The effect of colchicine on synapsis and chiasma formation in microsporocytes of Lilium

Microsporocytes of Lilium that are exposed to colchicine as late as early zygotene show reduced chiasma frequencies and the presence of univalents at Division I. These effects are preceded at pachytene by the appearance of pairing gaps (light microscopy) and by a relatively high ratio of uncomplexed lateral elements/synaptonemal complexes (EM). Chiasma formation thus appears to be reduced by failures in synapsis. Unlike the behavior of wheat, colchicine can disrupt chiasma formation in Lilium after cells have entered meiosis.     

Cytogenetics of chromosome pairing in wheat

Meiotic chromosome pairing in Triticum aestivum is controlled by genetic systems promoting and reducing pairing. The pairing of homoeologous chromosomes is prevented principally by the activity of a single locus (Ph) distally located on the long arm of chromosome 5B. In certain hybrids, supernumerary chromosomes (B chromosomes) from Aegilops species can compensate for the absence of chromosome 5B preventing or reducing homoeologous pairing. Temperature-dependent variants and colchicine sensitivity have been used to show that there are at least two stages in the G1 of meiosis at which the occurrence of meiotic pairing is determined. Wheat may differ from lily in the detailed organization of meiosis.     

Disruption of topoisomerase II perturbs pairing in drosophila cell culture

Homolog pairing refers to the alignment and physical apposition of homologous chromosomal segments. Although commonly observed during meiosis, homolog pairing also occurs in nonmeiotic cells of several organisms, including humans and Drosophila. The mechanism underlying nonmeiotic pairing, however, remains largely unknown. Here, we explore the use of established Drosophila cell lines for the analysis of pairing in somatic cells. Using fluorescent in situ hybridization (FISH), we assayed pairing at nine regions scattered throughout the genome of Kc167 cells, observing high levels of homolog pairing at all six euchromatic regions assayed and variably lower levels in regions in or near centromeric heterochromatin. We have also observed extensive pairing in six additional cell lines representing different tissues of origin, different ploidies, and two different species, demonstrating homolog pairing in cell culture to be impervious to cell type or culture history. Furthermore, by sorting Kc167 cells into G1, S, and G2 subpopulations, we show that even progression through these stages of the cell cycle does not significantly change pairing levels. Finally, our data indicate that disrupting Drosophila topoisomerase II (Top2) gene function with RNAi and chemical inhibitors perturbs homolog pairing, suggesting Top2 to be a gene important for pairing.     

Colchicine induced asynapsis and c-meiosis in pollen mother cells of cultivated anthers of rye

Rye anthers were cultured in medium with colchicine for periods of 12–24 h. Unaffected PMCs showed normal meiotic pairing in the same anthers in which other PMCs showed several degrees of asynapsis (or desynapsis), c-meiosis or anaphase irregularities. Pairing was affected only in cells which were at mid zygotene at the time of treatment initiation. Chiasma terminalization and release was possible in the absence of a functional spindle. Possibly, spindle function is more sensitive to colchicine than pairing. Variation within anthers can be adscribed to gradients in stage and development. The possibility of differences in penetration of colchicine must be recognized.     

Tetrahymena meiosis: Simple yet ingenious

The presence of meiosis, which is a conserved component of sexual reproduction, across organisms from all eukaryotic kingdoms, strongly argues that sex is a primordial feature of eukaryotes. However, extant meiotic structures and processes can vary considerably between organisms. The ciliated protist Tetrahymena thermophila, which diverged from animals, plants, and fungi early in evolution, provides one example of a rather unconventional meiosis. Tetrahymena has a simpler meiosis compared with most other organisms: It lacks both a synaptonemal complex (SC) and specialized meiotic machinery for chromosome cohesion and has a reduced capacity to regulate meiotic recombination. Despite this, it also features several unique mechanisms, including elongation of the nucleus to twice the cell length to promote homologous pairing and prevent recombination between sister chromatids. Comparison of the meiotic programs of Tetrahymena and higher multicellular organisms may reveal how extant meiosis evolved from proto-meiosis.     

Meiosis and fertility of reciprocal triploid hybrids of Lolium multiflorum with Festuca pratensis

Diploid and tetraploid forms of Lolium multiflorum and Festuca pratensis were crossed under controlled conditions and after embryo rescue all four combinations of autoallotriploid hybrids were obtained. Male and female fertility and chromosome pairing at metaphase I of meiosis were studied in several plants from each hybrid combination. The hybrids with two genomes of L. multiflorum and one of F. pratensis (genomic formulae LmLmFp and FpLmLm) were male and female fertile while the hybrids with two genomes of F. pratensis and one of L. multiflorum had a reduced fertility (FpFpLm) or were completely sterile (LmFpFp). Chromosome pairing at metaphase I varied among hybrid combinations depending on their genomic composition. LmLmFp and FpLmLm hybrids had similar patterns of pairing (1.83I + 5.29II + 2.85III and 2.22I + 5.22II + 2.75III, respectively) but they differed from those of FpFpLm (3.65I + 4.65II + 2.68III) and especially from LmFpFp (4.78I + 5.87II + 1.49III). Conventional analysis of meiosis failed to explain the differences in chromosome behaviour and fertility/sterility levels between the autoallotriploid hybrids with two Lolium or two Festuca genomes.     

Functional analysis of maize RAD51 in meiosis and double-strand break repair

In Saccharomyces cerevisiae, Rad51p plays a central role in homologous recombination and the repair of double-strand breaks (DSBs). Double mutants of the two Zea mays L. (maize) rad51 homologs are viable and develop well under normal conditions, but are male sterile and have substantially reduced seed set. Light microscopic analyses of male meiosis in these plants reveal reduced homologous pairing, synapsis of nonhomologous chromosomes, reduced bivalents at diakinesis, numerous chromosome breaks at anaphase I, and that >33% of quartets carry cells that either lack an organized nucleolus or have two nucleoli. This indicates that RAD51 is required for efficient chromosome pairing and its absence results in nonhomologous pairing and synapsis. These phenotypes differ from those of an Arabidopsis rad51 mutant that exhibits completely disrupted chromosome pairing and synapsis during meiosis. Unexpectedly, surviving female gametes produced by maize rad51 double mutants are euploid and exhibit near-normal rates of meiotic crossovers. The finding that maize rad51 double mutant embryos are extremely susceptible to radiation-induced DSBs demonstrates a conserved role for RAD51 in the repair of mitotic DSBs in plants, vertebrates, and yeast.     

Colchicine-induced heritable modification of chiasma frequency and distribution in perennial ryegrass (Lolium perenne).

One-week-old seedlings of inbred lines of perennial ryegrass (Lolium perenne L.) were given a 3-h period of treatment with 0.2% colchicine, and this treatment was found to have heritable effects on chromosome behaviour at meiosis. The mean pollen mother cell chiasma frequency was reduced, and there was a corresponding increase in the number of univalents per cell and also changes in cell and bivalent variances. Patterns of chiasma distribution within bivalents were also modified. The effects were seen in the generation in which the treatment was given and they were also observed in the following sexual generation.     

Differential chromosome pairing affinities at meiosis in polyploid sugarcane revealed by molecular markers

Chromosome pairing at meiosis is an essential feature in cell biology, which determines trait inheritance and species evolution. Complex polyploids may display diverse pairing affinities and offer favorable situations for studying meiosis. The genus Saccharum encompasses diverse forms of polyploids with predominantly bivalent pairing. We have focused on a modern cultivar of sugarcane, R570, and taken advantage of a particular single copy probe (BNL 12.06) revealing 11 alleles by restriction fragment length polymorphism (RFLP). As for other cultivars, R570 is highly polyploid (2n=ca. 115) and indirectly derived from interspecific hybridization between Saccharum officinarum (2n=80, x=10) and S. spontaneum (2n=40-128, x=8). Here we determined the doses of the various BNL12.06 RFLP alleles among 282 progeny of R570 and estimated the mutual pairing frequencies among the corresponding homo- or homoeologous chromosomes using a maximum likelihood method. The result is an atypical picture, with pairing frequencies ranging from 0 to 40% and differential affinities leading to the identification of several chromosome subsets. This example illustrates the unsystematic meiotic behavior in a complex polyploid. It highlights a continuous range of pairing affinities between chromosomes and pinpoints a strong role of individual chromosome features, partly related to their ancestral origin, in the determination of these affinities.     

融安金柑雌雄配子发育相关性及2n配子诱导研究

以融安金柑为试验材料,观察了2n花粉自然发生率、雌雄配子发育过程及减数分裂行为,以不同秋水仙素浓度及处理时间对花蕾进行2n配子诱变.结果显示:(1)融安金柑2n花粉自然发生率为0.31％;(2)花蕾纵径为0.228～0.270cm时,为花粉母细胞减数分裂Ⅰ前期;花蕾纵径为0.274～0.318 cm时,大孢子处于减数分...     

异细胞质八倍体小黑麦的获得及其细胞遗传

改变小黑麦细胞质有可能增加减数分裂的稳定性,提高小黑麦的结实率与籽粒饱满度。作者以不同细胞质的“中国春”小麦与黑麦杂交,F_1幼苗用秋水仙素加倍获得双二倍体、或以八倍体小黑麦为父本与F_1杂交,获得异细胞质八倍体小黑麦(Triticale 8x)8个品系。实验结果表明:细胞质不同的“中国春”小麦与黑麦杂交结实率差异显著,出苗率亦不同,F_1株型多为两亲的中间型,花药不开裂,个别组合出现雄蕊雌化现象,有的组合表现生长弱性,减数分裂中期Ⅰ常出现1至数个末端交叉的棒状二价体,其数量在不同组合间差异显著,表明异细胞质对染色体配对有影响。D类细胞质对改进八倍体小黑麦的结实率可能有一定的作用。     

The effects of cytochalasin B and colchicine on the morphogenesis of mitochondria in Drosophila hydei during meiosis and early spermiogenesis

Summary Morphogenesis of mitochondria in male germ cells in cultivated cytocysts begins in early prophase I at which time mitochondria thicken and become ordered along the spindle apparatus during meiosis. At the end of the second meiotic division they aggregate to form the Nebenkern.In the presence of colchicine or cytochalasin B mitochondria are able to begin differentiation, although the correct course of meiosis is not guaranteed. In medium supplemented with colchicine they undergo normal thickening but do not aggregate, in a pattern known from untreated cultures. This may indicate that microtubules are involved in the aggregation process of mitochondria as colchicine is known to inhibit microtubule formation. Moreover, in cell cultures treated with cytochalasin B mitochondrial aggregation does occur; it is concluded that microfilaments, which are sensitive to cytochalasin B, do not play a detectable role in the aggregation of mitochondria.     

普通小麦与簇毛麦双二倍体的合成,育性及细胞遗传学研究

通过杂种幼胚无性系培养获得大量再生植株F_1,经秋水仙碱处理,合成了普通小麦与簇毛麦属间双二倍体(AABBDDVV)。其形态特征除株高、穗长、小穗数,籽粒大小和育性明显增加,生育期延长外,分别与各自的再生植株F_1相似。双二倍体的体细胞染色体数目变化范围为48—56。花粉母细胞减数分裂中期Ⅰ 2n=28Ⅱ的细胞占56.82％,每个细胞平均有27.10个二价体,1.44个单价体,0.08个三价体,0.03个四价体。经过连续两代单穗单株选择,结实率由15.91％提高到36.52％。     

The effect of colchicine on synaptonemal complex formation in Allium ursinum

Interference of colchicine with meiotic chromosome pairing in the wild garlic, Allium ursinum, was studied using a whole-mount spreading technique for synaptonemal complexes. Colchicine was found to cause (i) pairing suppression (arrest of leptotene) and (ii) deficient pairing initiation at zygotene in connection with morphologically anomalous, malfunctioning pairing initiation sites. Both of these phenomena could be responsible for the reduction of chiasma frequency by colchicine previously reported in the literature.     

Pairing centers recruit a Polo-like kinase to orchestrate meiotic chromosome dynamics in C. elegans

Faithful segregation of homologous chromosomes during meiosis requires pairing, synapsis, and crossing-over. In C.?elegans, homolog pairing and synapsis depend on pairing centers (PCs), special regions near one end of each chromosome that interact with the nuclear envelope (NE) and cytoplasmic microtubules. Here, we report that PCs are required for nuclear reorganization at the onset of meiosis. We demonstrate that PCs recruit the Polo-like kinase PLK-2 to induce NE remodeling, chromosome pairing, and synapsis. Recruitment of PLK-2 is also required to mediate a cell cycle delay and selective apoptosis of nuclei containing unsynapsed chromosomes, establishing a molecular link between these two quality control mechanisms. This work reveals unexpected functions for the conserved family of Polo-like kinases, and advances our understanding of how meiotic processes are properly coordinated to ensure transmission of genetic information from parents to progeny.     

The Synaptonemal Complex Protein Zip1 Promotes Bi-Orientation of Centromeres at Meiosis I

In meiosis I, homologous chromosomes become paired and then separate from one another to opposite poles of the spindle. In humans, errors in this process are a leading cause of birth defects, mental retardation, and infertility. In most organisms, crossing-over, or exchange, between the homologous partners provides a link that promotes their proper, bipolar, attachment to the spindle. Attachment of both partners to the same pole can sometimes be corrected during a delay that is triggered by the spindle checkpoint. Studies of non-exchange chromosomes have shown that centromere pairing serves as an alternative to exchange by orienting the centromeres for proper microtubule attachment. Here, we demonstrate a new role for the synaptonemal complex protein Zip1. Zip1 localizes to the centromeres of non-exchange chromosomes in pachytene and mediates centromere pairing and segregation of the partners at meiosis I. Exchange chromosomes were also found to experience Zip1-dependent pairing at their centromeres. Zip1 was found to persist at centromeres, after synaptonemal complex disassembly, remaining there until microtubule attachment. Disruption of this centromere pairing, in spindle checkpoint mutants, randomized the segregation of exchange chromosomes. These results demonstrate that Zip1-mediated pairing of exchange chromosome centromeres promotes an initial, bipolar attachment of microtubules. This activity of Zip1 lessens the load on the spindle checkpoint, greatly reducing the chance that the cell will exit the checkpoint delay with an improperly oriented chromosome pair. Thus exchange, the spindle checkpoint, and centromere pairing are complementary mechanisms that ensure the proper segregation of homologous partners at meiosis I.     

SUPERNUMERARY CHROMOSOMES IN TRADESCANTIA II. EFFECT OF COLCHICINE ON MEIOTIC ISOCHROMOSOME CONFIGURATIONS

The effect of colchicine on meiotic pairing and configuration frequencies of three homologous isosupernumerary chromosomes was investigated. In the absence of colchicine, the three isochromosomes displayed a high degree of interchromosomal pairing and chiasma formation. As a consequence, a high frequency of bivalents and trivalents were observed at diakinesis-metaphase I. The unique structure of isochromosomes enables them to pair intrachromosomally (i.e., foldback pairing) yet the preferential occurrence of interchromosomal pairing suggests that all six arms of the three isosupernumerary chromosomes were in close association prior to or upon initiation of synapsis. Supernumerary chromosomes in microsporocytes treated during presynapsis or early synapsis with colchicine exhibited a significant reduction (P < 0.001) in the number of bivalents and trivalents at diakinesis. However, there was no reduction in overall chiasma frequency among supernumeraries due to the induction of increased intrachromosomal pairing and chiasma formation. A colchicine-sensitive association or alignment of homologues preceding effective pairing has been demonstrated in standard chromosomes of a number of plant species. This study provides the first evidence to indicate that at least certain supernumerary chromosomes may display presynaptic association as well. The results also support the strongly held contention that colchicine is not directly preventing or inhibiting the actual formation of chiasmata, since no reduction in chiasma frequency was observed in the isochromosomes.     

Colchicine action at meiotic prophase revealed by SC-spreading

In the present paper earlier findings on the interference of colchicine with presynaptic alignment and synaptonemal complex formation in Allium ursinum are corroborated and several other direct or indirect consequences of colchicine treatment, like deformations and decomposition of lateral elements and a high incidence of non-homologous associations are reported. Mechanisms of meiotic alignment and pairing are discussed with respect to their susceptibility to the known cytological and biochemical activities of colchicine.     

The effect of B chromosomes on homoeologous pairing in species hybrids

The effect of B chromosomes on meiosis is described in the diploid and tetraploid interspecific hybrid Lolium multiflorum x Lolium perenne. Although the parental species are very closely related, the presence of B chromosomes in the diploid hybrid reduced both chiasma frequency and the number of bivalents at meiosis by a small but significant amount. However at the tetraploid level the presence of B chromosomes did not seem to alter the pairing pattern and chiasma frequency in any way. The use of B chromosomes to stabilize meiosis in amphiploids of this type between closely related outbreeding species is therefore ruled out.     

Structural and metabolic correlation for Bacillus megaterium ACBT03 in response to colchicine biotransformation

This study aims to evaluate the effects of colchicine on metabolic and structural changes in Bacillus megaterium ACBT03, enduring colchicine bioconversion. Electron microscopy examination of cells adapted to different concentrations of colchicine for its bioconversion to pharmacologically active 3-demethylated colchicine, endowed changes in cell shape, decreased cell wall and plasma membrane thickness. In line with microscopic studies, lipid and membrane protein contents were drastically reduced in bacterial cells adapted to higher concentrations of colchicine and resulting into decrease in cell membrane thickness. More numbers of polyhydroxybutyrate (PHB) rich inclusion bodies were found inside the colchicine adapted cells and presence of higher amount of PHB, a carbon source for generation of redox potential, indicates that it might be responsible for activation of P450 BM-3 enzyme and plays significant role in colchicine demethylation. The presence of dense ribosome like bodies in colchicine adapted cells showed higher biosynthesis of P450 BM-3. Reduction in cell wall and cell membrane thickness, presence of more inclusion bodies and ribosome like masses in colchicine adapted cells were some of the key interlinked phenomena responsible for colchicine bioconversion. This is the first study which reports that colchicine demethylation process severely affects the structural and metabolic functions of the bacteria.