Polytene chromosome maps in four species of tsetse flies Glossina austeni, G. pallidipes, G. morsitans morsitans and G. m. submorsitans (Diptera: Glossinidae): a comparative analysis

Photographic polytene chromosome maps from pupal trichogen cells of four tsetse species, Glossina austeni, G. pallidipes, G. morsitans morsitans and G. m. submorsitans were constructed and compared. The homology of chromosomal elements between the species was achieved by comparing banding patterns. The telomeric and subtelomeric chromosome regions were found to be identical in all species. The pericentromeric regions were found to be similar in the X chromosome and the left arm of L1 chromosome (L1L) but different in L2 chromosome and the right arm of L1 chromosome (L1R). The L2 chromosome differs by a pericentric inversion that is fixed in the three species, G. pallidipes, G. morsitans morsitans and G. m. submorsitans. Moreover, the two morsitans subspecies appeared to be homosequential and differ only by two paracentric inversions on XL and L2L arm. Although a degree of similarity was observed across the homologous chromosomes in the four species, the relative position of specific chromosome regions was different due to chromosome inversions established during their phylogeny. However, there are regions that show no apparent homology between the species, an observation that may be attributed to the considerable intra—chromosomal rearrangements that have occurred following the species divergence. The results of this comparative analysis support the current phylogenetic relationships of the genus Glossina.     

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.     

A cytogenetical study of Glossina fuscipes fuscipes including a comparison of the polytene chromosome maps with those of Glossina austeni (Diptera,Glossinidae)

The male meiotic sequence is described for the tsetse fly Glossina fuscipes fuscipes together with the polytene chromosome maps and all principal cytological markers. The diploid chromosome number is 2n=6 and includes a pair of large submetacentric autosomes (L₁), a shorter pair of metacentric autosomes (L₂), and an X and Y which constitute a heteromorphic pair. Male meiosis is normally achiasmate although evidence is presented which suggests that chiasmata do form in about 1% of males. A detailed comparison between the polytene chromosomes of this species and Glossina austeni indicates that although they must have had a common ancestor, G. austeni is genetically more closely related to morsitans group tsetses.     

Differential sex chromosome replication and dosage compensation in polytene trichogen cells of Lucilia cuprina (Diptera: Calliphoridae)

Non banded sex chromosome elements have been identified in polytene trichogen cells of Lucilia cuprina using Y-autosome translocations, C-banding and Quinacrine fluorescence. The X chromosome is an irregular granular structure while the much smaller Y chromosome has both a dense darkly stained and a loosely organised segment. The X and Y chromosomes are underreplicated in polytene cells but comparison of C- and Q-banding characteristics of sex chromosomes in diploid and polytene tissues indicates that selective replication of non C-banding material occurs in both the sex chromosomes. Brightly fluorescing material in the Y chromosome is replicated to such an extent that it consists of half the polytene element, while the C-banding material, which makes up most of the diploid X chromosome, is virtually unreplicated. Differential replication also occurs in autosomes. In XXY males, and in males carrying a duplication of the X euchromatic region, a short uniquely banded polytene chromosome is formed. It is suggested that in males carrying two doses of X euchromatin a dosage compensation mechanism operates in which genes in one copy are silenced by forming a banded polytene chromosome.     

Initial phases of DNA synthesis in Drosophila melanogaster

Replication patterns of the X chromosomes and autosomes in D. melanogaster male and female larvae during the discontinuously labeled initial and end phases of DNA synthesis were compared. In female larvae X and autosomes behaved correspondingly during all the replication stages. In males, however, the X chromosome shows a differential replication behavior from that of the autosomes already during the discontinuously labeled initial stage.—In those nuclei of both sexes, in which the autosomes correspond in their initial replication patterns, significantly more labeled regions are to be found over the male X than over the female X. The complementary behavior during the end phases (Berendes, 1966), i.e. the reverse of that above, leads to an earlier completion of the replication cycle in most of the labeled regions of the male X chromosome. The differential replication revealed in the autoradiograms is interpreted as a consequence of the polytene structure in giant chromosomes.     

The JIL-1 kinase regulates the structure of Drosophila polytene chromosomes

The JIL-1 kinase localizes to interband regions of Drosophila polytene chromosomes and phosphorylates histone H3 Ser10. Analysis of JIL-1 hypomorphic alleles demonstrated that reduced levels of JIL-1 protein lead to global changes in polytene chromatin structure. Here we have performed a detailed ultrastructural and cytological analysis of the defects in JIL-1 mutant chromosomes. We show that all autosomes and the female X chromosome are similarly affected, whereas the defects in the male X chromosome are qualitatively different. In polytene autosomes, loss of JIL-1 leads to misalignment of interband chromatin fibrils and to increased ectopic contacts between nonhomologous regions. Furthermore, there is an abnormal coiling of the chromosomes with an intermixing of euchromatic regions and the compacted chromatin characteristic of banded regions. In contrast, coiling of the male X polytene chromosome was not observed. Instead, the shortening of the male X chromosome appeared to be caused by increased dispersal of the chromatin into a diffuse network without any discernable banded regions. To account for the observed phenotypes we propose a model in which JIL-1 functions to establish or maintain the parallel alignment of interband chromosome fibrils as well as to repress the formation of contacts and intermingling of nonhomologous chromatid regions. Electronic Supplementary Material Supplementary material is available for this article at and accessible for authorised users     

The interrelation of the polytene chromosomes of generative tissues in Drosophila melanogaster]

The principles of three dimensional organization of primary and secondary orders polytene chromosomes in ovarian nurse cells of Drosophila melanogaster were elucidated. Contrary to somatic tissues, no joining of chromosome arms into local chromocentre was discovered. The chromosomes are separated in the nuclear space and are attached to the nuclear envelope by the centromeric (and the XL arm--by the telomeric) sites, the arms of autosomes (especially primary polytene chromosomes) being separated in the area of attachment. Polytenized XR arm of the X chromosomes were discovered. The architecture of chromosomes discovered in ovarian nurse cells is tissue-specific and differs considerably from the organization of polytene chromosomes of somatic tissues.     

Microsatellite diversities and gene flow in the tsetse fly,Glossina morsitans s.l

Tsetse flies occupy discontinuous habitats and gene flow among them needs to be investigated in anticipation of area-wide control programs. Genetic diversities were estimated at six microsatellite loci in seven Glossina morsitans submorsitans Newstead (Diptera: Glossinidae) populations and five microsatellite loci in six G. m. morsitans Westwood populations. Nei's unbiased diversities were 0.808 and 76 alleles in G. m. submorsitans and 0.727 and 55 alleles in G. m. morsitans. Diversities were less in three laboratory cultures. Matings were random within populations. Populations were highly differentiated genetically. Populations were strongly subdivided, as indicated by fixation indices (F(ST)) of 0.18 in G. m. morsitans and 0.17 in G. m. submorsitans. 35% of the genetic variance in G. m. submorsitans was attributed to differences between populations from The Gambia and Ethiopia. All available genetic evidence suggests that genetic drift is much greater than gene flow among G. morsitans s.l. populations.     

Characterization and chromosomal distribution of a tandemly repeated DNA sequence from the Australian sheep blowfly,Lucilia cuprina

In the course of making a Lucilia cuprina genomic DNA library, a ladder of bands was seen in partial Sau3A digests. Complete digestion reduced this ladder to predominantly monomer units of approximately 190 bp. Nine independently isolated copies of this repeat were cloned and sequenced. Only two of these isolates are identical in sequence, the most divergent being 71% homologous. This satellite DNA occurs in all three wildtype strains tested, and, for the single case examined, in the embryonic, larval, pupal, and adult DNA. It represents approximately 3%–4% of the genome. Data obtained from in situ chromosome hybridizations indicate that this sequence is concentrated around the centromeric regions of the autosomes and over most of the sex chromosomes. Labelling is much stronger in mitotic compared with polytene chromosomes showing directly that this centromeric satellite DNA is grossly under-replicated during polytenization. This under-replication is even more pronounced on the sex chromosomes compared with the autosomes.by A. Bird The EMBL accession numbers are: X57584 L.C.SAT TRS 188-1; X57585 L.C.SAT TRS 188-13; X57586 L.C.SAT TRS 188-14; X57587 L.C.SAT TRS 188-15; X57588 L.C.SAT TRS 188-19; X57589 L.C.SAT TRS 188-16; X57590 L.C.SAT TRS 188-21; X57591 L.C.SAT TRS 188-7     

The COOH-terminal domain of the JIL-1 histone H3S10 kinase interacts with histone H3 and is required for correct targeting to chromatin

The JIL-1 histone H3S10 kinase in Drosophila localizes specifically to euchromatic interband regions of polytene chromosomes and is enriched 2-fold on the male X chromosome. JIL-1 can be divided into four main domains including an NH(2)-terminal domain, two separate kinase domains, and a COOH-terminal domain. Our results demonstrate that the COOH-terminal domain of JIL-1 is necessary and sufficient for correct chromosome targeting to autosomes but that both COOH- and NH(2)-terminal sequences are necessary for enrichment on the male X chromosome. We furthermore show that a small 53-amino acid region within the COOH-terminal domain can interact with the tail region of histone H3, suggesting that this interaction is necessary for the correct chromatin targeting of the JIL-1 kinase. Interestingly, our data indicate that the COOH-terminal domain alone is sufficient to rescue JIL-1 null mutant polytene chromosome defects including those of the male X chromosome. Nonetheless, we also found that a truncated JIL-1 protein which was without the COOH-terminal domain but retained histone H3S10 kinase activity was able to rescue autosome as well as partially rescue male X polytene chromosome morphology. Taken together these findings indicate that JIL-1 may participate in regulating chromatin structure by multiple and partially redundant mechanisms.     

A photographic map of Drosophila madeirensis polytene chromosomes.

A photographic map of salivary gland polytene chromosomes of Drosophila madeirensis has been constructed showing homologies and differences with respect to the standard gene arrangement of D. subobscura. Only two paracentric inversions in the X chromosome and some slight minor dissimilarities of one or two bands in the autosomes differentiate the chromosomes of these species.     

The evolution of unusual chromosomal systems in sciarid flies: intragenomic conflict and the sex ratio

A model is presented for the evolution of the sciarid chromosomal system. In this model, a driving X chromosome caused female-biased sex ratios. The drive was exploited by maternal autosomes that segregated with the X at spermatogenesis. Genes in mothers converted some of their XX daughters into sons by eliminating a paternal X from the embryonic soma. L chromosomes were derived from X chromosomes and favored male-biased sex ratios. An X' chromosome arose that suppressed the effects of L chromosomes. The 1:1 sex ratio is a stalemate between the X' and X'X mothers causing all-female broods and the L chromosomes in XX mothers causing all-male broods. Any element (such as an L chromosome) that is preferentially transmitted through one sex will be selected to bias the sex ratio towards this sex.     

A Yeast Artificial Chromosome Clone Map of the Drosophila Genome

We describe the mapping of 979 randomly selected large yeast artificial chromosome (YAC) clones of Drosophila DNA by in situ hybridization to polytene chromosomes. Eight hundred and fifty-five of the clones are euchromatic and have primary hybridization sites in the banded portions of the polytene chromosomes, whereas 124 are heterochromatic and label the chromocenter. The average euchromatic clone contains about 211 kb and, at its primary site, labels eight or nine contiguous polytene bands. Thus, the extent as well as chromosomal position of each clone has been determined. By direct band counts, we estimate our clones provide about 76% coverage of the euchromatin of the major autosomes, and 63% coverage of the X. When previously reported YAC mapping data are combined with ours, euchromatic coverage is extended to about 90% for the autosomes and 82% for the X. The distribution of gap sizes in our map and the coverage achieved are in good agreement with expectations based on the assumption of random coverage, indicating that euchromatic clones are essentially randomly distributed. However, certain gaps in coverage, including the entire fourth chromosome euchromatin, may be significant. Heterochromatic sequences are underrepresented among the YAC clones by two to three fold. This may result, at least in part, from underrepresentation of heterochromatic sequences in adult DNA (the source of most of the clones analyzed), or from clone instability.     

Polytene chromosome maps of the melon fly Bactrocera cucurbitae (Diptera: Tephritidae).

Standard photographic maps of the polytene chromosomes are presented for the melon fly Bactrocera cucurbitae, a serious pest of fleshy fruits and vegetables. Five larval salivary gland polytene chromosomes (10 polytene arms) were isolated, and their characteristic features and landmarks have been recognized. Banding patterns of each of the polytene arms are presented, where variation in band intensity and puffs appear to reflect fundamental differences in chromosomes. The whole polytene genome has been typically mapped by dividing it into 100 sections and the subsections were lettered. The mitotic chromosomes of larval brain ganglia are also examined, five pairs of autosomes and an XX/XY sex chromosome pair. In addition, a heterochromatic mass corresponding to the sex chromosomes are observed in the polytene nuclei of salivary gland tissue. This investigation showed that B. cucurbitae has excellent cytological material for polytene chromosome analysis and proved to be very useful for obtaining more detailed genetic information on the pest's natural populations.     

Cattle-tsetse contact in relation to the daily activity patterns of Glossina morsitans submorsitans in The Gambia

Abstract. The daily flight activity patterns of one of the main vectors of animal trypanosomiasis in West Africa, Glossina morsitans submorsitans , were assessed using four different methods. Results from all the methods showed that there was some flight activity nearly every hour in all seasons but they differed in the level of contact between grazing cattle herds and G.m.submorsitans. In the late dry season, trap data indicated that there was negligible activity from midday to late afternoon, whereas observations of tsetse contact with cattle herds or hand-net collections on herd followings showed no fall in attack rates on the cattle by G.m.submorsitans.
Differences between trap and animal-baited collection data may be attributable to the type of G.m.submorsitans sampled by each method. Male G.m.submorsitans captured by traps were more fat depleted than those caught on ox-baited flyrounds or by hand-net collections on herd followings. All methods showed that male G.m.submorsitans were most fat depleted in the late dry season and least in the early dry season. It was concluded that the traps were mainly sampling the spontaneous flights of G.m.submorsitans. Hunger and endogenous rhythms increase the likelihood of spontaneous flights towards dusk, particularly in conditions such as those at midday in the very hot, late dry season. However, the presence of cattle herds in infested habitats probably activated nearby G.m.submorsitans and the continual movement through the grazing areas ensured contact with tsetse throughout grazing.
The data indicated that strategic management of herd grazing times cannot eliminate the risk of trypanosomiasis transmission occurring, irrespective of the harshness of the dry season climate. An assessment of the level of this risk could only be measured suitably by collecting tsetse using animal-baited methods, not from trap data.     

Radiation-induced and transposon-induced chromosome damage in Drosophila: translocations and transmission distortion

The possible interaction between X-ray- and transposon-induced chromosome damage was monitored in the P-M system of hybrid dysgenesis in Drosophila melanogaster. One- to two-day-old F1 dysgenic males originating from a cross between M strain females and P strain males were irradiated with 5.5 Gy (550 rad) or used as controls to monitor X-Y translocations and transmission ratio distortion. Two 3-day sperm broods were sampled for the former and two 4-day broods for the latter to detect damage induced in the most radiosensitive cells. F1 nondysgenic males derived from M female to M male crosses (controls) were treated identically. X-Y chromosome translocations induced by P element mobility alone declined sharply with a decrease in temperature (18 versus 21 degrees C) and they were significantly reduced with aging of hybrid males from brood 2, 4-8 days of age, to brood 3, 7-11 days of age. No significant increase in translocations was observed when X irradiation and P-M dysgenesis were combined, showing no interaction between damages induced by the two mutator systems. In contrast, interaction was observed in transmission ratio distortion which was significantly increased by X irradiation of hybrid males derived from both reciprocal M X P and P x M crosses. The preferential elimination of P element-bearing autosomes occurred when either spermatocytes or spermatids were irradiated. An aging effect was also observed, resulting in less distortion in 9- to 14-day-old dysgenic males compared to 5- to 10-day-old hybrids.     

The distribution of transposable elements within and between chromosomes in a population of Drosophila melanogaster. II. Inferences on the nature of selection against elements.

Data were collected on the distribution of nine families of transposable elements among a sample of autosomes isolated from a natural population of Drosophila melanogaster, by means of in situ hybridization of biotinylated probes to polytene chromosomes. There is no general tendency for elements to accumulate at the tips of chromosomes. Elements tend to be present in excess of random expectation in the euchromatin proximal to the centromeres of the major autosomes, and on chromosome four. There is considerable heterogeneity between different families in the extent of this excess. The overall abundance of element families is inversely related to the extent to which they accumulate proximally. The level of proximal accumulation for the major autosomes is similar to that on the fourth chromosome, but less than that for the X chromosome. There is an overall deficiency of elements in the mid-section of the X compared with the mid-sections of the major autosomes, with considerable heterogeneity between families. The magnitude of this deficiency is positively related to the extent to which elements accumulate proximally. No such deficiency is seen if the proximal regions of the X and autosomes are compared. There is a small and non-significant excess of elements in third chromosomes carrying inversions. There is some between-year heterogeneity in element abundance. The implications of these findings are discussed, and it is concluded that they generally support the hypothesis that transposable element abundance is regulated primarily by the deleterious fitness consequences of meiotic ectopic exchange between elements. If this is the case, such exchange must be very infrequent in the proximal euchromatin, and the elements detected in population surveys of this kind must be inserted into sites where they have negligible mutational effects on fitness.     

Cytogenetic analysis of Malpighian tubule and salivary gland polytene chromosomes of Bactrocera oleae (Dacus oleae) (Diptera: Tephritidae).

Photomaps of the Malpighian tubule and the salivary gland polytene chromosomes of Bactrocera oleae (Dacus oleae) are presented and compared with those of the fat body. Five polytene chromosomes (10 polytene arms) corresponding to the five autosomes of the mitotic nuclei, as well as a heterochromatic mass corresponding to the sex chromosomes, are observed in the nuclei of the three somatic tissues. The most prominent features of each polytene chromosome, the reverse tandem duplications, as well as the rather unusual ectopic pairing of the telomeric regions of different chromosome arms, are described. The constancy of the banding pattern based on the analysis of the three larval tissues is discussed.