The induction of orientational instability and bivalent interlocking at meiosis

The administration of 40° C heat-treatments was found to induce bivalent orientational instability and interlocking at male meiosis in the locust Locusta migratoria. Only the longest members of the complement showed orientational instability and these usually possessed single distally sited chiasmata, with near-maximal intercentromeric distances. An effect on the stability of spindle fibre microtubule association, or attachment to the chromosome, is considered to be a possible explanation of the behaviour found. Bipolar orientation was generally achieved prior to anaphase I so that chromosome segregation was usually normal. Diamphitelic bivalents provided the most common exception to this rule. They sometimes lagged at anaphase, with the separation of half-bivalents and the production of structures indistinguishable from lagging univalents. The bivalent interlocking also involved the longest members of the complement. Most combinations of rod/rod, rod/ring and ring/ring types of interlocking were found. Usually only two bivalents were interlocked in any one cell, although occasionally three were found interlocked. All types appeared to involve an effect on the regulation of chromosome pairing, although at least one of the cells found showed interlocking caused by the metaphase orientational instability. In most cells, interlocked bivalents showed stable orientation and this usually involved the unipolar orientation of each bivalent's two centromeres. Such configurations provide concrete support for the importance of physical tension in the maintenance of metaphase orientational stability. They lead to double non-disjunction at anaphase I. Interlocked bivalents showed normal congression to a mid-equatorial position with no tendency for the re-adjustment of arm ratios to equalise centromere distances from the poles. This behaviour is discussed in relation to spindle fibre dynamics and it is concluded that no hypothesis of congression currently available can satisfactorily explain all that we know of the behaviour of univalents, bivalents, multivalents and interlocked bivalents.     

Genetic induction of bivalent interlocking in common wheat

The frequency of interlocking bivalents at first meiotic metaphase of common wheat Triticum aestivum L., which is normally very low, is significantly increased by raising the dosage (from two to three, four and six) of the Ph1 gene, located on the long arm of chromosome 5B (5BL). In several cells more than three bivalents were interlocked in one chain configuration indicating involvement of non-homoeologous bivalents. Plants with reduced dose (one or zero) of Ph1 also exhibited an increased frequency of interlocking but to a lesser extent than those with high gene dosage. However, chains of more than three interlocked bivalents were never observed in these plants, suggesting that with one or zero doses of Ph1 interlocking is restricted to homoeologous bivalents only. Chromosomal arm 5BS affected interlocking in an opposite manner to 5BL; namely, two and four doses of 5BS markedly reduced interlocking frequency. The modification in the frequency of interlocking bivalents by these genetic manipulations represents the first successful attempt to affect interlocking by genetic means. The results are explained on the basis of the hypothesis that this gene system controls somatic and premeiotic association of both homologous and homoeologous chromosomes.     

Meiosis in Mesostoma ehrenbergii ehrenbergii (Turbellaria,Rhabdocoela)

At metaphase I of meiosis in spermatocytes of Mesostoma ehrenbergii ehrenbergii [2n=10] three bivalents and four univalents form. The same two chromosome pairs always form the univalents. Analysis of metaphase I, anaphase I and metaphase II configurations in fixed testis material suggested that the distribution of the four univalents is not a random process but the correct segregation of one member of each pair to each pole is actively achieved before the end of metaphase I. In live preparations of testis material univalents were observed to move between the poles of metaphase I cells, eventually reaching the correct segregation. All cells observed to enter anaphase I had the correct segregation of univalents. It is proposed that the univalent movement during metaphase I is directed towards obtaining the correct segregation of univalents before the cells enter anaphase.     

Induced Bivalent Interlocking and the Course of Meiotic Chromosome Synapsis

WHEN chromosomes pair at meiosis the bivalents so formed do not normally interlock. Heat-treatments can, however, induce bivalent interlocking in the locust Locusta migratoria. Only the longest bivalents interlock and usually only two are found per cell; two “rod” bivalents, with single chiasmata, two “ring” bivalents, each with two or three chiasmata, or one “rod” and one “ring” bivalent (Fig. 1a, b and c). The nature of this interlocking and the metaphase orientational and congressional properties of interlocked bivalents are analysed in detail elsewhere¹.     

The spatial arrangement ofAllium triquetrum chain quadrivalents at metaphase I as viewed by confocal microscopy

We examined the three-dimensional arrangement of bivalents and, in particular, a chain of four chromosomes (chain quadrivalent) in the metaphase I spindle of pollen mother cells ofAllium triquetrum by confocal microscopy. Firstly, we show by optical sectioning and three-dimensional image reconstruction that the cooriented pairs of centromeres of all seven bivalents lie virtually parallel to each other in the metaphase I spindle, parallel to the long axis of the spindle. Secondly, we like-wise show that the four centromeres of the chain quadrivalent are aligned in the metaphase I spindle in, essentially, atwo-dimensional array, not in a three-dimensional array, as proposed by some other authors. This two-dimensionality has its basis, we argue, in the principle that poleward directed spindle forces minimise centromere-to-pole distances and therefore align pairs of centromeres connected to opposite poles most axially (vertically) in the spindle. These distances are minimised for the quadrivalent as a whole only when it lies in two dimensions, i.e. in aplane parallel to the spindle axis.     

Chromosomal interchange inEleutherine plicata Herb

Chromosome behaviour at metaphase I and anaphase I of meiosis inEleutherine plicata Herb. (2n=14) is studied. Cells with chromosome associations comprising an association of four long chromosomes, in addition to five bivalents were observed more frequently than those with seven bivalents. it is concluded that the ring of four is due to a segmental interchange between the two long non-homologous chromosome pairs. The ring of four at anaphase I showed delayed disjunction, bridge formation and irregular separation of chromosomes in a number of cells while the behaviour of the other bivalents was normal.     

Cytogenetics of Crotalaria VI. Chiasma frequency and position,and univalent behaviour in a (partially) asynaptic mutants of C. juncea

In Crotalaria juncea (n=8) a plant exhibiting partial asynapsis was isolated in the M₁ of a combined treatment of 50 kR gamma rays +0.2% EMS. The majority (48.14%) of PMCs at diplotene, diakinesis and metaphase I had 16 univalents. The bivalents in the asynaptic mutant were always rod-shaped with one terminal chiasma. In comparison, controls had on the average 7.08 ring bivalents. The asynapsis is genetically controlled, monofactorially recessive, and it is concluded that chromosome pairing is interrupted at a very early stage. There is a possible correlation between the number of bivalents and the arrangement of the univalents at metaphase I. When there were less than four bivalents, the univalents tended to be polar, and when there were more than four, the univalents were more equatorial in arrangement. The arrangement of univalents was random and apparently not influenced by the bivalents, when their number (4) was exactly half the zygotic number.     

Chiasma distribution in asynaptic Locusta migratoria

The formation of chiasmata in six full sib male partially asynaptic individuals of Locusta migratoria has been studied. The mean chiasma frequency per cell was 2.3 both at diplotene and metaphase I. Chiasmata tended to be distributed evenly among the bivalents. The frequency and distribution of the chiasmata in each type of bivalent (L, M, or S) depended on the level of asynapsis and on interference between the bivalents. Short bivalents were the most affected by interference, while M bivalents associated independently of L and S bivalent behaviour.     

Morphology of the axial structures in the neo-XY sex bivalent of Pycnogaster cucullata (Orthoptera) by silver impregnation

A simple method, using silver impregnation after 2 x SSC pretreatment allowed us to demonstrate axial structures (cores) in the metaphase I bivalents of the neo-XY race of Pycnogaster cucullata under bright-field microscopy. Axial structures can also be shown in DNA-depleted metaphase I bivalents, suggesting that DNA is not essential to demonstrate these elements. Specifically differentiated regions, which coincide with kinetochores, secondary constrictions (including the NOR), and chiasmata were also found. These regions have a characteristic morphology and therefore may be utilized for cytogenetic analysis. The simultaneous visualization of all these regions allowed us to establish their spatial relationships and hence the basic structural organization of the neo-XY sex bivalent in this species.     

Biosystematic studies on the genus Isoetes (Isoetaceae) in Japan. II. Meiotic behavior and REPRODUCTIVE MODE OF EACH CYTOTYPE

Meiotic behaviors and reproductive modes of Japanese Isoetes were studied. The hexaploid (2n = 66) and the octaploid (2n = 88) of I. japonica consistently formed 33 and 44 bivalents, respectively, at diakinesis and/or metaphase I in both micro- and megaspore mother cells. The tetraploid (2n = 44) of I. sinensis formed 22 bivalents and its hexaploid made 33 bivalents in both types of spore mother cells. At diakinesis and/or metaphase I of microspore mother cells in I. asiatica with 2n = 22, 11 bivalents were detected. Because behaviors of meiosis in all cytotypes mentioned above were quite regular and plants yielded normal-appearing spores, they should reproduce sexually. Aneuploids of I. japonica with 2n = 87 formed 43 bivalents and one univalent, and I. sinensis with 2n = 65 formed 32 bivalents and one univalent in microspore mother cells. Meiosis of both cytotypes was almost regular and yielded microspores of normal appearance. In the heptaploid (2n = 77) of I. japonica, a configuration of 22 bivalents and 33 univalents was detected in micro- and megaspore mother cells, and various irregularities were observed throughout the meiotic divisions. Therefore, the genomic formula of the heptaploid is symbolized as AABBCDE, and the heptaploid is a sterile F, hybrid between the hexaploid (AABBCC) and the octaploid (AABBDDEE) of I. japonica. Since diploid and even-numbered polyploids regularly formed bivalents and odd-numbered ones displayed irregularities, allopolyploidy should act as a significant speciation mechanism in this genus.     

Diploidisation ofLotus corniculatus L. (Fabaceae) by elimination of multivalents

Lotus corniculatus L. (Fabaceae) is a natural tetraploid of probably hybrid origin, which regularly forms bivalents at metaphase I of meiosis. Whole-mount surface-spreading of synaptonemal complexes (SCs) under the electron microscope reveals that diploidisation of this spccies is achieved not by exclusive pairing of homologues during meiotic prophase, but by the elimination of multivalents in favour of bivalents before metaphase I. Observations show that 43% of multivalents are eliminated between zygotene and pachytene, presumably by dissolution and reassembly of SCs between homologous chromosomes. A further 63% are eliminated between pachytene and diakinesis, with a commensurate increase in the number of univalents. Elimination ensures few multivalents reach first metaphase and effectively diploidises this tetraploid.     

THE QUESTION OF POST-REDUCTION IN SPHAGNUM

The 19 spatially distinct chromosomal units at first meiotic metaphase in sporophytically diploid species of Sphagnum have usually been considered to be bivalents, but one investigator (Sorsa, 1956) has interpreted them as chromosomes from dissociated bivalents and meiosis as post-reductional. The present studies on diploid S. squarrosum (Pers.) Crome establish the chromosome number on the basis of the following evidence: there are in addition to m-chromosomes, 19 pairs of chromosomes in early prophase, 19 bivalents at diakinesis, 19 chromosomes in each of the two sets at second metaphase, 19 daughter chromosomes in each of the four sets at late second anaphase, and 19 chromosomes in gametophytic mitoses. The 19 bodies at first meiotic metaphase in diploid species are true bivalents in loose secondary association, which has led to their erroneous interpretation as chromosomes of dissociated bivalents. The gametic chromosome number in sporophytically diploid Sphagnum is therefore, without doubt, n = 19, and this evidence negates the claim for post-reduction in Sphagnum.     

CHROMOSOME MICROMANIPULATION : III. Spindle Fiber Tension and the Reorientation of Mal-Oriented Chromosomes

Kinetochore reorientation is the critical process ensuring normal chromosome distribution. Reorientation has been studied in living grasshopper spermatocytes, in which bivalents with both chromosomes oriented to the same pole (unipolar orientation) occur but are unstable: sooner or later one chromosome reorients, the stable, bipolar orientation results, and normal anaphase segregation to opposite poles follows. One possible source of stability in bipolar orientations is the normal spindle forces toward opposite poles, which slightly stretch the bivalent. This tension is lacking in unipolar orientations because all the chromosomal spindle fibers and spindle forces are directed toward one pole. The possible role of tension has been tested directly by micromanipulation of bivalents in unipolar orientation to artificially create the missing tension. Without exception, such bivalents never reorient before the tension is released; a total time "under tension" of over 5 hr has been accumulated in experiments on eight bivalents in eight cells. In control experiments these same bivalents reoriented from a unipolar orientation within 16 min, on the average, in the absence of tension. Controlled reorientation and chromosome segregation can be explained from the results of these and related experiments.     

Metaphase I bound arms frequency and genome analysis in wheat-Aegilops hybrids

Summary Meiotic associations of different wheat-Aegilops variabilis and wheat-Ae. kotschyi hybrid combinations with low and high homoeologous pairing were analyzed at metaphase I. Five types of pairing involving wheat and Aegilops genomes were identified by using C-banding. A genotype that seems to promote homoeologous pairing has been found in Ae. variabilis var. cylindrostachys. Its effect is detectable in the low pairing hybrids but not in the high ones. Pairing affinity has been analyzed on the basis of metaphase I associations in the low and high homoeologous pairing hybrids, and in bivalents and multivalents in the high pairing hybrids. The results indicate that the amount of bound arms of each type of identifiable association relative to the total associations formed (relative contribution) was not maintained, either between the different levels of pairing (low and high) or between different meiotic configurations (bivalents and multivalents). These findings seem to indicate that quantifications of genomic relationships based on the amount of chromosome pairing at metaphase I must be carefully done in this type of hybrid combinations.     

The fate of multivalents during meiotic prophase in the hybrid Gibasis consobrina x G. karwinskyana Rafin. (Commelinaceae)

The chromosomes of the two closely related diploid species, Gibasis consobrina and G. karwinskyana (Commelinaceae; 2n=2x=10), are morphologically alike, yet form few chiasmate associations at metaphase I in the f₁ hybrid. During meiotic prophase, however, synaptonemal complexes join the majority of the chromosomes of the complement in complex multiple pairing configurations. The F₁ hybrid between different tetraploid genotypes of the same two species similarly forms multivalents during meiotic prophase, which are subsequently eliminated in favour of strictly homologous bivalents before metaphase I. One quadrivalent comprising interchange chromosomes inherited from one of the parents, usually persists to first metaphase. Evidently the resolution of multivalents to bivalents at first metaphase, which accounts for diploidisation, is not attributable to the elimination of multivalents per se, but of multivalents comprising chromosomes of limited homology.     

Cytogenetics of chromosome rearrangements in Tribolium castaneum.

The karyotype of the red flour beetle, Tribolium castaneum, was reexamined and improved by restriction enzyme banding with HpaII. After this treatment, each of the 10 chromosomes were identified in spermatogonial metaphase cells and 3 of the 8 autosomal bivalents and the XY pair were identified in spermatocyte metaphase I nuclei. Based on centromere position, relative length, and banding pattern, probable correlations between some of the mitotic chromosomes and some of the metaphase I bivalents were ascertained. Thus improved, the karyotypes of beetles harboring genetically defined translocations were investigated. Spermatocyte metaphase I nuclei were most informative, as normal chromosome pairing was visibly disrupted by rearrangements. Bivalents associated with each rearrangement were identified. Results demonstrated that each of the five best defined T. castaneum linkage groups corresponds to a different chromosome and established correspondence between bivalents and linkage groups 1-4. The relevance of these findings is discussed with regard to Tribolium genetics and evolution.     

mei-41 is required for precocious anaphase in Drosophila females

This paper reports on a new role for mei-41 in cell cycle control during meiosis. This function is revealed by the requirement of mei-41 for the precocious anaphase observed in crossover-defective mutants. Normally in Drosophila oocytes, tension on the meiotic spindle causes a metaphase I arrest. This tension results because crossovers, and the resulting chiasmata, hold homologs together that are being pulled by kinetochore microtobules toward opposite spindle poles. In the absence of tension, such as in a recombination-defective mutant, metaphase arrest is not observed and meiosis proceeds through the two divisions. Here we show that in some recombination-defective mutants, the precocious anaphase requires the mei-41 gene product. For example, metaphase arrest is not observed in mei-218 mutants because of the severe reduction in crossing over. In mei-41 mei-218 double mutants, however, metaphase arrest was restored. The effect of mei-41 is dependent on double-strand break formation. Thus, in mutants that fail to initiate meiotic recombination the absence of mei-41 has no effect. Received: 15 October 1999; in revised form: 9 December 1999 / Accepted: 13 December 1999     

Bivalent behavior in Drosophila melanogaster males containing the In(1)sc 4Lsc8RX chromosome

The sex chromosome bivalent was examined in Drosophila melanogaster males possessing the In(1)sc ^4Lsc^8R X chromosome. Three-dimensional reconstructions from electron micrographs of serially cut thin sections were made. A large proportion of the kinetochores of In(1)sc ^4Lsc^8R/Y bivalents did not face opposite poles during metaphase I and anaphase I. This suggests that In(1)sc ^4Lsc^8R/Y bivalents may have difficulty achieving bipolar stability. Delay in achieving bipolar stability could contribute to the nondisjunctional behavior found in In(1)sc ^4Lsc^8R/Y males.     

Relationship between pachytene synapsis, metaphase I associations, and transmission of 2B and 4B chromosomes in rye.

2B rye plants selected for high (H) or low (L) B transmission rate were studied at pachytene and metaphase I of meiosis to determine the relationship between synapsis, bivalents at metaphase I, and B transmission rate. The results show that the 2 B chromosomes (Bs) form bivalents at pachytene in both the H and L lines, whereas the frequency of bivalents at metaphase I is much higher in the H than in the L line. This demonstrates that B transmission is mainly related to the proper association of Bs at metaphase I, as well as that synapsis of the 2 Bs in the L line is normal, but the bivalent is not consolidated by a chiasma in most cases. Crosses were made between 2B plants of the H and L lines in all combinations (H x H, H x L, L x H, and L x L) to obtain 4B plants. Similarly, bivalent formation at pachytene and metaphase I was studied. The results show that 4B plants of the H x H and L x L classes differ significantly at pachytene and metaphase I since the former forms more bivalents. The heterozygous 4 Bs of the H x L and L x H classes show intermediate values. The relation H x H > H x L > L x H > L x L was consistently found for the variables transmission rate, bivalents at pachytene, bivalents at metaphase I, and B mean chiasma frequency. A maternal effect was also found. Our data suggest that there are two separate mechanisms acting upon synapsis and chiasma formation in H and L B chromosomes: (i) there is variable efficiency of the control of synapsis at early stages of meiosis; and (ii) there is variable efficiency of the control of the number of chiasmata.     

Ultrastructure of meiosis in the hollyhock rust fungus,Puccinia malvacearum II. Metaphase I—Telophase I

Summary The three-dimensional structure of the spindle pole body (SPB) and meiotic spindle during early metaphase I through telophase I inPuccinia malvacearum is analyzed ultrastructurally from serial sections. During early metaphase I the spindle rotates from the perpendicular to a position oblique to the longitudinal axis and parallel to the sagittal plane of the cell. Tubular cisternae are present within the spindle at this stage. The half middle piece (MP) subtends a collateral disc (co-disc) which is inserted eccentrically within each SPB. The SPB, co-disc and half MP at opposite poles are in mirror image. During the transition from early metaphase I to full metaphase I, the spindle orients parallel to the lateral wall of the promycelium and the half MPs are lost. The co-discs partially detach from each discoid SPB and maintain this relation until the end of interphase I. Co-discs become further differentiated as they attach to the subtending sheath-like extension of the nuclear envelope previously occupied by the half MPs. Microvesicles within the nucleoplasm are specific to mid metaphase I. A metaphase plate is absent. The 14 bivalents, which are directly connected to each polar SPB by 2 to 3 kinetochore MTs, are spread over nearly the entire length of the central spindle. The first anaphasic movement involves asynchronous shortening of the kinetochore MTs while the second consists of extensive pole-to-pole elongation. Astral MTs first appear at early metaphase I and become most numerous at anaphase I. An intact nuclear envelope constricts against the central spindle at either end of the interzonal region. Concurrently, centripetal growth of the nuclear envelope under each SPB results in their gradual externalization by the end of telophase I. The sibling nuclei are cut off by constriction of the nuclear envelope at either end of the interzonal region. These meiotic stages inP. malvacearum are compared with those in other basidiomycetes and ascomycetes.