Non-homologous Associations Research Articles

Agent-based modeling of nuclear chromosome ensembles identifies determinants of homolog pairing during meiosis.

During meiosis, pairing of homologous chromosomes (homologs) ensures the formation of haploid gametes from diploid precursor cells, a prerequisite for sexual reproduction. Pairing during meiotic prophase I facilitates crossover recombination and homolog segregation during the ensuing reductional cell division. Mechanisms that ensure stable homolog alignment in the presence of an excess of non-homologous chromosomes have remained elusive, but rapid chromosome movements appear to play a role in the process. Apart from homolog attraction, provided by early intermediates of homologous recombination, dissociation of non-homologous associations also appears to contribute to homolog pairing, as suggested by the detection of stable non-homologous chromosome associations in pairing-defective mutants. Here, we have developed an agent-based model for homolog pairing derived from the dynamics of a naturally occurring chromosome ensemble. The model simulates unidirectional chromosome movements, as well as collision dynamics determined by attractive and repulsive forces arising from close-range physical interactions. Chromosome number and size as well as movement velocity and repulsive forces are identified as key factors in the kinetics and efficiency of homologous pairing in addition to homolog attraction. Dissociation of interactions between non-homologous chromosomes may contribute to pairing by crowding homologs into a limited nuclear area thus creating preconditions for close-range homolog attraction. Incorporating natural chromosome lengths, the model accurately recapitulates efficiency and kinetics of homolog pairing observed for wild-type and mutant meiosis in budding yeast, and can be adapted to nuclear dimensions and chromosome sets of other organisms.

Dispersive forces and resisting spot welds by alternative homolog conjunction govern chromosome shape in Drosophila spermatocytes during prophase I

The bivalent chromosomes that are generated during prophase of meiosis I comprise a pair of homologous chromosomes. Homolog pairing during prophase I must include mechanisms that avoid or eliminate entanglements between non-homologous chromosomes. In Drosophila spermatocytes, non-homologous associations are disrupted by chromosome territory formation, while linkages between homologous chromosomes are maintained by special conjunction proteins. These proteins function as alternative for crossovers that link homologs during canonical meiosis but are absent during the achiasmate Drosophila male meiosis. How and where within bivalents the alternative homolog conjunction proteins function is still poorly understood. To clarify the rules that govern territory formation and alternative homolog conjunction, we have analyzed spermatocytes with chromosomal aberrations. We examined territory formation after acute chromosome cleavage by Cas9, targeted to the dodeca satellite adjacent to the centromere of chromosome 3 specifically in spermatocytes. Moreover, we studied territory organization, as well as the eventual orientation of chromosomes during meiosis I, in spermatocytes with stable structural aberrations, including heterozygous reciprocal autosomal translocations. Our observations indicate that alternative homolog conjunction is applied in a spatially confined manner. Comparable to crossovers, only a single conjunction spot per chromosome arm appears to be applied usually. These conjunction spots resist separation by the dispersing forces that drive apart homologous pericentromeric heterochromatin and embedded centromeres within territories, as well as the distinct chromosomal entities into peripheral, maximally separated territories within the spermatocyte nucleus.

Bivalent individualization during chromosome territory formation in Drosophila spermatocytes by controlled condensin II protein activity and additional force generators.

Reduction of genome ploidy from diploid to haploid necessitates stable pairing of homologous chromosomes into bivalents before the start of the first meiotic division. Importantly, this chromosome pairing must avoid interlocking of non-homologous chromosomes. In spermatocytes of Drosophila melanogaster, where homolog pairing does not involve synaptonemal complex formation and crossovers, associations between non-homologous chromosomes are broken up by chromosome territory formation in early spermatocytes. Extensive non-homologous associations arise from the coalescence of the large blocks of pericentromeric heterochromatin into a chromocenter and from centromere clustering. Nevertheless, during territory formation, bivalents are moved apart into spatially separate subnuclear regions. The condensin II subunits, Cap-D3 and Cap-H2, have been implicated, but the remarkable separation of bivalents during interphase might require more than just condensin II. For further characterization of this process, we have applied time-lapse imaging using fluorescent markers of centromeres, telomeres and DNA satellites in pericentromeric heterochromatin. We describe the dynamics of the disruption of centromere clusters and the chromocenter in normal spermatocytes. Mutations in Cap-D3 and Cap-H2 abolish chromocenter disruption, resulting in excessive chromosome missegregation during M I. Chromocenter persistence in the mutants is not mediated by the special system, which conjoins homologs in compensation for the absence of crossovers in Drosophila spermatocytes. However, overexpression of Cap-H2 precluded conjunction between autosomal homologs, resulting in random segregation of univalents. Interestingly, Cap-D3 and Cap-H2 mutant spermatocytes displayed conspicuous stretching of the chromocenter, as well as occasional chromocenter disruption, suggesting that territory formation might involve forces unrelated to condensin II. While the molecular basis of these forces remains to be clarified, they are not destroyed by inhibitors of F actin and microtubules. Our results indicate that condensin II activity promotes chromosome territory formation in co-operation with additional force generators and that careful co-ordination with alternative homolog conjunction is crucial.

BRUCE preserves genomic stability in the male germline of mice.

BRUCE is a DNA damage response protein that promotes the activation of ATM and ATR for homologous recombination (HR) repair in somatic cells, making BRUCE a key protector of genomic stability. Preservation of genomic stability in the germline is essential for the maintenance of species. Here, we show that BRUCE is required for the preservation of genomic stability in the male germline of mice, specifically in spermatogonia and spermatocytes. Conditional knockout of Bruce in the male germline leads to profound defects in spermatogenesis, including impaired maintenance of spermatogonia and increased chromosomal anomalies during meiosis. Bruce-deficient pachytene spermatocytes frequently displayed persistent DNA breaks. Homologous synapsis was impaired, and nonhomologous associations and rearrangements were apparent in up to 10% of Bruce-deficient spermatocytes. Genomic instability was apparent in the form of chromosomal fragmentation, translocations, and synapsed quadrivalents and hexavalents. In addition, unsynapsed regions of rearranged autosomes were devoid of ATM and ATR signaling, suggesting an impairment in the ATM- and ATR-dependent DNA damage response of meiotic HR. Taken together, our study unveils crucial functions for BRUCE in the maintenance of spermatogonia and in the regulation of meiotic HR-functions that preserve the genomic stability of the male germline.

45S rDNA sites in meiosis of Lolium multiflorum Lam.: variability, non-homologous associations and lack of fragility

In this study, we evaluated the behavior of 45S ribosomal DNA (rDNA) sites during the meiosis of Lolium multiflorum. The reason to study it in this species is that 45S rDNA sites are usually visualized as gaps in mitotic metaphase chromosomes and were initially denominated fragile sites (FSs). In different species, FSs were related to rearrangements that alter the karyotype and affect the chromosome pairing in meiosis. However, our findings show that the chromosome pairing in L. multiflorum is regular and, as in mitosis, the number of sites is variable. In diakinesis with five sites, one of the bivalents was in hemizygous state while, in diakinesis with seven sites, one of the bivalents had three conspicuous signals, two in syntheny in one of the homologous. Only four cells had gaps in the region of the 45S rDNA. Owing to the lower number of signals observed at the initial stages of meiosis, it is assumed that they are involved both in homologous and non-homologous associations and that they might assist the chromosome pairing. Regarding segregation, only meiocytes with five and six 45S rDNA signals were observed, and they were characterized by the segregation of 2/3 signals in the poles of anaphases I up to metaphases II; 2/2 and 3/3 in anaphases II and telophases II; and also 2/2 and 4/4 in the nuclei of tetrads, unlike the number of 45S signals expected. The numerical non-equivalence of sites among nuclei at later stages of meiosis is explained by the presence of chromosomes with hemizygous sites.

Immunodeficiency, centromeric heterochromatin instability of chromosomes 1, 9, and 16, and facial anomalies: the ICF syndrome

Instability of the heterochromatic centromeric regions of chromosomes 1 associated with immunodeficiency was found in a 3 and half months old girl. The case was referred to Department of Genetics, Global Reference Laboratory, Metropolis Healthcare Ltd, Mumbai with the suspicion of Downs Syndrome for chromosomal karyotyping. This patient had facial anomalies in addition to combined immunodeficiency and chromosomal instability. Stretching of the heterochromatic centromeric regions of chromosomes 1 and homologous and non-homologous associations of these regions were the most common cytogenetic findings in this patient. Multi-branched configurations and whole arm deletions of chromosomes 1 were also found. Comparing clinical and chromosomal data we conclude that the patient was suffering from immunodeficiency, centromeric heterochromatin instability and facial syndrome. The chromosomal karyotyping report was showing instability around vicinity of chromosome 1 and various abnormalities around vicinity of both chromosomes 1 were found in form of random breakages of chromosome 1, fragile sites, deletions/duplications of small and long arm, extra copies of chromosome 1 with rosette formations, exchange of arms and partial aneuploidies of chromosome 1. Further, the investigations regarding the immune status revealed that the level of IgM (5.98 mg/dl), IgA (<6.16mg/dl) and IgG (92.10 mg/dl) subgroup of immunoglobulin was very low. The results were consistent with The Immunodeficiency, Centromeric region instability, Facial anomalies (ICF) syndrome. Second sample from the patient for molecular studies could not be collected and performed since the patient failed to survive after 3 and half months.

Meiotic Cohesin SMC1β Provides Prophase I Centromeric Cohesion and Is Required for Multiple Synapsis-Associated Functions

Cohesin subunit SMC1β is specific and essential for meiosis. Previous studies showed functions of SMC1β in determining the axis-loop structure of synaptonemal complexes (SCs), in providing sister chromatid cohesion (SCC) in metaphase I and thereafter, in protecting telomere structure, and in synapsis. However, several central questions remained unanswered and concern roles of SMC1β in SCC and synapsis and processes related to these two processes. Here we show that SMC1β substantially supports prophase I SCC at centromeres but not along chromosome arms. Arm cohesion and some of centromeric cohesion in prophase I are provided by non-phosphorylated SMC1α. Besides supporting synapsis of autosomes, SMC1β is also required for synapsis and silencing of sex chromosomes. In absence of SMC1β, the silencing factor γH2AX remains associated with asynapsed autosomes and fails to localize to sex chromosomes. Microarray expression studies revealed up-regulated sex chromosome genes and many down-regulated autosomal genes. SMC1β is further required for non-homologous chromosome associations observed in absence of SPO11 and thus of programmed double-strand breaks. These breaks are properly generated in Smc1β−/− spermatocytes, but their repair is delayed on asynapsed chromosomes. SMC1α alone cannot support non-homologous associations. Together with previous knowledge, three main functions of SMC1β have emerged, which have multiple consequences for spermatocyte biology: generation of the loop-axis architecture of SCs, homologous and non-homologous synapsis, and SCC starting in early prophase I.

Together yes, but not coupled: new insights into the roles of RAD51 and DMC1 in plant meiotic recombination

The eukaryotic recombinases RAD51 and DMC1 are essential for DNA strand-exchange between homologous chromosomes during meiosis. RAD51 is also expressed during mitosis, and mediates homologous recombination (HR) between sister chromatids. It has been suggested that DMC1 might be involved in the switch from intersister chromatid recombination in somatic cells to interhomolog meiotic recombination. At meiosis, the Arabidopsis Atrad51 null mutant fails to synapse and has extensive chromosome fragmentation. The Atdmc1 null mutant is also asynaptic, but in this case chromosome fragmentation is absent. Thus in plants, AtDMC1 appears to be indispensable for interhomolog homologous recombination, whereas AtRAD51 seems to be more involved in intersister recombination. In this work, we have studied a new AtRAD51 knock-down mutant, Atrad51-2, which expresses only a small quantity of RAD51 protein. Atrad51-2 mutant plants are sterile and hypersensitive to DNA double-strand break induction, but their vegetative development is apparently normal. The meiotic phenotype of the mutant consists of partial synapsis, an elevated frequency of univalents, a low incidence of chromosome fragmentation and multivalent chromosome associations. Surprisingly, non-homologous chromosomes are involved in 51% of bivalents. The depletion of AtDMC1 in the Atrad51-2 background results in the loss of bivalents and in an increase of chromosome fragmentation. Our results suggest that a critical level of AtRAD51 is required to ensure the fidelity of HR during interchromosomal exchanges. Assuming the existence of asymmetrical DNA strand invasion during the initial steps of recombination, we have developed a working model in which the initial step of strand invasion is mediated by AtDMC1, with AtRAD51 required to check the fidelity of this process.

C-heterochromatin during synapsis and recombination in grey cockroach Nauphoeta cinerea spermatogenesis

The behaviour of large distal C-heterochromatic blocks in spermatogenesis of grey cockroach Nauphoeta cinerea was investigated by light and electron microscopy. Heterochromatic blocks of some autosomes are involved into nonhomologous association with formation of chromocenter in early meiotic prophase I. Fluorescent in situ hybridization with ribosomal DNA-probe revealed the signal on only two pairs of middle chromosomes, which did not participate in the chromocenter formation, therefore appearance of ectopic contacts was not caused by the nucleolus formation. The analysis showed that heterochromatin in the chromocenter was eliminated spatially and functionally from main meiotic events. Heterochromatin does not participate in the bouquet formation, initiation of homologous pairing, and recombination events. Heterochromatic polymorphism results in asymmetric synaptonemal complexes with different degree of synaptic adjustment. Meiotic axis of sex univalent (XO -- sex male determination) is split in different regions irrespective of heterochromatin localization.

A new look on the role of the diffuse stage in problems of plant and animal meiosis

Especially in plant hybrids and polyploids, the diffuse stage occurring between pachytene and diplotene can be a source of the following meiotic defects: (1) difficulties in stainability or fixability of diplotene or diakinesis chromosomes, (2) desynchronisation of meiotic stages in the anther. (3) timing upset of meiosis, defects in spiralisation, (4) the irregular occurrence of nonhomologous multivalent associations or stickiness in diakinesis or metaphase I, (5) normal anaphase configurations after a strongly aberrant diakinesis stage, (6) irregular occurrence of anaphase 1 bridges of uncertain origin, (7) variable chromosomal behaviour in different cells, anthers and buds in the same plant. The literature on plants where the diffuse stage was reported is reviewed. As similar defects are found in meiosis of many other plants, the diffuse stage is supposed to be widespread in plant meiosis but as it has an interphase-like structure it is overlooked in squash preparations. The aberrations are believed to be caused by disturbances of the folding process between the diffuse stage and diakinesis, due to a different constitution of chromosomes from different parents or to internal unbalance of chromosome sets in haploids. polyploids and hybrids. Presumably, spontaneous breaks in the chromatin fibers (23 nm) and exchanges sometimes occur during prophase. As the chromatin fibers are the real functional chromatids, the exchanges are regarded as chromatid aberrations.

Genetic Regulation of Meiotic Cross-Overs between Related Genomes inBrassica napusHaploids and Hybrids

Although the genetic regulation of recombination in allopolyploid species plays a pivotal role in evolution and plant breeding, it has received little recent attention, except in wheat (Triticum aestivum). PrBn is the main locus that determines the number of nonhomologous associations during meiosis of microspore cultured Brassica napus haploids (AC; 19 chromosomes). In this study, we examined the role played by PrBn in recombination. We generated two haploid x euploid populations using two B. napus haploids with differing PrBn (and interacting genes) activity. We analyzed molecular marker transmission in these two populations to compare genetic changes, which have arisen during meiosis. We found that cross-over number in these two genotypes was significantly different but that cross-overs between nonhomologous chromosomes showed roughly the same distribution pattern. We then examined genetic recombination along a pair of A chromosomes during meiosis of B. rapa x B. napus AAC and AACC hybrids that were produced with the same two B. napus genotypes. We observed significant genotypic variation in cross-over rates between the two AAC hybrids but no difference between the two AACC hybrids. Overall, our results show that PrBn changes the rate of recombination between nonhomologous chromosomes during meiosis of B. napus haploids and also affects homologous recombination with an effect that depends on plant karyotype.

Measuring differentiation among populations at different levels of genetic integration

BackgroundMost genetic studies of population differentiation are based on gene-pool frequencies. Population differences for gene associations that show up as deviations from Hardy-Weinberg proportions (homologous association) or gametic disequilibria (non-homologous association) are disregarded. Thus little is known about patterns of population differentiation at higher levels of genetic integration nor the causal forces.ResultsTo fill this gap, a conceptual approach to the description and analysis of patterns of genetic differentiation at arbitrary levels of genetic integration (single or multiple loci, varying degrees of ploidy) is introduced. Measurement of differentiation is based on the measure Δ of genetic distance between populations, which is in turn based on an elementary genic difference between individuals at any given level of genetic integration. It is proven that Δ does not decrease when the level of genetic integration is increased, with equality if the gene associations at the higher level follow the same function in both populations (e.g. equal inbreeding coefficients, no association between loci). The pattern of differentiation is described using the matrix of pairwise genetic distances Δ and the differentiation snail based on the symmetric population differentiation ΔSD. A measure of covariation compares patterns between levels. To show the significance of the observed differentiation among possible gene associations, a special permutation analysis is proposed. Applying this approach to published genetic data on oak, the differentiation is found to increase considerably from lower to higher levels of integration, revealing variation in the forms of gene association among populations.ConclusionThis new approach to the analysis of genetic differentiation among populations demonstrates that the consideration of gene associations within populations adds a new quality to studies on population differentiation that is overlooked when viewing only gene-pools.

Cytological characteristics of chromosome behaviour during female meiosis in Sphinx ligustri L. (Sphingidae, Lepidoptera)

Nokkala, S. 1987. Cytological characteristics of chromosome behaviour during female meiosis in Sphinx ligustri L. (Sphingidae, Lepidoptera). - Hereditas 106:169–179. Lund, Sweden. ISSN 0018–0661. Received May 26,1986 The behaviour of chromosomes was studied in female meiosis of Sphinx ligustri L. (Sphingidae, Lepidoptera) by using improved cytological methods. The haploid chromosome number is n = 28. Throughout pro-phase from pachytene until metaphase I, bivalents consist of two chromosomes aligned in parallel, i. e., meiosis is achiasmatic. All bivalents appear to be homomorphic, thus the ZW bivalent could not be identified. In early vitellogenesis, bivalents begin to associate nonhomologously via telomeres, forming a simple chain consisting of all 28 bivalents. After secondary modifications, a more complex, branched chain is produced. The branched chain configuration of bivalents is retained throughout the time when yolk is produced, and it breaks down during prometaphase. At metaphase I, only short chains are remaining. Based on present knowledge on chromosome behaviour in female meioses in different ovary types, it is suggested that the formation of the nonhomologous associations of biVdlentS is a prerequisite for karyosome or karyosphere formation, the karyosome being meiotically a resting stage. The effects of nonhomologous associations of bivalents on segregation are discussed.

Telomeric and interstitial telomeric sequences in holokinetic chromosomes of Lepidoptera: telomeric DNA mediates association between postpachytene bivalents in achiasmatic meiosis of females.

Telomeres, besides their main role in the protection and maintenance of chromosome ends, have several other vital functions in the cell cycle. We studied their role in the achiasmatic meiosis of female Lepidoptera, insects with holokinetic chromosomes. By fluorescence in-situ hybridization (FISH) with the insect telomeric probe, (TTAGG)n, we mapped the distribution of telomeric and interstitial telomeric sequences (ITS) in female meiotic chromosomes of two species, Orgyia antiqua with a reduced chromosome number (2n = 28) and Ephestia kuehniella mutants, possessing a radiation-induced chromosome fusion in the genome (2n = 59). In addition to the strong typical telomeric signals, O. antiqua displayed weaker hybridization signals in interstitial sites of pachytene bivalents. The observed ITS most probably reflect remnants of chromosomal rearrangements and support the hypothesis that the Orgyia karyotype had arisen by multiple fusions of ancestral chromosomes. On the other hand, the absence of ITS in the chromosome fusion of Ephestia indicated the loss of telomeres before the two original chromosomes fused. When the telomeric probe was amplified by enzymatic reaction with tyramid, the number of ITS observed increased in Orgyia, and a few ITS were also observed in several chromosomes of Ephestia but not in the fused chromosome. This suggests that the genomes of both species also contain ITS other than those originating from chromosome fusions. The analysis of female meiotic prophase I revealed non-homologous associations of postpachytene bivalents mediated by telomeric DNA, which were not observed in the pachytene stage. Surprisingly, in early postpachytene nuclei the telomeric associations also involved ITS, whereas later postpachytene nuclei displayed chains of bivalents interconnected only by true telomeres. This finding favours a hypothesis that telomeric associations between bivalents play a role in chromosome segregation in the achiasmatic meiosis of female Lepidoptera.

Synaptic behaviour and morphological modifications of the X and Y chromosomes during pachytene in three species of Ctenomys (Rodentia, Caviomorpha, Ctenomyidae).

Synaptic behaviour and the progression of morphological differentiation of the XY chromosome pair during pachytene was studied for the first time in three species of the South American subterranean rodents of the genus Ctenomys (tuco-tucos). In general, synapsis progression in the sex pair could be subdivided into four substages: (i) initial partial synapsis of the X and Y chromosome axes and beginning of the differentiation of the unsynapsed regions; (ii) complete or almost complete synapsis of the Y axis accompanied with morphological differentiation of the unsynapsed region of the X chromosome; (iii) a novel stage exclusive to Ctenomys perrensi consisting in a retraction of the free X axis, associated with the formation of a homogeneous and dense structure along the synaptic region, which leads to the achievement of full synapsis between sex chromosomes; or (iv) an increase in morphological complexity involving extreme splitting of the XY pair. The implications of the peculiar synaptic behaviour displayed by sex chromosomes in C. perrensi, a species complex highly polymorphic for Robertsonian translocations, are discussed in relation to both the triggering of the pachytene checkpoint and the avoidance of non-homologous associations between sex chromosomes and the asynaptic pericentromeric regions of trivalents in translocation heterozygotes.

Sex-dependent synaptic behaviour in triploid turbot, Scophthalmus maximus (Pisces, Scophthalmidae).

A surface-spreading synaptonemal complex (SC) technique was used to analyse spermatocytes and oocytes of triploid turbot (Scophthalmus maximus) in order to visualise the process of chromosome synapsis. The most conspicuous characteristic of triploid oocytes is that, in the trivalents, the lateral elements of the SC were frequently associated in threes, either completely along the length of the trivalent, or partially, forming a variety of forked structures. In these nuclei, synapsis usually occurred among homologous chromosomes and the number of bivalents observed was significantly higher than that expected under the assumption of random chromosome association among all partners. However, the frequency of trivalents was very low in triploid spermatocytes, triple synapsis being also scarce. In these nuclei chromosomes that were excluded from homologous synapsis become engaged in random SC formation, and, therefore a considerable number of non-homologous associations are produced. The causes of the synaptic differences observed in triploid males and females of turbot and their possible relation to the sterility displayed by these animals are discussed.

Chromatin insulator elements block the silencing of a target gene by the Drosophila polycomb response element (PRE) but allow trans interactions between PREs on different chromosomes.

Polycomb response elements (PREs) can establish a silenced state that affects the expression of genes over considerable distances. We have tested the ability of insulator or boundary elements to block the repression of the miniwhite gene by the Ubx PRE. The gypsy element and the scs element interposed between PRE and miniwhite gene protect it against silencing but the scs' is only weakly effective. When the PRE-miniwhite gene construct is insulated from flanking chromosomal sequences by gypsy elements at both ends, it can still establish efficient silencing in some lines but not others. We show that this can be caused by interactions in trans with PREs at other sites. PRE-containing transposons inserted at different sites or even on different chromosomes can interact, resulting in enhanced silencing. These trans interactions are not blocked by the gypsy insulator and reveal the importance of nonhomologous associations between different regions of the genome for both silencing and activation of genes. The similarity between the behavior of PREs and enhancers suggests a model for their long-distance action.

6 Chromosome Rearrangements in Neurospora and Other Filamentous Fungi

Knowledge of fungal chromosome rearrangements comes primarily from N. crassa, but important information has also been obtained from A. nidulans and S. macrospora. Rearrangements have been identified in other Sordaria species and in Cochliobolus, Coprinus, Magnaporthe, Podospora, and Ustilago. In Neurospora, heterozygosity for most chromosome rearrangements is signaled by the appearance of unpigmented deficiency ascospores, with frequencies and ascus types that are characteristic of the type of rearrangement. Summary information is provided on each of 355 rearrangements analyzed in N. crassa. These include 262 reciprocal translocations, 31 insertional translocations, 27 quasiterminal translocations, 6 pericentric inversions, 1 intrachromosomal transposition, and numerous complex or cryptic rearrangements. Breakpoints are distributed more or less randomly among the seven chromosomes. Sixty of the rearrangements have readily detected mutant phenotypes, of which half are allelic with known genes. Constitutive mutations at certain positively regulated loci involve rearrangements having one breakpoint in an upstream regulatory region. Of 11 rearrangements that have one breakpoint in or near the NOR, most appear genetically to be terminal but are in fact physically reciprocal. Partial diploid strains can be obtained as recombinant progeny from crosses heterozygous for insertional or quasiterminal rearrangements. Duplications produced in this way precisely define segments that cover more than two thirds of the genome. Duplication-producing rearrangements have many uses, including precise genetic mapping by duplication coverage and alignment of physical and genetic maps. Typically, fertility is greatly reduced in crosses parented by a duplication strain. The finding that genes within the duplicated segment have undergone RIP mutation in some of the surviving progeny suggests that RIP may be responsible for the infertility. Meiotically generated recessive-lethal segmental deficiencies can be rescued in heterokaryons. New rearrangements are found in 10% or more of strains in which transforming DNA has been stably integrated. Electrophoretic separation of rearranged chromosomal DNAs has found useful applications. Synaptic adjustment occurs in inversion heterozygotes, leading progressively to nonhomologous association of synaptonemal complex lateral elements, transforming loop pairing into linear pairing. Transvection has been demonstrated in Neurospora. Beginnings have been made in constructing effective balancers. Experience has increased our understanding of several phenomena that may complicate analysis. With some rearrangements, nondisjunction of centromeres from reciprocal translocation quadrivalents results in 3:1 segregation and produces asci with four deficiency ascospores that occupy diagnostic positions in linear asci. Three-to-one segregation is most frequent when breakpoints are near centromeres. With some rearrangements, inviable deficiency ascospores become pigmented. Diagnosis must then depend on ascospore viability. In crosses between highly inbred strains, analysis may be handicapped by random ascospore abortion. This is minimized by using noninbred strains as testers.

Meiotic behaviour of 11 primary trisomics in diploid potato and its consequences for the transmission of the extra chromosome

Meiosis was studied in 11 primary trisomics (2n = 2x + 1 = 25) from diploid Solanum tuberosum L. ssp. tuberosum Hawkes (2n = 2x = 24) and from interspecific Solanum hybrids. The three homologous chromosomes were associated in a trivalent in 90% of more than 175 pollen mother cells analysed at pachytene. Trivalents showing a two by two pairing and partner exchanges at pachytene along with incomplete triple synapsis were frequently observed. Foldback pairing, predominantly observed in the heterochromatic parts of the chromosomes, occurred in 28.3% of the trivalents. Nonhomologous association of chromosome segments was observed in 29.1% of the trivalents. Up to six telomeres were associated homologously in 46.2% of the trivalents. Genotypic differences with respect to trivalent formation at metaphase I occurred in almost all the trisomic types and was significant for the trisomics for chromosomes 4, 7, and 9. The coefficient of realization of a trivalent (CRT) at metaphase I in the 11 primary trisomics varied from 0.20 to 0.80, and was positively correlated with the absolute length of the extra chromosome (r = 0.61, p ≤ 0.05) and with the absolute, as well as the relative, length of the euchromatic segments of the extra chromosome (r = 0.70, p ≤ 0.05). There was no apparent relationship between the CRT and the distribution pattern at anaphase I or anaphase II. The rate of female transmission of the extra chromosome varied from 10.0 to 44.6% among different trisomics. For chromosomes 4, 7, and 9, this rate differed significantly among genotypes of the same trisomic.

ICF syndrome with variable expression in sibs.

We describe a new familial case of ICF syndrome (immunodeficiency, centromeric instability, facial anomalies) in a woman of 29 years and in her brother of 30 years. The proband showed mental retardation, facial anomalies, recurrent respiratory infections, combined deficit of IgM and IgE immunoglobulin classes, and paracentromeric heterochromatin instability of chromosomes 1, 9, and 16. The brother had minor signs of the syndrome and had an apparently normal phenotype. Their parents were healthy and non-consanguineous. Chromosome anomalies consisted of homologous and non-homologous associations, chromatid and isochromatid breaks, deletions of whole arms, interchanges in the paracentromeric region, and multibranched configurations of chromosomes 1, 9, and 16. CD bands and fluorescence in situ hybridisation with alphoid DNA sequence probes specific for the centromeres of chromosomes 1 and 16 showed that the centromere was not directly implicated in the formation of multibranched configurations. These cases indicate the autosomal recessive mode of inheritance and the variable expressivity of the ICF syndrome.