Sequence homogenization and chromosomal localization of VicTR-B satellites differ between closely related Vicia species

Satellite sequences of the VicTR-B family are specific for the genus Vicia (Leguminosae), but their abundance varies among the species, being the highest in Vicia sativa and Vicia grandiflora. In this study, we have sequenced multiple randomly cloned VicTR-B fragments from these two species and analyzed their sequence variability, periodicity, and chromosomal localization. We have found that V. sativa VicTR-B sequences are homogeneous with respect to their nucleotide sequences and periodicity (monomers of 38 bp), whereas V. grandiflora repeats are considerably more variable, occurring in at least four distinct sequence subfamilies. Although the periodicity of 38 bp was conserved in most of the V. grandiflora sequences, one of the subfamilies was composed of higher-order repeats of 186 bp, which originated from a pentamer of the basic repeated unit. Individual VicTR-B subfamilies were preferentially located in either intercalary or subtelomeric regions of chromosomes. Interestingly, two V. grandiflora subfamilies with the highest similarity to V. sativa VicTR-B sequences were located in intercalary heterochromatic bands, showing similar chromosomal distribution as the majority of VicTR-B repeats in V. sativa. The other two V. grandiflora subfamilies showing a considerable divergence from V. sativa sequences were found to be accumulated at subtelomeric regions of V. grandiflora chromosomes.Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.Communicated by I. Schubert     

Cytotaxonomy of four species in the Simulium perflavum species group (Diptera: Simuliidae) from Brazilian Amazonia

Polytene chromosomes of four members of the Simulium perflavum species group in Brazil are described, and a standard map for the species group is presented. Simulium rorotaense Floch & Abonnenc, S. perflavum Roubaud and an undescribed species (S.‘X’) are chromosomally conservative, each representing a single species with a unique preimaginal habitat. Chromosomal, morphological and ecological evidence indicates that S. maroniense Floch & Abonnenc, previously considered synonymous with S. rorotaense, is a good species. Independent morphological and chromosomal analyses yielded 99.4% agreement in separating larvae of S. rorotaense and S. maroniense. The two species can be distinguished by gill morphology or by a subterminal inversion on the long arm of chromosome III. Simulium maroniense consists of at least four cytotypes, each with different sex chromosomes and autosomal polymorphism profiles and associated with a particular landscape type, altitude, temperature regime or geographical location. Simulium rorotaense and S. maroniense share one unique inversion, as do S. perflavum and S. ‘X’, indicating two pairs of sister species. The anthropogenic S. perflavum probably dispersed into Central Amazonia sometime after the mid-1970s.     

Comparative cytogenetic analysis of eleven species of subfamilies Neoplecostominae and Hypostominae (Siluriformes: Loricariidae)

The family Loricariidae with about 690 species divided into six subfamilies, is one of the world’s largest fish families. Recent studies have shown the existence of several problems in the definition of natural groups in the family, which has made the characterization of the subfamilies and even of some genera quite difficult. With the main objective of contributing for a better understanding of the relationships between loricariids, cytogenetic analysis were conducted with two species of Neoplecostominae and nine species of Hypostominae that, according to morphological and molecular data, may belong to a new monophyletic unit. The results obtained showed a marked chromosomal conservation with the presence of 2n = 54 chromosomes and single interstitial Ag-NORs in all species analyzed. Considering that Neoplecostominae is the primitive sister-group of all other loricariids, with exception of Lithogeneinae, this karyotypic structure may represent the primitive condition for the family Loricariidae. The cytogenetic characteristics partaken by the species of Neoplecostominae and Hypostominae analyzed in the present study reinforce the hypothesis that the species of both these subfamilies might belong to a natural group.     

Localization of Shaw-related K+ channel genes on mouse and human chromosomes

Four related genes, Shaker, Shab, Shaw, and Shal, encode voltage-gated K⁺ channels in Drosophila. Multigene subfamilies corresponding to each of these Drosophila genes have been identified in rodents and primates; this suggests that the four genes are older than the common ancestor of present-day insects and mammals and that the expansion of each into a family occurred before the divergence of rodents and primates.In order to define these evolutionary relationships more precisely and to facilitate the search for mammalian candidate K⁺ channel gene mutations, we have mapped members of the Shaw-homologous gene family in humans and mice. Fluorescence in situ hybridization analysis of human metaphase chromosomes mapped KCNC2 (KShIIIA, KV3.2) and KCNC3 (KShIIID, KV3.3) to Chromosome (Chr) 19q13.3-q13.4. Inheritance patterns of DNA restriction fragment length variants in recombinant inbred strains of mice placed the homologous mouse genes on distal Chr 10 near Ms15-8 and Mdm-1. The mouse Kcnc1 (KShIIIB, NGK2-KV4, KV3.1) gene mapped to Chr 7 near Tam-1.These results are consistent with the hypothesis that the generation of the mammalian KCNC gene family included both duplication events to generate family members in tandem arrays (KCNC2, KCNC3) and dispersion of family members to unlinked chromosomal sites (KCNC1). The KNCN2 and KCNC3 genes define a new synteny group between humans and mice.     

Identification and chromosomal location of four subfamilies of the rubisco small subunit genes in common wheat

Summary Three different 3 noncoding sequences of wheat rubisco small subunit (SSU) genes (RbcS) were used as probes to identify the gene members of different RbcS subfamilies in the common wheat cultivar Chinese Spring (CS). All genes of the wheat RbcS multigene family were previously assigned to the long arm of homoeologous group 5 and to the short arm of homoeologous group 2 chromosomes of cv CS. Extracted DNA from various aneuploids of these homoeologous groups was digested with four restriction enzymes and hybridized with three different 3 noncoding sequences of wheat SSU clones. All RbcS genes located on the long arm of homoeologous group 5 chromosomes were found to comprise a single subfamily, while those located on the short arm of group 2 comprised three subfamilies. Each of the ancestral diploid genomes A, B, and D has at least one representative gene in each subfamily, suggesting that the divergence into subfamilies preceded the differentiation into species. This divergence of the RbcS genes, which is presumably accompanied by a similar divergence in the 5 region, may lead to differential expression of various subfamilies in different tissues and in different developmental stages, in response to different environmental conditions. Moreover, members of one subfamily that belong to different genomes may have diverged also in the coding sequence and, consequently, code for distinguishable SSU. It is assumed that such utilization of the RbcS multigene family increases the adaptability and phenotypic plasticity of common wheat over its diploid progenitors.     

A comparative study of the chromosomal polymorphs in the incipient species of the Drosophila paulistorum complex

Ninety-six geographical strains distributed among the incipient species of the Drosophila paulistorum complex were examined cytologically, and the results obtained were correlated with available data on hybridization tests and chromosomal analysis. The complex was found to contain more than sixty-three different inversions, out of which thirty-two were 3rd chromosome configurations. This placed Drosophila paulistorum among the most chromosomally polymorphic species in the genus. The species differs from D. willistoni, in that a great number of inversions is concentrated in one of the chromosomes, as opposed to approximately equal distribution of inversions in the chromosomes of willistoni. — The data obtained in the course of this investigation seem to support the idea that either massive populations become isolated and then form new species, or that the newly forming species tend to retain some of their ancestral polymorphs which might present them with heterotic effects, gradually replacing them with more successful combinations as speciation progresses.The work reported in this article has been carried under Contract No. AT (30-1)-3096, U. S. Atomic Energy Commission.     

Chromosomes of three freshwater stingrays (Rajiformes Potamotrygonidae) from the Rio Negro basin, Amazon, Brazil

Potamotrygonidae is the representative family of South American freshwater elasmobranchs. It is a monophyletic group containing 20 species grouped into three genera. Three species belonging to two genera of this family were collected from the middle Negro River, Amazonas, Brazil, and studied cytogenetically: Paratrygon aiereba, Potamotrygon motoro and Potamotrygon orbignyi. Paratrygon aiereba presented 2n = 90 chromosomes and 4M+2SM+10ST+74A. Both species of Potamotrygon presented 2n = 66 chromosomes and differed in their chromosomal formulas: P. motoro had 18M+12SM+10ST+26A and P. orbignyi had 22M+10SM+8ST+26A. No sex heteromorphism was detected. The Fundamental Number (FN) was 106 for the three species. A system of multiple NORs was found in the three species, but with interspecific differences in terms of location and position of the active Ag-NORs sites. Paratrygon aiereba presented only four sites on the short arms of two chromosomal pairs, both in terminal regions. Potamotrygon motoro presented seven sites, on the long and short arms, all in terminal regions of non-homologous chromosomes; P. orbignyi presented eight sites on the long arms, all in terminal regions, of non-homologous chromosomes. The constitutive heterochromatin was in pericentromeric regions of all chromosomes, and no significant interspecific difference was found in relation to this marker.     

Karyotypes and meiotic mechanisms of some eumastacid grasshoppers from East Africa,Madagascar, India and South America

Five species of the subfamily Thericleinae from East Africa show various types of cryptochiasmatic meiosis in the male, in which the chiasmata are not visible in prophase and are only revealed in the course of first metaphase. Several of these species have very large chromosomes. Two species belonging to the subfamily Miraculinae, from Madagascar have 2 n=25 acrocentric chromosomes, the highest number known in the Eumastacidae. Their meiosis is of the normal type. — Eighteen species of the subfamily Pseudoschmidtiinae, also from Madagascar have, for the most part, small chromosomes and a very uniform karyotype, of 2 n=21 acrocentrics. A species of Xenomastax has acquired an X₁X₂Y sex chromosome mechanism as a result of two successive chromosomal fusions. A fusion beween autosomes has reduced the chromosome number to 2 n=19 in the genus Tetefortina. The male meiosis of the Pseudoschmidtiinae is quite orthodox. — Two species of Eumastacids belonging to the subfamily Mastacideinae, from South India, show 2 n=21 acrocentrics. The male meiosis is quite orthodox, with a rather high chiasma frequency. — Three South American species of Eumastacidae, belonging to the subfamilies Paramastacinae, Parepisactinae and Eumastacinae respectively, have karyotypes which seem to be very different from those of the Old World subfamilies that have been studied. Meiosis is normal.The cytological evidence thus confirms the view of systematists that the evolutionary divergence of the subfamilies of Eumastacidae is very considerable, and presumably ancient. As far as we can tell, each subfamily may be characterized by a typical or primitive karyotype. There have been fewer evolutionary fusions of chromosomes in the Thericleinae and Pseudoschmidtiinae than in the Morabinae and no chromosomal dissociations are known to have occurred in the two former subfamilies.Affectionately dedicated to Sally Hughes-Schrader, whose work has enlarged our cytogenetic horizons so greatly, on her 75th birthday, January 25, 1970.Supported by Public Health Service Grant No. GM-07212 from the Division of General Medical Sciences, U.S. National Institutes of Health and by a grant from the Australian Research Grants Committee.     

Cytogenetic Analysis of Experimental Hybrids in Species of Triatominae (Hemiptera-Reduviidae)

A cytogenetic analysis was performed in experimental hybrids between species of Chagas disease transmitting bugs with remarkable differences in the amount and distribution of heterochromatin. Using C-banding technique, we identified the parental species chromosomes and analysed the meiotic behaviour in the male hybrids between Triatoma platensis and T. infestans, T. platensis and T. delpontei, and T. infestans and T. rubrovaria. The two former hybrids have an entirely normal meiotic behaviour despite the extensive differences in C-banded karyotypes observed in the parental species, indicating that heterochromatin differences between homeologous chromosomes are not a barrier that influences meiotic synapsis and recombination. On the contrary, the experimental hybrids between T. infestans and T. rubrovaria show failures in pairing of homeologous chromosomes that lead to the production of abnormal spermatids and hybrid sterility. Our data suggest that karyotypic repatterning within triatomines has involved at least two different pathways. Among closely related species, chromosomal changes have largely involved addition or deletion of heterochromatic regions. In more distant species, chromosomal rearrangements (i.e. inversions and translocations) have also arisen. Hybridisation data also allow to hypothesize about the origin and divergence of this taxonomic group, as well as the mechanisms that maintain species isolation.     

The first karyogram of a Bromeliaceae species: an allopolyploid genome

The Bromeliaceae family has been traditionally distributed in the subfamilies Bromelioideae, Tillandsioideae and Pitcairnioideae. However, phylogenetic studies have provided other classifications, highlighting the need for analyses in order to characterize the genome of different species from this family. In this sense, the present work aimed to determine nuclear 2C-value and base composition, characterize the chromosomes and establish the karyogram of Pitcairnia flammea. Flow cytometry yielded 2C = 1.44 pg, AT = 64.28 % and GC = 35.72 % for this species, indicating its relatively small genome size. Despite reduced length and morphological similarity of the chromosomes, P. flammea metaphases presented well-spread chromosomes, with well-defined primary constriction, without chromatin damage and cytoplasmic background. These aspects allowed morphometric chromosomal characterization and assembly of the first karyogram of a Bromeliaceae species. The karyogram displayed 2n = 50 chromosomes, of which all were submetacentric. Karyomorphological analysis revealed grouped pairs of cytogenetically identical chromosomes (2–3, 4–5, 6–9, 10–17, 18–19, 20–23 and 24–25), plus one isolated chromosome (1), not identical to any other. This result suggests an allopolyploid origin for the P. flammea genome. Thus, the present investigation contributed with karyotype data for taxonomic and evolutionary aspects of this group.     

Karyotypic studies in Solanum section Lasiocarpa (Solanaceae)

Mitotic chromosomes of 13 species included in Solanum sect. Lasiocarpa were studied. All species have 2n = 24. The chromosome numbers of S. stagnale, S. felinum, and S. repandum are reported for the first time. Statistical analyses of chromosomes, genome length, and centromere position yielded estimates of karyotype composition and asymmetry. A generalized karyotype of the section shows that most of the chromosomes are metacentric (73%) or submetacentric (25.6%). Two pairs of subtelocentric chromosomes are present only in S. sessiliflorum. Satellites are quite common, characterizing chromosomes for ten species and are attached to the short arms of either m or sm chromosomes. They could be seen in 10% to 70% of the cells. Only S. pseudolulo bears two satellited chromosomes, one of them attached to a long chromosome arm. Although the section is chromosomally quite homogeneous, species can be distinguished by karyotype formula, presence of satellites in a particular chromosome pair, and total chromosome length. There are no indications of major chromosomal rearrangements within the section. Using cytological data exclusively, cluster analysis indicates S. sessiliflorum is isolated in the section. Solanum candidum and S. vestissimum are somewhat isolated as well and have unique karyological traits. Solanum pectinatum has a peculiar karyotype, but in the phenogram it is not particularly separated. Karyotype features suggest that morphological differentiation was not always followed by chromosomal divergence. The origin of the domesticated S. quitoense is no further elucidated by our data.     

Isolation of a series of chromosomal variants (2n=10 to 21) from an open-pollinated population of interspecific hybrids between Coix gigantea and Coix aquatica (Poaceae)

A series of chromosomal variants has been isolated from an open-pollinated progeny of interspecific hybrids between aneuploids of Coix gigantea (2n=18–24) and Coix aquatica (2n=10). The interspecific hybrids (2n=14, 15 and 16) produced several types of gametes not only with different chromosome numbers but also comprised of varied permutations and combinations of gigantea and aquatica chromosomes. This was evident when the open-pollinated progeny obtained from these hybrids was screened chromosomally. Two such open-pollinated experimental progenies were studied in two successive years (1983 and 1984) and plants with from 2n=10 to 2n=21 chromosomes were isolated. Chromosomal configurations at diakinesis in all the variants revealed frequent pairing between the gigantea and aquatica chromosomes. This indicated that the two species are phylogenetically closely related. Restoration of pure parental species from the F₁ hybids and chromosomal variants through genomic segregation and spontaneous back-cross are unique and noteworthy features.     

An extraordinarily stable karyotype of the woody Populus species revealed by chromosome painting

The karyotype represents the basic genetic make‐up of a eukaryotic species. Comparative cytogenetic analysis of related species based on individually identified chromosomes has been conducted in only a few plant groups and not yet in woody plants. We have developed a complete set of 19 chromosome painting probes based on the reference genome of the model woody plant Populus trichocarpa. Using sequential fluorescence in situ hybridization we were able to identify all poplar chromosomes in the same metaphase cells, which led to the development of poplar karyotypes based on individually identified chromosomes. We demonstrate that five Populus species, belonging to five different sections within Populus, have maintained a remarkably conserved karyotype. No inter‐chromosomal structural rearrangements were observed on any of the 19 chromosomes among the five species. Thus, the chromosomal synteny in Populus has been remarkably maintained after nearly 14 million years of divergence. We propose that the karyotypes of woody species are more stable than those of herbaceous plants since it may take a longer period of time for woody plants to fix chromosome number or structural variants in natural populations.     

Comparative cytogenetics of six Indo‐Pacific moray eels (Anguilliformes: Muraenidae) by chromosomal banding and fluorescence in situ hybridization

A comparative cytogenetic analysis, using both conventional staining techniques and fluorescence in situ hybridization, of six Indo‐Pacific moray eels from three different genera (Gymnothorax fimbriatus, Gymnothorax flavimarginatus, Gymnothorax javanicus, Gymnothorax undulatus, Echidna nebulosa and Gymnomuraena zebra), was carried out to investigate the chromosomal differentiation in the family Muraenidae. Four species displayed a diploid chromosome number 2n = 42, which is common among the Muraenidae. Two other species, G. javanicus and G. flavimarginatus, were characterized by different chromosome numbers (2n = 40 and 2n = 36). For most species, a large amount of constitutive heterochromatin was detected in the chromosomes, with species‐specific C‐banding patterns that enabled pairing of the homologous chromosomes. In all species, the major ribosomal genes were localized in the guanine‐cytosine‐rich region of one chromosome pair, but in different chromosomal locations. The (TTAGGG)_n telomeric sequences were mapped onto chromosomal ends in all muraenid species studied. The comparison of the results derived from this study with those available in the literature confirms a substantial conservation of the diploid chromosome number in the Muraenidae and supports the hypothesis that rearrangements have occurred that have diversified their karyotypes. Furthermore, the finding of two species with different diploid chromosome numbers suggests that additional chromosomal rearrangements, such as Robertsonian fusions, have occurred in the karyotype evolution of the Muraenidae.     

BoS: A Large and Diverse Family of Short Interspersed Elements (SINEs) in Brassica oleracea

Short interspersed elements (SINEs) are nonautonomous non-LTR retrotransposons that populate eukaryotic genomes. Numerous SINE families have been identified in animals, whereas only a few have been described in plants. Here we describe a new family of SINEs, named BoS, that is widespread in Brassicaceae and present at ∼2000 copies in Brassica oleracea. In addition to sharing a modular structure and target site preference with previously described SINEs, BoS elements have several unusual features. First, the head regions of BoS RNAs can adopt a distinct hairpin-like secondary structure. Second, with 15 distinct subfamilies, BoS represents one of the most diverse SINE families described to date. Third, several of the subfamilies have a mosaic structure that has arisen through the exchange of sequences between existing subfamilies, possibly during retrotransposition. Analysis of BoS subfamilies indicate that they were active during various time periods through the evolution of Brassicaceae and that active elements may still reside in some Brassica species. As such, BoS elements may be a valuable tool as phylogenetic makers for resolving outstanding issues in the evolution of species in the Brassicaceae family.     

Fissions,fusions, and translocations shaped the karyotype and multiple sex chromosome constitution of the northeast‐Asian wood white butterfly,Leptidea amurensis

Previous studies have shown a dynamic karyotype evolution and the presence of complex sex chromosome systems in three cryptic Leptidea species from the Western Palearctic. To further explore the chromosomal particularities of Leptidea butterflies, we examined the karyotype of an Eastern Palearctic species, Leptidea amurensis. We found a high number of chromosomes that differed between the sexes and slightly varied in females (i.e. 2n = 118–119 in females and 2n = 122 in males). The analysis of female meiotic chromosomes revealed multiple sex chromosomes with three W and six Z chromosomes. The curious sex chromosome constitution [i.e. W_1–3/Z_1–6 (females) and Z_1–6/Z_1–6 (males)] and the observed heterozygotes for a chromosomal fusion are together responsible for the sex‐specific and intraspecific variability in chromosome numbers. However, in contrast to the Western Palearctic Leptidea species, the single chromosomal fusion and static distribution of cytogenetic markers (18S rDNA and H3 histone genes) suggest that the karyotype of L. amurensis is stable. The data obtained for four Leptidea species suggest that the multiple sex chromosome system, although different among species, is a common feature of the genus Leptidea. Furthermore, inter‐ and intraspecific variations in chromosome numbers and the complex meiotic pairing of these multiple sex chromosomes indicate the role of chromosomal fissions, fusions, and translocations in the karyotype evolution of Leptidea butterflies.     

CYTOGENETIC ANALYSIS OF CHROMOSOMAL INTERMEDIATES FROM A HYBRID ZONE BETWEEN TWO CHROMOSOME RACES OF THE SCELOPORUS GRAMMICUS COMPLEX (SAURIA,PHRYNOSOMATIDAE)

Underdominance for chromosomal rearrangements is the central assumption of several models of chromosomally based speciation including the cascade model, proposed for the Sceloporus grammicus complex. Several cytotypes of the S. grammicus complex hybridize at localities in central México. A hybrid zone between two of the most chromosomally divergent races (= cytotypes) of S. grammicus (F5, 2n = 34 and FM2, 2n = 44–46) was examined to assess the meiotic effects of heterozygosity at multiple chromosomes. Meiosis was examined in males heterozygous for “simple” Robertsonian fissions at chromosomes 1, 3, 4, and 6 and/or a pericentric inversion at chromosome 4. Analysis of synaptonemal complexes and chromosomal configurations at diakinesis showed trivalent formation in fission heterozygotes and heterosynapsis (lack of reverse-loop formation) in an inversion heterozygote. Analysis of metaphase II configurations revealed primarily balanced segregation and low levels of nondisjunction regardless of chromosomal background. The lack of underdominance associated with “simple” fission heterozygosity in this narrow hybrid zone contradicts the key premise of most chromosomally based models of speciation.     

The structure,amount and chromosomal localisation of defined repeated DNA sequences in species of the genus Secale

The structure, copy number and chromosomal location of arrays of four families of highly repeated sequences have been investigated in representative species of the genus Secale. The four unrelated families, previously characterised in Secale cereale, have repeating units of 480, 610, 630 and 120 base pairs respectively. The following general conclusions can be drawn in addition to detailed knowledge of the sequence content of heterochromatin in each accession studied: (1) Every species is unique in its complement or chromosomal distribution or both of the four highly repeated sequence families. S. montanum and S. cereale accessions studied here show the same complement of repeated sequences, but they differ substantially in the amounts they contain of the 610 and 630 base pair (bp) families, and in the distribution over the chromosomes of the 480 bp family. The structure of the repeating unit is also different in many members of the 480 bp family in S. montanum. — (2) The substantial differences between species in the amounts of the most highly repeated DNA sequences exist in the absence of any such conspicuous differences in most other repeated sequences which were detected as fluorescent bands after restriction enzyme digestion and gel electrophoresis. — (3) Each of the different highly repeated families can exist independently of the other families, though all the families have telomeric sites. Also, in the outbreeding species, heteromorphisms are frequent, and are particularly conspicuous in hybridisation detecting the 480 bp sequence family. — (4) The association of the highly repeated sequences with heterochromatin, discussed in the accompanying paper is generally true for other species in the genus, and the lower amounts of heterochromatin in other Secale species compared to S. cereale are associated with lower amounts of specific families of highly repeated DNA sequences. — (5) Analysis of highly repeated sequence families is likely to provide an easy method of identification of new accessions of Secale.     

Chromosomes of sixteen European species ofLongitarsus flea-beetles (Coleoptera,Chrysomelidae)

Sixteen species ofLongitarsus have been chromosomally surveyed, showing a continuous range of even numbers from 2n=26 to 2n=32 chromosomes. Among the total of twenty-three known species, about 40% display a 14+Xy male karyotypic formula, the possible modal and most primitive one for the genus. The current taxonomy of species groupings is in good agreement with the chromosome numbers in some cases, but not in others. Also, there is no interrelationship between chromosome numbers and foodplant selection. The number of large bivalents at metaphase I is generally negatively correlated with the diploid value, suggesting the possible role of centric fusions coupled to shifts in the amount of chromatin as the main chromosomal changes in the evolution ofLongitarsus. The karyotypes of a few studied species are composed of metacentric chromosomes, some of them of rather large size, and a minute y-chromosome. A possible example of polymorphism for the chromosome number inL. nigrofasciatus is reported and briefly discussed.