Relation between the size of C-segments and corresponding euchromatic regions of human chromosomes 1, 9 and 16 during their mitotic condensation

The nature of associations between the length of C-segments and the corresponding euchromatic regions of chromosomes 1, 9, and 16 in the process of their mitotic condensation has been studied. Their statistically significant linear nature in the range of chromosome 2 condensation from 11 to 4 micron has been established. Within the interval of 6.5-8.5 micron the above association is less significant, at the same time minimal variability of C-segment length is observed as compared to other stages of mitotic condensation. It is recommended to define the absolute size of C-segments in chromosomes 1, 9 and 16 by measuring their dimensions in metaphase plates with chromosome 2 length from 6.5 to 8.5 micron. The regressional correction of the results of C-segment measurements or approximation of values depending on the statistical significance of linear regression equation coefficient has been demonstrated.     

Three euchromatic DNA sequences under-replicated in polytene chromosomes of Drosophila are localized in constrictions and ectopic fibers

We examined three regions of under-represented euchromatic DNA sequences (histone, Ubx, and 11 A), for their possible correlation with euchromatic constrictions in polytene chromosomes of Drosophila melanogaster. Cloned sequences were hybridized to filters and to chromosomes prepared for light microscopy. Under-represented sequences hybridized to DNA within constrictions and in ectopic fibers. In contrast, adjacent sequences that were fully endoreplicated in the Ubx and 11A regions in polytene cells hybridized to sites just adjacent to their respective constrictions. For one region (Ubx), sequences under-represented in salivary gland cells were fully endoreplicated in fat body cells. For this particular region, the morphology of the polytene chromosomes differs between these two cell types in that the specific constriction is absent at this region in fat body polytene chromosomes, thus strengthening the correlation between under-representation and chromosome constrictions. Although all three sequences are in regions that have been classified by others as intercalary heterochromatin, we detect no common functional or sequence organizational feature for these examples of under-represented DNA. We suggest that the lower efficiencies of the replication origins, or special regions of termination at these sites, are the primary cause of the under-replication, and that this under-replication is sufficient to confer the properties of intercalary heterochromatin.     

Differential chromosome contraction at the pachytene stage of meiosis in alfalfa (Medicago sativa L.)

Using the length of the total chromosome complement as a measure of the pachytene stage of a cell, most of the variation from cell to cell in chromosome lengths can be accounted for. Significant regression equations were obtained for chromosome and arm lengths upon the cell stage and these provide estimates of the relative contraction rates of the chromosomes. The regression lines for chromosomes 1 and 2 were quadratic whereas they were linear for the remaining six chromosomes. The contraction rates were different for each chromosome as well as for the short and long arms of chromosomes 1 to 5. The relative contraction rates for the heterochromatic short arms of chromosomes 3, 4 and 5 were very low and therefore arm ratios as well as relative lengths of chromosomes could vary with the cell stage examined. The differences in chromosome numbering systems reported by alfalfa researchers are mainly attributed to the pachytene stages observed and the small numbers of observations per study.     

Classification of qh regions in human chromosomes 1, 9, and 16 by C-banding

Summary We present a classification for secondary constriction (qh) regions with C-banding technique in chromosomes 1,9, and 16 by means of comparing them to the short arm of chromosome 16. It is simple and convenient and can be used routinely. It can be incorporated into the modified Paris nomenclature system.     

Karyotypes and C-banding patterns in species of Cyphomandra Mart, ex Sendtner (Solanaceae)

The orcein and C-banded karyotypes of 11 species of Cyphomandra (Solanaceae) were described. All species were diploid with 2n = 2x = 24. The chromosomes were large, ranging from 4 to 10 u.m iⁿ length, and in each complement were largely metacentric or submetacentric with few subtelocentrics. There was a significant negative correlation between chromosome length and arm ratio within a complement as well as between taxa. In general, chromosomes of the larger complements were more symmetrical in terms of both relative chromosome length and arm ratio, implying that similar amounts of DNA had been added to or taken away from every chromosome are of each complement during evolutionary divergence. Two pairs of non-homologous chromosomes were seen to contain subterminal secondary constrictions in most species. The two Brazilian species studied differed from those of Andean origin in the location of one of these secondary constrictions, suggesting a major evolutionary divergence between these two groups of specieS. Non-homologous chromosomes were difficult to distinguish from one another without the aid of C-banding, due to a continuum in the distribution of chromosome lengths and arm ratios. Telomeric and interstitial bands were shown in all species but not all chromosomes in each complement were banded. There were no centromeric bandS. Nuclear DNA amount and the length, but not proportional length, of C-bands were correlated in each specieS. One species ( C. Luteoalba (Pers.) Child, section Cyphomandropsis ) was unique in its banding pattern, providing further evidence for the delimitatation of this species and perhaps section from other Cyphomandra taxa.     

Karyotype Analysis of Speirantha gardenii (Hook.) Baill.

This paper reports the chromosome numbers and karyotype analysis of Speirantha gardenii, which is endemic to China. The material was collected from Huang Shan, Anhui. It is a diploid species. Its somatic chromosome is 2n=38=22m+6sm+10st. The 9th pair is submedian centromere chromosomes, but it has two constrictions. The secondary constriction is on the short arm near centromere. Of the 19 chromosome pairs,secondary constriction is present only in this pair.     

Chromosome analysis and DNA homology in threePicea species,P. mariana,P. rubens,andP. glauca (Pinaceae)

A detailed karyotype analysis was made on the somatic complement ofPicea rubens andP. glauca. B-chromosomes were observed in someP. glauca populations. The karyotypes are generally asymmetrical with most of the chromosomes having median to median-submedian centromeres.Picea glauca chromosomes 2, 3, 7, and 8 have secondary constriction on their short arm and chromosome 10 has a secondary constriction on the long arm. Chromosome 3 was the most easily identifiable, as it has two secondary constrictions located on the short arm. InP. rubens, all the chromosomes but chromosomes 8 and 9 have one to four distinctive secondary constrictions. In general, the diagrammatic comparisons show a high degree of similarity amongP. mariana, P. rubens, andP. glauca. GenomicP. mariana probe strongly hybridized to dots of genomic DNA fromP. rubens andP. glauca indicating that there is a high sequence homology among these three species. The synchronizing agent, hydroxyurea was used at different concentrations to enhance the mitotic index of cell suspensions derived from embryogenic cultures. Hydroxyurea at 1.25 mM increased significantly the mitotic index. An increase of hydroxyurea from 1.25 mM to 5 mM and 10 mM resulted in a steady decrease of mitotic index.     

Comparison of areas of quinacrine mustard fluorescence and modified Giemsa staining in human metaphase chromosomes

The methods of quinacrine mustard fluorescence and modified Giemsa staining were compared in view of the structural details revealed in human mitotic chromosomes derived from the peripheral blood of normal healthy humans. Over the chromatids both techniques produced a crossbanding pattern where larger segments of heavy staining in the latter technique and the fluorescing bands in the former occurred at similar locations. The centromeric heterochromatin, intensely stained with Giemsa was, however, negative in fluorescence, except for chromosome no. 3 and less often no. 6. The regularly occurring secondary constrictions in chromosomes 1, 9, and 16 behaved generally like areas of centromeric heterochromatin. The area of secondary constriction in the Y chromosome as also that of chromosome 9 in the ASG modification of the Giemsa technique was both non-fluorescent and non-staining.     

Detection of nucleolus organizer regions in chromosomes of human,chimpanzee, gorilla,orangutan and gibbon

Nucleolus organizer regions were detected by the Ag-AS silver method in fixed metaphase chromosomes from human and primates. In the human, silver was deposited in the secondary constriction of a maximum of five pairs of acrocentric chromosomes: 13, 14, 15, 21 and 22. The chimpanzee also had five pairs of acrocentric chromosomes stained, corresponding to human numbers 13, 14, 18, 21 and 22. A gibbon had a single pair of chromosomes with a secondary constriction, which corresponded to the nucleolus organizer region. In each case the Ag-AS method detected the sites which have been shown by in situ hybridization to contain the ribosomal RNA genes. An orangutan had eight pairs of acrocentric chromosomes stained with Ag-AS, probably corresponding to human numbers 13, 14, 15, 18, 21 and 22, plus two others. Two gorillas had silver stain over two pairs of small acrocentric chromosomes and at the telomere of one chromosome 1. The larger gorilla acrocentric chromosomes had no silver stain although they all had secondary constrictions and entered into satellite associations.     

Molecular topography of the secondary constriction region (qh) of human chromosome 9 with an unusual euchromatic band.

Heterochromatin confined to pericentromeric (c) and secondary constriction (qh) regions plays a major role in morphological variation of chromosome 9, because of its size and affinity for pericentric inversion. Consequently, pairing at pachytene may lead to some disturbances between homologous chromosomes having such extreme variations and may result in abnormalities involving bands adjacent to the qh region. We encountered such a case, where a G-positive band has originated de novo, suggesting a maternal origin from the chromosome 9 that has had a complete pericentric inversion. In previously reported cases, the presence of an extra G-positive band within the 9qh region has been familial, and in the majority of those cases it was not associated with any clinical consequences. Therefore, this anomaly has been referred to as a "rare" variant. The qh region consists of a mixture of various tandemly repeated DNA sequences, and routine banding techniques have failed to characterize the origin of this extra genetic material. By the chromosome in situ suppression hybridization technique using whole chromosome paint, the probe annealed with the extra G-band, suggesting a euchromatic origin from chromosome 9, presumably band p12. By the fluorescence in situ hybridization technique using alpha- and beta-satellite probes, the dicentric nature was further revealed, supporting the concept of unequal crossing-over during maternal meiosis I, which could account for a duplication of the h region. The G-positive band most likely became genetically inert when it was sandwiched between two blocks of heterochromatin, resulting in a phenotypically normal child. Therefore, an earlier hypothesis, suggesting its origin from heterochromatin through so-called euchromatinization, is refuted here.(ABSTRACT TRUNCATED AT 250 WORDS)     

A Technique for C-Banding Plant Chromosomes with Wright Stain

A technique is presented for C-banding plant chromosomes with a modified Wright stain. This procedure consistently produces brightly stained, well defined telomeric and interstitial heterochromatic bands, identifiable centromeric constrictions, and lightly stained euchromatic areas on chromosomes of rye.     

Chromosome nucleolus organizer regions of the African green monkey

The silver staining Ag-I for detection of nucleolar organizer regions (NOR) in the African green monkey chromosomes was used. NOR were situated on the secondary constrictions of one pair of homologous marker chromosomes and consisted of two parts. A block of condensed chromatin was seen in the middle of the despiralized portion corresponding to the secondary constriction. The data suggest that the stalks of satellites of chromosomes, rather than satellites themselves, are regions of nucleolar organizer.     

Variability and familial transmission of constitutive heterochromatin of human chromosomes evaluated by the method of linear measurement

Summary Using the method of linear measurement, the lengths of constitutive heterochromatin of chromosomes 1, 9, 16, and Y were determined in 125 unrelated individuals, and in 30 members of ten families. The method used eliminates the variations in the C-band length due to different degrees of contraction of chromosomes in different mitoses, and enables the size of heterochromatin blocks to be expressed. It was found that the distribution of C-band lengths in the group of 125 individuals was normal, i.e., Gaussian, for all four classes of chromosomes measured. On the basis of length distribution and by computing the P₁, P₁₀, P₉₀ and P₉₉ percentiles, the actual numerical limits could be proposed for the five-step evaluation of heterochromatin length according to the Paris Conference (1971), Supplement (1975), for chromosomes 1, 9, 16, and in a preliminary way also for Y. When applying the proposed limits to data obtained in the present study, 165 C-band variants could be identified among the 125 individuals.In ten families, C-block lengths of the chromosomes transmitted from parents to progeny could be determined in 63 cases. The mean difference in C-band length of transmitted chromosomes, as measured in parents and in children, was 0.46×10^-7 m. An analysis was carried out to detect the factors upon which the magnitude of this difference depends, and to define what differences are attributable to methodological errors. The results revealed that the difference rises slightly with the increasing length of the measured C block. Three degrees, defined by concrete ranges of difference in C-block length, were proposed for expressing the probability that the compared chromosomes had been transmitted.The study further attests to the effectiveness of the method of constitutive heterochromatin measurement for paternity testing. In our set of ten families, the comparison of C-band lengths of chromosomes 1, 9, 16, and Y led to rejection of paternity in 64% of unrelated individuals; excluding the Y chromosome, the percentage decreased to 61. As many as 47% of the individuals were rejected by a difference higher than two units (i.e., transmission of the compared chromosome highly improbable).     

A technique for C-banding plant chromosomes with Wright stain

A technique is presented for C-banding plant chromosomes with a modified Wright stain. This procedure consistently produces brightly stained, well defined telomeric and interstitial heterochromatic bands, identifiable centromeric constrictions, and lightly stained euchromatic areas on chromosomes of rye.     

The quantitative analysis of polymorphism on human chromosomes 1, 9, 16 and Y

Summary The generalized characteristic of the C-segment lengths on chromosomes 1, 9, 16, and Y is suggested for a study of population heterogeneity. For this purpose, the concept of the distance D is introduced, taking into account the individual C-segment lengths, the mean lengths and standard deviations of C-segment lengths in a group of subjects, as well as the coefficients of correlation of the C-segment lengths on the said chromosomes.It is demonstrated that distance D may be employed to study the relevance of the given subject to the group studied, the relation to the mean characteristics within the group, and selection of subjects' pairs with almost identical C-segment lengths on respective chromosomes.In the study of such problems as zygosity of twins, family analysis, etc., along with the absolute C-segment lengths, it is recommended to employ the relative C-segment lengths on chromosomes 1, 9, 16, and Y, calculated as a part of the sum total of their absolute lengths.     

Somatic chromosomes of higher diptera

Summary The chromosome complements of eighty brain cells ofHylemya antiqua have been studied. The eighty cells were found in thirty-three larvae. Total complement length (TCL) is not randomly distributed among the larvae. Because there is an inverse correlation between chromosome length and width, it appears that in the cells studied the different chromosome lengths are partly expressions of different stages of metaphase contraction. It is suggested that synchronous division of cells still occurs in late larvae.The length of each chromosome arm is highly correlated with that of every other arm. It is possible that the correlations are complete but that inadequate technique causes the departures from completeness which are observed. The chromosome lengths are corrected slightly for distortions, but the corrections make very little difference in the correlation coefficients. There is a high value for the correlation between the correlation of two arm lengths and the sum of the two arm lengths. This is to be expected if the perfect correlation between all arm lengths is being obscured by errors of drawing and measurement.The autosomal arms have very similar coefficients of variation. The arm ratios (length of long arm divided by short arm) are not correlated with TCL or with each other, and arm ratio is randomly distributed among the larvae. The sex chromosomes have a smaller coefficient of variation than the autosomes, so that they are relatively large in small cells and relatively small in large cells.Twenty-two cells inHylemya fugax were measured. The autosomes also showed a high correlation between arm lengths. An entirely heterochromatic autosomal arm showed the same phenomenon of a low coefficient of variation which was shown by the heteropycnotic sex chromosomes inH. antiqua. The low variability of heterochromatic regions accompanied by an apparently non-random distribution of the TCL may produce an erroneous picture of the species complement when dealing with small numbers.It is suggested that for simplicity in using cytological observations of this sort for taxonomic purposes, the technique of measuring the percent TCL of a chromosome plus its arm ratio be replaced by the percent TCL of each arm plus the average length difference between the arms of each chromosome pair in units of percent TCL.     

Die intraspezifische Variabilität des heterochromatischen Armes eines Chromosoms bei der GattungCarabus L. (Coleoptera)

The chromosome complement ofC. auronitens Fabr. is 2n _♂=26+XY. One autosomal pair—called A-chromosomes—is relatively long.A-chromosomes consist of a euchromatic and a heterochromatic arm. Labelling of mitotic chromosomes with³H-thymidine shows that replication of the heterochromatic arm continues when it has ended in the euchromatic arm. In males and females the length of the heterochromatic arm varies intraindividually. In 47 of 99 males the heterochromatic arms were heteromorphic. Calculations of the quotient length of the euchromatic/length of the heterochromatic arm have shown that at least 6 different types of the A-chromosome exist. These types differ from each other in the number of heterochromatic sections separated by constrictions. The longest heterochromatic arm observed consisted of 8 such sections. The genetic significance of the heterochromatin in the genus ofCarabus is at present unknown (Zusammenfassung see p.305).      

Karyotypes of the Genus Fritillaria from South Anhui

This paper reports chromosome numbers and karyotypes of five species of the genus Fritillaria from south Anhui. The origin of the material used in this work is provided in Table 1, micrographs of mitotic metaphase in Plate 1,2, and the parameters of chromosomes in Table 2. Except F. thunbergii Miq., the karyotypes and chromosome numbers of all the species in this paper were studied for the first time. The results are shown as follows: 1. Fritillaria qimenensis D. C. Zhang et J. Z. Shao Collected from Qimen, Anhui, it has the karyotype formula 2n = 24+4Bs = 3m+lsm+8st (2sc)+12t (2sc)+4Bs (Plate 1:1, 2). The chromosomes range in length 8.72-19.13μm, with the ratio of the longest to the shortest 2.19. Therefore, the karyotype belongs to Stebbins’ (1971) 3B. The secondary constrictions are found on the long arms of 7th and 10th pairs. All the five B-chromosomes are of terminal centromeres. The two chromosomes of the second pair show heteromorphy (Fig. 1, E) with arm ratios 1.86 and 1.56 respectively. 2. Fritillaria monantha Miq. var. tonglingensis S. C. Chen et S. F. Yin Collected from Tongling, Anhui, this species is shown to have three chromosome numbers, 2n =24+5Bs, 2n=24+2Bs and 2n=24. This paper reports 2 cytotypes: Type I: 2n = 24+5Bs = 4m+8st (2sc) +12t (2sc) +5Bs (Plate 1: 3, 4). The chromosomes range in length from 10.40 to 22.19μm, with the ratio of the longest to the shortest 2.13. It belongs to 3B of stebbins’(1971) karyotypic symmetry. The secondary constrictions are found on the short arms of 7th and the long arms of 9th chromosome pairs. The metacentric B-chromosomes and the small satellites located on the short arms are major characters of this cytotype. Type II: 2n=24=2m+2sm+8st(2sc)+12t(2sc) (Plate 1:5, 6). The chromosomes range in length from 13.84 to 29.81μm, with the ratio of the longest to the shortest 2.15. The karyotype belongs to Stebbins’3B. The secondary constrictions are found on the long arms of 5th and 10th pairs. No B-chromosomes are found. 3. Fritillaria xiaobeimu Y. K. Yang, J. Z. Shao et M. M. Fang Collected from Ningguo, Anhui, it has karyotype formula 2n = 24 = 2m+2sm+10st (4sc) + 10t (Plate 2:7, 8). The chromosomes range in length from 13.86 to 26.27μm, with the ratio of the longest to the shortest 1.89. The karyotype belongs to stebbins’3A. The secondary constrictions are found on the long arms of 7th and 9th pairs. 4. Fritillaria ningguoensis S. C. Chen et S. F. Yin Collected from Ningguo, Anhui, it is of karyotype formula 2n = 24 = 2m+2sm+8st (2sc) +12t (Plate 2: 9, 10). The chromosomes range in length from 9.11 to 23.23μm, with the ratio of the longest to the shortest 2.55. The karyotype belongs to Stebbins’3B. The secondary constrictions are only found on the long arms of the 10 th pair. 5. Fritillaria thunbergii Miq. Collected from Ningguo, Anhui, it is of karyotype formula 2n = 24 = 2m+2sm+8st(2sc) +12t(2sc)(Plate 2:11, 12). The chromosomes range in length from 8.83 to 19.85μm, with the ratio of the longest to the shortest 2.25. The karyotype belongs to stebbins’3B. There are secondary constrictions on the long arms of 5th and 7th pairs. The karyotype of the Ningguo material is similar to that of the Huoqiu (Anhui) material reported by Xu Jin-lin et al. (1987), but it is obviously different from 2n=2m(sc)+2sm+4st(2sc)+16t (2sc) reported byZhai et al. (1985) for the material from Xingjiang, Northwest China.     

The chromosome complement of the Rhesus monkey (Macaca mulatta) determined in kidney cells cultivated in vitro

Summary The chromosome complement of male and female Rhesus monkey has been investigated in kidney cells cultivatedin vitro for 3 to 6 days. The chromosome number is 42. The Y chromosome of the heterogametic male is the smallest element in the complement, and it is acrocentric. The X chromosome ranks eigth in decreasing order of size and typically has an arm ratio of 1.4. The autosomes form a graded size series of metacentric chromosomes, 3–15μ long in early metaphase, and with arm ratios from 1.1 to 3.3. Chromosome IX carries a large secondary constriction near the centromere; it is presumed to be the main nucleolar chromosome. A smaller secondary constriction is found consistently in the long arm of chromosome I. The X chromosome and chromosome XXI appear to be dimorphic in the limited population studied, the alternative forms differing in arm ratios but not in total length. An idiogram of the haploid chromosome complement is presented incorporating measurements of 10 completely analyzed nuclei, five from male monkeys and five from females. On the basis of relative length, arm ratio, and occurrence of secondary constrictions, most chromosomes of the complement can be individually identified. Supported in part by grants from the National Cancer Institute of Canada; the National Institutes of Health of the United States, Public Health Service; and the National Foundation for Infantile Paralysis.