Establishment and characterization of India's first marine fish cell line (SISK) from the kidney of sea bass (Lates calcarifer)

A continuous cell line (SISK) from kidney of sea bass, Lates calcarifer, has been established and characterized. The cell line was maintained in Leibovitz' L-15 supplemented with 15% fetal bovine serum. This cell line has been subcultured more than 100 times over a period of 2 years. The SISK cell line consists of predominantly of epithelial-like cells. These cells showed strong positive for epithelial markers such as cytokeratin 19 and pancytokeratin. The cells were able to grow at temperature between 25 and 32 °C with optimum temperature of 28 °C. The growth rate of sea bass kidney cells increased as the FBS proportion increased from 2% to 20% at 28 °C with optimum growth at the concentrations of 15% or 20% FBS. The distribution of chromosome number was 30 to 56 with a modal peak at 48 chromosomes. Polymerase chain reaction products were obtained from SISK cells and tissues of sea bass with primer sets of microsatellite markers of sea bass. Five fish viruses were tested on this cell line to determine its susceptibility to these viruses and this was found to be susceptible to MABV NC1 and nodavirus, and the infection was confirmed by RT-PCR and CPE. This suggests that the SISK cell line has good potential for the isolation of various fish viruses. This cell line has been shown to be susceptible to bacterial extracellular products. The SISK cell line is the India's first marine fish cell line.

Mitochondrial and chloroplast DNA-based phylogeny of Pelargonium (Geraniaceae)

Overall phylogenetic relationships within the genus Pelargonium (Geraniaceae) were inferred based on DNA sequences from mitochondrial(mt)-encoded nad1 b/c exons and from chloroplast(cp)-encoded trnL (UAA) 5' exon-trnF (GAA) exon regions using two species of Geranium and Sarcocaulon vanderetiae as outgroups. The group II intron between nad1 exons b and c was found to be absent from the Pelargonium, Geranium, and Sarcocaulon sequences presented here as well as from Erodium, which is the first recorded loss of this intron in angiosperms. Separate phylogenetic analyses of the mtDNA and cpDNA data sets produced largely congruent topologies, indicating linkage between mitochondrial and chloroplast genome inheritance. Simultaneous analysis of the combined data sets yielded a well-resolved topology with high clade support exhibiting a basic split into small and large chromosome species, the first group containing two lineages and the latter three. One large chromosome lineage (x = 11) comprises species from sections Myrrhidium and Chorisma and is sister to a lineage comprising P. mutans (x = 11) and species from section Jenkinsonia (x = 9). Sister to these two lineages is a lineage comprising species from sections Ciconium (x = 9) and Subsucculentia (x = 10). Cladistic evaluation of this pattern suggests that x = 11 is the ancestral basic chromosome number for the genus.

Deficiency of the Cytoskeletal Protein SPECC1L Leads to Oblique Facial Clefting

Genetic mutations responsible for oblique facial clefts (ObFC), a unique class of facial malformations, are largely unknown. We show that loss-of-function mutations in SPECC1L. are pathogenic for this human developmental disorder and that SPECC1L is a critical organizer of vertebrate facial morphogenesis. During murine embryogenesis, Speed 1 1 is expressed in cell populations of the developing facial primordial, which proliferate and fuse to form the face. In zebrafish, knockdown of a SPECC1L homolog produces a faceless phenotype with loss of jaw and facial structures, and knockdown in Drosophila phenocopies mutants in the integrin signaling pathway that exhibit cell-migration and -adhesion defects. Furthermore, in mammalian cells, SPECC1L colocalizes with both tubulin and actin, and its deficiency results in defective actin-cytoskeleton reorganization, as well as abnormal cell adhesion and migration. Collectively, these data demonstrate that SPECC1L functions in actin-cytoskeleton reorganization and is required for proper facial morphogenesis.

Recurrent Deletion of ZNF630 at Xp11.23 Is Not Associated With Mental Retardation

ZNF630 is a member of the primate-specific Xp11 zinc finger gene cluster that consists of six closely related genes, of which ZNF41, ZNF81, and ZNF674 have been shown to be involved in mental retardation. This suggests that mutations of ZNF630 might influence cognitive function. Here, we detected 12 ZNF630 deletions in a total of 1,562 male patients with mental retardation from Brazil, USA, Australia, and Europe. The breakpoints were analyzed in 10 families, and in all cases they were located within two segmental duplications that share more than 99% sequence identity, indicating that the deletions resulted from non-allelic homologous recombination. In 2,121 healthy male controls, 10 ZNF630 deletions were identified. In total, there was a 1.6-fold higher frequency of this deletion in males with mental retardation as compared to controls, but this increase was not statistically significant (P-value = 0.174). Conversely, a 1.9-fold lower frequency of ZNF630 duplications was observed in patients, which was not significant either (P-value = 0.163). These data do not show that ZNF630 deletions or duplications are associated with mental retardation. (C) 2010 Wiley-Liss, Inc.

Complex meiotic configuration of the holocentric chromosomes: the intriguing case of the scorpion Tityus bahiensis

Mitotic and meiotic chromosomes of Tityus bahiensis were investigated using light (LM) and transmission electron microscopy (TEM) to determine the chromosomal characteristics and disclose the mechanisms responsible for intraspecific variability in chromosome number and for the presence of complex chromosome association during meiosis. This species is endemic to Brazilian fauna and belongs to the family Buthidae, which is considered phylogenetically basal within the order Scorpiones. In the sample examined, four sympatric and distinct diploid numbers were observed: 2n = 5, 2n = 6, 2n = 9, and 2 = 10. The origin of this remarkable chromosome variability was attributed to chromosome fissions and/or fusions, considering that the decrease in chromosome number was concomitant with the increase in chromosome size and vice versa. The LM and TEM analyses showed the presence of chromosomes without localised centromere, the lack of chiasmata and recombination nodules in male meiosis, and two nucleolar organiser regions carrier chromosomes. Furthermore, male prophase I cells revealed multivalent chromosome associations and/or unsynapsed or distinctly associated chromosome regions (gaps, less-condensed chromatin, or loop-like structure) that were continuous with synapsed chromosome segments. All these data permitted us to suggest that the chromosomal rearrangements of T. bahiensis occurred in a heterozygous state. A combination of various factors, such as correct disjunction and balanced segregation of the chromosomes involved in complex meiotic pairing, system of achiasmate meiosis, holocentric nature of the chromosomes, population structure, and species dispersion patterns, could have contributed to the high level of chromosome rearrangements present in T. bahiensis.

Lapanthus (Bromeliaceae, Bromelioideae): A New Genus from the Southern Espinhaco Range, Brazil

In this paper we describe a new genus of Bromehaceae, Lapanthus, restricted to the southern portion of the Espinhaco Range, Minas Germs State, in southeastern Brazil Two new combinations to accommodate species previously described in the genera Orthophytum and cryptanthus and one new synonym are proposed Lapanthus has morphological affinities with both Cryptanthus and Orthophytum but nevertheless differs by the combination of margins of the petals ciliate, presence of lanceolate petal appendages and free stamens, and also by molecular data Cryptanthus and Orthophytum have petals entire along the margins, and the filaments of the most internal whorl are adnate to the petals Lapanthus stands out by having a pair of lanceolate petal appendages, which are almost completely adnate to the petals In Orthophytum, however, appendages are cupuhform or sacciform and they are totally absent in the genus Cryptanthus Lapanthus and Orthophytum present meiotic and mitotic chromosome numbers equal to n=25 and 2n=50, 100 and 150 respectively, while Cryptanthus presents meiotic and mitotic chromosome numbers n=17 and 2n=34, 36, 54 respectively, and this difference is considered to be an autapomorphic feature of Cryptanthus Descriptions of the genus and species, identification keys, illustrations, photographs of living specimens, and taxonomic comments are provided

Caracterização morfológica, citogenética e molecular de uma população de tangerineiras híbridas de 'Clementina fina' (Citrus clementina Hort. ex Tan.) e 'Montenegrina' (Citrus deliciosa Ten.)

A produção citrícola se encontra dispersa por todos os continentes e no Brasil, os citros são a produção frutícola de maior volume de produção. A produção de citros de mesa, como as tangerinas, possibilita ao produtor obter maior valor pelo seu produto. O mercado consumidor é ávido por novas variedades e para tanto, um programa de melhoramento deve estar sempre em busca de genótipos que atendam ao mercado consumidor, bem como a cadeia produtiva. Na Estação Experimental Agronômica da Universidade Federal do Rio Grande do Sul, está localizada uma população de tangerineiras híbridas oriundas do cruzamento da tangerineira ‘Clementina Fina’ (Citrus clementina Hort. ex Tan.) e ‘Montenegrina’ (Citrus deliciosa Ten.) a qual foi caracterizada neste estudo, avaliando-se características morfológicas de acordo com os descritores propostos pelo International Board for Plant Genetic Resources, além da identificação da época de maturação, viabilidade de pólen, número cromossômico e caracterização molecular, utilizando marcadores do tipo microssatélites. Através da análise morfológica foi possível distinguir todas as 96 plantas avaliadas, porém não foi possível agrupar a F1 em grupos distintos de cada um dos genitores. A época de maturação de frutos das plantas se concentra entre a primeira quinzena de abril até a primeira quinzena de agosto. Todas as plantas analisadas apresentaram um alto grau de viabilidade de pólen, variando entre 79,04 e 98,08 %. Todas as plantas avaliadas são diplóides com um número cromossômico de 2n=18. Utilizando 12 pares de primers de microssatélites foi possível diferenciar 90 acessos do estudo, e agrupar a F1 em indivíduos mais próximos do genitor feminino e do genitor masculino. O PIC (Conteúdo de Informação de Polimorfismo) dos primers variou de 0,27 a 0,65. Não foi possível estabelecer uma relação entre a caracterização utilizando marcadores morfológicos e a caracterização utilizando marcadores moleculares.

Parâmetros citogenéticos de espécies comerciais de carangidae (perciformes), com vistas a sua empregabilidade na conservação biológica e piscicultura marinha

Worldwide, families Carangidae and Rachycentridae represent one of the groups most important commercial fish, used for food, and great potential for marine aquaculture. However, the genetic bases that can underpin the future cultivation of these species, cytogenetic between these aspects are very weak. The chromosomal patterns have provided basic data for the exploration of biotechnological processes aimed at handling chromosomal genetic improvement, such as induction of polyploidy, androgenesis and ginogenesis, as well as obtaining monosex stocks and interspecific hybridizations. This paper presents a comprehensive cytogenetic survey in 10 species, seven of the family Carangidae and the monotypic family Rachycentridae. Classical cytogenetic analysis and in situ mapping of multigene sequences were employed, and additionally for the genus Selene and morphotypes of Caranx lugubris, comparisons were made using geometric morphometrics. In general, conservative species exhibit a marked chromosome number (2n=48). Although present in large part, different karyotypic form, retain many characteristics typical of chromosomal Order Perciformes, the high number of elements monobrachyal, Ag-NORs/18S rDNA sites and heterochromatin simply reduced, preferably centromeric. The main mechanisms involved in karyotypic diversification are the pericentric inversions, with secondary action of centric fusions. In addition to physical mapping and chromosome detail for the species are presented and discussed patterns of intra-and interspecific diversity, cytotaxonomic markers. This data set provides a better understanding of these patterns caryoevolutyonary groups and conditions for the development of protocols based on Biotechnology for chromosomal manipulation Atlantic these species

Morfo-anatomia de plântulas e número cromossômico de Cybistax antisyphilitica (Mart.) Mart. (Bignoniaceae)

Plântulas de Cybistax antisyphilitica (ipê-verde), espécie arbórea que ocorre no Estado de São Paulo, Brasil, foram estudadas morfoanatomicamente e citogeneticamente. As plântulas são eudicotiledôneas, fanerocotiledonares e epigéias. Suas sementes apresentam alas hialinas, assimétricas e de textura papirácea. A raiz é axial com tecido epidérmico irregular. O hipocótilo é verde, glabro e apresenta os elementos do xilema em diferenciação. Os cotilédones são verdes, foliáceos, reniformes, com mesofilo heterogêneo, epiderme pilosa e feixes vasculares colaterais. Os eófilos são glabros, peciolados, de filotaxia oposta, dorsiventrais, hipoestomáticos e possuem mesofilo heterogêneo e assimétrico. Não há diferenças anatômicas significativas entre os eófilos e os metáfilos. O sistema vascular do pecíolo dos metáfilos dispõe-se em forma de ferradura. A espécie apresenta número cromossômico mitótico 2n = 40 com comprimento cromossômico geral médio de 1,042 µm ± 0,140 e amplitude variando de 0,58 µm até 1,60 µm.

Aspectos morfológicos de frutos e sementes e caracterização citogenética de Crotalaria lanceolata E. Mey. (Papilionoideae - Fabaceae)

Frutos, sementes e plântulas de Crotalaria lanceolata, conhecida popularmente como guizo-de-cascavel, chocalho-de-cobra, xique-xique ou feijão-de-guizo, planta tóxica infestante que ocorre no Estado de São Paulo, foram estudadas morfologicamente e citogeneticamente. Os frutos são secos, deiscentes, polispérmicos e do tipo legume. As sementes são reniformes e o embrião é constituído de eixo embrionário e dois cotilédones. A testa pode apresentar variadas tonalidades de castanhos. A germinação é epígea e fanerocotiledonar. A espécie apresenta número cromossômico diplóide 2n = 16 com formulação cariotípica 12M + 4SM e comprimento cromossômico médio geral de 3,340 ± 0,689.

Caracterização citogenética em Schlumbergera truncata (Haworth) Moran e Schlumbergera × buckleyi (T. Moore) Tjaden (Cactaceae)

O número cromossômico diplóide de Schlumbergera truncata e Schlumbergera x buckleyi, de indivíduos com diferentes tipos de coloração de pétalas, foi determinado usando-se pontas de raízes. A utilização de 8-hidroxiquinoleína 0,003 M à 36 °C por 3 horas possibilitou melhor separação cromossômica. Técnica de bandeamento C e de coloração Giemsa permitiram o estudo cariológico dessas espécies. O híbrido Schlumbergera × buckleyi (rósea) apresenta 2n = 22 cromossomos com fórmula cariotípica 16 M + 6 SM. Schlumbergera truncata, apresentando pétalas nas cores vermelha, branca e pink, possui 2n = 22 cromossomos, formulação cariotípica idêntica à de Schlumbergera × buckleyi, enquanto a planta com flores de coloração amarelada mostrou 2n = 34 cromossomos. A classificação cromossômica foi baseada no índice centromérico. Nas plantas que apresentam coloração vermelha, branca, pink e rósea nas pétalas, o melhor período de obtenção de metáfases corresponde ao período de florescimento. Schlumbergera truncata com flores amareladas apresenta dois picos anuais de divisão mitótica. Esses resultados dão suporte à um melhor entendimento da biologia no gênero Schlumbergera e auxiliam na classificação taxonômica nos casos onde apenas as características fenotípicas não são suficientemente confiáveis para a classificação das plantas no mesmo táxon.

Cytogenetic characterization of the strongly electric Amazonian eel, Electrophorus electricus (Teleostei, Gymnotiformes), from the Brazilian rivers Amazon and Araguaia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Occurrence of ZZ/ZW sex chromosomes in Thoracocharax stellatus fish (Characiformes, Gasteropelecidae) from the Araguaia River, south America

The karyotypic and chromosomal characteristics of the hatchetfish Thoracocharax stellatus from the Araguaia River, Brazil (Araguaia-Tocantins basin) were analyzed using Giemsa, AgNO3, and CMA(3) fluorescent staining, and C-banding. The diploid chromosome number was 54 and the karyotypes of females and males were composed of six metacentrics, six submetacentrics, six subtelocentrics and 36 acrocentrics. Two impaired acrocentric chromosomes were detected in the female karyotype. C-banding showed heterochromatic blocks at several chromosomes and an entirely heterochromatic acrocentric chromosome in females that was lacking in the male karyotype. This discovery indicated a heteromorphic sex chromosome system of the ZZ/ZW type. Ag-staining and CMA(3) fluorescence revealed one major chromosome pair bearing the NORs with the presence of additional signals in some metaphases. Both heterochromatic segments associated with Ag-NORs and the W chromosome were positively stained by CMA(3). Considering the present data and previous findings it is hypothesized that the occurrence of ZW sex chromosome system is widespread in the genus Thoracocharax.

Comparative Cytogenetic Analysis of the Genus Symphysodon (Discus Fishes, Cichlidae): Chromosomal Characteristics of Retrotransposons and Minor Ribosomal DNA

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Three sympatric karyomorphs in the fish Astyanax fasciatus (Teleostei, Characidae) do not seem to hybridize in natural populations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)