Structure of chromosomes

The number of gyres, mode, and direction of coils, centromere, secondary constriction, and telomere were studied in the human somatic chromosomes in terms of spiral structure, employing the newly developed technique involving a hypotonic treatment. Results obtained are: 1. the number of gyres is most probably constant in analogous chromosome from different cells; 2. the direction of coils is random; 3. each ehromatid is accepted as a unit of chromonema coiling; 4. the centromere seems to be a regionally elongated portion of the chromonema, without showing any differentiated structure; 5. chromonemata at secondary constrictions usually appear as uncoiled or less tightly coiled entities, and 6. two different types of the telomere structure were observed, one being a pointed-end type, while the other is a rounded-end type (Summary see p. 424). Supported by a grant from the National Institute of General Medical Sciences, U.S.P.H.S. (GM 14185).     

X-ray diffraction from isolated metaphase chromosomes

Metaphase chromosomes were isolated from Chinese hamster fibroblasts. They were found to give the same characteristic diffraction pattern in the wet state as isolated nucleohistone but when dry did not give the reflections normally associated with dry nucleohistone. No higher structural order than that associated with the nucleohistone supercoil was detected.     

Demonstration of membranous patches on isolated chromosomes

High resolution scanning electron microscopy of isolated Chinese hamster ovary metaphase chromosomes revealed “membranous patches” at telomeric and juxtatelomeric regions of the chromosomes. The “membranous patches” remained bound to the chromosomes during centrifugation through dense sucrose, but not after treatment with detergents. These membrane fragments on isolated purified chromosomes may represent a component that binds the chromosome to the inner portion of the nuclear envelope up to late stages of prophase. These chromosome associated membranous patches may represent sites of reformation of the nuclear envelope at telophase.     

The composition and structure of isolated chromosomes

1. The preparation of isolated chromosomes from liver, kidney, and pancreas has been described. 2. It has been shown that there is no gross cytoplasmic contamination in these preparations. 3. In a microscopic study of isolated chromosomes the same chromosomes have been found in different tissues of the same organism. Since individuality is one of the main characteristics of chromosomes, there can be little doubt that the preparations do, in fact, contain isolated chromosomes. 4. A quantitative study of staining with crystal violet shows that this basic dye competes with histone for the phosphoric acid groups of the DNA in chromosomes. The displacement of histone by protamine has been demonstrated. 5. Preparation of histone-free chromosomes has been described. Removal of histone does not affect the microscopic appearance of chromosomes. 6. The non-histone or residual protein has been prepared from histone-free chromosomes. The quantity of residual protein in a preparation of chromosomes is correlated with the amount of cytoplasm in the cells from which the chromosomes were prepared. 7. The microscopic appearance of chromosomes depends upon the association of DNA with residual protein. 8. Evidence has been given that in a chromosome there are two DNA-containing nucleoproteins; in one DNA is combined with histone, and in the other it is combined with residual protein.     

Structure and dynamics of interphase chromosomes

During interphase chromosomes decondense, but fluorescent in situ hybridization experiments reveal the existence of distinct territories occupied by individual chromosomes inside the nuclei of most eukaryotic cells. We use computer simulations to show that the existence and stability of territories is a kinetic effect that can be explained without invoking an underlying nuclear scaffold or protein-mediated interactions between DNA sequences. In particular, we show that the experimentally observed territory shapes and spatial distances between marked chromosome sites for human, Drosophila, and budding yeast chromosomes can be reproduced by a parameter-free minimal model of decondensing chromosomes. Our results suggest that the observed interphase structure and dynamics are due to generic polymer effects: confined Brownian motion conserving the local topological state of long chain molecules and segregation of mutually unentangled chains due to topological constraints.     

Arrangement of chromosomes in the interphase nucleus of plants

Chromosomal arrangement in the interphase nucleus has two main aspects: (1) arrangement of chromosomes with respect to nuclear polarity and to other nuclear components, and (2) arrangement of chromosomes with respect to one another. The latter aspect consists of two main types of spatial relationships; (a) relationships between different members of one chromosomal set, (b) relationships between different chromosomal sets. Data concerning various aspects of chromosomal arrangement in the interphase nucleus are presented and discussed and the genetic control as well as subcellular mechanisms which are involvled in nuclear organization, are elucidated. Evidence is presented indicating that, in common wheat, the gene system that determines the specific pattern of chromosomal arrangement in the nucleus is operating via the microtubular elements of the spindle system. The significance of ordered arrangement of chromosomes in the nucleus for the regularity of genetic activity and chromosomal behavior, is pointed out.Supported in part by a grant from the Stiftung Volkswagenwerk AZ I/34 075/76     

Scanning electron microscopy of isolated Chironomus polytene chromosomes

Individual polytene chromosomes have been isolated from Chironomus stigmaterus for scanning electron microscope observations. The three dimensional ultrastructure of these chromosomes consists of a series of chromatin strands extended in the interbands and more tightly coiled or folded in the banded regions. The nucleolus is observed to be a dense disc or doughnut shaped structure surrounding the chromosome while the Balbiani Rings appear as diffuse regions consisting of both fibrillar and granular elements.     

Uptake of isolated plant chromosomes by plant protoplasts

For mass isolation of plant metaphase chromosomes, cultured cells of wheat (Triticum monococcum) and parsley (Petroselinum hortense) were synchronized by hydroxyurea and colchicine treatment. This synchronization procedure resulted in high mitotic synchrony, especially in suspension cultures of parsley in which 80% of the cells were found to be at the metaphase stage. Mitotic protoplasts isolated from these synchronized cell cultures served as a source for isolation of chromosomes. The described isolation and purification method yielded relatively pure chromosome suspension. The uptake of the isolated plant chromosomes into recipient wheat, parsley, and maize protoplasts was induced by polyethylene-glycol treatment. Cytological studies provided evidences for uptake of plant chromosomes into plant protoplasts.Abbreviations PEG polyethylene glycol - HU hydroxyruea - C colchicine - HUC hydroxyurea and colchicine - CIM chromosome isolation medium - TCM Tris chromosome medium     

Flow microfluorometric analysis of isolated Chinese hamster chromosomes

Isolated Chinese hamster chromosomes were analysed by flow microfluorometry after staining with ethidium bromide. Peaks corresponding to the expected DNA content of each of the autosomes were observed. Examination of chromosomes partially fractionated by zonal centrifugation on sucrose gradients verified the identity of the chromosomes in the various peaks of the whole karyotype. The significance of these observations to karyotype analysis is discussed.     

Genetics of dioecy and causal sex chromosomes in plants

Dioecy (separate male and female individuals) ensures outcrossing and is more prevalent in animals than in plants. Although it is common in bryophytes and gymnosperms, only 5% of angiosperms are dioecious. In dioecious higher plants, flowers borne on male and female individuals are, respectively deficient in functional gynoecium and roecium. Dioecy is inherited via three sex chromosome systems: XX/XY, XX/X0 and WZ/ZZ, such that XX or WZ is female and XY, X0 or ZZ are males. The XX/XY system generates the rarer XX/X0 and WZ/ZZ systems. An autosome pair begets XY chromosomes. A recessive loss-of-androecium mutation (ana) creates X chromosome and a dominant gynoecium-suppressing (GYS) mutation creates Y chromosome. The ana/ANA and gys/GYS loci are in the sex-determining region (SDR) of the XY pair. Accumulation of inversions, deleterious mutations and repeat elements, especially transposons, in the SDR of Y suppresses recombination between X and Y in SDR, making Y labile and increasingly degenerate and heteromorphic from X. Continued recombination between X and Y in their pseudoautosomal region located at the ends of chromosomal arms allows survival of the degenerated Y and of the species. Dioecy is presumably a component of the evolutionary cycle for the origin of new species. Inbred hermaphrodite species assume dioecy. Later they suffer degenerate-Y-led population regression. Cross-hybridization between such extinguishing species and heterologous species, followed by genome duplication of segregants from hybrids, give rise to new species.     

Serratia species isolated from plants

Using the selective caprylate-thallous agar medium, the presence ofSerratia species was systematically examined in 623 plant samples. A total of 167Serratia strains was isolated from these plant samples and identified to species and biogroups. Uniform and characteristicSerratia populations were found in figs and coconuts: (i)Serratia ficaria was recovered from most figs collected in California, Tunisia, and France; various biotypes ofS. marcescens also were found in figs; (ii) onlyS. marinorubra was recovered from coconuts bought on two continents. From plants other than figs and coconuts, representatives were isolated of all eightSerratia species we presently recognize—with a large preponderance ofS. liquefaciens andS. proteamaculans. These other plant samples fell into threeSerratia-prevalence groups: (i) vegetables-mushrooms-mosses-decaying plant material (53.8% of these samples were positive forSerratia); (ii) grasses (23.7% positive); and (iii) trees and shrubs-small plants (8.4% positive). PigmentedS. marcescens biotypes were rarely isolated from plants (except from figs). Of theS. marcescens biogroups most frequently encountered in nosocomial and iatrogenic infections of man, A3 and A4 were isolated from plants in this study, but A5/8 and TCT were not.     

B chromosomes and genome size in flowering plants.

B chromosomes are extra chromosomes found in some, but not all, individuals within a species, often maintained by giving themselves an advantage in transmission, i.e. they drive. Here we show that the presence of B chromosomes correlates to and varies strongly and positively with total genome size (excluding the Bs and corrected for ploidy) both at a global level and via a comparison of independent taxonomic contrasts. B chromosomes are largely absent from species with small genomes; however, species with large genomes are studied more frequently than species with small genomes and Bs are more likely to be reported in well-studied species. We controlled for intensity of study using logistic regression. This regression analysis also included effects of degree of outbreeding, which is positively associated with Bs and genome size, and chromosome number, which is negatively associated with Bs and genome size, as well as variable ploidy (more than one ploidy level in a species). Genome size, breeding system and chromosome number all contribute independently to the distribution of B chromosomes, while variable ploidy does not have a significant effect. The genome size correlates are consistent with reduced selection against extra DNA in species with large genomes and with increased generation of B sequences from large A genomes.     

Absence of true interchromosomal connectives in microsurgically isolated chromosomes

Mitotic chromosomes of the Indian muntjac were isolated from cultured fibroblast-like cells by microsurgery. The entire complement of seven chromosomes could be obtained with the radial array of chromosomes on the mitotic spindle intact. The center of the radial array was occupied by a fibrous network which stained with tubulin antiserum. This network was absent when cells were treated with colchicine or vinblastine sulfate prior to chromosome isolation, and probably represents a remnant of the mitotic spindle. Most isolated chromosomes were connected to the spindle by fibers attached to the centromeres. Such fibers did not stain for DNA and were resistant to DNases but sensitive to proteases. No interconnections were found to run from chromosome to chromosome except for occasional artifactual adhesions resulting from collisions between chromosomes which occurred during micromanipulation. We therefore found no evidence that chromosomes of the Indian muntjac are interconnected at mitosis.