Retroelements: tools for sex chromosome evolution

Many eukaryotic taxa inherit a heteromorphic sex chromosome pair. It is a generally accepted hypothesis that the sex chromosome pair is derived from a pair of homologous autosomes that has developed after the occurrence of a sex differentiator in an evolutionary process into two structurally and functionally different partners. In most of the analyzed systems the occurrence of the dominant sex differentiator is paralleled by the suppression of recombination within and close by that region. The recombinational isolation can spread in an evolutionary selection process from neighboring regions finally over the whole chromosome. Suppression of recombination strongly biases the distribution of retrotransposons in the genome. Our results and that from others indicate that the major force driving the evolution of Y chromosomes are retrotransposons, remodeling euchromatic chromosome structures into heterochromatic ones. In our model, intact or already eroded retrotransposons become trapped due to their inherent transposition mechanisms in non-recombining regions. The massive accumulation of retrotransposons interferes strongly with the activity of genes. We hypothesize that Y chromosome degeneration is a stepwise evolutionary process: (1) Massive accumulation of retrotransposons occurs in the non-recombining regions. (2) Heterochromatic nucleation centers are formed as a consequence of genomic defense against invasive parasitic elements; the established nucleation centers become epigenetically inherited. (3) Spreading of heterochromatin from the nucleation centers into flanking regions induces in an adaptive process gene silencing of neighbored genes that could either be still intact or in an already eroded condition, e.g., showing point mutations, deletions, insertions; the retroelements should be subjects to the same forces of deterioration as the genes themselves. (4) Constitutive silenced genes are not committed to the same genetic selection pressure as active genes and therefore more exposed to the decay process. (5) Gene dosage balance is reestablished by the parallel evolution of dosage compensation mechanisms. The evolving secondary sex chromosomes, neo-X and neo-Y, of Drosophila miranda are revealed to be a unique and potent model system to catch the evolutionary Y deterioration process in progress.     

Biased distribution of repetitive elements: a landmark for neo-Y chromosome evolution in Drosophila miranda

It is generally assumed that the sex chromosomes developed from a pair of homologs. Over evolution, the proto-Y chromosome, with a very short differential segment, matured in its final stage into a heterochromatic and, for the most part, genetically eroded Y chromosome. The constraints on the evolution of the proto-Y chromosome have been speculated upon since the sex chromosomes were discovered. Several models have been suggested. Drosophila miranda has proved to be a unique and potent model system to study Y-chromosome evolution. We use selected test genes distributed along the neo-Y chromosome as entry gates to analyze the molecular mechanisms involved in the process of Y-chromosome evolution. Here, we report our findings on the Krüppel gene (Kr), which is located distally on the neo-sex chromosome pair.     

Evolution of DNA in Heterochromatin: the Drosophila Melanogaster Sibling Species Subgroup as a Resource

The Drosophila melanogasterspecies subgroup is a closely-knit collection of eight sibling species whose relationships are well defined. These species are too close for most evolutionary studies of euchromatic genes but are ideal to investigate the major changes that occur to DNA in heterochromatin over short periods during evolution. For example, it is not known whether the locations of genes in heterochromatin are conserved over this time. The 18S and 28S ribosomal RNA genes can be considered as genuine heterochromatic genes. In D. melanogasterthe rRNA genes are located at two sites, one each on the X and Y chromosome. In the other seven sibling species, rRNA genes are also located on the sex chromosomes but the positions often vary significantly, particularly on the Y. Furthermore, rDNA has been lost from the Y chromosome of both D. simulansand D. sechellia, presumably after separation of the line leading to present-day D. mauritiana.We conclude that changes to chromosomal position and copy number of rDNA arrays occur over much shorter evolutionary timespans than previously thought. In these respects the rDNA behaves more like the tandemly repeated satellite DNAs than euchromatic genes. This revised version was published online in July 2006 with corrections to the Cover Date.     

The Genetic Identification of a Heterochromatic Segment on the X Chromosome of DROSOPHILA MELANOGASTER

An autosomal euchromatic maternal-effect mutant, abo (= abnormal oocyte), interacts with, or regulates the activity of, the heterochromatin of the sex chromosomes of Drosophila melanogaster. It is shown that this interaction or regulation with the X chromosome involves a specific heterochromatic locus or small region that maps to the distal penultimate one-eighth of the basal X-chromosome heterochromatic segment.     

Molecular Cytogenetic Characterization of the Dioecious Cannabis sativa with an XY Chromosome Sex Determination System

Hemp (Cannabis sativa L.) was karyotyped using by DAPI/C-banding staining to provide chromosome measurements, and by fluorescence in situ hybridization with probes for 45 rDNA (pTa71), 5S rDNA (pCT4.2), a subtelomeric repeat (CS-1) and the Arabidopsis telomere probes. The karyotype has 18 autosomes plus a sex chromosome pair (XX in female and XY in male plants). The autosomes are difficult to distinguish morphologically, but three pairs could be distinguished using the probes. The Y chromosome is larger than the autosomes, and carries a fully heterochromatic DAPI positive arm and CS-1 repeats only on the less intensely DAPI-stained, euchromatic arm. The X is the largest chromosome of all, and carries CS-1 subtelomeric repeats on both arms. The meiotic configuration of the sex bivalent locates a pseudoautosomal region of the Y chromosome at the end of the euchromatic CS-1-carrying arm. Our molecular cytogenetic study of the C. sativa sex chromosomes is a starting point for helping to make C. sativa a promising model to study sex chromosome evolution.     

Die Speicheldrüsenchromosomen der StechmückeCulex Pipiens L. IV. Der Chromosomale Geschlechtsdimorphismus

In the salivary gland chromosomes of the mosquitoCulex pipiens L. band 10 C 3 of the small chromosome I in female larvae always shows a homozygous heterochromatic balloon, while male larvae are heterozygous for this structure, the homologous band being euchromatic. In males, heterozygous for a male-linked reciprocal translocation, the chromosome with the distinct euchromatic band is always involved in the segmental interchange with an autosome. It is concluded that the euchromatic state of band 10 C 3 represents the male-determining allelomorph, and the heterochromatic state the female-determining allelomorph (M and m, respectively). C. pipiens thus appears to be an excellent example of diplogenotypic determination of sex where the sexes are different in one pair of allelomorphs only. At the chromosomal level, this sexual dimorphism is expressed as a heterochromatic versus an euchromatic state of a single band. The significance of this observation is discussed in connection with the problem of the evolution of sex-determining mechanisms within the Nematocera.     

C-banding and non-homologous associations

A chromosome complement formed by 16 autosomes and an Xy_p sex chromosome system was found in Epilachna paenulata Germar (Coleoptera: Coccinellidae). All autosomes were metacentric except pair 1 which was submetacentric. The X and the Y chromosomes were also submetacentric but the Y was minute. The whole chromosome set carried large paracentric heterochromatic C-segments representing about 15% of the haploid complement length. Heterochromatic segments associated progressively during early meiotic stages forming a large single chromocenter. After C-banding, chromocenters revealed an inner networklike filamentous structure. Starlike chromosome configurations resulted from the attachment of bivalents to the chromocenters. These associations were followed until early diakinesis. Thin remnant filaments were also observed connecting metaphase I chromosomes. Evidence is presented that, in this species, the Xy_p bivalent resulted from an end-to-end association of the long arms of the sex chromosomes. The parachute Xy_p bivalent appeared to be composed of three distinct segments: two intensely heterochromatic C-banded corpuscles formed the canopy and a V-shaped euchromatic filament connecting them represented the parachutist component. The triple constitution of the sex bivalent was interpreted as follows: each heterochromatic corpuscle corresponded to the paracentric C-segment of the X and Y chromosomes; the euchromatic filament represented mainly the long arm of the X chromosome terminally associated with the long arm of the Y chromosome. The complete sequence of the formation of the Xy_p bivalent starting from nonassociated sex chromosomes in early meiotic stages, and progressing through pairing of heterochromatic segments, coiling of the euchromatic filament, and movement of the heterochromatic corpuscles to opposite poles is described. These findings suggest that in E. paenulata the Xy_p sex bivalent formation is different than in other coleopteran species and that constitutive heterochromatic segments play an important role not only in chromosome associations but also in the Xy_p formation.     

Reversion of XO to XY sex chromosome mechanism in a phasmid

Summary The sex chromosomes of the male phasmid Isagoras schraderi Rehn comprise an X and a Y, — each with a submedian kinetochore, and one euchromatic and one heterochromatic arm. At meiosis X and Y form an unequal sex bivalent in which the euchromatic arms are terminally associated. Relatively recent reversion from the XO-XX mechanism characteristic of the Phasmidae is indicated by the presence of the euchromatic arm in both X and Y. The diploid number of the male is 34.Unequal autosomal bivalents are found at meiosis in two other species of Isagoras — Isagoras subaquiles Rehn and Isagoras sp. — and in Pseudophasma menius Westwood. The chromosome complements of these species are described.     

Early origins of the X and Y chromosomes: lessons from tilapia

Differentiated sex chromosome pairs in diverse species display certain common characteristics, normally comprising one largely heterochromatic genetically inactive chromosome and one euchromatic genetically active chromosome (e.g. the mammalian Y and X respectively). It is widely accepted that dimorphic sex chromosomes evolved from homologous pairs of autosomes. Although the exact mechanisms through which the pair diverged are not fully understood, an initial suppression of recombination in the sex-determining region is required by all of the major theories. Here we address the question of the mechanism by which this initial suppression of recombination occurs. Our model postulates that the stochastic, de novo accumulation of heterochromatin in the sex determining region can delay pairing of the sex chromosomes in meiosis, resulting in a decrease in recombination. Data to support this model is presented from the cichlid fish, Oreochromis niloticus. Although such a decrease would in most circumstances be evolutionarily disadvantageous, if the region concerned included the major sex determining gene and other gene(s) with sex-specific functions, then this would be selectively advantageous and could trigger the process(es) which, ultimately, lead to the differentiation of the sex chromosomes.     

Distribution of heterochromatin on the mitotic chromosomes of Musca domestica L. in relation to the activity of male-determining factors

In the housefly, male sex is determined by a dominant factor, M, located either on the Y, on the X, or on any of the five autosomes. M factors on autosome I and on fragments of the Y chromosome show incomplete expressivity, whereas M factors on the other autosomes are fully expressive. To test whether these differences might be caused by heterochromatin-dependent position effects, we studied the distribution of heterochromatin on the mitotic chromosomes by C-banding and by fluorescence in situ hybridization of DNA fragments amplified from microdissected mitotic chromosomes. Our results show a correlation between the chromosomal position of M and the strength of its male-determining activity: weakly masculinizing M factors are exclusively located on chromosomes with extensive heterochromatic regions, i.e., on autosome I and on the Y chromosome. The Y is known to contain at least two copies of the M factor, which ensures a strong masculinizing effect despite the heterochromatic environment. The heterochromatic regions of the sex chromosomes consist of repetitive sequences that are unique to the X and the Y, whereas their euchromatic parts contain sequences that are ubiquitously found in the euchromatin of all chromosomes of the complement. Received: 20 February 1998; in revised form: 11 May 1998 / Accepted: 23 May 1998     

Chromosomal Analysis of Neohydrocoptus jaechi (Wewalka) and Canthydrus diophthalmus (Reiche & Saulcy) (Coleoptera: Noteridae)

Neohydrocoptus jaechi (Wewalka) is recorded for the first time in Egypt. It is very similar to N. angolensis (Peschet), but there are aedeagal differences. The karyotype consists of 6 pairs of autosomes and sex chromosomes which are neo-XY (♂) and neo-XX (♀). C-banding shows that the sex chromosomes of Canthydrus diophthalmus (Reiche & Saulcy) are not the small pair suggested by Bilton (1992), but are one of the largest pairs, with the Y-chromosome totally heterochromatic. The implications of these findings are considered in the light of Belkaceme's (1991) work on the phylogeny of Noteridae. Neohydrocoptus is placed among the group of more primitive genera, outside the closely-knit group of higher Noteridae, while Canthydrus is placed right at the top of the higher Noteridae. The neo-XY sex chromosomes of Neohydrocoptus could be a primitive condition for the family, while the system found in Canthydrus is derivable from that of Synchortus Sharp (Saleh Ahmed et al., 1997) (one of Belkaceme's basal genera of the higher Noteridae) by loss of the mainly heterochromatic X₂-chromosome, leaving the more normal X₁-chromosome and the Y, which is almost entirely heterochromatic in Synchortus and completely so in Canthydrus.     

Chromosome relationships and meiotic mechanisms of certain morsitans group tsetse flies and their hybrids

There are three pairs of euchromatic components, the L₁ and L₂ autosomes and the X chromosome pair respectively, which are found in both G. austeni and the three forms of G. morsitans. Each species/sub-species also includes in its complement a group of heterochromatic autosomes (S) which have various morphologies and differ in number both within and between the species/sub-species. Several lines of evidence are outlined which point to these being supernumerary B chromosomes. Male meiosis is normally achiasmate and only L₁ and L₂ autosomes pair completely. X-Y association is restricted to a small pairing segment the position of which on the X is constant for all the species/sub-species. It is located in one of two positions on the Y chromosome according to the species/sub-species. The S chromosomes behave as hereditary univalents at first anaphase while the sex bivalent can undergo distance pairing best exemplified in G. austeni and G. submorsitans. A Y structural mutant line gives some indication of the size of the pairing segment and demonstrates that survival and maleness is possible even when two-thirds of the chromosome is missing. Meiotic and polytene chromosome studies connected with hybridisation experiments designed to test the sterility factor as a potential means of tsetse control assist in establishing the evolutionary relationship of the subspecies.     

Evolution of the large genome in Capsicum annuum occurred through accumulation of single-type long terminal repeat retrotransposons and their derivatives

Although plant genome sizes are extremely diverse, the mechanism underlying the expansion of huge genomes that did not experience whole‐genome duplication has not been elucidated. The pepper, Capsicum annuum, is an excellent model for studies of genome expansion due to its large genome size (2700 Mb) and the absence of whole genome duplication. As most of the pepper genome structure has been identified as constitutive heterochromatin, we investigated the evolution of this region in detail. Our findings show that the constitutive heterochromatin in pepper was actively expanded 20.0–7.5 million years ago through a massive accumulation of single‐type Ty3/Gypsy‐like elements that belong to the Del subgroup. Interestingly, derivatives of the Del elements, such as non‐autonomous long terminal repeat retrotransposons and long‐unit tandem repeats, played important roles in the expansion of constitutive heterochromatic regions. This expansion occurred not only in the existing heterochromatic regions but also into the euchromatic regions. Furthermore, our results revealed a repeat of unit length 18–24 kb. This repeat was found not only in the pepper genome but also in the other solanaceous species, such as potato and tomato. These results represent a characteristic mechanism for large genome evolution in plants.     

Sex chromosome evolution in frogs—helpful insights from chromosome painting in the genus Engystomops

The differentiation of sex chromosomes is thought to be interrupted by relatively frequent sex chromosome turnover and/or occasional recombination between sex chromosomes (fountain-of-youth model) in some vertebrate groups as fishes, amphibians, and lizards. As a result, we observe the prevalence of homomorphic sex chromosomes in these groups. Here, we provide evidence for the loss of sex chromosome heteromorphism in the Amazonian frogs of the genus Engystomops, which harbors an intriguing history of sex chromosome evolution. In this species complex composed of two named species, two confirmed unnamed species, and up to three unconfirmed species, highly divergent karyotypes are present, and heteromorphic X and Y chromosomes were previously found in two species. We describe the karyotype of a lineage estimated to be the sister of all remaining Amazonian Engystomops (named Engystomops sp.) and perform chromosome painting techniques using one probe for the Y chromosome and one probe for the non-centromeric heterochromatic bands of the X chromosome of E. freibergi to compare three Engystomops karyotypes. The Y probe detected the Y chromosomes of E. freibergi and E. petersi and one homolog of chromosome pair 11 of Engystomops sp., suggesting their common evolutionary origin. The X probe showed no interspecific hybridization, revealing that X chromosome heterochromatin is strongly divergent among the studied species. In the light of the phylogenetic relationships, our data suggest that sex chromosome heteromorphism may have occurred early in the evolution of the Amazonian Engystomops and have been lost in two unnamed but confirmed candidate species.Subject terms: Cytogenetics, Evolutionary genetics     

Giant sex chromosomes retained within the Portuguese lineage of the field vole (<Emphasis Type="Italic">Microtus agrestis</Emphasis>)

The field vole (Microtus agrestis) is characterised by extremely large blocks of heterochromatin on both the X and Y chromosome. Some other Microtus also have blocks of heterochromatin on their sex chromosomes but not as extensive and always of independent origin from the heterochromatic expansion found in M. agrestis. Coupled with evidence of geographic variation in large heterochromatic blocks within other species (e.g. in the western hedgehog Erinaceus europaeus), it might be expected that field voles would show substantial variation in size and disposition of the sex chromosome heterochromatin. In fact, only minor variation has been described up to now. Those studies conducted previously were largely on field voles from central and northern Europe. Here, we describe the karyotype of field voles from Portugal, of interest because recent molecular studies have shown field voles from western Iberia to be a separate evolutionary unit that might be considered a cryptic species, distinct from populations further to the east. The two Portuguese field voles (one female, one male) that we examined also had essentially the same karyotype as seen in other field voles, including the giant sex chromosomes, but with small differences in the structure of the Y chromosome from that described previously. The finding that field voles throughout Europe show relatively little variation in their giant sex chromosomes is consistent with molecular data which suggest a recent origin for this complex of species/near-species.     

The Comstockiella system of chromosome behavior in the armored scale insects (Coccoïdea: Diaspididae)

Summary The Comstockiella chromosome system occurs in the armored scale insects and the closely allied palm scales. During development of the males, the paternal chromosome set becomes heterochromatic and remains so until spermatogenesis. With the exception of one chromosome, the heterochromatic complement loses its differential aspect during early spermatogenesis and its members pair with their euchromatic homologues There is but one division during which the two components of each bivalent separate to opposite poles. Both division products form sperm.One pair of chromosomes, the D pair, always shows differential behavior. The D pair usually does not form a bivalent. The heterochromatic homologue, D^H, divides equationally and is eliminated by anaphase lagging or telophase ejection; its daughter halves remain as pycnotic residues during the early phases of spermiogenesis. The euchromatic homologue, D^E, also divides equationally to contribute to both of the telophase nuclei. Compensation for the division of the D^E univalent may occur during either the early or late phases of spermatogenesis.In some species the D pair is a fixed entity, analogous to the sex chromosomes in this regard. In other species, more than one pair may be elected to the D role, but only one at a time, and always the same one within each cyst.Taxonomic evidence indicates the Comstockiella system was derived from the lecanoid system, previously known from the work of the Schraders and others. In the lecanoid system, the paternally derived heterochromatic set divides equationally, along with the euchromatic set, during the first spermatogenic division. During the second spermatogenic division, the two sets are segregated from each other. The two euchromatic derivatives form sperm while the heterochromatic derivatives persist for a while as pycnotic residues. Both the lecanoid and Comstockiella systems occur in some species often in the same testis, but only one of the two systems within any one cyst.The discussion is devoted to an analysis of the mode of inheritance expected in the Comstockiella system and its evolutionary derivation. The Comstockiella system may have been derived in a step-by-step fashion from the lecanoid. The two systems differ by four processes which occur at spermatogenesis in the Comstockiella but not the lecanoid system; these are (1) deheterochromatization, (2) chromosome pairing, (3) compensation for the extra division of the D^E chromosome, and (4) lagging or ejection to eliminate the D^H chromosome.In addition, the residual genetic effects of the heterochromatic set may have undergone considerable change before the lecanoid system could evolve into a Comstockiella. Once the evolutionary step were otherwise possible, mechanistic features would aid and abet the emergence of the new system even though it lacked immediate selective advantage.The variable-D aspect of some examples of the Comstockiella system cannot be readily understood in terms of known examples of chromosome behavior; an admittedly highly speculative hypothesis is offered in an attempt to explain the situation.The diaspidid system, in which the paternal chromosomes are eliminated at late cleavage, is believed on taxonomic grounds to have stemmed from the Comstockiella, and forms the final stage of the four-step evolutionary sequence. Necessary changes for the derivation of the diaspidid system from the Comstockiella are discussed.This work was begun during the tenure of a Guggenheim Memorial Fellowship, 1956–57, and has subsequently been supported in part by grants from the National Science Foundation (G-4497 and G-9772).     

Hoechst 33258 banding of Drosophila nasutoides metaphase chromosomes

Hoechst 33258 banding of D. nasutoides metaphase chromosomes is described and compared with Q and C bands. The C band positive regions of the euchromatic autosomes, the X and the Y fluoresce brightly, as is typical of Drosophila and other species. The fluorescence pattern of the large heterochromatic chromosome is atypical, however. Contrary to the observations on other species, the C negative bands of the large heterochromatic chromosome are brightly fluorescent with both Hoechst 33258 and quinacrine. Based on differences in the various banding patterns, four classes of heterochromatin are described in the large heterochromatic chromosome and it is suggested that each class may correspond to an AT-rich DNA satellite.     

First report on C-banding,fluorochrome staining and NOR location in holocentric chromosomes of Elasmolomus (Aphanus) sordidus Fabricius, 1787 (Heteroptera,Rhyparochromidae)

In spite of various cytogenetic works on suborder Heteroptera, the chromosome organization, function and its evolution in this group is far from being fully understood. Cytologically, the family Rhyparochromidae constitutes a heterogeneous group differing in chromosome numbers. This family possesses XY sex mechanism in the majority of the species with few exceptions. In the present work, multiple banding techniques viz., C-banding, base-specific fluorochromes (DAPI/CMA₃) and silver nitrate staining have been used to cytologically characterize the chromosomes of the seed plant pest Elasmolomus (Aphanus) sordidus Fabricius, 1787 having 2n=12=8A+2m+XY. One pair of the autosomes was large while three others were of almost equal size. At diplotene, C-banding technique revealed, that three autosomal bivalents show terminal constitutive heterochromatic bands while one medium sized bivalent was euchromatic. Microchromosomes (m-chromosomes) were positively heteropycnotic. After DAPI and CMA₃ staining, all the autosomal bivalents showed equal fluorescence, except CMA₃ positive signals, observed at both telomeric heterochromatic regions of one medium sized autosomal bivalent. Silver nitrate staining further revealed that this chromosome pair carries Nucleolar Organizer Regions (NORs) at the location of CMA₃ positive signals. The X chromosome showed a thick C-band, positive to both DAPI /CMA₃ while Y, otherwise C-negative, was weakly positive to DAPI and negative to CMA₃, m-chromosomes were DAPI bright and CMA₃ dull.