Variation of B Chromosome Associated with Tissue Culture in Wheat-rye Cross

In vitro variation of B chromosomes was studied by examining the callus cells derived from the immature embryos from a cross of Chinese Spring wheat ( Triticum aestivum L.) and Fin 7416 rye ( Secale cereale L.) carrying two B chromosomes. In 40-d-old callus cells, the numbers of B chromosomes ranged from one to four in 65.6% of the cells observed. The distribution of B chromosome numbers was associated with the ploidy levels of the normal chromosomes (A chromosomes). The frequency of the cells with high numbers of B chromosomes (i.e., three or four B chromosomes) in the amphiploid cells with 56 A chromosomes was greater than those in the haploid cells with 28 A chromosomes. Although structural changes in the rye A chromosomes were observed, cytological observation and genomic in situ hybridization demonstrated that the rye B chromosomes were conserved in morphological appearance following tissue culture.     

甘蓝型油菜与诸葛菜属间杂种的减数分裂观察

在甘蓝型油莱与诸葛菜属间杂种细胞减数分裂中,观察到两种类型的染色体分离方式。第一种,细胞内的３１条染色体呈两组分离,其中１９条染色体至细胞一极,１２条染色体至另一极;且在两极的染色体进行分裂,产生４个核,两个具有１９条染色体,另两个具有１２条染色体。故杂种细胞通过这种染色体分离方式,可同时产生具有１９与１２条染色体的两类配子。杂种细胞内发生的这种染色体行为,极有可能与来自两个亲本种的染色体组相关;即在细胞两极的１９与１２条染色体分别来自甘蓝型油菜与诸葛菜。第二种,在后期Ｉ至末期Ｉ,花粉母细胞内以６个二价体形式出现的１２条染色体被落后在赤道板附近,而被排斥在末期核之外。这些落后二价体很有可能来自诸葛菜。因此,在杂种细胞减数分裂中,来自两个亲本种的染色体组被分开。文中还对染色体组分开的遗传意义等进行了讨论。     

Late replicating X chromosomes in human triploidy.

The status of X-chromosome replication was studied in twenty-seven 69,XXY and nine 69,XXX human triploids in which the parental origin of the additional haploid set was known from the study of chromosome heteromorphisms. Among the 69,XXY triploids, fourteen had no late replicating X, two had one late replicating X in all cells examined, and eleven had two populations of cells, one with late replicating X chromosome, and one without any. Among the 69,XXX triploids, four had a single late replicating X, and five had two populations of cells, one with one late replicating X, and one with two late replicating X chromosomes. There was no correlation between the parental origin of the triploidy and the type of X-chromosome inactivation. However the number of late replicating X chromosomes was significantly lower in cultures grown from fetal tissue when compared with those grown from extra-embryonic tissue. In cultures derived from extra-embryonic tissue there was a significant correlation between the gestational age of the sample and the proportion of late replicating X chromosomes. The older the specimen, the greater the number of late replicating X chromosomes.     

Double fertilization in maize: the two male gametes from a pollen grain have the ability to fuse with egg cells

In flowering plants, two male gametes from a single pollen grain fuse with two female gametes, the egg and central cells, to form the embryo and endosperm, respectively. The question then arises whether the two male gametes fuse randomly with the egg and central cells. We investigated this question using two nearly isogenic maize lines with supernumerary B chromosomes (TB10L18) or without (r-tester). B chromosomes regularly undergo non-disjunction at the second pollen mitosis, producing one sperm cell with zero B chromosomes and one with two. We first confirmed earlier studies showing an excess of transmission of the B chromosomes to the embryo rather than to the endosperm. We then tested the possibility of a directed fertilization. For TB10L18 pollen, we could demonstrate the existence of a size dimorphism between the two sperm cells, correlated to the content in B chromosomes, as detected by fluorescence in situ hybridization (FISH). However, no directed fusion of B chromosome containing sperm to egg cells could be detected when using in vitro fertilization. The absence of directed fusion in vitro could also be demonstrated for control lines. We conclude that both male gametes have the capacity to fuse with the egg cell in maize, although sexual reproduction results in a preferential transmission of supernumerary B chromosomes.     

X chromosomes alternate between two states prior to random X-inactivation

Early in the development of female mammals, one of the two X chromosomes is silenced in half of cells and the other X chromosome is silenced in the remaining half. The basis of this apparent randomness is not understood. We show that before X-inactivation, the two X chromosomes appear to exist in distinct states that correspond to their fates as the active and inactive X chromosomes. Xist and Tsix, noncoding RNAs that control X chromosome fates upon X-inactivation, also determine the states of the X chromosomes prior to X-inactivation. In wild-type ES cells, X chromosomes switch between states; among the progeny of a single cell, a given X chromosome exhibits each state with equal frequency. We propose a model in which the concerted switching of homologous X chromosomes between mutually exclusive future active and future inactive states provides the basis for the apparently random silencing of one X chromosome in female cells.     

Studies on the activity of the X chromosomes in female teratocarcinoma cells in culture

Embryonal carcinoma cells derived from murine teratocarcinomas are able to differentiate into the same variety of tissue types as early embryonic cells. Because embryonal carcinoma cells resemble those of the embryo at a stage before X chromosome inactivation has occurred in females embyronal carcinoma cells containing two X chromosomes were examined to determine whether both are genetically active. The specific activities of X-linked enzymes were measured in embryonal carcinoma cells containing either one or two X chromosomes. The activities in both cell types were similar, suggesting that only one X chromosome was active in the female cells. Further support for this conclusion came from experiments in which azaguanine-resistant mutants were recovered with similar frequencies from embryonal carcinoma cell lines containing one and two X chromosomes. Late replication of an X chromosome DNA was detected in one embryonal carcinoma cell line with two X chromosomes but not in another. This suggests that cells of these two lines were arrested at different developmental stages, and that late DNA replication may not be a necessary adjunct of X inactivation. Evidence is presented which suggests that X chromosome reactivation does not occur during differentiation of the cells in vitro.     

Centric-fusion translocation and whole-arm heterochromatin in the karyotype of the blue fox (Alopex lagopus L.): synaptonemal complex analysis.

Conventional observations of mitotic chromosomes from two male blue foxes, revealing a centric-fusion translocation and whole-arm heterochromatin, were verified by synaptonemal complex analysis. This analysis revealed that the centric fusion had been preceded by a conspicuous loss of chromosome material in the two one-armed chromosomes involved, but the chromosomal origin of the centric-fusion kinetochore could not be established. The nontranslocated chromosomes of the trivalent, which in all cells but one were in cis configuration, had reached by early pachytene a stage in which almost complete homologous pairing and nonhomologous association or pairing of the free ends of the chromosomes could be observed. In later stages, complete pairing of the nontranslocated chromosomes with the corresponding arms of the centric-fusion translocation was seen occasionally. One to six autosomal bivalents demonstrated unpaired heterochromatic arms in early pachytene, and the heterochromatic chromosome arms were sometimes unpaired even in late pachytene. Some of them showed a distinct size heteromorphism in late zygotene and early pachytene. In most late-pachytene cells, however, the heteromorphic chromosomes were completely length-adjusted. Only a small fraction of the cells showed pairing interference between nonhomologous chromosomes.     

Accessory Chromosomes in a tree--Ficus krishnae

Accessory chromosomes in a tree, Ficus krishnae are reportedfor the first time. Besides the normal complement of 2n = 26;1–2 small euchromatic accessory chromosomes were encountered.The frequency of accessory chromosomes in somatic cells was30 per cent. In meiotic cells one accessory chromosome was presentin 54 per cent of cells while two were observed in 9 per centof the pollen mother cells.     

Chromatin diminution and a chromosomal mechanism of sexual differentiation in Strongyloides papillosus

Eggs obtained from feces of rabbits infected with Strongyloides papillosus were squashed and the karyotypes were determined. They contained cells with either two long and two medium sized chromosomes (2L2M), or one long, three medium and one short chromosome (L3MS). Two types of parasitic female gonad could be distinguished on the basis of oocyte chromosome morphology at prometaphase of the maturation division. All the oocytes in a gonad contained either two upaired long chromosomes and two unpaired medium sized chromosomes, or two unpaired medium sized chromosomes and two unpaired chromosomes segmented into beads in one region. At the maturation division in mitotic parthenogenesis the beads appear to be lost from one of the chromosomes. This generates a medium sized and a shorter chromosome, which together with the undiminished chromosomes make up the L3MS karyotype. Animals with beaded oocyte chromosomes lay eggs that develop into males. It is suggested that males are heteromorphic for the long homologue due to chromatin diminution, that occurs in the maturation division of mitotic parthenogenesis.     

TRANSMISSION AND SIGNIFICANCE OF B CHROMOSOMES IN ANTHURIUM WAROCQUEANUM

Somatic and meiotic chromosomes of one plant of Anthurium warocqueanum J. Moore and its selfed offspring were analyzed. The parent showed 2n = 30 + 3B in both somatic cells and pollen mother cells. The B chromosomes divided normally in somatic cells, but meiotic associations of Bs varied. Three configurations of three B chromosomes were observed at metaphase I of parent meiosis: one trivalent, one bivalent and one univalent, or three univalents. The number of B chromosomes in offspring ranged from 0 to 6, indicating their transmission from both male and female gametes. Offspring with two B chromosomes appeared in greatest frequency. It was hypothesized that both male and female gametes of the 3 B parent frequently contained one B chromosome through the normal distribution of the bivalent Bs at meiosis and the elimination of the univalent B chromosome due to lagging. Examination of pollen mother cells of offspring also revealed irregular behavior of B chromosomes. With a high number of B chromosomes, normal A chromosome bivalent formation seemed to be reduced. No phenotypic effects of B chromosomes were observed.     

Chromosomal changes in three cell strains of the tasmanian rat-kangaroo,Potorous tridactylis,cultured in vitro (Marsupialia)

The chromosomal changes occurring in three strains of cells from Potorous tridactylis, one derived from testis and two of kidney tissue, were followed during the in vitro life of the strains.One kidney cell strain was a slow growing one and died after 23 passages showing aneuploidy with very aberrant metaphases. The strain derived from testis showed aneuploidy after a period of growth retardation, about 50% of the aneuploid cells having 18, 19 or 20 chromosomes. In these cells the chromosomes 1, 2 and 4 were always present in triplicate and the cells always had two X-chromosomes. The second kidney strain showed aneuploidy after a period of growth retardation, cells with 22 and 23 chromosomes being the most frequent ones, but in different proportions. As the number of aneuploid cells gradually decreased, diploid cells appeared in the population. Their number also decreased and a new population of aneuploid cells arose, having 23 chromosomes, missing one chromosome nr. 5 from the tetraploid complement. Then again the cell strain returned to diploidy but as the frequency of these diploid cells decreased, the strain died out.The work was carried out, in part, under the association between Euratom and the University of Leiden, contract No. 052-64-I-BIAN, and it also received support from the Foundation for Basic Medical Research (FUNGO).     

Synaptonemal complex analysis of a three-breakpoint translocation in a subfertile bull.

Somatic chromosome analysis of a subfertile Brown Swiss bull demonstrated a three-breakpoint translocation involving chromosomes 1, 8, and 9 in G- and R-banded karyotypes. Based on standard bovine chromosome nomenclature, the translocation was defined as t(1;8;9)(q43;q13;q26). Synaptonemal complex analysis of the chromosome aberration by electron microscopy revealed a hexavalent configuration in 52 of 53 pachytene cells. Twenty-seven cells (51%) had a completely paired hexavalent configuration showing distinctly nonhomologous pairings between normal and/or translocated chromosomes involved in the exchanges. Thirteen cells showed a hexavalent configuration with centrally unpaired chromosome segments but with completely paired terminal arms. In 13 cells (including one at zygotene) the translocation chromosomes formed an open hexavalent, and in one cell there were two completely paired trivalents. Thirty-two cells at diakinesis-MI demonstrated 28 configurations, including one large hexavalent. Testicular histology, testis size, and seminal characteristics were normal.     

A developmental study of constitutive heterochromatin in Microtus agrestis

In the vole, Microtus agrestis, the constitutive heterochromatin is largely restricted to the giant sex chromosomes but varies in its degree of condensation in various cell types. In the cleavage embryos and fibroblasts it formed one or two long and extended heterochromatic fibers, in hepatocytes it formed two large and diffuse masses and in neurons, spermatogonia and oogonia it formed two large and compact masses. The basic patterns of all differentiated cells were essentially unchanged throughout development.—At all stages of development and in cells of all types, mitotic nuclei displayed two large heteropycnotic chromosomes in prophase and persistent condensation in telophase. Apposition and delayed separation of chromatids of the giant chromosomes was also observed in metaphase and anaphase, respectively. During the first meiotic prophase of spermatocytes and oocytes, the giant chromosomes were also heteropycnotic.—The results strongly suggest that constitutive heterochromatin is localized in the same chromosomes throughout development and represents a specific entity.     

Cytogenetic and biochemical investigations on fibroblast cultures and clones with one and two active X chromosomes of a 69,XXY triploidy

Bromodeoxyuridine replication patterns showed that fibroblasts from a 69,XXY triploidy carried either one or two early replicating X chromosomes. The activity of alpha-galactosidase A measured in single cells fell into two classes with a ratio of 1:2. Dilute plating produced clones of both types with the activity of alpha-galactosidase A corresponding to the number of active X chromosomes. To our knowledge, this is the first report on clones of a triploidy with different numbers of active X chromosomes, and on a gene-dosage effect of an X-linked gene using triploid cells with one active X as control.     

Observations on dicentrics in living cells

Summary In previously irradiated endosperm cells of Haemanthus katherinae studied in vitro by means of micro-cinematography, two-kinetochore chromatids and dicentric chromosomes have been observed. Breaking of such dicentric chromatids and chromosomes has been analysed. Behaviour of some of the dicentric chromosomes during anaphase deserves special attention: interlocking dicentrics cut one through another and rejoin in a few minutes. In this way from a metaphase interlocking dicentric, two sister anaphase dicentrics are formed. Interlocked dicentrics can also uncoil and not break at all. In this case no activity was observed in one kinetochore of one dicentric in later stages of anaphase (two kinetochores were active in one dicentric and only one in its sister). Analysis of chromosome movements in two-kinetochore chromatids and dicentrics is also presented.     

Formation and division of binucleated cells in kidney cell cultures of microtus agrestis

Summary Epithelial kidney cell cultures of Microtus agrestis contain 10 to 25% binucleated cells. Observations of living cells under the phase contrast microscope showed that binucleated cells can arise by nuclear mitosis without cytoplasmic division. When binucleated cells divide the two nuclei are highly synchronized as they enter mitosis. In mitosis the chromosomes of both nuclei combine to a common metaphase plate leading to polyploid cells. In one case a tripolar spindle was seen after formation of a metaphase by the chromosomes of the two nuclei of a binucleated cell. This tripolar mitosis resulted in one binucleated and one mononucleated cell. The DNA-content (Feulgen photometry) and the distribution of heterochromatic bodies of the nuclei were corresponding to a tetraploid, a triploid and a haploid chromosome set. This suggests the possibility of somatic segregation of complete haploid sets.Supported by the Deutsche Forschungsgemeinschaft.     

Germ line-restricted supernumerary (B) chromosomes in Eptatretus okinoseanus.

Cytogenetic studies were performed on two types of Japanese hagfish (Eptatretus okinoseanus) that eliminate about 45% (type A) and 55% (type B) of their DNA from presumptive somatic cells during the differentiation of somatic cells. The observations revealed inter- and intraindividual variations in the number of chromosomes in germ cells of both types of hagfishes. Although the modal number of chromosomes in the germ cells was 54 in both types, the percentage of cells with the modal number was rather low (38.6% [51/132] in five specimens of type A and 22.7% [25/110] in eight specimens of type B). In addition, one of seven type B specimens clearly had a modal number of 62 chromosomes. Another specimen of type B had a bimodal distribution of chromosome numbers, with peaks of 54 and 59 chromosomes. The observation of interindividual variations was supported by data on the amount of DNA in germ cells of type B specimens. However, these variations were rarely observed in somatic cells. These results suggest that supernumerary (B) chromosomes are maintained in germ cells and are eliminated together with some other chromosomes and/or chromatin from somatic cells.     

A Factual Analysis of Chromosomal Movement in Barbulanympha

SYNOPSIS. The role of eleven different types of achromatic figures in chromosomal movement of Barbulanympha is analyzed. When only one pole is present, no chromosomes are ever connected with it, and hence they do not move. The chromosomes go through their usual life cycle including pairing, but remain in the parent nucleus, which, of course, does not divide. When two poles are present with only one pole near the nucleus, the poles, which are the distal ends of the elongate centrioles, do not cooperate in the formation of a central spindle and the chromosomal behavior is just as if there were only one pole—no movement. The same is true when more than two poles are present with only one near the nucleus.
Unless a central spindle is present, movement of chromosomes never occurs. However, when many central spindles are present, sister chromosomes may separate and move to poles which are not directly connected by a central spindle. In other words, sisters may separate without moving along a central spindle.
In binucleate cells with one central spindle the chromosomes of one nucleus move to the poles, but those of the other do not. Movement always occurs in the nucleus that has its nuclear membrane depressed by the central spindle. When two or more central spindles are present, the chromosomes of both nuclei may move to the poles.     

Meiotic self-pairing of the Psalidodon (Characiformes,Characidae) iso-B chromosome: A successful perpetuation mechanism

B chromosomes are non-essential additional genomic elements present in several animal and plant species. In fishes, species of the genus Psalidodon (Characiformes, Characidae) harbor great karyotype diversity, and multiple populations carry different types of non-essential B chromosomes. This study analyzed how the dispensable supernumerary B chromosome of Psalidodon paranae behaves during meiosis to overcome checkpoints and express its own meiosis-specific genes. We visualized the synaptonemal complexes of P. paranae individuals with zero, one, or two B chromosomes using immunodetection with anti-medaka SYCP3 antibody and fluorescence in situ hybridization with a (CA)₁₅ microsatellite probe. Our results showed that B chromosomes self-pair in cells containing only one B chromosome. In cells with two identical B chromosomes, these elements remain as separate synaptonemal complexes or close self-paired elements in the nucleus territory. Overall, we reveal that B chromosomes can escape meiotic silencing of unsynapsed chromatin through a self-pairing process, allowing expression of their own genes to facilitate regular meiosis resulting in fertile individuals. This behavior, also seen in other congeneric species, might be related to their maintenance throughout the evolutionary history of Psalidodon.     

Budding yeast mitotic chromosomes have an intrinsic bias to biorient on the spindle

BACKGROUND: Chromosomes must biorient on the mitotic spindle, with the two sisters attached to opposite spindle poles. The spindle checkpoint detects unattached chromosomes and monitors biorientation by detecting the lack of tension between two sisters attached to the same pole. After the spindle has been depolymerized and allowed to reform, budding yeast sgo1 mutants fail to biorient their sister chromatids and die as cells divide. RESULTS: In sgo1 mutants, chromosomes attach to microtubules normally but cannot reorient if both sisters attach to the same pole. The mutants' fate depends on the position of the spindle poles when the chromosomes attach to microtubules. If the poles have separated, sister chromatids biorient, but if the poles are still close, sister chromatids often attach to the same pole, missegregate, and cause cell death. CONCLUSIONS: These observations argue that budding yeast mitotic chromosomes have an intrinsic, geometric bias to biorient on the spindle. When the poles have already separated, attaching one kinetochore to one pole predisposes its sister to attach to the opposite pole, allowing the cells to segregate the chromosomes correctly. When the poles have not separated, the second kinetochore eventually attaches to either of the two poles randomly, causing orientation errors that are corrected in the wild-type but not in sgo1 mutants. In the absence of spindle damage, sgo1 cells divide successfully, suggesting that kinetochores only make stable attachments to microtubules after the cells have entered mitosis and separated their spindle poles.