Chromosomal Variability among Allopatric Populations of Erythrinidae Fish Hoplias malabaricus: Mapping of Three Classes of Repetitive DNAs

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

Karyotype description of five species of Trichomycterus (Teleostei: Siluriformes: Trichomycteridae)

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

Cytogenetic characterization of the silverside fish Odontesthes regia (Humboldt, 1833) (Teleostei: Atheriniformes: Atherinopsidae) from Iquique, Chile

This paper describes the karyotype of Odontesthes regia by means of Giemsa staining, C-banding, to reveal the distribution of the constitutive heterochromatin, and by Ag-staining and fluorescent in situ hybridization (FISH), to locate ribosomal genes (rDNA). The chromosome diploid modal count in the species was 2n = 48. The karyotype is composed of one submetacentric pair (pair 1), 16 subtelocentric pairs (pairs 2 to 17), and 7 acrocentric pairs (pairs 18 to 24). With the exception of pair 1 it was not possible to classify the homologous chromosomes accurately because differences in chromosome size were too slight between adjacent pairs. The distribution of C-banded heterochromatin allowed for a more accurate matching of the majority of chromosomes of the subtelocentric series. Silver staining of metaphase spreads allowed for the identification of Nucleolus Organizer Regions (Ag-NOR) on pair 1. FISH experiments showed that 18S rDNA sequences were located, as expected, in the same chromosome pair identified as the Ag-NOR-bearing one.

松属单维管束亚属植物rDNA的染色体定位研究

松属植物的基因组十分庞大（大于20000Mbp），其中约90%是由重复序列组成的，我们对其结构和组成仍知之甚少。松属在系统分类上分为两个亚属:单维管束亚属和双维管束亚属。基因组大小研究发现单维管束亚属植物的基因组更大。rDNA作为一类有功能的多基因家族重复序列，其自身特性决定了它在基因组研究中的重要性。FISH技术为rDNA在染色体上物理定位提供了有力的工具。尽管现在对松属rDNA FISH已有不少报道，但主要集中在双维管束亚属，对单维管束亚属的研究几乎是空白。本研究选择5个单维管束亚属松属植物P. bungeana, P. koraiensis, P. armandii, P. wallichiana, P. strobus，进行rDNA FISH研究。旨在弄清18S-25S rDNA和5S rDNA在单维管束亚属植物染色体上的位点数目和分布模式。结合前人对松属双维管束亚属植物的工作，对单、双维管束亚属植物之间rDNA FISH结果进行比较，从而可以从整体上认识松属植物的18S-25S rDNA和5S rDNA在染色体上的分布式样。在此基础上进一步探讨18S-25S rDNA和5S rDNA这些重复序列在松属植物基因组结构和组成中的地位和作用。本研究主要结果如下： 1.rDNA FISH在松属染色体核型分析中的作用 本研究中5种松属单维管束亚属植物染色体数目均为2n=24，除最短一条染色体为亚中部着丝粒染色体外，其余11条均为中部着丝粒染色体，长度和臂比也十分接近，同源染色体的不容易鉴定，很难排出精确的核型。在我们的研究结果中，5个松属植物中，除了白皮松外，18S-25S rDNA和5S rDNA分布在12对染色体中的10对染色体上，这些位点可作为染色体标记，大大提高了同源染色体鉴定的准确度，但是染色体之间排序问题依然没有很好地解决。核型比较认为种间是否存在部分同源染色体关系也不是十分明确，仅Ⅺ号和Ⅻ号染色体有这种关系，这主要由于Ⅺ号和Ⅻ号染色体容易准确地鉴别出来。核型分析的精确仍有待增加标记来提高。 2．rDNA位点数目在松属两个亚属间的比较及其与基因组大小的关系 松属植物18S-25S rDNA位点通常为5-10个，5S rDNA位点为1-4个。其中单维管束亚属18S-25S rDNA位点通常为9-10个（除白皮松为4个外），5S rDNA位点为2- 4个;双维管束亚属为18S-25S rDNA位点通常为5-10个，5S rDNA位点通常为1-2个。而二倍体被子植物18S-25S rDNA位点通常为1-5个5S rDNA位点为1-3个。暗示18S-25S rDNA和5S rDNA位点数目多少和基因组大小还是有一定的相关性。因为松属植物的基因组比典型的二倍体被子植物大得多，单维管束亚属植物的的C-值又普遍比双维管束亚属植物的高。白皮松虽有些例外，18S-25S rDNA位点数目少，但信号强度大得多，代表拷贝数高，因此其基因组大小可以从rDNA拷贝数上得到解释。 3．18S-25S rDNA和5S rDNA位点在松属两个亚属之间的分布模式比较 18S-25S rDNA和5S rDNA位点在松属两个亚属染色体上的分布方式有明显不同，每个亚属均有两种分布形式，并形成各自稳定的分布模式。在单维管束植物中，18S-25S rDNA和5S rDNA位点或相邻分布于同一染色体同一臂上，5S rDNA位于臂的远端;或两位点分布于不同的染色体。而在双维管束植物中18S-25S rDNA和5S rDNA或相邻分布于同一染色体同一臂上，18S-25S rDNA在臂的远端;或两位点分布于同一染色体两条臂上。在两个亚属中，当18S-25S rDNA和5S rDNA位点位于同一条染色体臂上时，相对位置正好相反。这完全不同的rDNA分布模式的形成，可能与松属这两个亚属植物的物种形成和分化过程中染色体发生倒位或易位有关，暗示这两个亚属的基因组结构存在分化。但这各自的分布模式是否可以作为判断亚属的特征依据仍有待加大样本量证实。 4．rDNA 位点分布及变异具有系统学意义 rDNA FISH 结果符合分类中亲缘关系越近，分布模式越相似的原则，因而认为rDNA 位点在染色体上的分布模式，具有系统学意义。基于已知的松属植物rDNA FISH结果构建的系统关系，符合传统分类系统中对亚组划分。rDNA FISH结果与分子系统学的研究结果相比较认为，松属单维管亚属5种松中，以乔松和北美乔松关系最近，与同一个亚组的华山松稍远，与另一个亚组的红松更远。而白皮松作为一个特有的孑遗类群，系统位置比较特殊，分子系统学研究认为其处于基部的位置，本研究表明其rDNA位点有明显的特点：位点数目少，但信号强，反映了拷贝数多。那是否它就代表了祖先类群的位点分布模式，需要更多的基部类群的rDNA FISH结果支持。

Cytogenetic characterization of Rhomboplites aurorubens and Ocyurus chrysurus, two monotypic genera of Lutjaninae from Cubagua Island, Venezuela, with a review of the cytogenetics of Lutjanidae (Teleostei: Perciformes)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Classical and molecular cytogenetic characterization of Agonostomus monticola, a primitive species of Mugilidae (Mugiliformes)

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

New occurrence of a macro B-chromosome in Astyanax scabripinnis paranae (Pisces, Characiformes, Characidae)

Cytogenetic studies performed on 17 specimens (11 females and six males) of Astyanax scabripinnis paranae from the Cascatinha stream showed that this population has 2n = 50 chromosomes (8M + 22SM + 10ST + 10A), two chromosome pairs with NORs and conspicuous C-band positive blocks in the terminal position of the long arm of five chromosome pairs. Three females presented 2n = 51 chromosomes and the extra chromosome was a large metacentric similar in size and morphology to the first chromosome pair in the karyotype. This accessory chromosome was entirely heterochromatic in C-banded metaphases, which permitted its classification as a supernumerary chromosome. Some aspects related to the morphology of such macro B-chromosomes are discussed.

Cytogenetic studies of three species of Glandulocaudinae (Pisces, Characiformes, Characidae)

Cytogenetic studies of three species of the subfamily Glandulocaudinae showed that Mimagoniates microlepis presents 2n = 52 chromosomes (6M + 20SM + 18ST + 8A), Mimagoniates lateralis presents 2n = 52 chromosomes (6M + 20SM + 16ST + 10A) and Glandalocauda melanogenys presents 2n = 52 chromosomes (4M + 12SM + 22ST + 14A). The number of NOR-bearing chromosome pairs ranged from two to four. Differences in number and position of NORs at the species and population levels were detected, suggesting that several chromosome rearrangements occurred in the evolutionary process of this group. Some aspects related to the chromosome evolution of the Glandulocaudinae are discussed.

Nuclear DNA content of thirty species of neotropical fishes

The present paper reports nuclear DNA content in 30 Neotropical freshwater fish species and summarizes the data on other Neotropical species presented in the literature. Among Neotropical fishes, the nuclear DNA content ranges from 1.04 ± 0.09 pg/nucleus in Corydoras cf. simulatus (2n = 62) to 248.0 pg/nucleus in Lepidosiren paradoxa (2n = 38). A general analysis of the data obtained in the present study for each species showed that DNA measurements were practically constant at the individual level, while significant differences were observed among individuals of the same population. This observation was valid for all species analyzed and was more evident in those species that presented other karyotypic particularities such as sex chromosomes or supernumerary chromosomes. The importance of changes in nuclear DNA content in the evolutionary process of Neotropical fishes is discussed.

Cytogenetic analysis of species of the genera Acestrorhynchus, Oligosarcus and Rhaphiodon (Teleostei: Characiformes)

In the present study the karyotypes and the number and position of nucleolus organizer regions (NORs) of four species of Characiformes are described. The analyses showed that Acestrorhynchus lacustris had 2n=50 chromosomes (8M+34SM+6ST+2A), Oligosarcus longirostris had 2n=50 chromosomes (2M+20SM+10ST+18A), Oligosarcus cf. paranensis from Keller and Mourão rivers had 2n=50 chromosomes (2M+26SM+8ST+14A), and Rhaphiodon vulpinus had 2n=54 chromosomes (32M+12SM+8ST+2A). The number of NOR-bearing chromosome pairs ranged from one to three among the species studied. The results available show that there is no variation in the diploid number within the genera analyzed. However, clear differences related to the karyotypic formulae and the number and position of Ag-NORs were found even in the comparison between populations. The possible relationships between the genera analyzed and other Characiformes are discussed.

Chromosomal diversification of diploid number, heterochromatin and rDNAs in two species of Phanaeus beetles (Scarabaeidae, Scarabaeinae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Pseudoplatystoma metaense and P. orinocoense (Siluriformes: Pimelodidae) from the Orinoco basin, Venezuela: cytogenetic and molecular analyses

The genus Pseudoplatystoma includes catfish species distributed throughout the fresh waters of South America. These species are important fisheries resources and play a significant ecological role due to their piscivorous and migratory habits. The taxonomy of this genus is still debated: traditionally, only three species have been recognised, but recently, this number was raised to eight. The validity of these eight morphospecies, however, was not confirmed by two subsequent molecular phylogenetic studies, which identified either five or four main clades. In this study, we focused on the two morphospecies restricted to the Orinoco basin, P. metaense and P. orinocoense, which have been assigned to either the same or different clades in previous studies. We carried out cytogenetic analyses to describe their unknown karyotypes and to look for cytotaxonomic markers. We also analysed their mitochondrial sequences in order to assign the sampled specimens to the previously identified molecular clades. The two presumptive species show similar karyotypes (2n=56, 42 biarmed and 14 uniarmed chromosomes) and cytogenetic features in terms of the constitutive heterochromatin distribution and the number and location of minor and major ribosomal genes. Thus, no species-specific chromosome markers could be identified. The analysis of cytochrome b and cytochrome oxidase I mitochondrial genes (carried out by retrieving all the mtDNA Pseudoplatystoma sequences available in GenBank) distributed the sampled specimens into two distinct molecular clades and confirmed the need to re-evaluate, by parallel morphological and molecular analyses, the monophyly of some lineages.

Cayler Cardio-Facial Syndrome: An Uncommon Condition in Newborns

Introduction: Cayler cardio-facial syndrome is a rare syndrome associated with asymmetric crying faces with congenital heart disease. We report a newborn that was diagnosed as case of Cayler Cardio-facial syndrome based on clinical features and was confirmed with FISH analysis. Case Presentation: A term male baby, born to non-consanguineous couple through normal vaginal delivery was diagnosed to have asymmetric crying faces with deviation of angle of mouth to left side at the time of birth. The baby had normal faces while sleeping or silent. Mother was known case of hypothyroidism and was on treatment. Baby was diagnosed as case of Cayler Cardio-facial Syndrome and was investigated with echocardiogram, brain ultrasound, total body X-ray examination, X-ray of cervico-thoracic vertebral column and fundus examination. Echocardiogram showed muscular VSD, brain ultrasound was normal and fundus examination showed tortuous retinal vessels. Whole body X-ray and lateral X-ray of cervico-thoracic vertebral column were not suggestive of any skeletal abnormalities. The other associated malformation was right ear microtia. Baby FISH karyotype analysis showed deletion of 22q11.2 deletion. Baby was discharged and now on follow-up. Conclusions: Cayler syndrome is a rare syndrome which must be suspected if a baby has asymmetrical cry pattern and normal facies when baby sleeps. Patient must be evaluated with echocardiography to find out associated cardiac malformations. These infants should undergo FISH analysis for 22q11.2 deletion syndrome.

HETEROCHROMATIN AND KARYOTYPE REORGANIZATION IN FISH OF THE FAMILY ANOSTOMIDAE (CHARACIFORMES)

Mitotic chromosomes of four fish species of the family Anostomidae, belonging to the genera Leporinus, Leporellus, and Schizodon, were studied. With 2n = 54 meta- and submetacentric chromosomes, this family appears to be characterized by marked karyotypic stability. Although perceptible differences exist, mainly in the amount of constitutive heterochromatin present in the chromosomes of these species, these differences do not affect the structure and/or size of these chromosomes. Chromatin substitutions and/or modifications may have led, in one direction, to an increase in heterochromatin in some species and, in the opposite direction, to heterochromatin reduction in others. Whether these changes are accompanied by changes in the amount of euchromatin in the chromosomes is an open question. The nucleolar organizer regions, which may be located on different chromosomes in the various species, may also be indicators of reorganization of these karyotypes.

Reconstruction of the ancestral marsupial karyotype from comparative gene maps

BACKGROUND: The increasing number of assembled mammalian genomes makes it possible to compare genome organisation across mammalian lineages and reconstruct chromosomes of the ancestral marsupial and therian (marsupial and eutherian) mammals. However, the reconstruction of ancestral genomes requires genome assemblies to be anchored to chromosomes. The recently sequenced tammar wallaby (Macropus eugenii) genome was assembled into over 300,000 contigs. We previously devised an efficient strategy for mapping large evolutionarily conserved blocks in non-model mammals, and applied this to determine the arrangement of conserved blocks on all wallaby chromosomes, thereby permitting comparative maps to be constructed and resolve the long debated issue between a 2n=14 and 2n=22 ancestral marsupial karyotype. RESULTS: We identified large blocks of genes conserved between human and opossum, and mapped genes corresponding to the ends of these blocks by fluorescence in situ hybridization (FISH). A total of 242 genes was assigned to wallaby chromosomes in the present study, bringing the total number of genes mapped to 554 and making it the most densely cytogenetically mapped marsupial genome. We used these gene assignments to construct comparative maps between wallaby and opossum, which uncovered many intrachromosomal rearrangements, particularly for genes found on wallaby chromosomes X and 3. Expanding comparisons to include chicken and human permitted the putative ancestral marsupial (2n=14) and therian mammal (2n=19) karyotypes to be reconstructed. CONCLUSIONS: Our physical mapping data for the tammar wallaby has uncovered the events shaping marsupial genomes and enabled us to predict the ancestral marsupial karyotype, supporting a 2n=14 ancestor. Futhermore, our predicted therian ancestral karyotype has helped to understand the evolution of the ancestral eutherian genome.