Comparative cytogenetics of three species of Dichotomius (Coleoptera, Scarabaeidae)

Meiotic and mitotic chromosomes of Dichotomius nisus, D. semisquamosus and D. sericeus were analyzed after conventional staining, C-banding and silver nitrate staining. In addition, Dichotomius nisus and D. semisquamosus chromosomes were also analyzed after fluorescent in situ hybridization (FISH) with an rDNA probe. The species analyzed had an asymmetrical karyotype with 2n = 18 and meta-submetacentric chromosomes. The sex determination mechanism was of the Xy(p) type in D. nisus and D. semisquamosus and of the Xy (r) type in D. sericeus. C-banding revealed the presence of pericentromeric blocks of constitutive heterochromatin (CH) in all the chromosomes of the three species. After silver staining, the nucleolar organizer regions (NORs) were located in autosomes of D. semisquamosus and D. sericeus and in the sexual bivalent of D. nisus. FISH with an rDNA probe confirmed NORs location in D. semisquamosus and in D. nisus. Our results suggest that chromosome inversions and fusions occurred during the evolution of the group.     

Cytomolecular analysis of the male sex chromosome of Tropidacris cristata grandis (Thunberg, 1824) (Orthoptera: Romaleidae)

Many species of grasshopper have an XX/XO sex chromosome system, including Tropidacris cristata grandis (23, XX/XO). The X chromosome behaves differently from the autosomes, but little is known about its origin and molecular composition. To better understand the genomic composition and evolutionary processes involved in the origin of the sex chromosomes, we undertook an analysis of its meiotic behavior, heterochromatin distribution and microdissection in T. c. grandis. Analysis of meiotic cells revealed a difference in the behavior of the X chromosome compared to the autosomes, with different patterns of condensation and cellular arrangement. Heterochromatic terminal blocks were predominant. The chromosome painting revealed a bright block in the centromeric/pericentromeric region of the X chromosome and slight markings in the other regions. In the autosomes, the X chromosome probe hybridized in the centromeric/pericentromeric region, and hybridization signals on terminal regions corresponding to the heterochromatic regions were also observed. The results showed that the X chromosome contains a significant amount of repetitive DNA. Based on the hybridization pattern, it is possible that the autosomes and sex chromosomes of T. c. grandis have a similar composition of repetitive DNAs, which could mean that the X chromosome has an autosomal origin.     

Comparative Analysis (Hippotragini versus Caprini, Bovidae) of X-Chromosome's Constitutive Heterochromatin by in situ Restriction Endonuclease Digestion: X-Chromosome Constitutive Heterochromatin Evolution

The Bovidae X-chromosome shows a considerable variation, in contrast to the preservative autosomal conservatism. The X-chromosome variation is mostly a consequence of the constitutive heterochromatin (CH) variation; in what respect to its amount and position. This is especially common among the non-Bovinae subfamilies and tribes. In order to characterize the X-chromosome CH in non-Bovinae species--Hippotragini and Caprini tribes--we have used restriction endonuclease digestion on fixed chromosomes and sequential C-banding. With these techniques we were able to distinguish between the two X-chromosome types (Hippotragini and Caprini) CH, in what respect to its position and molecular nature. Moreover, we define at least, six subclasses of CH in both X-chromosome types analyzed. Evolutionary considerations were draw based on the results obtained. The technology used here for the analysis of the Bovidae X-chromosome CH showed to be more evolutionary informative than the classical approaches.     

Cytogenetic analysis of Australian tiger beetles (Coleoptera: Cicindelidae): chromosome number, sex-determining system and localization of rDNA genes

The chromosomes of four species of Australian tiger beetle are documented. The male meioformulae of three species of Cicindela (tribe Cicindelini) were determined as follow: Cicindela cardinalba Sumlin 1987, n = 10 + X₁ X₂ X₃Y; C. sp. ( saetigera group) and Cicindela gillesensis Hudson 1994, n = 11 + X₁ X₂ X₃Y. The male meioformula of Megacephala whelani Sumlin 1992 (tribe Megacephalini) is n = 12 + XY. Fluorescence in situ hybridization (FISH) was used to localize the 18S-28S ribosomal gene clusters on male mitotic and meiotic chromosomes and nuclei using a polymerase chain reaction-amplified ribosomal probe. Fluorescence in mitosis and meiotic first prophase stages in the three Cicindela species showed that rDNA genes are in two of the four chromosomes that form the sex vesicle. In M. whelani hybridization in mitosis and first metaphase stages indicate that rDNA genes are in three, medium- to small-sized, autosomal pairs. Silver staining of male meiotic nuclei reveals the presence of active nucleolar organizing regions (NORs) in the sex vesicle of the three species of Cicindela . The cytogenetic and evolutionary implications of these findings are discussed.     

Karyotypes,heterochromatin distribution and rDNA patterns in South American Grindelia (Asteraceae)

Grindelia is a genus with a complex evolutionary history with reticulate evolution. We studied the karyotype, fluorescent banding, and fluorescence in situ hybridization (FISH) using 18–5.8–26 S and 5 S ribosomal DNA probes to survey karyotypic diversity of South American Grindelia species. Chromosome basic numbers were x = 6 (with several ploidy levels: 2x, 4x, and 6x). All the Grindelia studied conserved the patterns of CG-rich heterochromatin and 18-5.8-26 S rDNA. The third m sat-chromosome pair was homeologous in Grindelia. Chromosome variation, although not always large, accompanied the evolutionary divergence of the taxa studied. The Grindelia studied formed two species groups: (1) G. globularifolia and G. pulchella var. pulchella, (2) G. buphthalmoides, G. cabrerae var. alatocarpa and var. cabrerae, G. chiloensis, G. orientalis, and G. prostrata. These groups do not show any morphological affinities and their phylogenetic relationships are not clearly resolved, suggesting that these groups have recently diverged.     

Karyotype of Callithrix mauesi (Callitrichidae,Primates) and its relations with those of C. emiliae and C. jacchus

A study was conducted on the most recently described marmoset species, Callithrix mauesi, and the results obtained were compared to those previously reported for the karyotypes of C. jacchus and C. emiliae. No mechanism of chromosome rearrangement differentiates the karyotypes of C. mauesi (2n = 44) and C. emiliae (2n = 44), which diverge from C. jacchus (2n = 46) by a Robertsonian translocation and a paracentric inversion. C. mauesi, like C. emiliae, presents telomeric constitutive heterochromatin in various chromosomes, forming large heterochromatic blocks in some. This does not occur in C. jacchus, which basically presents centromeric constitutive heterochromatin. The karyotype of C. mauesi differs from that of C. emiliae only by the amount and distribution of this telomeric constitutive heterochromatin. One of the chromosomes presenting a heterochromatic block in C. mauesi is chromosome X, a fact not previously reported in the Order Primates. The present chromosome data show that C. mauesi is closer to C. emiliae than to C. jacchus, in agreement with its inclusion in the C. argentata group. In the present paper, we describe for the first time, at the chromosome level, chimerism between fraternal twins of the same sex (XY/XY), with the heterochromatic block of pair 2 being the marker. © 1994 Wiley-Liss, Inc.     

Heterochromatin and rDNA patterns in Solanum species of the Morelloid and Dulcamaroid clades (Solanaceae)

The heterochromatin distribution and the position of 18-5.8-26S, and 5S rDNA loci were determined in 13 species of Solanum of the Morelloid and Dulcamaroid clades. The CMA/DAPI staining and FISH were employed. Two types of constitutive heterochromatin were determined: CMA⁺/DAPI^? associated to NOR and CMA⁺/DAPI^? distributed as terminal bands. In the Morelloid clade, CMA⁺/DAPI^? bands were found in five species while in the Dulcamaroid clade, only S. angustifidum presented this feature. In the Morelloid clade, two to four 18-5.8-26S rDNA loci occupied terminal positions and two rDNA 5S loci were found with variable positions (terminal, intercalary, and centromeric). In the Dulcamaroid clade, two terminal 18-5.8-26S rDNA loci were detected with the exception of S. salicifolium which possessed four such loci and two to four 5S rDNA loci. Solanum crispum is the only species possessing the 5S in synteny with 18-5.8-26S rDNA loci. Karyotype features chromosome banding pattern as well as the location of ribosomal genes which varied among the species, reflecting the chromosome differentiation and evolutionary divergence. The findings obtained contributed to the development of tools that can be used for establishing chromosomic homeologies among species and hence to clarify their taxonomic relationships.     

Variability of the 5S and 45S rDNA sites in Passiflora L. species with distinct base chromosome numbers

Cytologically, the species of Passiflora with known chromosome number can be divided into four groups: (1) 2n = 12, 24, 36; (2) 2n = 24; (3) 2n = 18, 72; and (4) 2n = 20. The base chromosome number proposed for the genus is x = 6, with x = 9, x = 10 and x = 12 being considered secondary base numbers. In the present study, variability of 5S and 45S rDNA sites was investigated in 20 species of these four groups to check the reliability of this hypothesis. In the group with x = 6, five diploid species (2n = 12) exhibit two 5S rDNA sites and two (P. capsularis, P. morifolia and P. rubra) or four (P. misera 2x and P. tricuspis) 45S rDNA sites. The hexaploid cytotype of P. misera had 12 45S rDNA sites and six 5S rDNA. A tetraploid species, P. suberosa, had ten 45S rDNA sites and four 5S rDNA sites, both in the same chromosomes as the 45S rDNA sites. In the group with x = 9, P. actinia, P. amethystina, P. edmundoi, P. elegans, P. galbana, P. glandulosa and P. mucronata displayed six 45S rDNA sites, whereas P. alata, P. cincinnata, P. edulis f. flavicarpa, P. edulis var. roxo and P. laurifolia had four sites. In this group, all species were diploid (2n = 18) and had only two 5S rDNA sites. Passiflora foetida, the only species with 2n = 20, had six 45S rDNA sites and four 5S rDNA sites. The species with x = 12 (2n = 24), P. haematostigma and P. pentagona, showed four 45S rDNA sites and two 5S rDNA. In general, the number and location of 5S and 45S rDNA sites were consistent with the hypothesis of x = 6 as the probable ancestral genome for the genus, while the groups of species with x = 9, x = 10 and x = 12 were considered to be of tetraploid origin with descending dysploidy and gene silencing of some redundant gene sites, mainly those of 5S rDNA.     

Physical mapping of rDNA genes in the ground beetle Carabus and related genera (Carabidae: Coleoptera)

The major ribosomal DNA (rDNA) loci were localized on meiotic and mitotic chromosomes and in interphase nuclei of 18 ground-beetle species belonging to three tribes of the supertribe Carabitae by fluorescence in situ hybridization (FISH), using a PCR-amplified 18S rDNA as a probe. Meiotic observations indicate that the 18S rDNA sequences are located on the largest autosomal bivalent in 12 species of Carabus , two species of Calosoma (both genera belonging to the tribe Carabini), and three sibling species of Ceroglossus chilensis (tribe Ceroglossini). The data suggest the occurrence of a conservative pattern in these three genera despite the chromosomal rearrangements that have led to karyotypes with higher chromosome numbers in Ceroglossus . A different result is found in Cychrus caraboides (tribe Cychrini), where ribosomal cistrons are located in two medium-sized autosomal pairs. Further species of Cychrini should be studied for corroborating the occurrence of molecular and karyotypical apomorphies in Cychrus with regard to the genera Carabus, Calosoma and Ceroglossus .     

Cytogenetic studies of four South American species of Paullinia L. (Sapindaceae)

Four South American species of Paullinia ( P. elegans , P. meliaefolia , P. pinnata , and P. rhomboidea ) were compared using conventional chromosome staining, C-Giemsa and C-chromomycin A₃/4',6-diamidino-2-phenylindole (C-CMA₃/DAPI) banding, and fluorescence in situ hybridization (FISH) with a 45S ribosomal DNA (rDNA) probe. All species showed a somatic complement of 2 n  = 24 chromosomes, agreeing with earlier records in some cases, and showing a tendency for the chromosome number to be conserved in this genus. The chromosome number of P. rhomboidea is a new report. The karyotypes differed in chromosome size and degree of karyotype asymmetry. The chromosomal band patterns and location of the 45S rDNA sites are reported for the first time in the genus. Terminal C-CMA₃ bands were associated with the 45S rDNA sites, but varied in number and size between the species. The occurrence of other C-Giemsa bands that were not revealed by CMA₃ suggests that more than one family of repetitive DNA may be involved in karyotype differentiation. The systematic implications of these results on the infrageneric relationships are discussed.   © 2007 The Linnean Society of London . Botanical Journal of the Linnean Society , 2007, 154 , 313–320.     

Molecular cytogenetic evidence of hybridization in the “purple corolla clade of the genus Capsicum” (C. eximium × C. cardenasii)

Abstract

A hybrid individual between two taxa from “the Purple Corolla clade of the genus Capsicum”, C. eximium × C. cardenasii (2n = 24) was found during a cytogenetic study of a population belonging to C. cardenasii cytotype 1, from Bolivia. 5S and 45S rDNA probes were located on mitotic chromosomes by fluorescent in situ hybridization. The hybrid haploid karyotype length was an average value of the two taxa; the hybrid presented 24 45S loci in the diploid complement, 18 45S sites belonged to C. cardenasii cytotype 2, and six came from C. eximium cytotype 2, although this cytotype of C. eximium has been mentioned only for Argentina. Whereas hybrids were previously reported in the purple flower group, their existence has not been cytologically corroborated. This finding constitutes the first molecular cytogenetic evidence of hybridization between two taxa from this group.     

C-Banding Patterns and Quantitative Karyotype Characteristics of Bulgarian Species of Crepis (Asteraceae)

Abstract: Giemsa C-banded idiograms were established from eight Crepis species of Bulgarian origin ( Crepis viscidula, C. paludosa, C. conyzaefolia, C. bithynica var. bithynica and var. fodorii, C. pulchra, C. sancta, C. setosa, and C. zacintha ) that had been previously studied for genome size and karyotype shape using classical methods. All taxa are diploids with descending chromosome numbers, x = 6, 5, 4, and 3. Satellites were always heterochromatic. Small but distinct centromeric bands were of general occurrence (except perhaps in C. paludosa ) and terminal bands were present at least in some chromosomes of the karyotypes. Higher amounts of heterochromatin in the form of terminal and intercalary bands were found in C. bithynica and C. zacintha, but banding patterns were not so specifically similar as to indicate common origin. There are a number of particular karyotypic characters which may turn out to be phylogenetically significant when corresponding information on many related taxa becomes available. Correlations of karyotypic parameters were calculated. In heterochromatin only percent heterochromatin, but not absolute heterochromatin amount, and genome size were significantly correlated. This is the result of a strong euchromatin variation but independent heterochromatin variation.     

Cytogenetic studies in South American species of Serjania (Sapindaceae: Paullinieae)

Abstract

Serjania Mill. (Paullinieae) is considered the most important neotropical genus of Sapindaceae due to species number and its widespread distribution. In this study, 14 species belonging to three sections were analyzed using conventional staining, C/CMA/DAPI banding, and fluorescence in situ hybridization (FISH) with a 18S-5.8S-26S rDNA probe. New chromosome counts are reported for Serjania crassifolia, Serjania platycarpa, and Serjania regnellii, all with 2n = 24, which is remarkably constant for Serjania. The karyotypes are moderately asymmetric, and variations observed in A1 and A2 indices show resemblances between S. platycarpa, Serjania hebecarpa, and S. crassifolia, and between Serjania communis, Serjania gracilis, and S. regnellii. The banding pattern was homogeneous in Serjania. C/DAPI bands (AT-rich sites) were not clearly evidenced, but changes in the number and position of GC-rich sites (CMA bands) were observed. These segments were associated with 18S-5.8S-26S rDNA sites. The significance of the results is discussed in relation to chromosomal data available for the genus and in regard to the infrageneric treatment of Serjania.     

Karyotype of Mesembryanthemum crystallinum (Aizoaceae) studied by chromosome banding,FISH with rDNA probes and immunofluorescence detection of DNA methylation

The karyotype of the common ice plant Mesembryanthemum crystallinum L. (Aizoaceae) was studied using Chromomycin A₃ (CMA)/4′,6-diamidino-2-phenylindole (DAPI) staining, fluorescence in situ hybridization with 5S and 18S–5.8S–25S rDNA probes, DAPI/C-banding and immunodetection of 5-methylcytosine. A single bright CMA-band was revealed on the satellite chromosome, whose location was coincided with a position of a site of 18S–5.8S–25S rRNA genes. A site of 5S rRNA genes was observed on one of the other chromosomes. Relatively large DAPI/C-bands were mainly localized in the pericentromeric regions of the chromosomes. DAPI/C-banding patterns allowed us to identify all the chromosomes in the karyotype of M. crystallinum. The methylation of euchromatic chromosome regions was weaker as compared with heterochromatic DAPI/C-bands, which were hypermethylated. The obtained results may provide opportunities for investigating, at the chromosomal level, the genomic changes occurring in M. crystallinum either under salinization or under the action of other stress factors.     

Morphology and Phylogenetic Position of Two New Gregarine Species (Apicomplexa: Eugregarinorida) Parasitizing the Lubber Grasshopper Taeniopoda centurio (Drury, 1770) (Insecta: Orthoptera: Romaleidae) in Mexico

Eugregarines are understudied apicomplexan parasites of invertebrates inhabiting marine, freshwater, and terrestrial environments. Most currently known terrestrial eugregarines have been described parasitizing the gut from less than 1% of total insect diversity, with a high likelihood that the remaining insect species are infected. Eugregarine diversity in orthopterans (grasshoppers, locusts, katydids, and crickets) is still little known. We carried out a survey of the eugregarines parasitizing the Mexican lubber grasshopper, Taeniopoda centurio, an endemic species to the northwest of Mexico. We described two new eugregarine species from the gut of the host: Amoebogregarina taeniopoda n. sp. and Quadruspinospora mexicana n. sp. Both species are morphologically dissimilar in their life‐cycle stages. Our SSU rDNA phylogenetic analysis showed that both species are phylogenetically distant to each other, even though they parasitize the same host. Amoebogregarina taeniopoda n. sp. clustered within the clade Gregarinoidea, being closely related to Amoebogregarina nigra from the grasshopper Melanoplus differentialis. Quadruspinospora mexicana n. sp. clustered within the clade Actinocephaloidea and grouped with Prismatospora evansi, a parasite from dragonfly naiads. Amoebogregarina taeniopoda n. sp. and Q. mexicana n. sp. represent the first record of eugregarines found to infect a species of the family Romaleidae.     

Cytogenetic studies in the Andropogon gayanus—A. tectorum complex (Graminae)

Cytogenetic studies in the Andropogon gayanus—A. tectorum complex (Granimae). Studies on the morphology, karyotype and meiotic behaviour of collections of Andropogon gayanus [2 n = 40), A. tectorum (2 n = 20,30) and their hybrids, obtained by both open pollination and controlled crosses, have been made. The chromosome number of 2 n = 30 for A. tectorum is new. Karyotypic analyses of the species and the preponderance of trivalents at diakinesis in the triploid and aneuploid hybrids show that considerable homology exists between the genomes of the two species. The evolutionary relationship between the two species and the probable origin of tetraploid A. gayanus are discussed.     

Cytogenetic analysis of three species of Pseudacteon (Diptera, Phoridae) parasitoids of the fire ants using standard and molecular techniques

Pseudacteon flies, parasitoids of worker ants, are being intensively studied as potentially effective agents in the biological control of the invasive pest fire ant genus Solenopsis (Hymenoptera: Formicidae). This is the first attempt to describe the karyotype of P. curvatus Borgmeier, P. nocens Borgmeier and P. tricuspis Borgmeier. The three species possess 2n = 6; chromosomes I and II were metacentric in the three species, but chromosome pair III was subtelocentric in P. curvatus and P. tricuspis, and telocentric in P. nocens. All three species possess a C positive band in chromosome II, lack C positive heterochromatin on chromosome I, and are mostly differentiated with respect to chromosome III. P. curvatus and P. tricuspis possess a C positive band, but at different locations, whereas this band is absent in P. nocens. Heterochromatic bands are neither AT nor GC rich as revealed by fluorescent banding. In situ hybridization with an 18S rDNA probe revealed a signal on chromosome II in a similar location to the C positive band in the three species. The apparent lack of morphologically distinct sex chromosomes is consistent with proposals of environmental sex determination in the genus. Small differences detected in chromosome length and morphology suggests that chromosomes have been highly conserved during the evolutionary radiation of Pseudacteon. Possible mechanisms of karyotype evolution in the three species are suggested.     

Comparative cytogenetic analysis of Cestrum (Solanaceae) reveals different trends in heterochromatin and rDNA sites distribution

Species of Cestrum L. (Solanaceae) exhibit large variability in the accumulation of repetitive DNA, although their species possess a stable diploid number with 2n = 16. In this study, we used chromosome banding and fluorescence in situ hybridization (FISH) to characterize the karyotypes and populations of two species, Cestrum nocturnum L. and C. mariquitense Kunth. We also performed a karyotype comparison using 16 idiograms, of which 4 were developed in this study and 12 were obtained from the literature. Cestrum nocturnum displayed more bands than C. mariquitense, but the latter exhibited greater interpopulational variation in the band patterns. There was a tendency for large bands to be located at intercalary/terminal regions and for small bands to be located at intermediate/proximal regions. The idiogram comparison revealed a large variation in the amount, distribution, and size of heterochromatic bands. FISH with rDNA probes revealed stability in the number and location of 5S sites, while 45S was more variable in size and number of sites. Although 45S rDNA always appeared in the subterminal regions, this DNA family exhibited a mobility among chromosome pairs. These data highlight the dynamic of repetitive DNA families in these genomes, as well as the contribution for intra- and interspecific karyotype differentiation in Cestrum.     

Cytogenetic analysis of Ctenostomini by C-banding and rDNA localization and its relevance to the knowledge of the evolution of tiger beetles (Coleoptera: Cicindelidae)

In this work, the first cytogenetic data on Neotropical Collyrinae is provided, by way of their karyotypes, C-banding and ribosomal genes (rDNA) localization using fluorescence in situ hybridization (FISH). The two species analysed, Ctenostoma (Procephalus) ornatum ornatum (male) and Ctenostoma (Euctenostoma) rugosum(female) showed, respectively, a diploid number of 17 and 18 chromosomes. C. ornatum ornatum has a multiple sex chromosome system ( n=7 + X₁X₂Y), and mitotic and meiotic metaphase cells showed rDNA gene labelling in the smallest autosomal pair. In this species, no C-bands were obtained, while C. rugosum seems to exhibit centromeric and/or interstitial C-bands in almost all chromosomes. The observation of a multiple sex chromosome system in Ctenostomini ensured the appearance of this characteristic in the hypothetical ancestral of Collyrinae and Cicindelini. The subfamily Collyrinae is not uniform in what concerns diploid chromosome number and rDNA gene localization, because C. ornatum ornatum possesses a lower chromosome number and autosomal rDNA genes when compared with the other Collyrinae species studied ( Neocollyris spp.). Independent events leading to the reduction in chromosome number might have taken place during the split of the Collyrinae into the tribes Ctenostomini and Collyrini.