Polymorphism of heterochromatic regions of flax chromosomes

The C-banding technique was used to study flax chromosomes (Linum usitatissimum L., 2n = 30). Heterochromatin was located mainly in pericentromeric regions of chromosomes. In spite of small size (1.5-3.5 microm), all 15 pairs of homologous chromosomes were identified on the basis of the C-banding pattern and morphology. An idiogram of C-banded chromosomes of L usitatissimum L. is presented. Polymorphism of chromosomal heterochromatic regions was studied in karyotypes of three flax samples: L usitatissimum L., accession K-603 (L usitatissimum var. usitatissimum), and accession K-594 (L. usitatissimum var. humile (Mill.)). A common C-banding pattern was observed in all forms studied, although there were some distinctions in the individual band size. The fibre flax (accession K-603) karyotype had the C-banding pattern similar to that of L usitatissimum L., but some intercalary and telomeric C-bands were somewhat larger, and a satellite (NOR) was observed in the short arm of chromosome I. In crown flax, (K-594) chromosomal C-banding pattern exhibited smaller pericentromeric and larger intercalary bands; telomeric bands were present on almost all chromosomes. Thus, the intraspecies polymorphism revealed in the chromosomal C-banding pattern makes possible the use of C-bands as chromosome markers in the studies of genetic and genomic polymorphism of this species.     

5-Methylcytosine in heterochromatic regions of chromosomes in Bovidae

Summary The centromeric regions of cattle, goat and sheep chromosomes bind anti-5-MeC as revealed by immunofluorescence technique, indicating concentration of 5-MeC at these heterochromatic regions. The centromere of the submetacentric X of cattle remains nearly unstained and so do the centromeres of the acrocentric X chromosomes in goat and sheep. The short arm of the cattle Y exhibits strong anti-5-MeC binding whereas the tiny Y chromosomes of goat and sheep contain no brightly fluorescent material.     

Parental origin of the extra chromosomes in polysomy X

Five polymorphic index markers were analyzed by polymerase chain reaction (PCR) to ascertain the parental origin of the extra X chromosomes in seven polysomic cases (one 49,XXXXX, three 49,XXXXY, two 48,XXXY, and one 48, XXYY). All four X chromosomes in 49, X polysomies were maternal in origin and the extra X chromosomes in 48 X polysomies were paternal. In each case the multiple X chromosomes were contributed by a single parent. Taken together with previously reported cases, these data support a single mechanism of sequential nondisjunction during either maternal or paternal gametogenesis as the cause of higher order sex chromosome polysomy.     

Translocations between Eu- and heterochromatic chromosomes,and spermatocytes lacking a heterochromatic set in male mealy bugs

In male mealy bugs the chromosomes of paternal origin become heterochromatic (H) early in embryogeny while those of maternal origin remain euchromatic (E). First instarPseudococcus obscurus Essig males which were irradiated with 3000 r of X-rays carried translocations between E and H chromosomes [T(E;H)'s] in their spermatocytes. In the T(E;H)'s the border between the E and H segments was usually quite sharp, but occasionally short E segments may have become either partially or completely heterochromatic. During the second meiotic division the E and H sets normally segregate to opposite poles. The T(E;H)'s often formed bridges in AII and TII, but in most of the cells they did succeed in reaching one of the poles. The segregation of the T(E;H)'s depended on the relative size of their E and H segments. When the E and H segments were of the same size, the T(E;H)'s segregated more often with the E chromosomes, even though the latter have been observed to be attached to their pole with fewer spindle fibers. Thirty-five of the 173 males analyzed had sectors in their testes which lacked an H set. The number of cysts per sector suggested that each sector was derived from a single irradiated cell. The karyotypes observed in some of the sectors indicated that the lack of an H set was the result of reversal of heterochromatization and not due to the loss of the H set and the endoduplication of the E set. Most of the cells lacking an H set divided normally during the first meiotic division. The second division, however, was abortive and resulted in the production of diploid sperm. Two possibilities for the origin of spermatocytes lacking an H set are considered: (i) that these spermatocytes resulted from X-ray induced reversal of heterochromatization in spermatogonia, and (ii) that these spermatocytes originated from presumptive cyst wall cells whose H set had undergone reversal prior to the irradiation.Supported by grant GB 6745 from the National Science Foundation, Washington, D. C.     

Somatic synapsis of eu- and heterochromatic regions of Drosophila melanogaster chromosomes

Quantitative cytogenetical analysis has been used to study the synapsis of D. melanogaster neuroblast mitotic chromosomes from normal females, flies with heterozygous deletions, duplications or inversions in the heterochromatic regions of chromosome 2 and in triploid females. In all these genotypes chromocentric fusion of heterochromatic regions of heterologous chromosomes is observed. Eu- and heterochromatic regions of homologous chromosomes are intimately paired at the same time during the cell cycle. The structural rearrangements lead to reduced frequencies of chromocentric association as well as of homologous synapsis compared with the frequencies in the wild-type. The results obtained are discussed with respect to the general problem of the homologous interaction of chromosomes and the significance of heterochromatin for these processes.     

Differential transmission of extra genome chromosomes in pentaploid blueberry

Summary Transmission of extra genome chromosomes by three Vaccinium ashei (2n=6x=72)/V. corymbosum (2n=4x=48) pentaploid hybrids backcrossed to the hexaploid species V. ashei was examined. Chromosome numbers were determined for 36 and 31 progeny representing 5x × 6x and 6x × 5x type crosses, respectively. Chromosome numbers ranged from hypopentaploid (2n=4x+11=59) to hexaploid with means of 2n=66.2 for 5x × 6x progeny and 2n=68.0 for 6x × 5x progeny, representing overall extra genome chromosome gains of 3.3% and 33.3%, respectively. Extra chromosome number distributions for both the 5x × 6x and x × 5x progeny deviated significantly from the theoretical distribution assuming random chromosome transmission and were also found to be heterogeneous. The 2n=5x+9=69 class predominated in 6x × 5x progeny, while a predominate class was lacking in the 5x × 6x progeny. Higher than expected frequencies of plants with chromosome numbers near the pentaploid and hexaploid levels were found in the 5x × 6x progeny, whereas the frequency was only greater at the hexaploid number in 6x × 5x progeny. Present and previous results (Vorsa et al. 1986) indicate that extra genome chromosome transmission in oddploids can be influenced by selection at both gametophytic (pollen) and post-zygotic stages. However, post-zygotic selection may involve two different mechanisms acting concurrently: 1) chromosome imbalance due to aneuploidy and/or 2) endosperm imbalance referring to maternal: paternal genome ratios deviating from 21. Such a mechanism could result in differential transmission rates of extra genome chromosomes in oddploids when crosses are made to differing ploidy levels, and to reciprocal differences as well.     

Preferential somatic pairing between homologous heterochromatic regions of human chromosomes.

The cytidine analog 5-azacytidine (5-azaC) induces an undercondensation of the heterochromatin in human chromosomes 1, 9, 15, 16, and Y when it is added in low concentrations to the late S-phase of growing lymphocyte cultures. In interphase nuclei, these heterochromatic regions are frequently somatically paired. The somatic pairing configurations are preserved up to metaphase stage in the 5-azaC-treated cultures and are thus susceptible to a direct microscopical examination. The statistical analysis of 1,000 somatic pairing configurations from 5-azaC-treated cells showed that the somatic pairing between the heterochromatic regions of homologous chromosomes is preferred over that between nonhomologous chromosomes.     

Direct FISH of 5S rDNA on tomato pachytene chromosomes places the gene at the heterochromatic knob immediately adjacent to the centromere of chromosome 1.

We describe a molecular cytogenetic procedure for high resolution physical mapping of DNA markers, an essential step toward construction of an integrated molecular-classical-cytological map. Tomato was selected as material because its pachytene chromosomes are amenable for study and because detailed molecular, classical, and cytological maps are available. Karyotyping of acetocarmine-stained pachytene chromosomes showing detailed cytogenetic landmarks was combined with direct FISH of the 5S rDNA gene. This enabled us to pinpoint the 5S rDNA gene to the first heterochromatic knob immediately adjacent to the centromere in the short arm of chromosome 1. Thus the position of the 5S rDNA gene on the molecular map was related to the position of the 5S rDNA on the cytogenetic map. The results also provide conclusive evidence of the location of a functional gene in the pericentric heterochromatic region, a rare event to date in plants. We conclude that karyotyping of pachytene chromosomes can be combined with FISH to map a DNA sequence to a cytogenetically defined region and to determine the chromatin origin of an expressed gene. Key words : direct fluorescence in situ hybridization, 5S rDNA, pachytene chromosomes, heterochromatic gene.     

An extra band in human 9qh+ chromosomes

Summary Chromosome analysis of G-banded cells from nine individuals showed that 9qh+ chromosomes have an extra band in the h region in approx. 3 to 50% of the cells.     

5-Methylcytosine in heterochromatic regions of chromosomes: chimpanzee and gorilla compared to the human

Fixed metaphase chromosomes of gorilla and chimpanzee were UV-irradiated to produce regions of single-stranded DNA and then treated with antibodies specific for the minor DNA base 5-methylcytosine (5 MeC). An indirect immunofluorescence technique was used to visualize sites of antibody binding. In the gorilla six pairs of autosomes contained major fluorescent regions, indicating localized regions of highly methylated DNA. These corresponded, with the exception of chromosome 19, to the major regions of constitutive heterochromatin as seen by C-banding. The Y chromosome also contained a highly fluorescent region which was located just proximal to the intense Q-band region. In the chimpanzee no comparable concentrations of highly methylated DNA were seen. Smaller regions of intense 5 MeC binding were present on perhaps six chimpanzee chromosomes, including the Y. Five of these corresponded to chromosomes which were highly methylated in the gorilla.--There is diversity among the human, gorilla and chimpanzee in both the size and location of concentrations of 5 MeC, supporting the idea that satellite DNA evolves more rapidly than DNA in the remainder of the chromosome.     

Kinetochore microtubule numbers of different sized chromosomes

For three species of grasshoppers the volumes of the largest and the smallest metaphase chromosome differ by a factor of 10, but the microtubules (MTs) attached to the individual kinetochores show no corresponding range in numbers. Locusta mitotic metaphase chromosomes range from 2 to 21 μm, and the average number of MTs per kinetochore is 21 with an SD of 4.6. Locusta meiotic bivalents at late metaphase I range from 4 to 40 μm(3), and the kinetochore regions (= two sister kinetochores facing the same spindle pole) have an average of 25 kinetochore microtubules (kMTs) with an SD of 4.9. Anaphase velocities are the same at mitosis and meiosis I. The smaller mitotic metaphase chromosomes of neopodismopsis are similar in size, 6 to 45 μm(3), to Locusta, but they have an average more kMTs, 33, SD = 9.2. The four large Robertsonian fusion chromosomes of neopodismopsis have an average of 67 MTs per kinetochore, the large number possibly the result of a permanent dicentric condition. Chloealtis has three pairs of Robertsonian fusion chromosomes which, at late meiotic metaphase I, form bivalents of 116, 134, and 152 μm (3) with an average of 67 MTs per kinetochore similar to Locusta bivalents, but with a much higher average of 42 MTs per kinetochore region. It is speculated that, in addition to mechanical demands of force, load, and viscosity, the kMT numbers are governed by cell type and evolutionary history of the karyotype in these grasshoppers.     

Two repeated DNA sequences from the heterochromatic regions of rye (Secale cereale) chromosomes

Rye DNA sequences renaturing with a C₀t <0.02 mol·sec/l, are largely undigested by the restriction enzyme HindIII. These HindIII-spared sequences are mostly located in telomeric heterochromatin. When digested with EcoRI* and cloned into the EcoRI site of pBR 325, these sequences yielded clones of two classes when hybridized to a probe of rapidly renaturing DNA. One class contains a DNA sequence which is a major constituent of the telomeric heterochromatic blocks, while the other is a minor component of the highly repeated DNA of the genome. The major component was sequenced, its chromosomal distribution mapped using wheat-rye addition lines and its distribution in meiotic prophase nuclei determined. The minor component is present in significant amounts in wheat as well as in rye and is localized at the terminal heterochromatic regions of three rye chromosomes but not in the major blocks of heterochromatin.     

Analysis of the inheritance of heterochromatic regions in human chromosomes 1, 9, 16 and Y

The inheritance of heterochromatic regions of chromosomes 1, 9, 16 and Y was studied in twelve families by means of measuring their C-segments. Maternal and paternal origin of chromosomes 1, 9 and 16 in the child was determined by two methods. The advantages and disadvantages of these methods and possibilities of their application are under discussion.     

Eu-heterochromatic rearrangements induce replication of heterochromatic sequences normally underreplicated in polytene chromosomes of Drosophila melanogaster

In polytene chromosomes of D. melanogaster the heterochromatic pericentric regions are underreplicated (underrepresented). In this report, we analyze the effects of eu-heterochromatic rearrangements involving a cluster of the X-linked heterochromatic (Xh) Stellate repeats on the representation of these sequences in salivary gland polytene chromosomes. The discontinuous heterochromatic Stellate cluster contains specific restriction fragments that were mapped along the distal region of Xh. We found that transposition of a fragment of the Stellate cluster into euchromatin resulted in its replication in polytene chromosomes. Interestingly, only the Stellate repeats that remain within the pericentric Xh and are close to a new eu-heterochromatic boundary were replicated, strongly suggesting the existence of a spreading effect exerted by the adjacent euchromatin. Internal rearrangements of the distal Xh did not affect Stellate polytenization. We also demonstrated trans effects exerted by heterochromatic blocks on the replication of the rearranged heterochromatin; replication of transposed Stellate sequences was suppressed by a deletion of Xh and restored by addition of Y heterochromatin. This phenomenon is discussed in light of a possible role of heterochromatic proteins in the process of heterochromatin underrepresentation in polytene chromosomes.     

Nucleotide sequence of an heterochromatic segment recognized by the antibodies to Z-DNA in fixed metaphase chromosomes.

The purpose of this work was to analyse at the molecular level the DNA recognized by the antibodies to Z-DNA in in situ experiments. Antibodies to Z-DNA interact strongly with R-band positive heterochromatic segments of fixed metaphase chromosomes of Cebus (Viegas-Pequignot et al., 1983). These segments are constituted of a satellite DNA the repeat unit of which is about 1520 base pairs long. The base sequence of the repeat unit has been determined. It contains a (AC)n rich region which, in vitro, adopts the Z conformation under topological constraints. Experiments with nuclei suggest that this sequence is not predominantly in the Z conformation in vivo. The polymorphic structure of the (AC)n rich region argues for an active recombination sequence.     

Asynchrony of DNA replication and mitotic spiralization along heterochromatic portions of Chinese hamster chromosomes

Sequence of DNA synthesis and mitotic chromosome spiralization along heterochromatic portions of the sex (X₁X₂) and of some marker chromosomes in cultured Chinese hamster cells were studied, employing two methods: study of segmentation pattern caused in chromosomes with colcemid, and autoradiography with tritiated thymidine. The heterochromatic portions of all chromosomes studied were characterized by striking internal asynchrony of DNA replication. In particular, they had segments that replicated relatively early. The short arm of the X₂ chromosome, heterochromatic in female somatic cells, had at least three such segments. Replication patterns of the long arms of the X₁ and X₂ chromosomes were different. In X₁ this arm contains several segments showing relatively early replication. The long arm of X₂ had no similar segments. The possible significance of the data obtained is discussed with regard to the problem of genetic inertness of heterochromatin. At the terminal stage of the S period, H³-thymidine seems to be incorporated into condensed chromatin of interphase nuclei. On the basis of the data obtained, it is proposed that during replication of heterochromatin consecutive despiralization of parts of it takes place.     

Destruction of specific heterochromatic chromosomes during spermatogenesis in the Comstockiella chromosome system (Coccoidea: Homoptera)

In male coccids with the Comstockiella chromosome system, the set of chromosomes of paternal orgin becomes heterochromatic (H) during early cleavage. Just prior to prophase I of spermatogenesis, some of the H chromosomes are destroyed; the rest are eliminated following meiosis. In this report a Comstockiella sequence is described from Dactylopius opuntiae (2n=10) in which one chromosome pair is about three times longer than the others. During prophase I the number of small H chromosomes present varied from cyst to cyst, but the long H chromosome was present in every cyst. These observations provide the best evidence to date that in the Comstockiella system a particular chromosome may always escape destruction. An analysis of Kitchin's (1975) data about the frequency of prophase I cysts with 1–4 H chromosomes in three species of Parlatoria with 2n = 8 suggested that in these species chromosomes of similar size may have very different probabilities of being destroyed. Evidence that in other species with the Comstockiella system a particular H chromosome is always retained is reviewed, and the possibility that in Ancepaspis tridentata the variation in the length of the H chromosome retained is due to the partial destruction of the longest chromosome is discussed.