The Role of the Ameiotic1 Gene in the Initiation of Meiosis and in Subsequent Meiotic Events in Maize

Understanding the initiation of meiosis and the relationship of this event with other key cytogenetic processes are major goals in studying the genetic control of meiosis in higher plants. Our genetic and structural analysis of two mutant alleles of the ameiotic1 gene (am1 and am1-praI) suggest that this locus plays an essential role in the initiation of meiosis in maize. The product of the ameiotic1 gene affects an earlier stage in the meiotic sequence than any other known gene in maize and is important for the irreversible commitment of cells to meiosis and for crucial events marking the passage from premeiotic interphase into prophase I including chromosome synapsis. It appears that the period of ameiotic1 gene function in meiosis at a minimum covers the interval from some point during premeiotic interphase until the early zygotene stage of meiosis. To study the interaction of genes in the progression of meiosis, several double meiotic mutants were constructed. In these double mutants (i) the ameiotic1 mutant allele was brought together with the meiotic mutation (afd1) responsible for the fixation of centromeres in meiosis; and with the mutant alleles of the three meiotic genes that control homologous chromosome segregation (dv1, ms43 and ms28), which impair microtubule organizing center organization, the orientation of the spindle fiber apparatus, and the depolymerization of spindle filaments after the first meiotic division, respectively; (ii) the afd1 mutation was combined with two mutations (dsy1 and as1) affecting homologous pairing; (iii) the ms43 mutation was combined with the as1, the ms28 and the dv1 mutations; and (iv) the ms28 mutation was combined with the dv1 mutation and the ms4 (polymitotic1) mutations. An analysis of gene interaction in the double mutants led us to conclude that the ameiotic1 gene is epistatic over the afd1, the dv1, the ms43 and the ms28 genes but the significance of this relationship requires further analysis. The afd gene appears to function from premeiotic interphase throughout the first meiotic division, but it is likely that its function begins after the start of the ameiotic1 gene expression. The afd1 gene is epistatic over the two synaptic mutations dsy1 and as1 and also over the dv1 mutation. The new ameiotic*-485 and leptotene arrest*-487 mutations isolated from an active ROBERTSON's Mutator stocks take part in the control of the initiation of meiosis.     

Chromosomes associate premeiotically and in xylem vessel cells via their telomeres and centromeres in diploid rice (<Emphasis Type="Italic">Oryza sativa</Emphasis>)

Studies of the meiosis of diploid plants such as Arabidopsis, maize and diploid progenitors of wheat have revealed no premeiotic association of chromosomes. Premeiotic and somatic association of chromosomes has only been previously observed in the anther tissues and xylem vessel cells of developing roots in polyploid plants such as hexaploid and tetraploid wheat, polyploid relatives of wheat and artificial polyploids made from the progenitor diploids of wheat. This suggested that this association was confined specifically to polyploids or was induced by polyploidy. However, we developed procedures for in situ hybridization on structurally well-preserved tissue sections of rice, and analysed two diploid rice species (Oryza sativa and O. punctata). Contrary to expectation, this has revealed that centromeres and telomeres also associate both in the xylem vessel cells of developing root and in undifferentiated anther cells in these diploids. However, in contrast to wheat and related polyploids, where the initial association in undifferentiated anthers is between either non-homologous or related chromosomes, and not homologous chromosomes, the initial association of rice chromosomes seems to be between homologues. Thus, in contrast to the diploid dicot model Arabidopsis, meiotic studies on the diploid model cereal, rice, will now need to take into account the effects of premeiotic chromosome association.Pilar Prieto and Ana Paula Santos are joint first authors.     

Effects of several meiotic mutations on female meiosis in maize

A modified enzyme digestion technique of ovary isolation followed by staining and squash preparation has allowed us to observe female meiosis in normal maize meiotically dividing megaspore mother cells (MMCs). The first meiotic division in megasporogenesis of maize is not distinguishable from that in mi-crosporogenesis. The second female meiotic division is characterized as follows: (1) the two products of the first meiotic division do not simultaneously enter into the second meiotic division; as a rule, the chalazal-most cell enters division earlier than the micropylar one, (2) often the second of the two products does not proceed with meiosis, but degenerates, and (3) only a single haploid meiotic product of the tetrad remains alive, and this cell proceeds with three rounds of mitoses without any intervening cell wall formation to produce the eight-nucleate embryo sac. This technique has allowed us to study the effects of five meiotic mutations (aml, aml-pral, afdl, dsy *-9101, and dvl) on female meiosis in maize. The effects of the two alleles of the aml gene (aml and aml-pral) and of the afdl and dsy *-9101mutations are the same in both male and female meiosis. The aml allele prevents the entrance of MMCs into meiosis and meiosis is replaced by mitosis; the aml-pral permits MMCs to enter into meiosis, but their progress is stopped at early prophase I stages. The afdl gene is responsible for substitution of the first meiotic (reductional) division by an equational division including the segregation of sister chromatid centromeres at anaphase I. The dsy * -9101 gene exhibits abnormal chromosome pairing; paired homologous chromosomes are visible at pachytene, but only univalents are observed at diakinesis and metaphase I stages. These mutation specific patterns of abnormal meiosis are responsible for the bisexual sterility of these meiotic mutants. The abnormal divergent shape of the spindle apparatus and the resulting abnormal segregation of homologous chromosomes observed in micro-sporogenesis in plants homozygous for the dv1 mutation have not been found in meiosis of megasporogenesis. Only male sterility is induced by the dv1 gene in the homozygous condition. © 1993 Wiley-Liss, Inc.     

The Rice OsRad21-4, an Orthologue of Yeast Rec8 Protein,is Required for Efficient Meiosis

In yeast, Rad21/Scc1 and its meiotic variant Rec8 are key players in the establishment and subsequent dissolution of sister chromatid cohesion for mitosis and meiosis, respectively, which are essential for chromosome segregation. Unlike yeast, our identification revealed that the rice genome has 4 RAD21-like genes that share lower than 21% identity at polypeptide levels, and each is present as a single copy in this genome. Here we describe our analysis of the function of OsRAD21-4 by RNAi. Western blot analyses indicated that the protein was most abundant in young flowers and less in leaves and buds but absent in roots. In flowers, the expression was further defined to premeiotic pollen mother cells (PMCs) and meiotic PMCs of anthers. Meiotic chromosome behaviors were monitored from male meiocytes of OsRAD21-4-deficient lines mediated by RNAi. The male meiocytes showed multiple aberrant events at meiotic prophase I, including over-condensation of chromosomes, precocious segregation of homologues and chromosome fragmentation. Fluorescence in situ hybridization experiments revealed that the deficient lines were defective in homologous pairing and cohesion at sister chromatid arms. These defects resulted in unequal chromosome segregation and aberrant spore generation. These observations suggest that OsRad21-4 is essential for efficient meiosis.     

mei-2, a mutagen-sensitive mutant of Neurospora defective in chromosome pairing and meiotic recombination

Summary A Neurospora crassa mutation, mei-2, affecting meiosis and mutagen sensitivity, was characterized for its effect on meiotic recombination and chromosome pairing. Results from homozygous mei-2 crosses involving distant markers on the same chromosome demonstrated a drastic reduction in meiotic recombination. However, mitotic recombination continued to occur. Cytological observations indicated that pairing of homologous chromosomes in zygotene was greatly reduced or absent, resulting in aberrant segregation at anaphase I and often at subsequent divisions as well. The few mature ascospores produced were frequently disomic for one or more chromosomes.     

Progress in studies of ZW10, a proper chromosome segregation protein

The conserved protein ZW10 is found in various organisms. It is localized on the kinetochores or spindle microtubules during cell division. ZW10 regulates not only the segregation of homologous chromosomes, each consisting of attached sister chromatids (during the first meiotic division), but also the separation of individual chromatids (during mitosis and the second meiotic division). ZW10 is required for proper chromosome segregation during both mitosis and meiosis. The effects of zwl0 mutations are similar for both equational and reductional divisions, giving rise to anaphases with lagging chromosomes and/or unequal numbers of chromosomes at the two poles. The localization of ZW10 is similar during mitosis, meiosis I, and meiosis II. In interphase the distribution of ZW10 changes; it is localized in the endoplasmic reticulum, Golgi apparatus, and in the cytosol and is involved in membrane trafficking between the endoplasmic reticulum and Golgi apparatus. ZW10 forms a subcomplex with RINT-1 and p31 which are involved in a larger complex comprising syntaxin 18, an endoplasmic reticulum-localized t-SNARE that is implicated in membrane trafficking. The text was submitted by the authors in English.     

Meiosis readiness in Lilium longiflorum “Croft”

It is proposed that anthers of Lilium longiflorum Croft approaching the end of premeiotic mitosis reach a state described as meiosis readiness after which cells in premeiotic prophase are unable to complete a mitotic division but despiralize to interphase and enter a meiotic division. Many of the laggard premeiotic cells begin despiralization before reaching an extremely contracted state of late prophase. Premeiotic despiralization is not, therefore, attributed to a deficiency in metaphase but to an inability of these cells to complete prophase. Premeiotic despiralization appears to be preceded by a slowing-down of prophase development. There is variation among anthers and anther regions in the onset of prophase retardation and meiosis readiness. It is suggested that meiosis readiness depends upon a gradual accumulation of meiosis-inducing substances in the cytoplasm of the premeiotic cells. It has not been determined whether the cells that undergo premeiotic despiralization give rise to the giant microsporocytes with shattered chromosomes observed at late prophase of meiosis.     

Cytological studies of the two-spotted spider mite Tetranychus urticae Koch (Tetranychidae,trombidiformes). I: Meiosis in eggs

Meiosis in eggs of Tetranychus urticae Koch is described. The two maturation divisions result in (a) a haploid female pronucleus consisting of three karyomeres; — (b) a divided first polar body in which the chromosomes change into karyomeres; — (c) a second polar body, entering a new mitosis which is blocked in metaphase. Irradiation of adult females produced chromosome fragments in the meiotic divisions. The fragments behave as intact chromosomes which proves that during meiosis a diffuse kinetochore is present. The meiotic divisions show the cytologically characteristic features of an inverted meiosis. The presence of such a meiosis is corroborated by observations on eggs heterozygous for chromosome mutations. In both maturation divisions the chromosomes are orientated equatorially. It is suggested that the equatorial orientation is brought about by chiasmata having terminalized to both ends of the dyads. It is argued that in organisms with holokinetic chromosomes during meiosis an axial orientation of the bivalents does not necessarily imply a normal meiosis but can also imply an inverted meiosis.This work was carried out with financial aid of the Institute for Atomic Sciences in Agriculture and the Ministry of Public Health and Environment.     

The cytological basis for reproductive variability in the Anoetidae (Sarcoptiformes: Acari)

Meiosis is described in a thelytokous strain of the anoetid mite Histiostoma feroniarum (Dufour) and in both sexes of the arrhenotokous strain of this species. Oogenesis in the thelytokous strain is accomplished by ameiotic mitosis with only one pseudo-maturation division. During this division one or more chromosomes may move to the poles precociously and while in this position can be mistaken for centrioles. Fourteen chromosomes are found at metaphase of the pseudo-maturation division and in cleaving eggs of this strain. In the arrhenotokous strain, male meiosis consists of a single mitotic division. Oogenesis is regular and 7 bivalents are observed at the first maturation division. Metaphases of the first cleavage division in fertilized eggs show 14 chromosomes and 7 chromosomes in unfertilized eggs.It is postulated that the thelytokous strain has arisen from the arrhenotokous strain. This assumption is in agreement with that suggested for several insect species previously reported. The evolution in the Acari and the variability in the modes of reproduction in this suborder are discussed in light of the findings in this paper on the Anoetidae.     

Chromosome engineering: power tools for plant genetics

The term "chromosome engineering" describes technologies in which chromosomes are manipulated to change their mode of genetic inheritance. This review examines recent innovations in chromosome engineering that promise to greatly increase the efficiency of plant breeding. Haploid Arabidopsis thaliana have been produced by altering the kinetochore protein CENH3, yielding instant homozygous lines. Haploid production will facilitate reverse breeding, a method that downregulates recombination to ensure progeny contain intact parental chromosomes. Another chromosome engineering success is the conversion of meiosis into mitosis, which produces diploid gametes that are clones of the parent plant. This is a key step in apomixis (asexual reproduction through seeds) and could help to preserve hybrid vigor in the future. New homologous recombination methods in plants will potentiate many chromosome engineering applications.     

Differential elongation of autosomal pachytene bivalents related to their DNA content in human spermatocytes

The establishment of the complete karyotype of human pachytene spermatocytes reveals differences in stretching of chromosomes between meiosis and mitosis. Bivalents or specific regions of bivalents which exhibit many R-bands are particularly elongated. In mitotic chromosomes, the DNA contained in such bands is known to be early replicating. The study of variations in the total length and the centromeric index of bivalent 1 suggests that differential elongation of pachytene bivalents is a premeiotic event, taking place during the last DNA replication.     

Choose your partner: Chromosome pairing in yeast meiosis

Premeiotic association of homologous chromosomes in the yeast, Saccharomyces cerevisiae has been shown, by means of fluorescent in situ hybridization (FISH)^(1,2). Time course and mutant studies show that the premeiotic associations are disrupted upon entry into meiosis, to be reestablished shortly before synapsis. The data are consistent with a model in which multiple, unstable interactions bring homologues together, prior to stable joining by recombination⁽³⁾.     

Potential Roles for Centromere Pairing in Meiotic Chromosome Segregation

One of the key differences between mitosis and meiosis is the necessity for exchange between homologous chromosomes. Crossing-over between homologous chromosomes is essential for proper meiotic chromosome segregation in most organisms, serving the purpose of linking chromosomes to their homologous partners until they segregate from one another at anaphase I. In several organisms it has been shown that occasional pairs of chromosomes that have failed to experience exchange segregate with reduced fidelity compared to exchange chromosomes, but do not segregate randomly. Such observations support the notion that there are mechanisms, beyond exchange, that contribute to meiotic segregation fidelity. Recent findings indicate that active centromere pairing is important for proper kinetochore orientation and consequently, segregation of non-exchange chromosomes. Here we discuss the implications of these findings for the behavior of meiotic chromosomes.     

PP2ACdc55 is required for multiple events during meiosis I

Protein phosphatase 2A (PP2A) is a heterotrimer consisting of A and B regulatory subunits and a C catalytic subunit. PP2A regulates mitotic cell events that include the cell cycle, nutrient sensing, p53 stability and various mitogenic signals. The role of PP2A during meiosis is less understood. We explored the role of Saccharomyces cerevisiae PP2A during meiosis. We show a PP2A^Cdc55 containing the human B/55 family B subunit ortholog, Cdc55, is required for progression through meiosis I. Mutant cells lacking Cdc55 remain mononucleated. They harbor meiotic gene expression, premeiotic DNA replication, homologous recombination and spindle pole body (SPB) defects. They initiate but do not complete replication and are defective in performing intergenic homologous recombination. Bypass alleles, which allow cells defective in recombination to finish meiosis, do not suppress the meiosis I defect. cdc55 cells arrest with a single SPB lacking microtubules, or duplicated but not separated SBPs containing microtubules. Finally, the premeiotic replication defect is suppressed by loss of Rad9 checkpoint function. We conclude PP2A^Cdc55 is required for the proper temporal initiation of multiple meiotic events and/or monitors these events to ensure their fidelity.     

The cytology of Brachycome

Over 1,000 plants of B. dichromosomatica have been counted. 10% of these carried one, two or three B chromosomes. The B chromosome is large, and though it is not heteropycnotic it condenses precociously at mitosis. It behaves regularly at mitosis, and when two are present they pair regularly at meiosis. Non-disjunction and preferential distribution of the B to the generative nucleus occurs at the first pollen grain mitosis, with very high frequency. This is corroborated by data from crosses, which also indicate that the transmission of the B through the female gamete is normal. — The frequency of B chromosomes in marginal populations of var. dichromosomatica is significantly higher than in central populations. In one population of var. alba the frequency of Bs increased significantly after two very dry seasons. It is suggested that both these cases of increased frequency were in response to a selective advantage of plants with Bs under arid conditions. — Plants with one B chromosome appear to be less fit than plants with 2 Bs. The combination of the calculated effects of the nondisjunction mechanism and the inferred relative fitness of the 0B, 1B and 2B plants, provides a reasonable explanation of the observed frequencies of the 0B, 1B and 2B plants in the populations studied.     

Holding chromatids together to ensure they go their separate ways

Association between sister chromatids is essential for their attachment and segregation to opposite poles of the spindle in mitosis and meiosis II. Sister-chromatid cohesion is also likely to be involved in linking homologous chromosomes together in meiosis I. Cytological observations provide evidence that attachment between sister chromatids is different in meiosis and mitosis and suggest that cohesion between the chromatid arms may differ mechanistically from that at the centromere. The physical nature of cohesion is addressed, and proteins that are candidates for holding sister chromatids together are discussed. Dissolution of sister-chromatid cohesion must be regulated precisely, and potential mechanisms to release cohesion are presented.     

THE ASSOCIATION OF A SINGLE B-CHROMOSOME WITH MALE STERILITY IN PLANTAGO CORONOPUS

Paliwal , Ripsudan L. (B. R. College, Agra, India.), and Beal B. Hyde . The association of a single B-chromosome with male sterility in Plantago coronopus. Amer. Jour. Bot. 46(6): 460–466. Illus. 1959.—Two species of Plantago showing male sterility have been studied cytogenetically. In P. ovata (n=4) the sterility appears to be cytoplasmic. In P. coronopus (n=5) all male-sterile plants contain a single extra chromosome which is largely heterochromatic, shorter, and not homologous with any of the other chromosomes. No male-fertile plants contain this B-chromosome. Meiosis is regular in the male-sterile lines. The accessory chromosome usually does not divide and moves to one pole at the first division of meiosis and divides regularly in the second division. Degeneration of all microspores occurs before pollen mitosis. Male-sterile plants are apomictic, but whether or not male-fertile plants are also apomictic has not yet been determined.     

Control of the meiotic cell division program in plants

While the question of why organisms reproduce sexually is still a matter of controversy, it is clear that the foundation of sexual reproduction is the formation of gametes with half the genomic DNA content of a somatic cell. This reduction in genomic content is accomplished through meiosis that, in contrast to mitosis, comprises two subsequent chromosome segregation steps without an intervening S phase. In addition, meiosis generates new allele combinations through the compilation of new sets of homologous chromosomes and the reciprocal exchange of chromatid segments between homologues. Progression through meiosis relies on many of the same, or at least homologous, cell cycle regulators that act in mitosis, e.g., cyclin-dependent kinases and the anaphase-promoting complex/cyclosome. However, these mitotic control factors are often differentially regulated in meiosis. In addition, several meiosis-specific cell cycle genes have been identified. We here review the increasing knowledge on meiotic cell cycle control in plants. Interestingly, plants appear to have relaxed cell cycle checkpoints in meiosis in comparison with animals and yeast and many cell cycle mutants are viable. This makes plants powerful models to study meiotic progression and allows unique modifications to their meiotic program to develop new plant-breeding strategies.     

Initiation of Meiotic Recombination in Ustilago maydis

A central feature of meiosis is the pairing and recombination of homologous chromosomes. Ustilago maydis, a biotrophic fungus that parasitizes maize, has long been utilized as an experimental system for studying recombination, but it has not been clear when in the life cycle meiotic recombination initiates. U. maydis forms dormant diploid teliospores as the end product of the infection process. Upon germination, teliospores complete meiosis to produce four haploid basidiospores. Here we asked whether the meiotic process begins when teliospores germinate or at an earlier stage in development. When teliospores homozygous for a cdc45 mutation temperature sensitive for DNA synthesis were germinated at the restrictive temperature, four nuclei became visible. This implies that teliospores have already undergone premeiotic DNA synthesis and suggests that meiotic recombination initiates at a stage of infection before teliospores mature. Determination of homologous recombination in plant tissue infected with U. maydis strains heteroallelic for the nar1 gene revealed that Nar⁺ recombinants were produced at a stage before teliospore maturation. Teliospores obtained from a spo11Δ cross were still able to germinate but the process was highly disturbed and the meiotic products were imbalanced in chromosomal complement. These results show that in U. maydis, homologous recombination initiates during the infection process and that meiosis can proceed even in the absence of Spo11, but with loss of genomic integrity.     

Inferences from genetical evidence on the course of meiotic chromosome pairing in plants.

Meiotic chromosome pairing is a process that is amenable to genetic and experimental analysis. The combined use of these two approaches allows for the process to be dissected into several finite periods of time in which the developmental stages of pairing can be precisely located. Evidence is now available, in particular in plants, that shows that the pairing of homologous chromosomes, as observed at metaphase I, is affected by events occurring as early as the last premeiotic mitosis; and that the maintenance of this early determined state is subsequently maintained by constituents (presumably proteins) that are sensitive to either colchicine, temperature or gene control. A critical assessment of this evidence in wheat and a comparison of the process of pairing in wheat with the course of meiotic pairing in other plants and animals is presented.