Male and female meiosis in diploid and colchitetraploid Phlox drummondii Hook. (Polemoniaceae)

Comparative studies on male and female meiosis in diploid (2n = 2x = 14) and colchitetraploid (2N = 4x = 28) ornamental Phlox drummondii reveal higher chiasma frequency in embryo-sac mother cells as compared with pollen mother cells in the diploid, and significant differences in the pairing properties of chromosomes in the pollen mother cells and embryo-sac mother cells of the colchitetraploid. Male meiosis in the colchicine-induced autotetraploid was abnormal with 51.70% of chromosomes associated in quadrivalents and tridents in 96% of the cells. This was followed by discordant anaphase I disjunction. In the female meiocytes the chromosomes formed 14 bivalents in 86.66% of the cells. Quadrivalents (1–3) appeared only in 13.34% of the cells. It is concluded that meiosis in male and female cells of the colchitetraploid is governed and regulated by different controlling systems of gene(s).     

Female meiosis in wild-type Arabidopsis thaliana and in two meiotic mutants

Female meiosis in Arabidopsis has been analysed cytogenetically using an adaptation of a technique previously applied to male meiosis. Meiotic progression was closely correlated with stages of floral development, including the length and morphology of the gynoecium. Meiosis in embryo sac mother cells (EMCs) occurs later in development than male meiosis, in gynoecia that range in size between 0.3 and 0.8 mm. The earliest stages in EMCs coincide with the second division to tetrad stages in pollen mother cells. However, the details of meiotic chromosome behaviour in EMCs correspond closely to the observations we have previously made in male meiosis. In addition, BrdU labelling coupled with an immunolocalisation detection system was used to mark the S phase in cells preceding their entry into prophase I. These techniques allow female meiotic stages of Arabidopsis to be analysed in detail, from the S-phase through to the tetrad stage, and are shown to be equally applicable to the analysis of female meiosis in meiotic mutants. Received: 3 April 2000 / Revision accepted: 2 August 2000     

Cytological studies of ameiotic and normal maize with reference to premeiotic pairing

This study reports cytological observations in maize plants homozygous for the recessive am allele which suppresses meiosis in both male and female meiocytes. The ultimate premeiotic mitosis in anthers from ameiotic plants is normal, but the resulting nuclei do not undergo meiosis. Instead, a synchronized mitosis occurs after which the cells degenerate. No evidence was found for a non-random association of homologous chromosomes in the premeiotic or ameiotic mitoses of homozygous ameiotic plants or in the premeiotic mitosis of normal sibs. These observations are in agreement with the classical view that synapsis of homologous chromosomes does not occur until zygotene.     

AtSPO11-1 is necessary for efficient meiotic recombination in plants

The Saccharomyces cerevisiae Spo11 protein catalyses DNA double-strand breaks (DSBs) that initiate meiotic recombination. The model plant Arabidopsis thaliana possesses at least three SPO11 homologues. T-DNA and ethyl-methane sulfonate mutagenesis allowed us to show that meiotic progression is altered in plants in which the AtSPO11-1 gene is disrupted. Both male and female meiocytes formed very few bivalents. Furthermore, no fully synapsed chromosomes were observed during prophase I. Later, in meiosis I, we observed that chromosomes segregated randomly, leading to the production of a large proportion of non-functional gametes. These meiotic aberrations were associated with a drastic reduction in meiotic recombination. Thus, our data show that initiation of meiotic recombination by SPO11- induced DSBs is a mechanism conserved in plants. Furthermore, unlike Drosophila and Caenorhabditis elegans, but like fungi, SPO11 is necessary for normal synapsis in plants.     

The novel gene HOMOLOGOUS PAIRING ABERRATION IN RICE MEIOSIS1 of rice encodes a putative coiled-coil protein required for homologous chromosome pairing in meiosis

We have identified and characterized a novel gene, PAIR1 (HOMOLOGOUS PAIRING ABERRATION IN RICE MEIOSIS1), required for homologous chromosome pairing and cytokinesis in male and female meiocytes of rice (Oryza sativa). The pair1 mutation, tagged by the endogenous retrotransposon Tos17, exhibited meiosis-specific defects and resulted in complete sterility in male and female gametes. The PAIR1 gene encodes a 492-amino acid protein, which contains putative coiled-coil motifs in the middle, two basic regions at both termini, and a potential nuclear localization signal at the C terminus. Expression of the PAIR1 gene was detected in the early stages of flower development, in which the majority of the sporocytes had not entered meiosis. During prophase I of the pair1 meiocyte, all the chromosomes became entangled to form a compact sphere adhered to a nucleolus, and homologous pairing failed. At anaphase I and telophase I, chromosome nondisjunction and degenerated spindle formation resulted in multiple uneven spore production. However, chromosomal fragmentation frequent in plant meiotic mutants was never observed in all of the pair1 meiocytes. These observations clarify that the PAIR1 protein plays an essential role in establishment of homologous chromosome pairing in rice meiosis.     

Identification and analysis of DYAD: a gene required for meiotic chromosome organisation and female meiotic progression in Arabidopsis

The dyad mutant of Arabidopsis was previously identified as being defective in female meiosis. We report here the analysis of the DYAD gene. In ovules and anthers DYAD RNA is detected specifically in female and male meiocytes respectively, in premeiotic interphase/meiotic prophase. Analysis of chromosome spreads in female meiocytes showed that in the mutant, chromosomes did not undergo synapsis and formed ten univalents instead of five bivalents. Unlike mutations in AtDMC1 and AtSPO11 which also affect bivalent formation as the univalent chromosomes segregate randomly, the dyad univalents formed an ordered metaphase plate and underwent an equational division. This suggests a requirement for DYAD for chromosome synapsis and centromere configuration in female meiosis. The dyad mutant showed increased and persistent expression of a meiosis-specific marker, pAtDMC1::GUS during female meiosis, indicative of defective meiotic progression. The sequence of the putative protein encoded by DYAD did not reveal strong similarity to other proteins. DYAD is therefore likely to encode a novel protein required for meiotic chromosome organisation and female meiotic progression.     

A puromycin-sensitive aminopeptidase is essential for meiosis in Arabidopsis thaliana

Puromycin-sensitive aminopeptidases (PSAs) participate in a variety of proteolytic events essential for cell growth and viability, and in fertility in a broad range of organisms. We have identified and characterized an Arabidopsis thaliana mutant (mpa1) from a pool of T-DNA tagged lines that lacks PSA activity. This line exhibits reduced fertility, producing shorter siliques (fruits) bearing a lower number of seeds compared with wild-type plants. Cytogenetic characterization of meiosis in the mutant line reveals that both male and female meiosis are defective. In mpa1, early prophase I appears normal, but after pachytene most of the homologous chromosomes are desynaptic, thus, by metaphase I a high level of univalence is observed subsequently leading to abnormal chromosome segregation. Wild-type plants treated with specific inhibitors of PSA show a very similar desynaptic phenotype to that of the mutant line. A fluorescent PSA-specific bioprobe, DAMPAQ-22, reveals that the protein is maximally expressed in wild-type meiocytes during prophase I and is absent in mpa1. Immunolocalization of meiotic proteins showed that the meiotic recombination pathway is disrupted in mpa1. Chromosome pairing and early recombination appears normal, but progression to later stages of recombination and complete synapsis of homologous chromosomes are blocked.     

New Insights into the Role of the Maize Ameiotic1 Locus

In maize the am1-1 mutant allele results in both the male and female meiocytes undergoing mitosis in place of the meiotic divisions. A second mutant allele am1-praI enables both the male and female meiocytes to proceed to the early zygotene stage of meiotic prophase I before being blocked. Here we report on three new alleles that allow all male meiocytes to undergo mitosis but in female meiocytes approximately one quarter (am1-2), one half (am1-485), or all (am1-489) of them are blocked at an abnormal interphase stage. Previous analysis has shown that am1-praI is dominant to am1-1 in male meiocytes. Cytological analysis of heteroallelic combinations in female meiocytes now indicates a dominance relationship of am1-praI > am1-1 > am1-2/am1-485 > am1-489. The evidence provided by the female phenotypes of the new mutant alleles suggest that, whereas the normal am1 allele is required for the meiocytes to proceed through meiosis, a partially functional allele may be required for their diversion into a mitotic division. The partial or complete blockage of mitosis in female meiocytes carrying the new am1 alleles rules out the possibility that the mitotic division of mutant meiocytes reflects a simple default pathway for cells that cannot initiate meiosis. This locus may have a dual function.     

Studies on male and female meiosis in Indian Allium

Male meiosis in autotetraploid Allium tuberosum (4×=32) is fairly regular, keeping in view its cytological status, with 81 percent of the chromosomes associated in quadrivalents and trivalents. About 5% of the cells have 32 univalents. Anaphase segregation is slightly irregular. While 48% of the pollen mitoses show 16 chromosomes, 87% of the mature pollen is viable as indicated by carmine or iodine staining. — Megaspore mother cells have 64 chromosomes associated in 32 bivalents at metaphase I. Anaphase segregation is normal. In three out of 56 cells studied multivalents, bivalents and univalents are observed as in male meiosis. — It is concluded that the species reproduces by pseudogamous parthenogenesis made possible by meiotic modification. This modification is almost perfect and almost completely specific for female meiosis. Slight effects are observed in male meiosis.     

Cytogenetic and nucleolar meiotic cycle analyses in Dysdercus imitator Blöte, 1931 (Pyrrhocoridae, Heteroptera) from Argentina

So far, only seven and five species of Dysdercus from the Old and New Worlds, respectively, have been cytogenetically analysed. They all have holokinetic chromosomes and a pre-reductional type of meiosis. In the present study the chromosome complement, male meiosis and nucleolar meiotic cycle of Dysdercus imitator were analyzed. During male meiosis several cytogenetic features are remarkable, namely the presence of a long diffuse stage after pachytene, the finding of one or two ring bivalents per cell in almost all specimens, and the presence of several prenucleolar bodies lasting up to telophase II. The origin and function of these prenucleolar bodies could be related to a particular physiological cycle of the meiocytes.     

Mitotic and meiotic chromosomes in Somatochlora metallica (Cordulidae,Odonata). The absence of localized centromeres and inverted meiosis

Spermatogonial metaphase chromosomes were examined in two dragonfly species, Somatochlora metallica (Cordulidae) and Aeshna grandis (Aeshnidae), and the behaviour of male meiotic chromosomes was studied in S. metallica. Both in S. metallica and A. grandis the male mitotic metaphase chromosomes from cells treated with colchicine consisted of two equidistantly aligned chromatids, showing no primary constriction. In meiosis the chromosomes of S. metallica males showed telokinetic activity during the first meiotic division, and kinetic activity was restricted in the middle parts of chromosomes during the second division. The kinetic behaviour of the chromosomes both in mitosis and meiosis showed that they were holocentric. One chiasma arises interstitially in each bivalent in S. metallica male meiosis. The chiasmata retain their interstitial position at metaphase I and do not terminalize. At metaphase I bivalents co-orient with homologous telomere regions towards the opposite poles. Thus genuine dyads segregate at the first anaphase. Meiosis in these male dragonflies is thus pre-reductional or conventional, not post-reductional or inverted, as has been previously proposed.     

Meiosis and the Neo- XY system of Dichroplus vittatus (Melanoplinae, Acrididae): a comparison between sexes

The origin of neo-XY sex systems in Acrididae is usually explained through an X-autosome centric fusion, and the behaviour of the neo-sex chromosomes has been solely studied in males. In this paper we analysed male and female Dichroplus vittatus. The karyotype comprises 2n = 20 chromosomes including 9 pairs of autosomes and a sex chromosome pair that includes a large metacentric neo-X and a small telocentric neo-Y. We compared the meiotic behaviour of the sex bivalent between both sexes. Mean cell autosomal chiasma frequency was low in both sexes and slightly but significantly higher in males than in females. Chiasma frequency of females increased significantly when the sex-bivalent was included. Chiasma distribution was basically distal in both sexes. Behaviour of the neo-XY pair is complex as a priori suggested by its structure, which was analysed in mitosis and meiosis of diploid and polyploid cells. During meiosis, orientation of the neo-XY is highly irregular; only 21% of the metaphase I spermatocytes show standard orientation. In the rest of cells, the alternate or simultaneous activity of an extra kinetochore in the distal end of the short arm (XL) of the neo-X, determined unusual MI orientations and a high frequency of non-disjunction and lagging of the sex-chromosomes. In females, the neo-XX bivalent had a more regular behaviour but showed 17% asynapsis in the XL arm which, in those cases orientated its distal ends towards opposite spindle poles suggesting, again, the activity of a second kinetochore. The dicentric nature and the unstable meiotic behaviour of the sex neo-chromosomes of D. vittatus suggest a recent origin of the sex determination mechanism, with presumable adaptive advantages which could compensate their potential negative heterosis. Our observations suggest that the origin of the neo-sex system was a tandem fusion of two original telocentric X-chromosomes followed by another tandem fusion with the small megameric bivalent and a further pericentric inversion of the neo-X. The remaining autosomal homolog resulted in the neo-Y chromosome. This revised version was published online in July 2006 with corrections to the Cover Date.     

The DUET gene is necessary for chromosome organization and progression during male meiosis in Arabidopsis and encodes a PHD finger protein

Progression through the meiotic cell cycle is an essential part of the developmental program of sporogenesis in plants. The duet mutant of Arabidopsis was identified as a male sterile mutant that lacked pollen and underwent an aberrant male meiosis. Male meiocyte division resulted in the formation of two cells instead of a normal tetrad. In wild type, male meiosis extends across two successive bud positions in an inflorescence whereas in duet, meiotic stages covered three to five bud positions indicating defective progression. Normal microspores were absent in the mutant and the products of the aberrant meiosis were uni- to tri-nucleate cells that later degenerated, resulting in anthers containing largely empty locules. Defects in male meiotic chromosome organization were observed starting from diplotene and extending to subsequent stages of meiosis. There was an accumulation of meiotic structures at metaphase 1, suggesting an arrest in cell cycle progression. Double mutant analysis revealed interaction with dyad, a mutation causing chromosome cohesion during female meiosis. Cloning and molecular analysis of DUET indicated that it potentially encodes a PHD-finger protein and shows specific expression in male meiocytes. Taken together these data suggest that DUET is required for male meiotic chromosome organization and progression.     

Disomic and ditelosomic alien chromosome additions in beet (Beta vulgaris), carrying an extra chromosome of B. procumbens or telosome of B. patellaris.

Alien chromosome transmission through the female germ line as well as meiosis in pollen mother cells were studied in disomic and ditelosomic alien chromosome additions of beet. Beta vulgaris, carrying an extra pair of chromosomes or telosomes of B. procumbens or B. patellaris, respectively. The alien chromosomes carried genes for resistance to the beet cyst nematode, Heterodera schachtii, and screening for this resistance was used to select plants with or without the alien chromosomes. A great variation for alien chromosome transmission was recorded and plants carrying two extra alien chromosomes were recovered in the backcross progenies of the disomic or ditelosomic additions. However, in these progenies the average frequencies of plants without alien chromosomes (86%) did not clearly differ from that in similar progenies of the original monosomic or monotelosomic chromosome additions, indicating that doubling the number of the alien chromosome did not enlarge their transmission to the next generation. The alien chromosomes fully paired at pachytene and desynapsed again before diakinesis, indicating decreased chiasma formation. At second metaphase nearly 60% of the cells had one extra chromosome, and the remaining cells carried two or no extra chromosomes in about equal proportions. The tetrads looked fully normal. The expected relation between the average number of alien chromosomes in the germ cells and in the plants of the progenies did not show up, indicating a strong selection favouring the female gametes without alien chromosomes.     

Cytogenetics of a new type of barley with 16 chromosomes

In the progeny of a trisomic type for chromosome 6, Purple, a 16-chromosome type was obtained, which had a pair of new metacentric chromosome 6 in excess. The new metacentric chromosome 6 was shorter than any of the 14 chromosomes of normal barley complement and showed a heteropycnotic nature at late prophase in somatic mitosis. At metaphase I in the plants with 14+one metacentric chromosome 6 (2n=15) the chromosome configuration was exclusively 7_II+1_I indicating that the extra metacentric chromosome 6 could not associate with the normal chromosome 6. At diakinesis and metaphase I in the new 16-chromosome plants most of the sporocytes showed 8_IIor 7_II+2_I. Neither tetravalents nor trivalents were observed at meiosis. The chromosome behaviour at anaphase I and later stages of meiosis was regular in general, resulted in a fairly high pollen fertility of about 61 per cent. Seed fertility however, was very low. The transmission rate of the new metacentric chromosome 6 through the pollen was extremely low in 16-chromosome plants. Possible origin of new basic number and B-chromosome in diploid level through trisomic condition was suggested (Summary see p. 138).Contribution No. 141 of the Department of Plant Science, University of Manitoba.     

Retinoblastoma protein is essential for early meiotic events in Arabidopsis

We have analysed the role of RBR (retinoblastoma related), the Arabidopsis homologue of the tumour suppressor Retinoblastoma protein (pRb), during meiosis. We characterise the rbr-2 mutation, which causes a loss of RBR in male meiocytes. The rbr-2 plants exhibit strongly reduced fertility, while vegetative growth is generally unaffected. The reduced fertility is due to a meiotic defect that results in reduced chiasma formation and subsequent errors in chromosome disjunction. Immunolocalisation studies in wild-type meiocytes reveal that RBR is recruited as foci to the chromosomes during early prophase I in a DNA double-strand-break-dependent manner. In the absence of RBR, expression of several meiotic genes is reduced. The localisation of the recombinases AtRAD51 and AtDMC1 is normal. However, localisation of the MutS homologue AtMSH4 is compromised. Additionally, polymerisation of the synaptonemal complex protein AtZYP1 is abnormal. Together, these data indicate that loss of RBR during meiosis results in a reduction of crossover formation and an associated failure in chromosome synapsis. Our results indicate that RBR has an important role in meiosis affecting different aspects of this complex process.     

Induction of cytomixis affects microsporogenesis in Sesamum indicum L. (Pedaliaceae)

Cytomixis was recorded during microsporogenesis in sesame (Sesamum indicum L.), a member of the family Pedaliaceae. The phenomenon of cytomixis was observed at various stages of meiosis in 0.5% sodium azide (SA) treated populations of Sesamum indicum L. Cytomixis was observed to occur through various methods, i.e. by forming cytoplasmic channels and direct fusion of pollen mother cells (PMCs), the former was more frequent than the latter. The migration of nuclear content involved all the chromatin/chromosomes or part of it from donor to recipient cell/cells. Some completely empty meiocytes were also observed. Stickiness, precocious movement, laggards, unorientation and micronuclei were observed in almost all the sets treated with various doses of SA. Increase in the doses of SA had a positive effect on the percentage of PMCs showing cytomixis and chromosomal abnormalities. The impact of cytomixis on meiotic behaviour, reduced pollen viability and heterogeneous sized pollen grains were observed.     

The resurgence of haploids in higher plants

The life cycle of plants proceeds via alternating generations of sporophytes and gametophytes. The dominant and most obvious life form of higher plants is the free-living sporophyte. The sporophyte is the product of fertilization of male and female gametes and contains a set of chromosomes from each parent; its genomic constitution is 2n. Chromosome reduction at meiosis means cells of the gametophytes carry half the sporophytic complement of chromosomes (n). Plant haploid research began with the discovery that sporophytes can be produced in higher plants carrying the gametic chromosome number (n instead of 2n) and that their chromosome number can subsequently be doubled up by colchicine treatment. Recent technological innovations, greater understanding of underlying control mechanisms and an expansion of end-user applications has brought about a resurgence of interest in haploids in higher plants.     

MULTIPOLAR MEIOSIS IN DIPLOID CRESTED WHEATGRASS,AGROPYRON CRISTATUM

A new model of spindle organizers is proposed: The spindle organizer in a higher plant is similar to the centriole of animal cells. It is a unit cell organelle which follows regular cell division cycles and is genome specific. Each genome carries its own spindle organizer. During fertilization, a male spindle organizer enters the egg cell. It may fuse with the female spindle organizer, or either one may degenerate. In a hybrid, both male and female spindle organizers may exist, and multipolar divisions separate different genomes into different groups. The same mechanism can be used to explain the formation of a polyhaploid from a polyploid. The chromosome behavior of an individual is believed to be an interaction of chromosome homology and the homology between chromosomes and their spindle organizers. This model is based on observations of multipolar meiosis which occurred in two cultures of diploid crested wheatgrass, Agropyron cristatum (L.) Gaertn. In these two cultures multipolar meiosis occurred at every stage after late diakinesis. The seven bivalents were separated into groups at late diakinesis. More than one equatorial plate was formed at metaphase I. Each micrometaphase plate behaved as an independent unit and had its own anaphase movement. Usually the chromosome complement separated into two groups with (4–3), (5–2), and (6–1) separations observed in about an equal number of cells. Cells with chromosomes divided into three or four groups were found less frequently. Multipolar meiosis may take place at either first or second division. Cell plates were formed across each spindle apparatus, cleaving each group of chromosomes into smaller micro-cells. At the “quartet” stage, 4- to 12-celled “quartets” were observed. Pollen stainability was measured at above 75% in both cultures. Stained pollen grains could be classified into two distinct size classes. Darkly stained, small pollen grains represented the result of multipolar meiosis and may have been viable. Multipolar cell divisions provided a mechanism which polyploids might reduce their ploidy level.