body and legs, with a black-striped pattern on the mid to posterior dorsal body. On both sides, five vertical black stripes occur between forelimb and hind limb, along the dorsal-ventral axis. The first stripe is interrupted. Stripes 2–5 are joined horizontally, describing a complex pattern. Stripes are present but less evident on the ventral body. Dark tail.  Behavioral characteristics:The head-bob display (nodding behavior) consists of repeated modules. Each module comprises three series of multiple head movements (“ups and downs”; Fig. 6a–c) and is completely executed within a time interval of 4–5 seconds. Frequency of movements performed in each series is high, with 4 to 6 movements per second. Two sub-series, separated by a few deciseconds, may be recognized within series 2. A fourth, additional series, similar to series 3, may be observed occasionally ( Fig. 6d).   Etymology.The new species is named in memory of Martha Rebecca Gentile, second daughter of the first author. Martha prematurely left this world. She was born dead, as consequence of a medical doctor’s negligence, on August 20th 2003.   Distribution.Thus far, this species is known to occur only on Volcan Wolf ( Fig. 1), the northernmost volcano of IslaIsabela (Galápagos National Park, Ecuador).   Remarks.The new species is easily distinguished from the other two congeneric species. The color pattern is typical of the new species and was never observed in any of the populations of the other two named species. The origin and the nature of the pink pigmentation deserve further investigation. Nevertheless, it is instructive to note that when we surgically removed one pink scale, blood flowed out of the tissue of the removed scale, which immediately lost its pink color. Traits i) and ii) in the diagnosis are more evident in males, whereas they are variable and generally less pronounced in females. Although in the Plaza Sur population of  C. subcristatusalmost flat dorsal head scales may be observed, such a trait never co-occurs in combination with the other traits characteristic of  C. marthae  sp. nov. Although the “head-bob” pattern is slightly different between  C. subcristatuspopulations in different islands (Gentile, unpublished data), the nodding behavior of  C. marthae  sp. nov.is very distinctive and characteristic. This is particularly relevant since it is exhibited in sympatry (syntopy) with  C. subcristatus. None of the other species of land iguanas or any marine iguanas show a similar pattern (see Carpenter, 1982, for a comparison).    Conolophus marthae  sp. nov.is distinct from the other two congeners by about 7% mtDNA genetic divergence, much higher than genetic divergence between  C. pallidusand  C. subcristatus(less than 2%, Gentile et al.2009). Twenty-four nucleotide sites of the control region and seventy-two nucleotide sites in cyt bgene sequences are diagnostically different and allow distinguishing between the new species and the other congeneric ones. The deep divergence is estimated to have started in a period when the Galápagos did not have their current configuration ( Gentile et al.2009). The absence of alleles shared with the other two species at the microsatellite locus CS7 and the presence of several private alleles at other loci ( Tzika et al., 2008; Gentile et al., 2009) indicate genetic isolation, even with the syntopic population of  C. subcristatus. Occasional hybridization between marine (  Amblyrhynchus cristatus Bell, 1825) and land iguanas (  C. subcristatus) may still occur on IslaPlaza Sur, generating a black, brow-striped F1 hybrid ( Rassmann et al.,1997).  Conolophus marthae  sp. nov.lacks in any of the adaptive traits exhibited by marine iguanas (shortened snout; laterally compressed tail; developed caudal crest; long, recurved claws) and genetic data ( Gentile et al.2009) provide strong evidence that  C. marthae  sp. nov.did not originate by hybridization between marine and land (yellow) iguanas. A total of 120 individuals of  Conolophus marthae  sp. nov.were observed and sampled in three field trips, in 2005, 2006, and 2009 (see the paragraph “Notes added in proofs”).  Besides the taxonomic implications,  C. marthae  sp. nov.is very important as it is the only evidence of deep divergence within the Galápagos land iguana lineage. In fact, the new species carries an ancient evolutionary legacy, being the only remnant of a lineage originated when the Galápagos archipelago did not have its present configuration.  Conolophus marthae  sp. nov.is a narrow endemism and its population size is small. Its inclusion in the Red List of the International Union for Conservation of Nature (IUCN) as "critically endangered" has been recommended ( Gentile et al.2009).