Meiotic behavior of two polyploid species of genus Pleurodema (Anura: Leiuperidae) from central Argentina

Polyploidy is an important evolutionary force but rare in vertebrates. However, in anurans, the genus Pleurodema has polyploid species, two of them tetraploid and one octoploid. The manner in which the chromosomes join in diakinesis can vary among species and, crucially, if they differ in their ploidy levels. In this work, we describe the meiotic configurations in two cryptic species from central Argentina, with different ploidy levels, Pleurodema kriegi (tetraploid) and P. cordobae (octoploid). A total of 306 diakineses from 19 individuals were analyzed. In meiosis, P. kriegi form 22 bivalents, whereas P. cordobae exhibits variation in meiotic figures. We discuss the possible allo- and autopolyploid origin of these species, and we consider that the autopolyploid origin of P. cordobae from P. kriegi might be the most feasible

Pleurodema kriegi Muller 1926

Re-description of <i>Pleurodema kriegi</i> (Müller, 1926) <p>Fig. 8</p> <p> <i>Paludicola kriegi</i> Müller, 1926.</p> <p> <i>Pleurodema kriegi</i> di Tada, Salusso and Martori, 1976.</p> <p> <b>Material. Neotype</b>. FML 20460, adult male (Fig. 8) near La Posta (31º36'46'' S, 64º52'29'' W, approximately 2151 m elevation), Pampa de Achala, Córdoba province, Argentina (Fig. 1), collected during the night of 17 January 2006 by Julián A. Valetti.</p> <p> <b>Referred specimens. (17 specimens).</b> FML 20459, 20461, 20463–20471, 20474-20476, fourteen adult males and FML 20462, 20472–20473, three adult females, all collected with the neotype.</p> <p> <b>Diagnosis.</b> A small species of <i>Pleurodema</i> (33.24 mm SVL), characterized by: (1) relatively small size; (2) snout short, canthus rostralis rounded in dorsal view and truncate in profile; (3) lumbar glands present, 1 ½ eye diameter; (4) round tympanic annulus, almost concealed; (5) tympanum length half of the eye diameter; (6) vomerine teeth absent, (7) comissural gland prominent; (8) brilliant red-orange spots on the groin and around lumbar glands; (9) ploidy level 2n= 4x =44; (10) axillary amplexus; (11) egg deposition in gelatinous nest attached to vegetation; (12) compound advertisement call with tri-pulsed pulse groups, average dominant frequency 1823 Hz.</p> <p> According to external morphology, <i>Pleurodema kriegi, Pleurodema bibroni</i> and the new species described below are cryptic and the properties of their advertisement calls are similar (see above). <i>Pleurodema kriegi</i> differs from the new allopatric species from Sierra de Comechingones by having tetraploid chromosomic complement (octoploid in the new species) and by having a lower pulse rate (55.9 pulses/sec in <i>Pleurodema kriegi</i> and 65.7 pulses/sec in the new species). <i>Pleurodema kriegi</i> and <i>Pleurodema bibroni</i> are the only tetraploid species of the genus. They are morphologically very similar and although Laurent (1975) found some morphological differences between these entities, an unequivocal distinction between these taxa is not possible. The advertisement call of <i>Pleurodema kriegi</i> is similar to <i>Pleurodema bibroni</i>, too. Barrio (1977) detected difference in the pulse rate between these taxa, although the temperatures of recording are different and this environmental variable can affect this temporal variable of the call; Kolenc <i>et al.</i> (2009) found no acoustic differences between these two species. Barrio and Rinaldi de Chieri (1970) found secondary constriction chromosomes 12 only in <i>Pleurodema kriegi.</i> Ultimately, <i>Pleurodema kriegi</i> and <i>P. bibroni</i> were considered as distinct species by Barrio (1977) based mostly on biogeographic and ecologic differences.</p> <p> <i>Pleurodema kriegi</i> is distinguished markedly from <i>P. tucumanum, P. nebulosum, P. guayapae, P. marmoratum</i> and <i>P. diplolister</i> by the presence of lumbar glands (lumbar glands absent in these species); from <i>P. bufoninum, P. borellii, P. cinereum</i> and <i>P. thaul</i> by the presence of advertisement call compound by tripulsed pulse groups (absent in <i>P. bufoninum,</i> Duellman & Veloso 1977; formed by a single pulse group in <i>P. borellii</i> and <i>P. cinereum</i>, McLister <i>et al</i>. 1991; and formed by pulse groups of 5-6 pulses in <i>P. thaul</i>, Barrio 1977); from <i>P. brachyops</i> by having lumbar glands bigger than eye diameter (smaller in <i>P. brachyops</i>), yellow lumbar glands with black central ocellus (black lumbar glands with whitish central blotchs in <i>P. brachyops</i>), tympanum size half of the eye diameter (smaller than half eye diameter in <i>P. brachyops</i>).</p> <p> <b>Description of the neotype.</b> The neotype (FML 20460, Fig. 8) is an adult male of 34.4 mm, body robust; head triangular, slightly wider than long; snout short, canthus rostralis rounded and vertically truncated. Eyes protuberant; eye diameter larger than interocular distance; interocular distance larger than internarinal interval. Round tympanic annulus almost concealed, approximately half the size of eye diameter. Commissural glands present, oval, approximately the same size of eye diameter. Vomerine teeth absent. Dark vocal sac. Fingers free; relative length of fingers: 3>4>1>2; two not-darkened palmar tubercles (Fig. 8). Femur length less than tibia length; sum of femur length and tibia length longer than foot length. Toes free with cutaneous edge and rudimentary interdigital basal membrane; two metatarsal tubercles, inner metatarsal tubercle pointed and outer metatarsal tubercle shovel-shaped (Fig. 8); relative length of toes: 4>5=3>2>1. Lumbar glands large, oval, one and a half times the size of eye diameter. Vocal sacs single, median and subgular. In life, dorsally brownish with large almost symmetrical dark spots. Dark transverse bands on upper surface of arms and legs. Yellow lumbar glands with a black central ocellus covering 60% of the gland. Brilliant red-orange spots on the groin and around lumbar glands. Dark palms and soles, with whitish palmar tubercles. Dark vocal sac with the rest of ventral body surface whitish and mild dotted dark. Iris gold with black reticulations. The neotype is ventrally very infected by the sub-cutaneous acari <i>Hannemania achalai</i> Alzuet and Mauri, 1987.</p> <p>Morphometric measures of neotype (mm): Snout-vent length 34.4; maximal head width 13.6; head length 11.5; snout-eye distance 5.0; internarinal distance 1.9; interocular distance 2.7; eye-narinal distance 2.7; rostronarinal distance 3.2; eye diameter 3.6; arm length 16.1; length of 3rd finger 4.6; femur length 15.0; tibia length 15.8; foot length (including tarsal length) 22.5; length of 3rd toe 5.2; length of 4th toe 9.1.</p> <p> <b>Distribution.</b> The species is currently known from Pampa de Achala, Córdoba province (Fig. 1) (see Barrio 1977, Kolenc et al. 2009) and cited to near Alpa Corral, Córdoba province by Ávila and Priotto (1995). However, the last record should be corroborated because the locality is to the south of the new species with octoploid chromosomic complement and the specimens could be octoploid, too.</p> <p> <b>Ecology.</b> <i>Pleurodema kriegi</i> males were acoustically active from the first rains of austral spring (November) to March from 21.00 hs. to 3.00 hr. (sunset time: 21.00 – 21.30 hr) <i>Pleurodema kriegi</i> breeds in temporary and semipermanent ponds with vegetation at the edges, or aquatic vegetation, and 10 to 40 cm deep (Fig. 9). The males emit their calls floating in the surface of the water or underground near the edge of ponds. The eggs are deposited in semisubmerged eggs-masses and adhered to the aquatic vegetation. The amplexus is axillary. This species was observed in sympatry with <i>Rhinella achalensis</i>, and in syntopy with <i>R. arenarum, Odontophrynus achalensis</i> and <i>Hypsiboas cordobae</i>.</p> <p> The tadpole of <i>Pleurodema kriegi</i> was recently redescripted by Kolenc <i>et al.</i> (2009) and a comparison with <i>Pleurodema bibroni</i> was performed.</p>Published as part of <i>Valetti, Julián A., Salas, Nancy E. & Martino, Adolfo L., 2009, A new polyploid species of Pleurodema (Anura: Leiuperidae) from Sierra de Comechingones, Córdoba, Argentina and redescription of Pleurodema kriegi (Müller, 1926), pp. 1-21 in Zootaxa 2073</i> on pages 10-13, DOI: <a href="http://zenodo.org/record/187133">10.5281/zenodo.187133</a&gt

Pleurodema cordobae Valetti, Salas & Martino, 2009, sp. nov.

<i>Pleurodema cordobae</i> sp. nov. <p>Fig. 10</p> <p> <b>Material. Holotype</b>. FML 20490, adult male (Fig. 11) from Estancia Los Tabaquillos (32º23'58.4'' S, 64º55'35.1'' W, approximately 2105 m elevation), Sierra de Comechingones, Córdoba province, Argentina (Fig. 1), collected during the night of 0 5 February 2006 by Adolfo L. Martino and Julián A. Valetti.</p> <p> <b>Paratypes (9 specimens).</b> FML 20492, adult female and FML 20484–20489, 20491, eight adult males, all collected with the holotype.</p> <p> <b>Diagnosis.</b> A small species of <i>Pleurodema</i> (35.41 mm SVL) with lumbar glands and chromosomic complement octoploid. The new species is distinguished from other species of the genus <i>Pleurodema</i> by the following combination of characters: (1) relatively small size; (2) snout short, canthus rostralis rounded in dorsal view and truncate in lateral view; (3) lumbar glands present, 1 ½ eye diameter; (4) round tympanic annulus, almost concealed; (5) tympanum length half of the eye diameter; (6) vomerine teeth absent, (7) comissural gland prominent; (8) brilliant red-orange spots on the groin and around lumbar glands; (9) ploid level 2n= 8x =88; (10) axillary amplexus; (11) egg deposition in gelatinous nest attached to vegetation; (12) compound advertisement call with tri-pulsed pulse groups, average dominant frequency 1787 Hz.</p> <p> According to external morphology, <i>Pleurodema kriegi, Pleurodema bibroni</i> and <i>Pleurodema cordobae</i> <b>sp. nov.</b> are cryptic and the properties of their advertisement calls are very similar. <i>Pleurodema cordobae</i> <b>sp. nov.</b> is the only octoploid species of the genus (<i>P. k r i e g i</i> and <i>P. bibroni</i> are tetraploid and the rest of species are diploid; Brum-Zorrilla & Sáez, 1968; Barrio & Rinaldi de Chieri, 1970; Veloso <i>et al</i>., 1972; Duellman & Veloso, 1977; Schmid <i>et al.</i>, 1993; Lourenço <i>et al.</i>, 2006). Morphometric comparisons indicate differences between <i>Pleurodema cordobae</i> <b>sp. nov.</b> and <i>P. k r i e g i</i> (Table 1). However, there are no morphometric measures that allow a precise identification between both <i>taxa</i>. The temporal variables of the advertisement call differ from <i>P. k r i e g i</i> (Table 2) and the pulse rate is higher than <i>P. k r i e g i</i>. The erythrocyte size of <i>Pleurodema cordobae</i> <b>sp. nov.</b> is significantly bigger than <i>P. k r i e g i</i> (Table 4) near 100 µm2.</p> <p> <i>Pleurodema cordobae</i> <b>sp. nov.</b> is distinguished markedly from <i>P. tucumanum, P. nebulosum, P. guayapae, P. marmoratum</i> and <i>P. diplolister</i> by the presence of lumbar glands; from <i>P. bufoninum, P. borellii, P. cinereum</i> and <i>P. thaul</i> by the presence of advertisement call compound by tri-pulsed pulse groups (absent in <i>P. bufoninum,</i> Duellman & Veloso 1977; formed by a single pulse group in <i>P. borellii</i> and <i>P. cinereum</i>, McLister <i>et al</i>. 1991; and formed by pulse groups of 5-6 pulses in <i>P. thaul</i>, Barrio, 1977); from <i>P. brachyops</i> by having lumbar glands bigger than eye diameter (smaller in <i>P. brachyops</i>), yellow lumbar glands with black central ocellus (black lumbar glands with whitish central blotchs in <i>P. brachyops</i>), tympanum size half of the eye diameter (smaller than half eye diameter in <i>P. brachyops</i>).</p> <p>Sixteen morphometric distances of the type series describe the quantitative morphological features of the new species (Table 5).</p> <p> <b>Description of the holotype.</b> The holotype (FML 20490, Fig. 11) is an adult male of 35.2 mm (Table 5), body robust; head triangular slightly wider than long; snout short, canthus rostralis rounded in dorsal view and truncated in lateral view. Eyes protuberant; eye diameter equal to interocular distance; interocular distance larger than internarinal interval. Round tympanic annulus almost concealed, approximately half the size of eye diameter. Commissural glands present, oval, approximately the same size of eye diameter. Vomerine teeth absent. Dark vocal sac. Fingers free; relative length of fingers: 3>4>1>2; two not-darkened palmar tubercles (Fig. 11). Femur length less than tibia length; sum of femur length and tibia length longer than foot length.</p> <p>Toes free with cutaneous edge and rudimentary interdigital basal membrane; two metatarsal tubercles (Fig. 11); relative length of toes: 4>5=3>2>1. Lumbar glands large, oval, one and a half times the size of eye diameter. Vocal sacs single, median and subgular. In life, dorsally brownish with large almost symmetrical dark spots. Dark transverse bands on upper surface of arms and legs. Yellow lumbar glands with a black central ocellus covering 60% of the gland. Brilliant red-orange spots on the groin and around lumbar glands. Dark palms and soles, with whitish palmar tubercles. Dark vocal sac, with the rest of ventral body surface being whitish and mild dotted dark. Iris gold with black reticulations.</p> <p> <b>TABLE 5.</b> Morphometric features of the type series of <i>Pleurodema cordobae</i> <b>sp. nov.</b> which were collected at Est. Los Tabaquillos, Córdoba. All data are given in [mm]. (1) snout-vent length (SVL); (2) maximal head width; (3) head length; (4) snout-eye distance; (5) internarinal distance; (6) interocular distance; (7) eye-narinal distance; (8) rostronarinal distance; (9) eye diameter; (10) arm length; (11) length of 3rd finger; (12) femur length; (13) tibia length; (14) foot length; (15) length of 3rd toe; (16) length of 4th toe. FML = Fundación Miguel Lillo, Tucumán, Argentina. M= male; F= female.</p> <p> <b>Distribution.</b> The species is currently known from <i>Terra typica</i>, Estancia Los Tabaquillos (Fig. 1) and two temporary ponds located 5 kilometers away from the former.</p> <p> <b>Ecology.</b> <i>Pleurodema cordobae</i> <b>sp. nov.</b> males were acoustically active from December to March (Austral summer) from 21.00 hs. to 4.00 hr. (sunset time: 21.00 – 21.30 hr) <i>Pleurodema cordobae</i> breeds in temporary and semipermanent ponds with vegetation at the edges and a depth of 20 to 30 cm (Fig. 12). The males emit their calls floating on the surface of the water near the edge of ponds (Fig. 13). The eggs are deposited in semisubmerged eggs-masses and adhered to the vegetation (Fig. 14). The amplexus is axillary. This species was observed in syntopy with <i>Rhinella achalensis</i>, <i>R. arenarum</i>, <i>Odontophrynus achalensis</i> and <i>Hypsiboas cordobae</i>.</p> <p> <b>Etymology.</b> The specific name (a noun in the genitive case) refers to the currently known geographical distribution of the new species.</p>Published as part of <i>Valetti, Julián A., Salas, Nancy E. & Martino, Adolfo L., 2009, A new polyploid species of Pleurodema (Anura: Leiuperidae) from Sierra de Comechingones, Córdoba, Argentina and redescription of Pleurodema kriegi (Müller, 1926), pp. 1-21 in Zootaxa 2073</i> on pages 13-18, DOI: <a href="http://zenodo.org/record/187133">10.5281/zenodo.187133</a&gt

Hannemania achalai Alzuet and Mauri 1987

<i>Hannemania achalai</i> Alzuet and Mauri, 1987 (Figures 1–3) <p> <i>H. achalai</i> Alzuet and Mauri, 1987: 114</p> <p> Diagnosis — SIF = 5B–B–3–2111.0000; fPp = B/B/BBB. Pc = 3; Gn = 2; fSc = PL> AL ≥ AM; PL/SB; fCx = 2.1.1; fSt = 0.1; DS = 53 – 75; VS = 49 – 69; NDV = 132; Ip = 765 – 865; AW = 50 – 60; PW = 65 – 80; SB = 20 – 30; ASB = 45 – 55; PSB = 20 – 25; SD = 75 – 80; AP = 15 – 20; AM = 30 – 40; AL = 30 – 40; PL = 60 – 70; S = 75 – 115; H = 40 – 45; D min = 30 – 45; D max = 50 – 65; Vmin = 30 – 35; V max = 35 – 45; pa = 280 – 320; pm = 240 – 275; pp = 250 – 290.</p> <p>Emended description (larva, n = 63)</p> <p>Idiosoma — 950 – 1215 long and 450 – 600 wide (Figures 1 A–B). Eyes 2 + 2, total number of idiosomal setae range from 102 to 144. Scutum with naso, 70 – 80 long and 70 – 85 wide (Figure 1E); diameter of sensillary bases 7 – 9, AMW = 9 – 11. Ventrally with only one pair of posterior sternal setae (Figure 1B).</p> <p> Gnathosoma — 95 – 110 long and 60 – 70 wide (Figures 1 C–D, 3A). Palp, five-segmented with following setal distribution: trochanter without setae, femur and genua with one branched 2009, coll. A. L. Martino, P. R. Grenat and M. Otero. 5 larvae (CNAC007188–007191) ex <i>Pleurodema cordobae</i> [884], ARGENTINA, Córdoba, Mal Paso, Pampa de Achala, - 31°49’52”, -64°51’40”, 2308 m a.s.l., 16 Feb. 2010, coll. A. L. Martino, J. A. Valetti, P. R. Grenat, M. Otero and F. G. Biolé. 11 larvae (CNAC007192–007197) same data except host [885]. 5 larvae (CNAC007198–007201) ex <i>Pleurodema cordobae</i> [898], ARGENTINA, Córdoba, Cerro Negro, -31°57’26.2”, -64°54’59.7”, 2323 m a.s.l., 5 Nov. 2011, coll. M. Otero. 12 larvae (CNAC007202–007206) ex <i>Pleurodema cordobae</i> [905], ARGENTINA, Córdoba, Los Tabaquillos, -32°23’58.4”, -64°55’35.1”, 2113 m a.s.l., 22 Nov. 2011, coll. J. A. Valetti, P. R. Grenat, M. Otero and A. L. Martino. 6 larvae (CNAC007207–007210) ex <i>Pleurodema kriegi</i> [759] ARGENTINA, Córdoba, La Posta, -31°36’43.7”, -64°52’26.7”, 2159 m a.s.l., 4 Dec. 2008, coll. J. A. Valetti, J. Marquez and M. Ammann. 6 larvae (CNAC007211–007213) ex <i>Pleurodema kriegi</i> [862], ARGENTINA, Córdoba, Pampa de Achala, La Trinidad, -31°44’13”, -64°50’58”, 2318 m a.s.l., 5 Jan. 2010, coll. J. A. Valetti, A. L. Martino and C. Casale. 12 larvae (CNAC007214–007217) ex <i>Pleurodema kriegi</i> [1633], ARGENTINA, Córdoba, Pampa de Achala, El Cóndor, -31°36’23”, -64°52’06”, 2178 m a.s.l., 16 Dec. 1981, coll. R. Martori.</p> <p>Remarks — All the specimens with 2 genualae I, except one specimen (CNAC007207) with 2 genualae on right leg and 1 genuala on left leg, and one specimen (CNAC007186) with 2 genualae on right leg and 3 genualae on left leg. All the specimens with 1 genuala III, except one specimen (CNAC007184) with 1 genuala on left leg and 2 genualae on right leg.</p> <p> We find that our specimens are conspecific with <i>H. achalai</i> based on the palpal setal formula, the palpal claw, and the number of specialized nude setae on legs: genualae I–III, tibialae I–III, tarsalae and pretarsalae I–II, and subterminala I.</p> <p>However, the specimens analysed in this study differ from the original description by Alzuet and Mauri (1987) in the following characters: 1) the shape of galeala which is sparsely branched instead nude as mentioned by Alzuet and Mauri (1987), and 2) the presence of 2 genualae I constantly, whereas in the type specimens the number of genualae I range 2–4 (Alzuet and Mauri 1987). These differences are considered here as intraspecific variation until type material can be examined.</p> <p> According to Alzuet and Mauri (1987) and Salazar Martínez <i>et al</i>. (2004) the type material is deposited at Museo de La Plata (MLP), Argentina, however, types were not seen for this study.</p>Published as part of <i>Paredes-León, Ricardo, Biolé, Fernanda G., Valetti, Julián A. & Martino, Adolfo L., 2018, A redescription of the chigger Hannemania achalai Alzuet and Mauri, 1987 (Acariformes: Prostigmata: Leeuwenhoekiidae) in frogs from Sierra Grande, Cordoba, Argentina, pp. 159-164 in Acarologia 1987 (1)</i> on pages 160-164, DOI: 10.24349/acarologia/20184234, <a href="http://zenodo.org/record/4487485">http://zenodo.org/record/4487485</a&gt

Are ploidy and age size-related? A comparative study on tetraploid Pleurodema kriegi and octoploid P. cordobae (Anura: Leptodactylidae) from Central Argentina

Size differences of anuran amphibian complexes with different ploidy levels are well known, whereas the larger species usually corresponds to a lower ploidy level. Neotropical Pleurodema kriegi (tetraploid) and P. cordobae (octoploid) differ in many morphometric variables and, in all cases, the species with the higher ploidy level corresponded to an increased body size. Although sexual size dimorphism (SSD) has been tested in P. kriegi without considering age-related differences in body size, SSD in P. cordobae is unknown. Here we determined individual age in order to analyse: (1) the intraspecific pattern of SSD of both species and (2) whether a larger body size and lower ploidy level relation observed in other polyploid anuran amphibians are reversed in a tetra-octoploid complex. Age and body size (snout-vent length) were correlated in both sexes and species. When the effect of age was controlled, significant a SSD were found. Intraspecific differences in demographic parameters were significantly higher than interspecific ones. We confirmed a female-biased SSD for both species and found the first case in which the larger species corresponds to a higher ploidy level. Different ploidy levels were an important factor determining age and body size under similar environmental conditions. Skeletochronology provided basic demographic data of P. kriegi and P. cordobae which are now available for comparative and conservation studies.Fil: Otero, Manuel Alejandro. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Ecología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Valetti, Julian Alonso. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Ecología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bionda, Clarisa de Lourdes. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Ciencias de la Tierra, Biodiversidad y Ambiente - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencias de la Tierra, Biodiversidad y Ambiente; ArgentinaFil: Salas, Nancy Edith. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Ecología; ArgentinaFil: Martino, Adolfo Ludovico. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Ecología; Argentin