Induced apogamous sporophytes inPteris dispar andP. semipinnata,and the meiotic behavior in their sporocytes

Spores ofPteris dispar andP. semipinnata were aseptically cultured in flasks for apogamous sporophyte induction. Calli or cell colonies similar to calli were induced in cultures supplemented with hormones. Sporophytic leaves subsequently developed from them in hormone-free medium and the young sporophytes were raised into plants with sporangia. Since the wild-type plants having 116 chromosomes are tetraploid, the sporophytic plants originating from spores would appear to be diploid (dihaploid). In induced sporophytes ofP. semipinnata, non-homologous chromosomes (58 univalents) were found during the meiotic process in sporocytes. InP. dispar, however, the meiotic cells showed many bivalent chromosomes (maximum 29ll). These results suggest thatP. semipinnata is allotetraploid, whereasP. dispar is autotetraploid.     

The effect of B chromosome number on chiasma frequency within and between individuals of Gibasis linearis (Commelinaceae)

Meiotic analysis of 21 plants from a single population of Gibasis linearis which contained from 0 to 6 apparently identical B chromosomes showed that the mean chiasma frequencies were significantly higher with increasing numbers of Bs. It was also found that those meiotic cells of a 5B plant which contained fewer than five B chromosomes showed a marked fall in chiasma frequency, demonstrating that the influence of B chromosomes on chiasma formation in A chromosomes acts at a cellular rather than a whole-anther level.     

Nuclear division in the red algae Chondria nidifica and Chondria tenuissima

Cytological investigations are reported for two Chondria species, the Pacific species Chondria nidifica Harvey and Chondria tenuissima (Goodenough et Woodward) C. A. Agardh from the shore of the Marmara Sea in Istanbul. Nuclear division during mitosis and meiosis has been followed in somatic cells and in tetrasporangial mother cells respectively of diploid tetrasporic plants. The spherical interphase nucleus stains densely, showing many chromatin granules. Mitotic nuclei in the apical groove show a large number of chromosomes at metaphase; the chromosome number has been estimated at diakinesis to be 40 in both C. nidifica and C. tenuissima. The meiotic nuclei of tetraspore mother cells in prophase contain several relatively large nucleolar-derivatives in both species. The nucleolar derivatives disappear completely before the chromosomes begin to differentiate. In meiotic prophase the tetraspore mother cell enlarges from its original diameter. The period of the second meiotic anaphase seems to be extremely short in comparison with other nuclear phases. When the chromosomes reach the poles, they spread and subsequently form a relatively compact mass at telophase. The spindle has not been observed in C. tenuissima. Photographs are presented of nucleoli and nucleolar-derivatives in mitotic and meiotic divisions.     

Production and cytogenetics of the intergeneric hybrids Brassica juncea×Orychophragmus violaceus and B. carinata×O. violaceus

 Intergeneric hybrids between Brassica juncea (2n=36), B. carinata (2n=34) and Orychophragmus violaceus (2n=24) were produced when B. juncea and B. carinata cultivars were used as female parents. The hybrids between B. juncea and O. violaceus had an intermediate morphology except for petal colour and were partially fertile. The hybrids between B. carinata and O. violaceus had a matroclinous morphology and were nearly fertile. Cytological analysis of the hybrids and their progenies gave the following results. (1) In the hybrids between B. juncea and O. violaceus, the somatic tissues of the roots, leaves and styles were mixoploid (2n=12–42), and cells with 24, 30 or 36 chromosomes were the most frequent. Based on the recorded numbers and behaviour of the mitotic and meiotic chromosomes, complete and partial separation of the parental genomes was proposed to have occurred during mitosis. This resulted in the occurrence of cells with possibly complete and incomplete complements of the parental species and cells with parental complements and some additional chromosomes from the other parent. (2)  Pollen mother cells (PMCs) possibly with both parental chromosome complements, only B. juncea chromosomes or a complete B. juncea complement with additional O. violaceus chromosomes were more competitive in entering meiosis. The majority of fertile gametes were deduced to have been produced by PMCs with a B. juncea complement with or without additional O. violaceus chromosomes. (3) The progeny plants from selfed hybrids between B. juncea and O. violaceus were morphologically either of a B. juncea, hybrid or variable type. Cytologically they were grouped into six types according to the frequencies of cells with various chromosome numbers. All of the plants except 2 which constituted two types, were mixoploids, composed of cells with various chromosome numbers, mainly in a certain serial range. (4) The hybrid plants between B. carinata and O. violaceus were mixoploids with chromosome numbers in the range of 12–34, and cells with 2n=34 were the most frequent. The main categories of PMCs with 17 bivalents at metaphase I and 17 : 17 segregations at anaphase I contributed to the high fertility of the hybrids and the fact that their progeny after selfing were mainly plants with 2n=34. Somatic and meiotic separation of the parental genomes was proposed to have occurred in the hybrids between B. carinata and O. violaceus. (5) Mitotic and meiotic elimination of what could be O. violaceus chromosomes might also have contributed to the observed mitotic and meiotic cell types in the two kinds of hybrids studied. Finally, the possible mechanisms behind these cytological observations and their potential in the production of Brassica aneuploids were discussed. Received: 4 February 1997/Accepted: 29 July 1997     

Meiotic behaviour of tetraploid wheats (<Emphasis Type="Italic">Triticum turgidum</Emphasis> L.) and their synthetic hexaploid wheat derivates influenced by meiotic restitution and heat stress

Meiotic restitution is considered to be a common mechanism of polyploidization in plants and hence is one of the most important processes in plant speciation. Meiotic behaviour of plant chromosomes is influenced by both genetic and environmental factors. In this study, the meiotic behaviour of cereal crops was investigated, which includes tetraploid wheat genotypes (with and without the meiotic restitution trait) and their derivates (synthetic hexaploid wheats and a doubled haploid (DH) line), grown at two planting dates in the field. In addition, two local landraces of emmer wheat (Triticum turgidum ssp. dicoccum), one wheat cultivar (Chinese spring), one DH triticale cultivar (Eleanor) and one rye accession were included. Immature spikes of mid-autumn and end-winter sowing plants were collected in April and May 2008, respectively, fixed in Carnoy’s solution and stained with hematoxylin. Pollen mother cells (PMCs) from anthers at different stages of meiotic process were analysed for their chromosomal behaviour and irregularities. Meiotic aberrations such as laggards, chromosome bridges, micronuclei, abnormal cytokines, chromatin pulling and meiotic restitution were observed and the studied genotypes were accordingly ranked as follows: triticale > synthetic hexaploid wheats > tetraploid wheats possessing meiotic restitution > tetraploid wheats lacking meiotic restitution > rye. The results indicated that the samples that had been planted in the autumn, thus experiencing an optimum temperature level at the flowering stage, exhibited less meiotic irregularities than winter planting samples that encountered heat stress at the flowering period.     

Absence of qualitative genes controlling interspecific pairing in rye B chromosomes

Summary The meiotic behaviour of hybrids between Secale cereale carrying B chromosomes and S. vavilovii has been studied in order to estimate the effects of B chromosomes on hybrid meiotic pairing. The possible effect of Bs on the meiotic pairing of the offspring from backcrosses with S. vavilovii has been studied also. The results obtained clearly indicate that no detectable differences existed in chromosome pairing of hybrids with or without B chromosomes. The hypothetical existence of epistatic genes on cereale genome masking the effect of Bs has been rejected after the results obtained in backcrosses. Therefore, lack of qualitative genes controlling interspecific pairing on rye B chromosomes has been concluded. A quantitative effect of B chromosomes was detected only when they were in alien cytoplasm.     

The effect of Secale cereale L. heterochromatin on wheat chromosome pairing

Metaphase-I chromosome association in PMCs of five F₁ hybrids 6x-triticale x T. turgidum (2n=5x=35 and genomes AABBR), and 13 plants from their backross or self offspring is reported. In wheat 18 chromosome arms and in rye 14 arms were recognized after C-banding and individually studied. Plants of backcross and F₂ showed variability for number and type of rye chromosomes, having in common the 28 durum wheat chromosomes (AABB). By testing meiotic association in plants with different rye chromosome constitutions, significant negative correlations were found. A clear negative effect of rye heterochromatin on pairing in wheat chromosomes is observed, the influence being more pronounced for large arms than for the short ones.     

Apogamous sporophyte formation in a fernPteris multifida and its characteristics

When spores of the fern,Pteris multifida, were aseptically cultured in the dark, sporophytic plants were apogamously induced. The plants have been subsequently grown in pots until the development of leaves with many sporangia for observations of meiotic characteristics in their sporocytes. The sporophytic plants originated from spores were estimated to be haploid, and the estimation was supported by abnormal meiosis in sporocytes and the absence of mature spores, but some chromosomes (n=58) formed bivalents in the meiotic process.     

Mutagens induced chromosomal damage in <Emphasis Type="Italic">Lablab purpureus</Emphasis> (L.) Sweet var. <Emphasis Type="Italic">typicus</Emphasis>

Cytological analysis with respect to meiotic behaviour is considered to be the one of the most dependable indices to estimate the potency of mutagens and to elucidate the response of various genotypes to a particular mutagen. Seeds of Lablab purpureus (L.) Sweet var. typicus cv. CO(Gb)14 were subjected to different doses/concentrations of gamma rays and EMS. The effects of different mutagenic treatments on meiosis were studied on treated and control plants. Various types of meiotic aberrations such as stickiness, clumping of chromosomes, laggards, ring chromosomes and precocious movements were observed in the mutagenic treatments. As increase in the concentration, the frequency of cells showing chromosomal aberrations shows a linear increase up to a certain level. However, the EMS treatments proved to be more effective in inducing meiotic aberrations as compared to gamma rays.     

CHROMOSOME NUMBERS IN POPULATIONS OF CLAYTONIA VIRGINICA FROM THE NEW YORK METROPOLITAN AREA

Buds of Claytonia virginica were examined from 9 locations in the metropolitan New York area. A large amount of meiotic irregularity was characteristic of the populations studied. Chromosome counts on pollen mother cells were higher than any reported previously for the taxon, which contains an extensive aneuploid series including at least 40 different numbers ranging from 2n = 12 to ca. 190. In one New Jersey population, plants having approximately 90 chromosomes were associated with those having only 16. All the observations indicated that Claytonia virginica is an unusual species with an ability to tolerate fluctuations in chromosome material without apparent effects on fertility and morphology.     

The cytology of Brachycome lineariloba

The frequencies and behaviour of two types of accessory chromosome in Brachycome lineariloba species A (n=2) are reported. These include a larger (ca. 4.0 macro-B chromosome, and a minute (<1.0 ) micro-B. Twelve per-cent of the 452 plants studied were carriers of one, two or more rarely three macro-B chromosomes. These were usually mitotically stable, but showed some meiotic irregularity when present in odd number. An equilibrium situation is inferred, and loss must be balanced against mechanisms tending to increase B frequency. A surplus of plants with two B chromosomes also suggests that mechanisms of doubling may operate. A possible source of the macro-B's is considered to be hybridisation between the several species of the B. lineariloba complex. — The micro-B chromosomes were found in 3.8% of two only of the three known cytological races of the species. They showed extreme irregularity in both mitotic and meiotic behaviour. Their relationship to the normal (A) chromosomes and to the macro-B's is obscure.     

Cytology of inbreds and f 1 hybrids of pearl millet

Summary The meiotic behaviour of chromosomes was studied in four inbred lines and their F ₁ hybrids of P. typhoides. The inbred lines showed a decrease in mean chiasma frequency when compared with the population plants, but differed from one another in their mean chiasma frequencies. Together with the decrease in mean chiasma frequency the inbreds showed variation in mean chiasma frequencies. The inbred lines showed a number of meiotic abnormalities such as extra chromosomes, extra fragments, desynapsis, persistent nucleoli and differential condensation of chromosomes. The F ₁ hybrids of these inbreds exhibited heterosis for chiasma frequency. All the F ₁'s had mean chiasma frequencies higher than the means of the respective participating parents. The F ₁'s, however, differed in the degree of heterosis exhibited. In the F ₁ hybrids, the variation in mean chiasma frequency, both between plants and between PMC's within plants, was significantly lower than that of the inbred lines. The effect of environment was studied in the inbred lines and their F ₁ hybrids. The mean chiasma frequencies of the inbred lines were significantly lower, and the variation in mean chiasma frequencies was greater, in the stress season. The mean chiasma frequencies of F ₁'s did not show any significant differences between the two seasons. The F ₁'s exhibited less variation in mean chiasma frequency than the inbred lines, showing that F ₁'s were developmentally more stable. The F ₁'s did not show any meiotic abnormalities in either season.     

TRANSMISSION AND SIGNIFICANCE OF B CHROMOSOMES IN ANTHURIUM WAROCQUEANUM

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

Meiotic pairing of the amphiploid Hordeum chilense X Triticum turgidum conv. durum studied by means of Giemsa C-banding technique

Summary The meiotic behaviour of the amphiploid Hordeum chilense X Triticum turgidum conv. durum using a C-banding staining method is studied. Nine pairs of chromosomes at metaphase-1 (4A, 7A and the seven of the B genome) were identified and the remaining wheat chromosomes (1A, 2A, 3A, 5A and 6A) and seven of the chilense (1 to 7 H ^ch chromosomes) were assigned to its particular genome. A similar mean number of univalents from parental genomes (wheat and wild barley) were found. No meiotic pairing between chilense and turgidum chromosomes was detected. Differences in the meiotic behaviour per chromosome and amongst genomes are explained on the basis of cytomorphological and heterochromatin characteristics.     

Meiotic division and fusion of nuclei in multinucleate cells from the induced fusion of meiotic protoplasts of liliaceous plants

Multinucleate protoplasts were produced from meiotic cells at the zygotene and pachytene stages in a lily andTrillium, and their meiotic divisions were followed during subsequent culture. In each multinucleate, a complete synchrony of nuclear division was maintained throughout the meiotic process, and chromosome behavior appeared normal up to the metaphase stage. In most dinucleates, chromosome segregation movement was organized in a common spindle, and the daughter nuclei at the telophase appeared to envelope each other in the newly formed nuclear membrane. The cell was divided into two daughter cells by a common cell plate. Trinucleates were similarly converted to two cells with a hexaploid number of chromosomes. Some of the di- and trinucleates subsequently completed the second meiotic division with the formation of typical tetrad configurations. In giant cells with more than several nuclei, chromosomes separated at random but reaggregated into one giant resting nucleus, with no later cytokinesis. The rate of meiotic development in multinucleates was relatively slower in cells which contained greater numbers of nuclei.     

A cytological approach to studying meiotic recombination and chromosome dynamics in Arabidopsis thaliana male meiocytes in three dimensions

During meiotic prophase I chromosomes undergo dramatic conformational changes that accompany chromosome condensation, pairing and recombination between homologs. These changes include the anchoring of telomeres to the nuclear envelope and their clustering to form a bouquet. In plants, these events have been studied and illustrated in intact meiocytes of species with large genomes. Arabidopsis thaliana is an excellent genetic model in which major molecular pathways that control synapsis and recombination between homologs have been uncovered. Yet the study of chromosome dynamics is hampered by current cytological methods that disrupt the three‐dimensional (3D) architecture of the nucleus. Here we set up a protocol to preserve the 3D configuration of A. thaliana meiocytes. We showed that this technique is compatible with the use of a variety of antibodies that label structural and recombination proteins and were able to highlight the presence of clustered synapsis initiation centers at the nuclear periphery. By using fluorescence in situ hybridization we also studied the behavior of chromosomes during pre‐meiotic G₂ and prophase I, revealing the existence of a telomere bouquet during A. thaliana male meiosis. In addition we showed that the number of telomeres in a bouquet and its volume vary greatly, thus revealing the complexity of telomere behavior during meiotic prophase I. Finally, by using probes that label subtelomeric regions of individual chromosomes, we revealed differential localization behaviors of chromosome ends. Our protocol opens new areas of research for investigating chromosome dynamics in A. thaliana meiocytes.     

Autosegregation of enzyme loci in agamospermous progenies of triploid plants of sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.)

The ratios of the phenotypic classes of glucosephosphate isomerase (GPI2) and malate dehydrogenase (MDH1 and MDH2) were studied in agamospermous progenies of triploid sugar beet plants. The ratio of the phenotypic classes of these enzymes corresponds to the calculations based on the assumption of polyteny of chromosomes carrying alleles of the enzyme loci accompanied by the loss of extra copies of the alleles in the first division of a cell entering embryogenesis. An increase in the gene dosage due to polyteny leads to the appearance in the progeny with a definite frequency of alleles that were absent in the original parental plant. The notions of “meiotic autosegregation” and “mitotic autosegregation” characteristic of meiotic and mitotic agamospermy are introduced, as well as the term locus polygenotype characterizing not only the allelic composition and the number of chromosomes, but also the number of chromatids carrying alleles of the marker locus in the cell before its entry into embryogenesis.     

Meiotic analysis of four cross-pollinated generations in a synthetic autotetraploid population of husk tomato (<i>Physalis ixocarpa</i>)

The cultivated husk tomato (Physalis ixocarpa) (2n = 2x = 24) is native from Mexico and Central America and shows a wide genetic variation. Presently, it is the fourth horticultural crop in cultivation surface in Mexico. The working team of this research previously developed an autotetraploid population by using colchicine. The objectives of the present work were to analyze the ploidy level and meiotic behavior of the subsequent generations (C3, C4, C5, C6) from the original (C2) composed only by plants with the duplicated genome from the Rendidora cultivar, and to determine pollen viability. As a diploid control the cultivar Rendidora of P. ixocarpa was used. Ploidy level was determined by flow citometry and meiotic analysis. For the meiotic study, the microsporocytes were prepared by the squash method, stained with carmin and analyzed in diakinesis. Pollen viability was evaluated through 0.01% Buffalo Black staining. The tetraploid condition prevailed through four cross-pollinating generations, maintaining a constant chromosome number 2n = 4x = 48. In diakinesis, the chromosomes of the diploid cultivar were associated into bivalents, whereas in tetraploid plants the chromosomes associated into univalents, bivalents and trivalents. Highly significant differences in bivalent pairing were detected between autotetraploid plants and between generations. Pollen viability did not show significant differences between generations and allowed reproduction. These results indicate that it is possible to develop an autotetraploid cultivar, because the polyploid state is naturally maintained and the plants are fertile. Furthermore, given the differences in bivalent pairing between plants and generations, a response to selection toward meiotic stability is expected.     

Meiosis in haploid barley — An interpretation of non-homologous chromosome associations

Detailed meiotic studies were conducted on ten haploid plants representing six different genotypes of barley (Hordeum vulgare, 2n=14). At pachytene stages the non-homologous chromosomes were observed to pair as intimately as homologous chromosomes in many cells. Foldback pairing, involving single chromosomes, and multivalent associations were common. At diplotene, up to 4 chiasmatalike structures were observed in paired chromosomes but it is not likely that they resulted from crossing over. At diakinesis the bivalent frequency mean was from 1 to 1.3 per cell whereas by metaphase I the paired associations were rare with a single rod bivalent being observed in 3 to 5% of the cells. The frequencies of various types of secondary associations at metaphase were also recorded. — The origin and significance of bivalents and secondary associations in haploids is reviewed and discussed. Caution is urged in the interpretation that low levels of chromosome pairing in haploids is evidence of homology. It is concluded that very little chromosome duplication is likely to be found within the haploid set of barley chromosomes and that the basic chromosome number is seven.     

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.