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.     

Meiotic chromosome interactions in inbred autotetraploid rye (Secale cereale).

Electron microscopy of whole-mount surface-spread synaptonemal complex complements and conventional light microscopy of chromosomes at first metaphase of meiosis were used to compare the relative frequencies of pairing configurations at the two stages in inbred autotetraploid rye (Secale cereale L.). Statistical tests showed significantly fewer multivalents at first metaphase than expectations based on random initiation of synapsis at each telomeric site within each group of four homologues. Direct observations of synaptic behaviour of chromosomes showed that this deviation is due primarily to a preponderance of bivalents during zygotene and pachytene. It is also the result of a significant drop in multivalent frequency from meiotic prophase to metaphase I, which is attributable both to a lack of chiasmata with which to consolidate multivalents and inhibition of chiasma formation in synaptonemal complex segments of multivalents that are nonhomologous.     

Meiosis in a triploid hybrid of <Emphasis Type="Italic">Gossypium</Emphasis>: high frequency of secondary bipolar spindles at metaphase II

Studies on meiosis in pollen mother cells (PMCs) of a triploid interspecific hybrid (3x = 39 chromosomes, AAD) between tetraploid Gossypium hirsutum (4n = 2x = 52,AADD) and diploid G. arboreum (2n = 2x = 26,AA) are reported. During meiotic metaphase I, 13 AA bivalents and 13 D univalents are expected in the hybrid. However, only 28% of the PMCs had this expected configuration. The rest of the PMCs had between 8 and 12 bivalents and between 12 and 17 univalents. Univalents lagged at anaphase I, and at metaphase II one or a group of univalents remained scattered in the cytoplasm and failed to assemble at a single metaphase plate. Primary bipolar spindles organized around the bivalents and multivalents. In addition to the primary spindle, several secondary and smaller bipolar spindles organized themselves around individual univalents and groups of univalents. Almost all (97%) of the PMCs showed secondary spindles. Each spindle functioned independently and despite their multiple numbers in a cell, meiosis I proceeded normally, with polyad formation. These observations strongly support the view that in plant meiocytes bilateral kinetochore symmetry is not required for establishing a bipolar spindle and that single unpaired chromosomes can initiate and stabilize the formation of a functional bipolar spindle.     

Meiotic studies of the F1 hybrid between rice bean (Vigna umbellata) and its wild relative V. minima

The interspecific F₁ hybrid V. umbellata x V. minima, together with the parental species was investigated cytogenetically. Both parents showed normal meiosis, with 11 bivalents at diakinesis. The hybrid was completely pollen sterile. The cytological causes underlying sterility in the hybrid include an array of meiotic abnormalities ranging from non-pairing of chromosomes through the presence of univalents, chain and ring multivalents and anaphase bridges to the formation of micronuclei, during microsporogenesis. The presence of chain and ring multivalents at diakinesis not only demonstrates the structural hybridity of the F₁ hybrid, but also suggests that small interchanges and inversions played a significant role in the speciation within the genus Vigna.     

Chromosome pairing and potential for intergeneric recombination in some hypotetraploid somatic hybrids of Lycopersicon esculentum (+) Solanum tuberosum.

Three somatic hybrids resulting from protoplast fusions of a diploid kanamycin-resistant line of tomato (Lycopersicon esculentum) and a dihaploid hygromycin-resistant transformant of a monohaploid potato (Solanum tuberosum) line were used for a cytogenetic study on chromosome pairing and meiotic recombination. Chromosome counts in root-tip meristem cells revealed two hypotetraploids with chromosome complements of 2n = 46 and one with 2n = 47. Electron microscope analyses of synaptonemal complex spreads of hypotonically burst protoplasts at mid prophase I showed abundant exchanges of pairing partners in multivalents involving as many as eight chromosomes. In the cells at late pachytene recombination nodules were found in multivalents on both sides of pairing partner exchanges, indicating recombination at both homologous and homoeologous sites. Light microscope observations of pollen mother cells at late diakinesis and metaphase I also revealed multivalents, though their occurrence in low frequencies betrays the reduction of multivalent number and complexity. Precocious separation of half bivalents at metaphase I and lagging of univalents at anaphase I were observed frequently. Bridges, which may result from an apparent inversion loop found in the synaptonemal complexes of a mid prophase I nucleus, were also quite common at anaphase I, though the expected accompanying fragments could be detected in only a few cells. Most striking were the high frequencies of first division restitution in preparations at metaphase II/anaphase II, giving rise to unreduced gametes. In spite of the expected high numbers of balanced haploid and diploid gametes, male fertility, as revealed by pollen staining, was found to be negligible.     

Interspecific hybrids of Antheraea roylei and A. pernyi — a cytogenetic reassessment

Summary A rare case of interspecific hybridization between the Indian oak feeding silkworm Antheraea roylei (n=31) and Chinese oak feeding silkworm A. pernyi (n=49) yielding fertile and vigorous offspring is reported. The F₁ and the backcross (A. roylei X A. pernyi X A. pernyi male individuals of the above cross and the F₂₃ and F₃₂ male offspring derived from an earlier cross between another race of A. roylei (n=30) and A. pernyi (n=49) were cytogenetically analysed in order to study their chromosome dynamics. The F₁ hybrids showed 18 trivalents and 13 bivalents in the first meiotic prophase and metaphase. The backcross individuals possessed either 9 trivalents and 31 bivalents or 49 bivalents, in Metaphase I cells. The F₂₃ and F₃₂ individuals were karyotypically alike and exhibited 49 bivalents. The following conclusions were drawn from the above observations: (a) in spite of allopatry and karyotypic divergence in number, a high degree of homology exists between the chromosomal complements of the two species; (b) A. pernyi possibly evolved from A. roylei, during the course of which 18 chromosomes of the latter underwent fission to give rise to the 36 chromosomes of the former. This is demonstrated by trivalent formation and pairing affinities in F₁ hybrids; (c) selection has favoured the elimination of large A. roylei chromosomes which participated in trivalent formation in successive generations of inbred hybrids to establish a stable Karyotype like that of A. pernyi.     

Cytological studies on meiotic configurations of intraspecific aneuploids ofCarex blepharicarpa (Cyperaceae) in Japan

Chromosome configurations at meiotic metaphase I ofCarex blepharicarpa were determined for 245 individuals collected from 11 localities in the Chugoku District of Japan. Nine intraspecific aneuploids, 2n=26–33 and 41, were found. The most common diploid number was 29, and found in 73 individuals. No clear geographical pattern was suggested by a distribution of these aneuploids. A consecutive series of chromosome numbers from 2n=26 to 32 was found in two populations from Okayama Prefecture. At meiotic metaphase I, univalents and multivalents were found, and one to three trivalents were observed in each aneuploid. Heteromorphic chain trivalents comprising large, medium and small chromosomes were prevalent. Homomorphic trivalents, which are thought to be originated from chromosome duplications caused by unreduced gametes, were found in only one individual with 2n=41. The high frequency of heteromorphic trivalents in this species indicates that most aneuploidy probably results from fission and/or fusion of chromosomes.     

Premiers stades de l'oogenèse de Rhabdophaga saliciperda (Cecidomyiidae,Diptera)

The females of Rhabdophaga saliciperda have in their somatic cells 8 chromosomes and the males 6. The type of sex determination is therefore: X₁X₁X₂X₂—♀; X₁X₂—♂. The cells of the germinal line have 46 chromosomes, but a variation of their number was observed. In the oogonia and spermatogonia the number of heterochromatic chromosomes may exceed the number of E chromosomes, i.e. 8. In the beginning of the growth stage of the oocytes an incorporation of somatic cells was observed. The nuclei of these somatic cells persist in the cytoplasm of the oocytes until the maturation divisions. The possibility of their participation in the reconstruction of the nucleus of the mature egg is envisaged. The metaphase of the I segmentation division has a complex character. During prophase of the first meiotic division the E chromosomes form 4 bunches of 6–8 chromosomes each. Some univalents may also be present. The 8 S chromosomes form 4 regular bivalents. The 4 groups of E chromosomes persist until metaphase I. During metaphase I a phenomenon of expulsion of the majority of E chromosomes from the metaphase spindle was observed. The 4 bivalents remain in the equatorial plain of the spindle with some E Chromosomes. After this expulsion 2 groups of chromosomes are formed. In connection with them 2 spindles develop. An irregular distribution of E chromosomes follows without their division. The bivalents are probably separated in regular manner. These 2 spindles correspond to the I maturation division. The II maturation division was not observed because of lack of respective stages.     

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 synaptic behaviour of the wild forms of Triticum turgidum and T. timopheevii.

Different wild allopolyploid species of Triticeae show extensive bivalent formation at zygotene while a considerable number of multivalents is present in cultivated polyploid wheats. To study the chromosome behaviour at early meiotic stages in wild forms of tetraploid wheats Triticum turgidum and T timopheevii (2n = 4x = 28) we have analysed the synaptic pattern in fully traced spread nuclei at mid- and late zygotene and at pachytene of wild accessions of these species. The mean number of synaptonemal complex (SC) bivalents at mid-zygotene ranged from 12.22 to 13.14 among the accessions studied indicating a strong restriction of synapsis initiation to homologous chromosomes. The mean of bivalents increased at pachytene because of the transformation of multivalents into bivalents. Ring bivalents observed at metaphase I support that SC bivalents were formed by homologous chromosomes. The average values of SC bivalents at mid-zygotene in the wild forms are much higher than the average values observed in the cultivated tetraploid wheats but similar to that of a mutant line of T turgidum with a duplication that includes Ph1, the major homoeologous pairing suppressor locus. These results suggest that the efficiency of the mechanism operating in the homologous recognition for synapsis is higher in wild wheat populations than in cultivated varieties. Apparently, a relatively detrimental modification of the pairing regulating genetic system accompanied the domestication of the wild wheat forms.     

Synaptonemal complex formation in hybrids of Lolium temulentum x Lolium perenne (L.)

Comparisons were made between two kinds of tetraploids derived from the hybrid Lolium temulentum x L. perenne. One hybrid behaves like an autotetraploid with multivalents at first metaphase of meiosis in pollen mother cells. The other behaves like an allotetraploid, in which pairing at first metaphase is restricted to bivalents comprised of strictly homologous chromosomes. The diploidisation of the latter form is controlled by determinants located on both the normal, A chromosomes and on supernumary B chromosomes. Reconstruction of synaptonemal complexes and their elements, from serial sections through pollen mother cell nuclei examined under the electron microscope, reveals that at zygotene pairing in both forms results in multivalent formation involving non-homologous as well as homologous chromosomes. The mechanism responsible for the diploidisation is, therefore, not based on a restriction of pairing at early meiosis to homologous chromosomes but on a correction or transformation of the multivalent chromosome associations to bivalents subsequent to zygotene. The transformation is not completed until late pachytene. In the multivalent-forming tetraploid a maximum of four chromosomes are associated at first metaphase. Yet configurations of a higher valency are found at zygotene. There is, therefore, a partial transformation of multivalents even in this autotetraploid form which restricts configurations at metaphase I to homologous and homoeologous chromosomes only. In both hybrids some homologous bivalents are not the product of resolution of multivalents but result from two-by-two pairing from the beginning of zygotene.     

A comparative study of male meiosis in Drosophila melanogaster and D. virilis

Male meiosis in D. melanogaster cytologically follows the usual pattern, whereas in D. melanogaster and in D. virilis oocytes the chromosomes clump into a karyosphere at early meiotic prophase and remain so up to metaphase I.Male meiosis in D. virilis spermatocytes has an intermediate character: a part of the chromatin clumps together in a karyosphere at early prophase, whereas the other part of the chromatin remains diffuse all through prophase. At the end of prophase, the diffuse chromatin becomes integrated into the karyosphere before metaphase I. During the meiotic divisions the chromosomes have the same clumped aspect as those in Drosophila oocytes and thus differ strikingly from the dividing chromosomes in D. melanogaster spermatocytes.In D. virilis spermatocytes the nucleolus exhibits changes during the meiotic prophase that may be related to synthetical activities. The DNA specific staining with the fluorochrome DAPI reveals the existence of extrachromosomal DNA in the later prophase. Other striking differences in meiotic events between the two Drosophila species concern the centrioles and spermiogenesis.     

Chromosome systems in the eriococcidae (Coccoidea Homoptera)

Spermatogenesis is described in two eriococcid species and the observations are compared to those previously reported. In Gossyparia spuria the diploid chromosome number is 28 in both males and females. In the female all the chromosomes are euchromatic. In most male tissues 14 of the chromosomes are euchromatic (E) and 14 are heterochromatic (H). Prior to the first meiotic division in males the number of H chromosomes was reduced. During prophase I all the cells showed 14 E chromosomes and from 1 to over 9 H chromosomes. The range of chromosome numbers in metaphase I was similar to that in prophase I. All the chromosomes divided in anaphase I, and, following differential uncoiling at interkinesis, the E and H groups of chromosomes segregated from each other at anaphase II. Only the E groups formed sperm. The presence of a variable number of H chromosomes and a haploid number of E chromosomes in spermatogenesis suggested the presence of the multiple-D variant of the Comstockiella chromosome system. In this system some of the H chromosomes become euchromatic prior to prophase I of spermatogenesis and pair with their E homologues. All the remaining H chromosomes are thus univalents, while among the E elements, some are univalents and the rest are bivalents. The observed reduction in the number of H chromosomes in the first meiotic division which was previously attributed to pairing among the H chromosomes, is now interpreted to be the result of the return of some of the H chromosomes to a euchromatic state and to their subsequent pairing with their E homologues. Spermatogenesis in Eriococcus araucariae was similar to that of G. spuria except that the reduction in the number of H chromosomes was not as extensive. The chromosome systems of the two species are compared to those of other eriococcids and the differences are briefly discussed.Supported by grant GB1585 from the National Science Foundation, Washington, D. C.     

Genome relationships between Hordeum procerum (6x) and H. lechleri (6x)

Investigations on the meiotic behaviour of chromosomes in interspecific hybrids (2n=6x=42) between Hordeum lechleri (6x) and H. procerum (6x) and in their component haploids have been utilized to assess the nature of pairing and the extent of genome homology between the two species. In the F₁ hybrids an average of 25 (60%) chromosomes associated at metaphase I, mostly as bivalents. A majority (60%) of the pollen mother cells (PMCs) in H. procerum haploids (2n=3x=21) displayed 21 univalents and even in the remainder, a maximum of two rod bivalents were formed resulting in an average of 0.52 bivalents per cell. In haploids of H. lechleri (2n=3x=21) however, 30% of chromosomes pair. The sum of the chromosomal associations in the component haploids represents only 17% of the complement, far below the observed frequency (60%) in the hybrids. Thus, the pairing displayed in hybrids between H. lechleri and H. procerum was mostly allosyndetic and suggestive of two genomes being common in these species.In haploid H. procerum 1/3 of the PMCs displayed a tripolar organisation of chromosomes leading to triad and hexad formation after divisions I and II respectively. The significance of hexad formation in the trihaploid H. procerum and a possible suppression of homoeologous pairing in H. procerum haploids are discussed.     

Quantitative analysis of diploid translocation heterozygotes: test of models and equations

Summary Equations have been derived for two different models of chromosome pairing and chiasmata distribution. The first model represents the normal condition and assumes complete synapsis of homologous bivalents and the arms of interchange quadrivalents. This is followed by a nonrandom distribution of chiasmata among bivalents and multivalents such that each bivalent or bivalent-equivalent always has at least one chiasma. Univalents occur only as part of a III, I configuration at diakinesis or metaphase I. The second model assumes that a hologenomic mutation is present in which all chromosomes of a genome are equally affected. Two different assumptions can be made for such a mutation, and both give the same results: (1) homologous or homoeologous chromosome arms may be randomly paired or unpaired, but synapsis always leads to a crossover; (2) homologous or homoeologous arms always pair, but chiasmata are randomly distributed among the arms. The meiotic configurations at diakinesis or metaphase I are the same for both assumptions. Meiotic configurations of normal diploid interchange heterozygotes show good agreement with numbers predicted by the equations for nonrandom chiasmata distribution among configurations. Inter-specific hybrids with supernumerary chromosomes produced meiotic configurations frequencies in agreement with predictions of equations for random chiasmata distribution, but a hybrid without supernumeraries fitted the nonrandom expectations.     

Chromosome number and secondary chromosomal associations in wild populations of Geranium pratense L. from the cold deserts of Lahaul-Spiti (India)

In this work we studied the meiotic chromosome number and details of secondary chromosomal associations recorded for the first time in Geranium pratense L. from the alpine environments in the cold deserts of Lahaul-Spiti (India). All the presently studied individuals of the species existed at 4x level (x = 14). The present chromosome count of n = 28 in the species adds a new cytotype to the already existing diploid chromosome count of 2n = 28 from the Eastern Himalayas and outside of India. Out of the six accessions scored presently four showed normal meiotic course. However, two accessions investigated from Mud, 3800 m and Koksar, 3140 m depicted abnormal meiotic course due to the presence of multivalents and univalents, and secondary associations of bivalents/chromosomes. The secondary chromosomal associations in the species existed among bivalents/chromosomes were noticed in the PMCs at prophase-I (diakinesis) and persisted till the separation of sister chromatids at M-II. The variation in the number of bivalents/chromosomes involved in the secondary associations at M-I (2–8) and A-I/M-II (2–12) has also been recorded. The occurrence of such secondary associations of bivalents/chromosomes in G. pratense which existed at 4x level indicated the secondary polyploid nature of the species.     

An original meiotic mutation in Paspalum regnellii

 Cytogenetic studies carried out over a period of 2 consecutive years on a native Brazilian accession of Paspalum regnellii (2n=40) revealed a meiotic mutation that has not been previously reported for any other species. Among 13 inflorescences investigated during the first collection year, three presented anomalous meiotic behavior starting from metaphase I. At the beginning of this phase, the chromosomes occupied the entire equatorial plate in a membrane-to-membrane arrangement, and the spindle fibers, which were clearly visible, did not converge towards the poles. Degeneration of spindle fibers occurred at the end of metaphase I. Chromosome segregation did not occur and the bivalents were left scattered at random in the cytoplasm. Remnants of chromosome fibers could be seen close to the centromere during this stage. The bivalents gave origin to micronuclei in telophase I, with extremely wide variations in number and size among cells. With the absence of spindle formation during meiosis II, metaphase and anaphase II were not observed. Second cytokinesis occurred in prophase II cells after the occurrence of first cytokinesis. The final product of meiosis was completely abnormal, with a predominance of polyads with microspores of different sizes that resulted in abortive pollen grains. In the affected inflorescences, all microsporocytes presented this anomaly, which caused total sterility. Received: 27 March 1997 / Revision accepted: 7 July 1997     

EXPERIMENTAL HYBRIDS IN EPILOBIUM (INCLUDING SECT. ZAUSCHNERIA) SPECIES WITH N = 15 (ONAGRACEAE)

Experimental hybrids involving the three diploid subspecies of Epilobium sect. Zauschneria formed 15 bivalents at meiotic metaphase I, as did experimental hybrids between the three other species of Epilobium (comprising sect. Cordylophorum) with n = 15. The gametic chromosome number of E. suffruticosum Nutt., n = 15, and its relationship to the other two species are reported for the first time. Although we have not obtained hybrids between the species of these two sections, their morphological similarities are impressive and they are surely closely related.     

Ultrastructural characterization of the sex chromosomes during spermatogenesis of spiders having holocentric chromosomes and a long diffuse stage

An ultrastructural study has been made of spermatogenesis in two species of primitive spiders having holocentric chromosomes (Dysdera crocata, XO and Segestria florentia X₁X₂O). Analysis of the meiotic prophase shows a scarcity or absence of typical leptotene to pachytene stages. Only in D. crocata have synaptonemal complex (SC) remnants been seen, and these occurred in nuclei with an extreme chromatin decondensation. In both species typical early prophase stages have been replaced by nuclei lacking SC and with their chromatin almost completely decondensed, constituting a long and well-defined diffuse stage. Only nucleoli and the condensed sex chromosomes can be identified. — In S. florentina paired non-homologous sex chromosomes lack a junction lamina and thus clearly differ from the sex chromosomes of more evolved spiders with an X₁X₂O male sex determination mechanism. In the same species, sex chromosomes can be recognized during metaphase I due to their special structural details, while in D. crocata the X chromosome is not distinguishable from the autosomes at this stage. — The diffuse stage and particularly the structural characteristics of the sex chromosomes during meiotic prophase are reviewed and discussed in relation to the meiotic process in other arachnid groups.