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Meiosis is the basis of the generative reproduction of eukaryotes. The crucial first step is homologous chromosome pairing. In higher eukaryotes, micrometer-scale chromosomes, micrometer distances apart, are brought together by nanometer DNA sequences, at least a factor of 1000 size difference. Models of homology search, homologue movement, and pairing at the DNA level in higher eukaryotes are primarily based on studies with yeast where the emphasis is on the induction and repair of DNA double-strand breaks (DSB). For such a model, the very large nuclei of most plants and animals present serious problems. Homology search without DSBs cannot be explained by models based on DSB repair. The movement of homologues to meet each other and make contact at the molecular level is not understood. These problems are discussed and the conclusion is that at present practically nothing is known of meiotic homologue pairing in higher eukaryotes up to the formation of the synaptonemal complex, and that new, necessarily speculative models must be developed. Arguments are given that RNA plays a central role in homology search and a tentative model involving RNA in homology search is presented. A role of actin in homologue movement is proposed. The primary role of DSBs in higher eukaryotes is concluded to not be in pairing but in the preparation of Holliday junctions, ultimately leading to chromatid exchange.

Chromosomal rearrangements, including reciprocal translocations, may prevent recombinational transfer of genes from a donor genotype to a recipient, especially when the gene is located in an interstitial segment. The effect of trisomy of chromosome arm 1RS on recombination was studied in translocation heterozygote T248W of rye ( Secale cereale ), involving chromosome arms 1RS and 6RS. (Pro)metaphase I configuration frequencies were analyzed. Crossing over, estimated as chiasma parameters, in five genetically different euploid heterozygotes was compared with those of 10 different single arm trisomics. The addition of 1RS greatly altered the crossing over pattern around the translocation break point, with a special increase in the interstitial segment of 6RS and adjoining regions, normally hardly accessible to recombination. Furthermore, there was considerable variation between plants of closely related genotypes. Heterogeneity widens the distribution of crossing overs, including segments normally not accessible to recombination, but decreases average recombination in other segments. The extra chromosome and abnormal segregants are eliminated by using the trisomic as the pollen parent.

Synaptic behaviour of the two tetraploids rye cultivars Gigantón (G) and Tetrapico (T) displaying significant differences in their quadrivalent frequencies at metaphase I was analyzed by electron microscopy in surface-spread prophase I nuclei. A different behaviour was observed between the two cultivars; the synaptonemal complex (SC) quadrivalents frequency being significantly higher in G than in T at prophase I. Moreover, the G SC quadrivalents had more synaptic partner exchanges (SPEs) and their location was more distal than the T SC quadrivalents. However, inverse findings were found at metaphase I, the quadrivalent frequency was higher in T than in G. The role that different factors, mainly the number and location of the SPEs and the frequency and distribution of chiasmata, could play in the evolution from prophase I to metaphase I in both cultivars is discussed.Key words: autotetraploid rye, synaptonemal complex, spreading.

The paradigm that meiotic recombination and chiasmata have the same basis has been challenged, primarily for plants. High resolution genetic mapping frequently results in maps with lengths far exceeding those based on chiasma counts. In addition, recombination between specific homoeologous chromosomes derived from interspecific hybrids is sometimes much higher than can be explained by meiotic chiasma frequencies. However, almost the entire discrepancy disappears when proper care is taken of map inflation resulting from the shortcomings of the mapping algorithm and classification errors, the use of dissimilar material, and the difficulty of accurately counting chiasmata. Still, some exchanges, especially of short interstitial segments, cannot readily be explained by normal meiotic behaviour. Aberrant meiotic processes involving segment replacement or insertion can probably be excluded. Some cases of unusual recombination are somatic, possibly premeiotic exchange. For other cases, local relaxation of chiasma interference caused by small interruptions of homology disturbing synaptonemal complex formation is proposed as the cause. It would be accompanied by a preference for compensating exchanges (negative chromatid interference) resulting from asymmetry of the pairing chromatid pairs, so that one side of each pair preferentially participates in pairing. Over longer distances, the pairing face may switch, causing the normal random chromatid participation in double exchanges and the relatively low frequency of short interstitial exchanges. Key words : recombination frequency, map length, chiasmata, discrepancy, chromatid interference.

American as well as British forms of the perennial plant Lathyrus palustris have 2n = 42 chromosomes with one group of 6 long submetacentric chromosomes, two groups of 6 medium-long subacrocentric chromosomes, three groups of 6 medium-long to medium-short submetacentric to subacrocentric chromosomes, and one group of 6 medium-short metacentric chromosomes. One haploid complement measures 45.8 μm at somatic metaphase. At meiotic first metaphase both forms show extensive multivalent formation. In 50 metaphase I cells of one plant of the British form there were 51 ring hexavalents, 80 chain and open-branched hexavalents of various forms, 2 quinquivalents, 41 ring quadrivalents, 58 chain quadrivalents, 20 branched quadrivalents of various forms, 1 trivalent, 128 ring bivalents, 280 open bivalents, and 9 univalents. It is concluded that L. palustris is most likely a natural autohexaploid with frequent multivalent pairing, many points of pairing initiation, several partner exchanges per multivalent, and many interstitial chiasmata. It may well be the first fully documented true natural autohexaploid reported. Pollen fertility was 56.2% and seed set 37%, apparently sufficient for a perennial plant species.Key words: Lathyrus, natural, autohexaploid, meiosis, multivalents.

Meiotic metaphase I configuration frequencies were determined in different tillers of genetically related plants of rye ( Secale cereale L.) heterozygous for reciprocal translocation T248W (between chromosome arms 1RS and 6RS) and with an additional (telocentric) arm 1RS. Seventeen different configurations could be recognized, grouped into three categories. Very different configuration frequencies were found not only between sister plants from the same parents but also between tillers of the same plant grown under identical conditions (climate chambers at 15 °C and 20 °C). The heterogeneity reflects variation in chromosome pairing and crossing over, and is variable and unpredictable. Anthers within florets were homogeneous. Between tiller heterogeneity is insufficient to explain differences between sister plants. It is ascribed to random somatic variation in the conditions of the chromatin which, at meiosis, govern chromosome pairing. During sexual differentiation, these conditions are fixed and subsequent cell lineages have the same pairing and crossing over characteristics. As homology search is an activity of DNA, this control of pairing and crossing over, consistent over long cell lineages, may be considered to be epigenetic even when no realistic suggestions concerning its character can be given.

Mathematical models are presented for estimating preferential pairing and chiasma parameters in amphidiploids and autotetraploids on the basis of diakinesis or metaphase I configuration frequencies and are compared with other approaches of estimating affinity. With a preferential pairing factor p, estimated from quadrivalent and trivalent frequencies, and estimated chiasmate association factors for the two arms in quadrivalents (a(qu) and b(qu) for arms A and B, respectively) and in bivalents (a(bi) and b(bi)) a perfect fit between observed and predicted configuration frequencies can often be obtained in amphidiploids of several plant species, including Solanaceae and Gramineae. Since several proven autotetraploids give very similar apparent preferential pairing estimates, the biological significance of such parameters as preferential pairing and affinity factors is considered limited. The same is true for pairing parameters estimated by optimizing fit of configuration frequencies expected on the basis of theoretical models to observed data.Key words: amphidiploid, diakinesis, metaphase, meiosis, model, preferential pairing, tetraploid.

In the tetraploid somatic hybrid between the diploid Lycopersicon species L. esculentum (tomato) and L. peruvianum, synaptonemal complexes formed quadrivalents in 73 of the 120 sets of four chromosomes (60.8%) in 10 cells studied in detail at pachytene. Of these, 43 had one pairing partner exchange, 22 had two, and 8 had three, very close to a Poisson distribution. The points of pairing partner exchange were concentrated at the middle of the two arms. The frequency per arm corresponded with physical arm length. There was a sharp drop around the centromere, and pericentric heterochromatin had a slightly lower probability of being involved in pairing partner exchange than euchromatin. The chromosomes align before pairing and there are several points of pairing initiation, with concentrations at or near the ends and the centromere. From zygotene to late pachytene the quadrivalent frequency decreased considerably. At late pachytene it was lower than expected with the observed high frequency of pairing partner exchange. Pairing affinity between species was only slightly lower than affinity within species, in spite of considerable genetic differentiation. The frequency of recombination nodules increased from early to late zygotene and then decreased strongly to full pachytene. There is a highly significant negative correlation between percent pairing and SC length. At metaphase I the frequency of quadrivalents was 0.444, and branched quadrivalents were rare, probably caused by interference and restriction of chiasma formation to distal euchromatin. Metaphase I quadrivalent frequency is a relatively good indication of pairing affinity in this material.Key words: Lycopersicon, tetraploid hybrid, synaptonemal complex, pairing partner exchange, diakinesis/metaphase I.

Genetically stable artificial autotetraploids with over 90% seed set were obtained by colchicine treatment of the solanaceous species Hyoscyamus niger L. (4x = 68). The tetraploids were vigorous and had earlier been shown to yield considerably more tropane alkaloid per individual than the source diploids. The tetraploids had high bivalent frequencies at meiosis, and high fertility as a result of the formation of genetically balanced gametes. There was a systematic and significant decrease in quadrivalent frequency and an increase in bivalent frequency in three subsequent generations tested (C0, C1, C2). Possible causes of high bivalent frequency are discussed. Seed fertility can probably be increased in advanced generations by further selection for fertility and maximization of heterozygosity.Key words: Hyoscyamus niger, autotetraploid, meiosis, bivalents, fertility, selection.

The association pattern of marked tetrasomes of Secale chromosome 1R at meiotic first metaphase was analyzed. Two of the four chromosomes were identical with terminal C-bands at both arms; the other two were also identical but lacked C-bands and were homologous or homeologous with the first two. Four different types of heterozygotes for 1R were studied: (i) autotetraploid hybrids between genetic variants within Secale cereale subsp. cereale, (ii) tetraploid hybrids between subspecies of Secale cereale, (iii) tetraploid hybrids between species of Secale, and (iv) autotetrasomes of S. cereale in a wheat background. Earlier observations that heterozygous associations (banded with unbanded) had consistently higher chiasma frequencies than homozygous associations were extended and confirmed. To analyze this phenomenon more closely, the possible relations between this correlation and several other meiotic phenomena were studied. For this analysis, three genetically different autotetraploid hybrids within S. cereale were selected that differed with respect to the relation between pairing type and chiasma frequency. Special attention was given to different patterns of interference and other meiotic phenomena in the two chromosome arms of chromosome 1R. No relations between such phenomena and the relation between pairing type and chiasma frequency could be established. A hypothesis is formulated assuming that long-distance homologue attraction is concentrated in a limited number of sites and that in different genotypes, different patterns of active sites are present. Moderately weak attraction sites can pair with strong homologous sites under favorable genetic conditions, but two weak sites cannot. Then, heterozygotes have more effective pairing initiation and consequently chiasma formation than homozygotes. Under less favorable conditions, only strong sites are effective, and then, homozygotes pair better, but the chiasma frequency is lower. A model of the forces involved in homologue attraction is presented.Key words: autopolyploids, preferential pairing, chiasma frequency, homologue attraction, Secale.

During a workshop with 13 participants, several aspects of multivalent orientation at meiotic (pro)metaphase were discussed in an attempt to resolve some of the most prominent controversies with respect to terminology, interpretation of observations, and the validity of hypotheses and theories. For several terms and concepts, descriptive definitions were formulated that are recommended for general use. In the analysis of the behaviour of the multivalent in meiosis preprometaphase shape and position as important factors in final orientation were discussed, as well as the first contact between spindle and kinetochores and the role of reorientation. Specific characteristics of different multivalents and expected frequencies of different orientation types were considered. Finally, a few remarks on data collection and analytical procedures were made Key words: meiosis, multivalents, orientation, workshop.

Different mathematical models for estimating pairing and recombination parameters for triploid hybrids with two-armed chromosomes are discussed. In most models all information on preferential pairing is contained in the ratio of trivalents and ring bivalents and can be estimated independently of the chiasmate association frequencies in the two chromosome arms. The single degree of freedom available only permits the estimation of ranges of the three possible pairing relations between the three genomes. Alternatively, a single parameter can be estimated when additional assumptions are made. When the ratio r between the frequencies of trivalents and ring bivalents, independent of arm length differences and with a theoretical maximum of 2, is about 1, the ranges of the frequencies of the three pairing combinations are wide. When r becomes smaller than 1, very soon the ranges become limited to values where one of the three is relatively large and positive and the other two negative and varying between equal to very different, depending on slight changes in the first. When the frequency of open bivalents is relatively high and the frequency of univalents low, there most probably is a difference in chiasmate association frequency between the two arms of the average chromosome and this difference can be quantified. When the number of univalents is only slightly higher than expected on the basis of the number of open bivalents, the reason may be that (quantifiably) more chiasmata are formed after bivalent pairing than after trivalent pairing within certain ranges of r, and certain ranges of the average chiasmate association frequency. When the excess of univalents is larger, this is best explained by a failure of entire chromosomes to find each other. This degree of pairing failure can be estimated. All models have been applied to the triploid hybrid between allotetraploid Trifolium repens and diploid T. nigrescens. Assuming that the two genomes of T. repens do not pair, which cannot be demonstrated with certainty, the two genomes pair with the nigrescens genome with frequencies of 0.828 and 0.171, respectively. Introgression then occurs into either genome but not with the same frequency. If the repens genomes pair, this would be caused by either genetic factors disturbing the normal pairing behaviour or the absence of strict homologues. Then the relative pairing frequency between T. nigrescens and one of the T. repens genomes would be 0.838 and on an average 0.081 between the two other combinations, with a possibly considerable but unknown difference. The high average chiasmate arm association frequency (0.650) suggests that affinity between the pairing genomes is not very low. The average two arms do not differ in chiasma frequency.Key words: triploid hybrids, preferential pairing, recombination, mathematical models, Trifolium.

Four rye plants heterozygous for translocation 248, involving chromosomes 1R and 6R, were used for a comparative study of synaptonemal complex formation at midprophase I and chromosome configurations at metaphase I. Synaptonemal complexes were obtained with a cell-spreading technique and studied with electron microscopy. The total length of the synaptonemal complexes in the 28 analyzable pachytene nuclei varied considerably, both within and among plants. The variation of synaptonemal complex lengths of the bivalents in a nucleus was partly stage dependent; i.e., it was greater at early than at late pachytene. In all but one pachytene nucleus, pairing in the quadrivalent was regular, and the four pairing arms were usually easy to identify. Most noticeable was the variation of pairing saturation at the breakpoint of the quadrivalent. Pairing in the breakpoint region was delayed with respect to the pairing in the bivalents. Variation in the arm lengths of the quadrivalent was the result of incomplete and nonhomologous pairing at the breakpoint as well as differential contraction rates among chromosome segments. It was shown that the completion of delayed pairing throughout pachytene is mainly long-arm pairing. The actual breakpoint was therefore not in the middle of the unpaired segments, but more distal. The analysis of metaphase I nuclei revealed that chiasma frequency in this material was higher than in similar material used in former studies. When one of the translocation segments lacked a chiasma, this was in most cases the short translocated segment 1RS, the terminal segment of the satellite of chromosome 1R. Positive chiasma interference was demonstrated between the interstitial and exchanged segment in 1RS. This agreed with the observation of a negative correlation in extent of pairing between these two segments. Other interference phenomena, which have been described for this translocation in other material, remained undetected because of lack of variation in chiasma formation owing to high chiasma frequency.Key words: Secale cereale, meiosis, translocation, synaptonemal complex, metaphase I.