Karyomorphology of the genusLycopodium sensu stricto and relationships among species

Karyomorphological comparisons were made of six species of JapaneseLycopodium sensu stricto. There were no marked differences at interphase and prophase among the six species.Lycopodium annotinum had 2n=68 and the formula of its metaphase karyotype was 18m(median centromeric chromosomes)+12sm(submedian)+12st(subterminal)+26t(terminal).Lycopodium casuarinoides had 2n=68=16m+10sm+18st+24t,L. clavatum 2n=68=22m+12sm+18st+16t, andL. obscurum 2n=68=10m+22sm+20st+16t. Each of these species, which belong to different sections, displayed several karyomorphological differences. Among themL. casuarinoides differs largely from the others in its mean chromosome length, ratio of the longest chromosome to the shortest, and frequency of m+sm chromosomes. BothLycopodium complanatum andL. nikoense, belonging to sectionComplanata, had a common karyotype 2n=46=10m+12sm+18st+6t. This section displayed a low differentiation in its karyotype. In the wholeLycopodium s.s., the ratios of m+sm in a complement varied from 38 to 50%, being higher among pteridophytes.     

Microsatellites loci isolated in the freshwater fish Brycon hilarii

We describe seven microsatellite loci, including tetra‐, tri‐ and dinucleotides, isolated from Brycon hilarii, which is a migratory fish inhabiting the Paraguay River basin (Brazil) and is highly valued for its meat quality. Three to eight alleles per locus were detected and the expected heterozygosity ranged from 0.31 to 0.81. Positive results were obtained with cross‐amplification in five other species of Brycon. These microsatellites provide a potential tool for wild population and aquaculture studies of B. hilarii and other species of the genus.     

Cytological studies on six cyprinid fishes

The chromosome complements of six cyprinid fishes were studied, using the routine air-drying Giemsa staining technique. The diploid chromosome number recorded is 2n = 50 (8m+18sm+14st+10t) with NF = 90 in Aspidoparia morar, 2n = 50 (8m+12sm+12st+18t) with NF = 82 in Crossocheilus latius latius, 2n = 50 (6m+12sm–16st+16t) with NF = 90 in Labeo pangusia, 2n = 70 (16m+6sm+16st–32t) with NF = 108 in Perilampus atpar, 2n = 48 (4m+6st+38t) with NF = 58 in Puntius chrysopterus and 2n = 50 (2m+2sm+4st+42t) with NF = 58 in P. tetrarupagus. Sex chromosomes are not identifiable in any of these species. A pair of marker chromosomes has been observed in all species excepting A. morar.     

Identification and characterization of microsatellites loci in Brycon opalinus (Cuvier, 1819) (Characiforme,Characidae, Bryconiae)

Brycon species are present in various basins in Brazil and were or still are part of commercial fisheries and aquaculture activities ( Ferreira et al. 1996 , Mendonça 1994 ). Despite the importance of this group of fish, natural populations of some Brycon species are endangered ( Lins et al. 1997 ). Here, we describe the characterization of seven microsatellite loci that will be useful for the genetic studies in natural and captive populations for these and other species of Brycon.     

Threatened fishes of the world: Brycon nattereri Günther, 1864 (Characidae)

Brycon nattereri (Ostariophysi: Characiformes: Characidae), a threatened South American freshwater fish, occurs in the Paraná, Tocantins and São Francisco river basins in central Brazil. It is a middle-sized (up to 50 cm SL), omnivorous species, which occurs in swift, clear-water rivers with well-preserved riparian vegetation. Main threats to the species are water pollution, dam building, and deforestation.     

The karyotypes of the thorny catfishes <Emphasis Type="Italic">Wertheimeria maculata</Emphasis> Steindachner, 1877 and <Emphasis Type="Italic">Hassar wilderi</Emphasis> Kindle, 1895 (Siluriformes: Doradidae) and their relevance in doradids chromosomal evolution

We studied the karyotypes of two doradids, the rare and endangered Wertheimeria maculata and a derived Amazonian species, Hassar wilderi. Cytogenetic characterization was assessed using conventional staining (Giemsa), C-banding, and NOR banding. Both species had 2n = 58 chromosomes but differed in their chromosome formulae, 24 m + 14sm + 8st + 12a for W. maculata and 32 m + 16sm + 10st for H. wilderi. In W. maculata heterochromatin was mainly telomeric, and three chromosomes had a fully heterochromatic arm; in H. wilderi heterochromatin was also predominantly telomeric and evident in many more chromosomes. Hassar wilderi also presented one pair of homologues with a fully heterochromatic arm. In both species, nucleolar organizer regions were restricted to one pair of subtelocentric chromosomes. Assuming a basal position for W. maculata, we hypothesized that underlying conserved diploid and NOR-bearing chromosome numbers, chromosomal evolution in doradids has involved pericentric inversions and an increase of heterochromatic blocks.     

Population responses of two omnivorous fish species to impoundment of a Brazilian tropical river

Brycon gouldingi and B. falcatus were the most abundant and widespread of five species of the genus captured in a sampling study encompassing 5 years (1995–2000) in the upper Rio Tocantins, where the river was impounded by the Serra da Mesa hydroelectric dam. Given that impoundments constitute a disturbance to which species will show demographic and biological responses, we assessed such responses of these two omnivorous fishes, whose genus is regarded as vulnerable to impoundments. Both species increased greatly in numbers during the initial reservoir filling period. Brycon gouldingi had higher abundance values than B. falcatus in all sampling months, except in the pre-impoundment period. Both species showed a significant weight increment relative to body length during the time they remained in the reservoir. No reproductive individuals of B. gouldingi were recorded in the study area, but those few of Brycon falcatus were found in both wet and dry seasons, suggesting a more opportunistic strategy of reproduction. Both species were omnivorous and overlapped broadly in diet. The proportions of dietary items, however, varied in relation to fish size class and water dynamics (lotic versus lentic sites). Terrestrial arthropods composed a larger proportion of the diet of smaller-sized individuals and those from lentic sites, whereas seeds and fruits contributed more to the diet of both larger-sized individuals and those from lotic environments. Except for fish and aquatic heteropterans, which were consumed mainly in the reservoir, all other food items were allochthonous, indicating the dependence of these two species of Brycon on the terrestrial environment. The two species responded similarly to the alterations imposed by the impoundment of the upper Tocantins, although reproduction/recruitment pattern was slightly different between species. As observed in other regions of Brazil, impoundments and deforestation appear to produce negative cumulative effects on species of Brycon. These impacts call for greater efforts to conserve and manage members of this diverse and widespread genus. Handling editor: S.M. Thomaz     

Hitting an Unintended Target: Phylogeography of Bombus brasiliensis Lepeletier, 1836 and the First New Brazilian Bumblebee Species in a Century (Hymenoptera: Apidae)

This work tested whether or not populations of Bombus brasiliensis isolated on mountain tops of southeastern Brazil belonged to the same species as populations widespread in lowland areas in the Atlantic coast and westward along the Paraná-river valley. Phylogeographic and population genetic analyses showed that those populations were all conspecific. However, they revealed a previously unrecognized, apparently rare, and potentially endangered species in one of the most threatened biodiversity hotspots of the World, the Brazilian Atlantic Forest. This species is described here as Bombus bahiensis sp. n., and included in a revised key for the identification of the bumblebee species known to occur in Brazil. Phylogenetic analyses based on two mtDNA markers suggest this new species to be sister to B. brasiliensis, from which its workers and queens can be easily distinguished by the lack of a yellow hair-band on the first metasomal tergum. The results presented here are consistent with the hypothesis that B. bahiensis sp. n. may have originated from an ancestral population isolated in an evergreen-forest refuge (the so-called Bahia refuge) during cold, dry periods of the Pleistocene. This refuge is also known as an important area of endemism for several animal taxa, including other bees. Secondary contact between B. bahiensis and B. brasiliensis may be presently prevented by a strip of semi-deciduous forest in a climate zone characterized by relatively long dry seasons. Considering the relatively limited range of this new species and the current anthropic pressure on its environment, attention should be given to its conservation status.     

Comparative karyomorphology and interrelationships ofSelaginella in Japan

Karyomorphological comparisons were made of 16 native and cultivated species ofSelaginella in Japan. The somatic chromosome numbers are 2n=16 inS. boninensis; 2n=18 inS. doederleinii, S. helvetica, S. limbata, S. lutchuensis, S. nipponica, S. selaginoides, S. tama-montana, andS. uncinata; 2n=20 inS. biformis, S. involvens, S. moellendorffii, S. remotifolia, andS. tamariscina; 2n=30 inS. rossii; and 2n=32 inS. heterostachys. The interphase nuclei of all species examined are uniformly assigned to the simple chromocenter type. The metaphase karyotype of 2n=16 (x=8) is 8 m (=median centromeric chromosomes)+8(st+t)(=subterminal and terminal). The group of the species having 2n=18 (x=9) is heterogeneous karyomorphologically: The karyotype ofS. nipponica is 2n=18=6 m+12(st+t),S. tama-montana 10 m+2 sm(=submedian)+6(st+t), andS. uncinata 6 m+7 sm+5(st+t). Although the remaining five species have the common karyotype 8 m+4 sm+6(st+t), the values of mean chromosome length are variable. Another group of the specles having 2n=20 (x=10) is homogeneous, since all species have the same karyotypes 8 m+4 sm+8(st+t) and have similar chromosome size. The karyotype of 2n=30 is 12 m+6 sm+12(st+t) and is suggested to be a triploid of x=10, and 2n=32=16m+16(st+t), a tetraploid of x=8. Thus, three kinds of basic chromosome numbers, x=8, 9, 10 are present in JapaneseSelaginella examined, and their karyomorphological relationships are discussed.     

Considerations on karyotype evolution in the genera Imparfinis Eigenmann and Norris 1900 and Pimelodella Eigenmann and Eigenmann 1888 (Siluriformes: Heptapteridae)

Heptapteridae is one of the fish families of the order Siluriformes with a wide distribution throughout the basins of the Neotropical region. The genera Imparfinis and Pimelodella comprise few species and/or populations with some chromosome information. Specimens of Imparfinis schubarti, Imparfinis mirini, and Pimelodella meeki from different sites located in the Paranapanema River Basin/PR/Brazil were cytogenetically analyzed. The two species of the genus Imparfinis exhibited 2n = 58 and FN = 116: I. schubarti, with a karyotypic formula of 30m + 28sm and I. mirini with a karyotype formula of 36m + 22sm. P. meeki presented a karyotype of 2n = 46 characterized by 26m + 14sm + 6st and FN = 92, confirming a variability in the 2n of the Heptapteridae family. Both Imparfinis species exhibited interstitial NORs in pair 1, coincident with a secondary constriction; P. meeki presented NORs located in the terminal position on the short arm of pair 17. All AgNORs were coincident with 18S rDNA probe and CMA₃ positive. P. meeki showed a small amount of heterochromatin rich in AT and GC bases. The heterochromatin in I. schubarti was CMA₃ positive. In I. mirini the heterochromatin was DAPI-positive. Furthermore, the long arm of one of the chromosomes of pair 19 revealed the presence of heterochromatic heteromorphism only in male individuals. After meiotic analyses, this heteromorphism could be easily identified in the pachytene and metaphase I stages, and was heteropyknotic and DAPI positive. This feature may be an indication of initial differentiation of sex chromosomes in I. mirini, increasing the great karyotypic variability within this family of fish.     

Differentiated ZZ/ZW sex chromosomes in Apareiodon ibitiensis (Teleostei,Parodontidae): cytotaxonomy and biogeography

Conventional and molecular chromosomal analyses were carried out on three populations of Apareiodon ibitiensis sampled from the hydrographic basins of the São Francisco River and Upper Paraná River (Brazil). The results reveal a conserved diploid number (2n = 54 chromosomes), a karyotype formula consisting of 50 m‐sm + 4st and a ZZ/ZW sex chromosome system that has not been previously identified for the species. C‐banding analysis with propidium iodide staining revealed centromeric and terminal bands located in the chromosomes of the specimens from the three populations and allowed the identification of heteromorphism of heterochromatin regions in the Z and W chromosomes. The number of 18S sites located through fluorescent in situ hybridization (FISH) varied between the populations of the São Francisco and Upper Paraná Rivers. The location of 5S rDNA sites proved comparable in one pair of metacentric chromosomes. Thus, the present study proposes a ZZ/ZW sex chromosome system for A. ibitiensis among the Parodontidae, and a hypothesis is presented regarding possible W chromosome differentiation stages in this species through DNA accumulation, showing geographical variations for this characteristic, possibly as a consequence of geographical reproductive isolation.     

Comparative karyology of species of the genera <Emphasis Type="Italic">Misgurnus</Emphasis> and <Emphasis Type="Italic">Cobitis</Emphasis> (Cobitidae) from the Amur River Basin in connection with their taxonomic relations and the evolution of karyotypes

Data are first provided on karyotypes of four species of Cobitidae from the Amur Basin: Misgurnus nikolskyi (2n = 50 = 10m + 4 sm + 36sta, NF = 64), Cobitis lutheri, (2n = 50 = 12m + 8sm + 30sta, NF = 70), C. choii (2n = 50 = 8m + 10sm + 8st + 24a, NF = 68), and C. melanoleuca (2n = 50 = 6m + 16sm + 28sta, NF = 72), and number of chromosomes in M. mohoity (2n = 50) and karyotypes of C. melanoleuca from the basin of the Don, Malyi and Bol’shoi Uzen rivers, and Selenga. These data are discussed in connection with problems of taxonomy of the studied genera, as well as the evolutionary relations of their karyotypes. On the basis of karyological differences between the populations of C. melanoleuca from different parts of the range, a new subspecies C. melanoleuca gladkovi subsp. nov. that inhabits waters of Europe is described.     

Karyology of four land-planarian species of the genus Bipalium from Japan

We have employed a new scale for characterizing chromosomal forms in the karyotypes of four species of Bipalium from five localities in Japan. Specimens of Bipalium nobile Kawakatsu et Makino, 1982, from Yokohama had a diploid chromosome number of 2x = 10 (2m + 2sm + 2sm + st & sm + 2sm); specimens of the same species from Toyonaka had this number as well but with slightly different chromosomal form (2m + 2sm + sm & st + 2st + m & sm). An undescribed species from Sanjô, Bipalium sp. 2, with two dorsal stripes and a yellow head crescent, had 2x = 10 (2m + 2sm + 2sm + 2sm + 2m); and another undescribed species from Chichijima Island, Bipalium sp. 3, with five dorsal stripes, had 2x = 10 (2m + 2sm + 2sm + 2sm + 2m). A non-sexual bipaliid tentatively identified as Bipalium kewense Moseley, 1878, from Chichijima Island had 2x = 18 (2m + 2m + 2m + 2sm + 2st + 2sm + 2sm + 2sm + 2sm).     

Megalictis,the Bone-Crushing Giant Mustelid (Carnivora,Mustelidae, Oligobuninae) from the Early Miocene of North America

We describe cranial and mandibular remains of three undescribed individuals of the giant mustelid Megalictis ferox Matthew, 1907 from the latest Arikareean (Ar4), Early Miocene mammal fauna of Nebraska, and Wyoming (USA) housed at the American Museum of Natural History (New York, USA). Our phylogenetic hypothesis indicates that Ar4 specimens assigned to M. ferox constitute a monophyletic group. We assign three additional species previously referred to Paroligobunis to Megalictis: M. simplicidens, M. frazieri, and “M.” petersoni. The node containing these four species of Megalictis and Oligobunis forms the Oligobuninae. We test the hypothesis that Oligobuninae (Megalictis and Oligobunis) is a stem mustelid taxon. Our results indicate that the Oligobuninae form the sister clade to the crown extant mustelids. Based on the cranium, M. ferox is a jaguar-size mustelid and the largest terrestrial mustelid known to have existed. This new material also sheds light on a new ecomorphological interpretation of M. ferox as a bone-crushing durophage (similar to hyenas), rather than a cat-like hypercarnivore, as had been previously described. The relative large size of M. ferox, together with a stout rostrum and mandible made it one of the more powerful predators of the Early Miocene of the Great Plains of North America.     

A revision of Habenaria section Macroceratitae (Orchidaceae) in Brazil

Habenaria sect.Macroceratitae from Brazil is revised, and seven species are recognized:H. bractescens, H. gourlieana, H. johannensis, H. longicauda, H. macronectar, H. nabucoi, and the newly describedH. paulistana. A main feature of these species is the presence of long, separated, involute stigmatic processes.Habenaria bractescens, H. gourlieana, H. johannensis, andH. macronectar are distributed mainly from central and southeastern Brazil to southern Brazil and southern South America, whereasH. longicauda andH. nabucoi are distributed mainly from west central, southeastern and northeastern Brazil to northern South America.Habenaria paulistana is restricted to the state of São Paulo.Habenaria bradei, H. juergensii, andH. sartoroides are lectotypified, andH. kleyi is neotypified. The identity ofH. fastor is discussed andH. nabucoi is recognized as the valid name for this species. Seven other species previously placed in sect.Macroceratitae are sufficiently distinct and are removed from the section.     

Cercosporoid hyphomycetes on malpighiaceous hosts from the Brazilian Cerrado: species of Pseudocercospora on hosts belonging to Byrsonima

Eleven new species of the genus Pseudocercospora were found on Byrsonima spp. from the Cerrado in Brazil: Pseudocercospora annellidica, P. byrsonimae-basilobae, P. byrsonimae-coccolobifoliae, P. byrsonimicola, P. byrsonimigena, P. campograndensis, P. grajauensis, P. mutabiliconidiophorum, P. pediformiconidiorum, P. planaltinensis, and P. subhyalina. The new species are described, illustrated and compared with hitherto described Pseudocercospora spp. on various hosts belonging to the Malpighiaceae, and all species concerned are keyed out.     

Karyotypic diversity and evolution of seven mahseer species (Cyprinidae) from India

Mahseer is a group of fish species that are well known as food and game fishes. The taxonomy of the mahseer species is confusing owing to the morphological variations and habitat adaptation. Detailed karyomorphological investigations have been carried out in seven species of mahseer, using karyotyping, Ag-NOR and fluorescent staining techniques. The basic diploid chromosome number (2n), in all mahseer species, was observed to be 100; however, the karyotype formula varied among the species, which were recorded as: 20m + 14sm + 22st + 44t (fundamental arm number, FN = 134) in Tor khudree ; 22m + 24sm + 24st + 30t (FN = 146) in Tor mussullah; 12m + 22sm + 14st + 52t (FN = 134) in Tor putitora ; 20m + 24sm + 24st + 32t (FN = 144) in Tor tor ; 20m + 30sm + 24st + 26t (FN = 150) in Tor chelynoides; 20m + 20sm + 20st + 40t (FN = 140) in Tor progeneius; and 20m + 18sm + 14st + 48t (FN = 138) in Neolissochilus hexagonolepis . Silver staining of the chromosomes revealed the presence of multiple nucleolar organizer regions (NOR) in these mahseer species. The highest number of NORs was observed in T. tor (four pairs of chromosomes), whereas the other six species possessed Ag-NOR signals on only two pairs of chromosomes. Although chromomycin A₃ (CMA₃) staining induced bright fluorescence signals on same Ag-NORs sites, with CMA₃, one additional signal was observed on the p arm of subtelocentric chromosomes in T. tor , T. chelynoides , T. progeneius and N. hexagonolepis , which may indicate the presence of inactive NOR in these species. The information on cytogenetic profile of these mahseer species is discussed in the light of cytotaxonomic implications and understanding the karyoevolution of these fish species.     

Karyotypes of two Antarctic fishes,Notothenia gibberifrons andNotothenia coriiceps neglecta

The chromosomes of two species of Antarctic fishes,Notothenia (Gobionotothen) gibberifrons andNotothenia (Notothenia) coriiceps neglecta, were prepared by the air-drying method at the Polish Antarctic Station “Henryk Arctowski” during the austral summer 1984–1985. ForN. (G.) gibberifrons the diploid number is2n = 46 consisting of 2 metacentric (m) pairs, 1 submetacentric (sm) pair and 20 telocentric (t) or subtelocentric (st) pairs. ForN. (N.) coriiceps neglecta the diploid number is 2n = 22 consisting of 9 m pairs, 1 sm pair and 1st pair. Some aspects of karyological evolution of these fishes are discussed.     

蟛蜞菊属和孪花菊属(菊科-向日葵族)的细胞学研究

研究了菊科向日葵族鳢肠亚族蟛蜞菊属(Sphagneticola O. Hoffm.)和孪花菊属(Wollastonia DC. ex Decne.)各2种植物的染色体数目和染色体形态。蟛蜞菊[S. calendulacea (L.) Pruski]的染色体数目为2n=50, 核型公式为2n=18m+30sm+2st,南美蟛蜞菊[S. trilobata (L.) Pruski]的染色体数目为2n=56, 核型公式为2n=24m+28sm+4st, 孪花菊[W. biflora (L.) DC.]的染色体数目为2n=30,核型公式为2n=24m+4sm+2st,山孪花菊[W. montana (Blume) DC.]的染色体数目为2n=74, 核型公式为2n=37m+31sm+6st。根据上述结果并结合以前的有关资料,推测蟛蜞菊属的染色体基数可能为x=14和x=25,而不应是x=15。该属的3个新世界热带种[S. brachycarpa (Baker) Pruski、S. gracilis (Richard) Pruski和南美蟛蜞菊]可能都基于x=14, 其中S. gracilis为二倍体(2n=2x=28), S. brachycarpa和南美蟛蜞菊为四倍体(2n=4x=56); 唯一的亚洲种(蟛蜞菊)可能是基于x=25的二倍体(2n=2x=50)。染色体资料不支持将山孪花菊(x=37)这一植物置于孪花菊属(x=15)中。     

两种羽叶菊属植物(菊科:千里光族)的细胞学研究及其系统学意义

该研究首次报道了滇羽叶菊和台湾刘寄奴的染色体数据。结果表明:两者的染色体数量都为48,核型公式均为2n=2x=36m+10sm+2st,与前人报道的刻裂羽叶菊的核型稍有不同。两者的染色体形态均由大到小逐渐变化,核型二型性不明显,但前者染色体明显比后者大。这说明羽叶菊属染色体基数确实应为x=24,且染色体大小在种间有较大差异。细胞学证据表明,该属与蒲儿根属中染色体基数为x=24的类群以及狗舌草属确实近缘。