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We studied twelve late Holocene organic deposits in West-Frisia, The Netherlands. Pollen, spores, non-pollen palynomorphs, mosses, other botanical macrofossils and insect remains were recorded for reconstructions of changing environmental conditions. Eastern West-Frisia was a cultivated landscape during the Bronze Age, but it became a freshwater wetland in the Late Bronze Age. In most of our sites, radiocarbon dates show that time transgressive inundation of soils preceded the climate shift at 850 cal BC for several centuries. We suggest that solar forcing of climate change may have delivered the final push to the inundation and depopulation of West-Frisia, which had already commenced several centuries before, due to sealevel rise. We did not find evidence for significant Bronze Age tree growth in West-Frisia before the inundations. Vegetation successions in the new wetlands developed from shallow mineral-rich freshwater to rich-fen vegetation. Subsequently poor fen vegetation with birch and pine developed, and the natural succession led to ombrotrophic raised bog vegetation. Complete successions from shallow, mineral-rich lakes to raised bog lasted between 1000 and 1500 calendar years. We hypothesize that medieval drainage and reclamation became possible only when the mires of West-Frisia had reached the raised bog stage. Reclamation of raised bogs by medieval farmers (drainage, eutrophication, peat digging) caused compaction, oxidation and loss of the upper part of the peat deposit. Seeds of salt-tolerant and salt-demanding plant species indicate that the medieval sites were inundated during storm surges with brackish or salt water, which triggered the farmers to build artificial mounds and, later, dikes. Under mounds and dikes, peat deposits remained protected against further decay. With our data we deliver a long-term perspective on contemporary ecosystem dynamics of freshwater wetlands, relevant for nature conservation and future climate change.

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.

Background: Cholangiocytes are the target cells of cholangiopathies including primary sclerosing cholangitis (PSC). Vimentin is an intermediate filament protein that has been found in various types of mesenchymal cells. The aim of this study is to evaluate the role of vimentin in the progression of biliary damage/liver fibrosis and whether there is a mesenchymal phenotype of cholangiocytes in the Mdr2−/− model of PSC. Methods: In vivo studies were performed in 12 wk. Mdr2−/− male mice with or without vimentin Vivo-Morpholino treatment and their corresponding control groups. Liver specimens from human PSC patients, human intrahepatic biliary epithelial cells (HIBEpiC) and human hepatic stellate cell lines (HHSteCs) were used to measure changes in epithelial-to-mesenchymal transition (EMT). Findings: There was increased mesenchymal phenotype of cholangiocytes in Mdr2−/− mice, which was reduced by treatment of vimentin Vivo-Morpholino. Concomitant with reduced vimentin expression, there was decreased liver damage, ductular reaction, biliary senescence, liver fibrosis and TGF-β1 secretion in Mdr2−/− mice treated with vimentin Vivo-Morpholino. Human PSC patients and derived cell lines had increased expression of vimentin and other mesenchymal markers compared to healthy controls and HIBEpiC, respectively. In vitro silencing of vimentin in HIBEpiC suppressed TGF-β1-induced EMT and fibrotic reaction. HHSteCs had decreased fibrotic reaction and increased cellular senescence after stimulation with cholangiocyte supernatant with reduced vimentin levels. Interpretation: Our study demonstrated that knockdown of vimentin reduces mesenchymal phenotype of cholangiocytes, which leads to decreased biliary senescence and liver fibrosis. Inhibition of vimentin may be a key therapeutic target in the treatment of cholangiopathies including PSC. Fund: National Institutes of Health (NIH) awards, VA Merit awards. Keywords: Ductular reaction, Fibroblast, Fibrosis, Senescence, Transforming growth factor beta 1

BACKGROUND: Following liver injury, mast cells (MCs) migrate into the liver and are activated in cholestatic patients. Inhibition of MC mediators decreases ductular reaction (DR) and liver fibrosis. TGF-β1 contributes to fibrosis and promotes liver disease. AIM: To demonstrate that reintroduction of MCs induces cholestatic injury via TGF-β1. METHODS: Wild-type, Kit(W-sh) (MC-deficient), and Mdr2(−/−) mice lacking l-histidine decarboxylase were injected with vehicle or PKH26-tagged murine MCs pretreated with 0.01% DMSO or the TGF-βR inhibitor, LY2109761 (TGF-βRi, 10 μM) three days prior to sac. Hepatic damage was assessed by H&E and serum chemistry. Injected MCs were detected in liver, spleen and lung by immunofluorescence (IF). DR was measured by CK-19 immunohistochemistry (IHC) and F4/80 staining coupled with qPCR for IL-1β, IL-33 and F4/80; biliary senescence evaluated by IF or qPCR for p16, p18 and p21. Fibrosis was evaluated by Sirius Red/Fast Green staining and IF for SYP-9, desmin and α-SMA. TGF-β1 secretion/expression was measured by EIA and qPCR. Angiogenesis was detected by IF for vonWillebrand Factor and VEGF-C qPCR. In vitro, MC TGF-β1 expression/secretion were measured after TGF-βRi treatment; conditioned medium was collected. Cholangiocytes and hepatic stellate cells (HSCs) were treated with MC conditioned medium and biliary proliferation/senescence was measured by MTS and qPCR; HSC activation evaluated for α-SMA, SYP-9 and collagen type-1a expression. RESULTS: MC injection recapitulates cholestatic liver injury characterized by increased DR; fibrosis/TGF-β1 secretion; and angiogenesis. Injection of MC-TGF-βRi reversed these parameters. In vitro, MCs induce biliary proliferation/senescence and HSC activation that was reversed with MCs lacking TGF-β1. CONCLUSION: Our novel study demonstrates that reintroduction of MCs mimics cholestatic liver injury and MC-derived TGF-β1 may be a target in chronic cholestatic liver disease.

Primary biliary cholangitis (PBC) primarily targets cholangiocytes and is characterized by liver fibrosis and biliary proliferation. Activation of the secretin (Sct)/secretin receptor (SR) axis, expressed only by cholangiocytes, increases biliary proliferation, liver fibrosis, and bicarbonate secretion. We evaluated the effectiveness of SR antagonist treatment for early-stage PBC. Male and female dominant-negative TGF-β receptor II (dnTGF-βRII) (model of PBC) and wild-type mice at 12 wk of age were treated with saline or the SR antagonist, Sec 5-27, for 1 wk. dnTGF-βRII mice expressed features of early-stage PBC along with enhanced Sct/SR axis activation and Sct secretion. dnTGF-βRII mice had increased biliary proliferation or senescence, inflammation, and liver fibrosis. In dnTGF-βRII mice, there was increased microRNA-125b/TGF-β1/TGF-β receptor 1/VEGF-A signaling. Human early-stage PBC patients had an increase in hepatobiliary Sct and SR expression and serum Sct levels. Increased biliary Sct/SR signaling promotes biliary and hepatic damage during early-stage PBC.—Kennedy, L., Francis, H., Invernizzi, P., Venter, J., Wu, N., Carbone, M., Gershwin, M. E., Bernuzzi, F., Franchitto, A., Alvaro, D., Marzioni, M., Onori, P., Gaudio, E., Sybenga, A., Fabris, L., Meng, F., Glaser, S., Alpini, G. Secretin/secretin receptor signaling mediates biliary damage and liver fibrosis in early-stage primary biliary cholangitis. FASEB J. 33, 10269–10279 (2019). www.fasebj.org.

Melatonin, a neuroendocrine hormone synthesized by the pineal gland and cholangiocytes, decreases biliary hyperplasia and liver fibrosis during cholestasis-induced biliary injury via melatonin-dependent autocrine signaling through increased biliary arylalkylamine N-acetyltransferase (AANAT) expression and melatonin secretion, downregulation of miR-200b and specific circadian clock genes. Melatonin synthesis is decreased by pinealectomy (PINX) or chronic exposure to light. We evaluated the effect of PINX or prolonged light exposure on melatonin-dependent modulation of biliary damage/ductular reaction/liver fibrosis. Studies were performed in male rats with/without BDL for 1 week with 12:12 h dark/light cycles, continuous light or after 1 week of PINX. The expression of AANAT and melatonin levels in serum and cholangiocyte supernatant were increased in BDL rats, while decreased in BDL rats following PINX or continuous light exposure. BDL-induced increase in serum chemistry, ductular reaction, liver fibrosis, inflammation, angiogenesis and ROS generation were significantly enhanced by PINX or light exposure. Concomitant with enhanced liver fibrosis, we observed increased biliary senescence and enhanced clock genes and miR-200b expression in total liver and cholangiocytes. In vitro, the expression of AANAT, clock genes and miR-200b was increased in PSC human cholangiocyte cell lines (hPSCL). The proliferation and activation of HHStecs (human hepatic stellate cell lines) were increased after stimulating with BDL cholangiocyte supernatant and further enhanced when stimulated with BDL rats following PINX or continuous light exposure cholangiocyte supernatant via intracellular ROS generation. Conclusion: Melatonin plays an important role in the protection of liver against cholestasis-induced damage and ductular reaction.

Ursodeoxycholic acid (UDCA) is used to treat biliary disorders; and, bile acids alter mast cell (MC) histamine release. MCs infiltrate Mdr2−/− mice liver (model of primary sclerosing cholangitis (PSC)). MC-derived histamine increases inflammation, hepatic stellate cell (HSC) activation and fibrosis. The objective was to determine the effects of UDCA treatment on MC infiltration, biliary damage, inflammation and fibrosis in Mdr2−/− mice and human PSC. Wild-type and Mdr2−/− mice were fed bile acid control diet or UDCA (0.5% wt/wt). Human samples were collected from control and PSC patients treated with placebo or UDCA (15 mg/kg/BW). MC infiltration was measured by immunhistochemistry and quantitative polymerase chain reaction (qPCR) for c-Kit, chymase, and tryptase. The HDC/histamine/histamine receptor (HR)-axis was evaluated by EIA and qPCR. Intrahepatic bile duct mass (IBDM) and biliary proliferation was evaluated by CK-19 and Ki-67 staining. Fibrosis was detected by immunostaining and qPCR for fibrotic markers. Inflammatory components were measured by qPCR. HSC activation was measured by SYP-9 staining. Inflammation was detected by qPCR for CD68. In vitro, MCs were treated with UDCA (40 μM) prior to HA secretion evaluation and coculturing with cholangiocytes or HSCs. BrDU incorporation and fibrosis by qPCR was performed. UDCA reduced MC number, the HDC/histamine/HR-axis, IBDM, HSC activation, inflammation, and fibrosis in Mdr2−/− mice and PSC patients. In vitro, UDCA decreases MC-histamine release, which was restored by blocking ASBT and FXRβ. Proliferation and fibrosis decreased after treatment with UDCA-treated MCs. We conclude that UDCA acts on MCs reducing histamine levels and decreases the inflammatory/hyperplastic/fibrotic reaction seen in PSC. Ursodeoxycholic acid (UDCA) is used to treat biliary disorders; and, bile acids alter mast cell (MC) histamine release. Following liver injury like primary sclerosing cholangitis in mice and humans, MCs infiltrate. MC-derived histamine increases biliary damage, fibrosis, and inflammation. UDCA treatment decreases these parameters via reduced MC activation.

Background Cholangiocytes are the target cells of cholangiopathies including primary sclerosing cholangitis (PSC). Vimentin is an intermediate filament protein that has been found in various types of mesenchymal cells. The aim of this study is to evaluate the role of vimentin in the progression of biliary damage/liver fibrosis and whether there is a mesenchymal phenotype of cholangiocytes in the Mdr2−/− model of PSC. Methods In vivo studies were performed in 12 wk. Mdr2−/− male mice with or without vimentin Vivo-Morpholino treatment and their corresponding control groups. Liver specimens from human PSC patients, human intrahepatic biliary epithelial cells (HIBEpiC) and human hepatic stellate cell lines (HHSteCs) were used to measure changes in epithelial-to-mesenchymal transition (EMT). Findings There was increased mesenchymal phenotype of cholangiocytes in Mdr2−/− mice, which was reduced by treatment of vimentin Vivo-Morpholino. Concomitant with reduced vimentin expression, there was decreased liver damage, ductular reaction, biliary senescence, liver fibrosis and TGF-β1 secretion in Mdr2−/− mice treated with vimentin Vivo-Morpholino. Human PSC patients and derived cell lines had increased expression of vimentin and other mesenchymal markers compared to healthy controls and HIBEpiC, respectively. In vitro silencing of vimentin in HIBEpiC suppressed TGF-β1-induced EMT and fibrotic reaction. HHSteCs had decreased fibrotic reaction and increased cellular senescence after stimulation with cholangiocyte supernatant with reduced vimentin levels. Interpretation Our study demonstrated that knockdown of vimentin reduces mesenchymal phenotype of cholangiocytes, which leads to decreased biliary senescence and liver fibrosis. Inhibition of vimentin may be a key therapeutic target in the treatment of cholangiopathies including PSC. Fund National Institutes of Health (NIH) awards, VA Merit awards.

To prevent unbalanced chromosome segregation, meiotic metaphase I – anaphase I transition is carefully regulated by delaying anaphase until all kinetochores are well oriented (anaphase checkpoint) in mammals and insects. In plants this has not yet been established. In heterozygotes of two reciprocal translocations of Secale cereale, with one chromosome replaced by its two telocentric arms, anaphase delay was correlated with the orientation of the kinetochores of the complex of five chromosomes. The terminal kinetochores of the half chromosomes were readily elongated and pole oriented. Chains of five chromosomes with all five kinetochores orienting on alternate poles where the first to start anaphase. Kinetochores of two adjacent chromosomes when oriented on the same pole were partly shielded and less well pole directed. Anaphase was delayed. Cells with this configuration accumulated during anther development. Kinetochores in metacentric chromosomes lacking chiasmata in one arm (in trivalents and bivalents) were slightly better pole oriented and delayed anaphase less. Release of chromatid cohesion as triggered by kinetochore stretch is apparently delayed by inadequate exposition and pole orientation of the kinetochores. It is a mild form of an anaphase checkpoint, in normal material synchronizing bivalent segregation.

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.

Telocentric trisomics (telotrisomics; one arm of a metacentric chromosome present in addition to two complete genomes) are used in theoretical studies of pairing affinities and chiasma formation in competitive situations and applied in genome analysis, gene localization, gene transfer, and breakage of close linkages. These applications require knowledge of the recombination characteristics of telotrisomics. Appropriate cytological and molecular markers and favorable chromosome morphology are not always available or applicable for quantitative analyses. We developed new mathematical models for extracting the maximum information from simple metaphase I observations. Two types of telotrisomics of the short arm of chromosome 1R of rye ( Secale cereale ), including several genotypes, were used as test material. In simple telotrisomics, pairing between morphologically identical complete chromosomes was more frequent than pairing between the telocentric and either of the normal chromosomes. In the telocentric substitution, morphologically identical telocentrics paired less frequently with each other than either one with the normal chromosome. Pairing partner switch was significant. Interaction between the two arms was variable. Variation within plants was considerable. Telotrisomics without markers are suitable for analyzing pairing preferences, for gene localization and gene transfer, and for breaking tight linkages, but less so for genome analysis.

somatic (diplontic) selection would sufficiently rapidly elim inate heterozygous cell lineages after tissue differentiation has started. An alternative explanation is elimination of one of the genomes, followed by chromosome doubling of the remaining (single) genome, resulting in a homozygous diploid. There are numerous examples of genome separation in hybrids (Sybenga 1992), in which centromere behaviour is species dependent, and in which the centromeres of one species are activated later than those of the other species. The late centromeres fail to reach the metaphase plate simultaneously with the early centromeres, and the central space of the anaphase‐telophase group is occupied by chromosomes of the early species. Only the periphery is then available for the late chromosomes. This is reflected by the position of the chromosomes at interphase or mitosis. In extreme cases the late chromosomes do not reach telophase, and are eliminated. This occurs especially in young embryos where rapid divisions require well synchronized centromere activity. This phenomenon is used for producing haploids of one species from hybrids with another species. Spontaneous chromosome doubling is rare, but when the haploids are artificially doubled, homozygous diploids result. In a number of insects, imprinting of the chromosomes of one parent has a comparable effect: one set of chromosomes is eliminated. This mechanism of elimination of one specific genome can not have operated in the particular case considered here, as homozygosity for markers of both parents is recorded. Random loss of (initially) single chromosomes with equivalent centromeres from diploid hybrid nuclei, first re sulting in genetic instability and consequently in rapid loss of other chromosomes until a balanced haploid set remains, is common in some imperfect fungi (Pontecorvo 1958). The haploid state is the natural somatic state, and a series of un balanced mitoses does not present a threat to the very restricted differentiation. If this would occur in hybrid plants, chromosome number doubling would result in homozygous diploid sectors. However, in plants, the diploid state is the natural state and mitosis is normally very stable when the

Meiosis was studied in one hypoploid (trisomic, 2n = 4x - 1 = 27) of an induced tetraploid (2n = 28) of Lathyrus odoratus and two hyperploids (single pentasomic, 2n = 4x + 1 = 29 and double pentasomic, 2n = 4x + 1 + 1 = 30, respectively) of an induced tetraploid of L. pratensis. Multivalent frequencies in the aneuploids were high, but the numbers of chiasmata were not sufficient to maintain all pairing configurations till metaphase I. Interstitial chbdsmata were relatively frequent. Chiasma frequencies were considerably reduced in all three aneuploids, compared with the eutetraploids. The two pentasomes (sets of five homologous chromosomes) of the double pentasomic were both present as a quinquivalent more frequently than expected when they would behave independently. The single pentasomic and the trisomic did not deviate much from expected, although, like in the eutetraploids, the multivalent frequencies were considerably higher than corresponds with the assumption that pairing initiation is restricted to the chromosome ends. The conclusion is that aneuploids of Lathyrus at the tetraploid level tend to behave similar to euploids, except that the chiasma frequency is considerably reduced. When maintained in the population their progeny will contain even more aneuploids than the euploids, not only because the aneuploidy will be perpetuated, but also because of lower chiasma frequencies leading to univalent formation. J. Sybenga, Fazantenlaan 4, NL-4351 RV Veere, The Netherlands

Chromosome pairing and chiasma formation were studied in natural and induced tetraploids (2n = 28) of Lathyrus odoratus (induced), Lathyrus pratensis (natural and induced), Lathyrus sativus (induced), and Lathyrus venosus (natural), as well as in triploids of L. pratensis and diploids of L. odoratus, L. pratensis, and L. sativus. All natural tetraploids appeared to be autotetraploids and their meiotic metaphase I behaviour was very similar to that of the induced autotetraploids, with average numbers of pairing partner switches exceeding 4 or even 5. Multivalent frequencies were high, but the numbers of chiasmata were not much higher than necessary to maintain the configurations. Interstitial chiasmata were common, but not predominant. Fertility was reduced, but sufficient for predominantly vegetatively reproducing species. The triploids of L. pratensis had an even higher multivalent frequency than the tetraploids, but still produced some viable progeny at or close to the tetraploid level, suggesting that in mixed populations of diploids and tetraploids, triploids can contribute to gene flow between the ploidy levels. There was no significant correlation between chiasma frequency and ring bivalent frequency in the diploids and multivalent frequency in the corresponding tetraploids. In the tetraploids, chiasma frequency and multivalent frequency were negatively correlated.Key words: Lathyrus, natural, induced, autotetraploid, triploid, meiosis.

When polyploid hybrids with closely related genomes are propagated by selfing or sib-breeding, the meiotic behaviour will turn into essentially autopolyploid behaviour as soon as the affinity between the genomes is sufficient to permit occasional homoeologous pairing. An allopolyploid will only be formed when the initial differentiation is sufficient to completely prevent homoeologous pairing (in some cases enhanced by specific genes), or when segregational dysgenesis prevents transmission of recombined chromosomes. A new polyploid hybrid may be considered a segmental allopolyploid and may show reduced multivalent formation as a result of preferential pairing between the least differentiated genomes. An established polyploid is either an autopolyploid or an allopolyploid. In exceptional cases it is thinkable that a stable segmental allopolyploid arises, in which some sets of chromosomes are well differentiated and behave as in an allopolyploid, whereas other sets are not well differentiated and behave as in an autopolyploid. No clear cases have been found in the literature so far. Key words : chromosome, pairing affinity, quadrivalent frequency, segmental allopolyploidy.

Cytological tester sets include series of aneuploids (nullisomics, monosomics, trisomics of different types, tetrasomics), series of rearranged chromosomes (translocations, inversions, duplications, deficiencies) and series of chromosomes recognizable by specific microscopically visible markers (C-or other banding, molecular markers). In rye, only a few (mainly tertiary and telocentric) monosomics and no viable nullisomics have been found. Several sets of primary trisomics and some telocentric sets, usually not fully complete, have been developed, but few are still available for gene localization. A few tertiary trisomics have been derived from translocation heterozygotes. Extensively used are different sets of additions of rye chromosomes to wheat. A relatively widely distributed set of marked chromosomes is the Wageningen translocation tester set, complemented with translocations from different other institutions. A disadvantage of rye translocations is insufficient heterozygote semisterility. Series of otherwise rearranged chromosomes have not been reported. Sets of lines with chromosomes conspicuously differing from the standard C-banding pattern have been obtained. Molecular markers are available for most rye chromosome, but lack of heterozygosity, necessary for classification afterin situ hybridization is a restriction for use as cytological testers. In the cases of most translocations, C-banding andin situ molecular markers, each separate plant in a segregating population must be screened cytologically, whereas with aneuploid markers or with translocations having sufficient heterozygote semisterility, analyzing segregations is sufficient.

The segregation of the 75K gamma secalin locus (Sec-2) in combination with five interchanges (reciprocal translocations) and two marker genes was analyzed. The translocations involved chromosome arms 1RL, 1RS, 2RL, 2RS, 4RL, 5RL, 5RS, 6RL and 6RS. The gene loci were both on 2R, but the arm was not known. Although the Sec-2 locus was expected to be on chromosome 2RS, no linkage between Sec-2 and any of the markers was found. This is concluded to be the result of exceptionally frequent recombination between Sec-2 and the break point of one of the translocations, which is the only marker in 2RS.

The effects of developmental stage of the anther, ageing of the plant, inbreeding and season on meiotic segregation of an interchange chain quadrivalent were studied in pearl millet, Pennisetum americanum (L.) Leeke. The frequency of adjacent orientation (and segregation) increased with developmental stage of the anther, independent of other factors. Plant age and degree of inbreeding had no demonstrable effects, but there was an indication that high temperature favoured adjacent orientation. Chromosome contraction as measured by change in chromosome length appeared to be negligible during metaphase-anaphase. Therefore, increased adjacent orientation cannot readily be explained by metaphase reorientation resulting from the straightening of chromosomes caused by an increase in their rigidity. It is probable that the unoriented or “mal-oriented” quadrivalents observed regularly at early metaphase I continue to straighten out prior to their delayed orientation. When they finally orientate late in metaphase, their orientation will more likely be adjacent than alternate.

Combined genetic and cytological analysis of positive and negative interference in an interchange heterozygote of rye ( Secale cereale L.) 1

The original identification of the chromosomes involved in each of the lines of the act of primary trisomics of winter rye variety ‘Esto’ does not correspond with recent results of gene localization studies. Using known morphological marker genes, N-banding and test crossing with the standard translocation tester set, a more precise identification was possible. In the nomenclature of the Triticinae, the lines can be designated as follows: A = 7R; B = 5R; C = 2R; D = 3R; E = 4R; F = 6R; G = 1R.

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.

The kinetic activity of chromosomes is either distributed evenly along the length of the chromosome or localized in a single centromere. Within the group of organisms with evenly distributed (holokinetic) activity great variation occurs, which has its consequences for chromatid segregation. Three major types, with some overlap, can be distinguished. They form a gradient of increasing specialization. Arguments are presented for the hypothesis that the monokinetic chromosomes are nothing but the end point in the same gradient and do not form a fundamentally different category. These arguments include: concentration of kinetic activity at meiosis in some types of holokinetic chromosomes, and ‘atavistic’ traits in monokinetic chromosomes pointing to a holokinetic origin. Possible reasons for the wide distribution of monokinetic chromosomes in the more evolved taxa are given.

The objective of plant breeding is the production of a genotype or an adapted combination of genotypes that meet specific performance requirements. The means to realize this objective are the introduction and manipulation of genetic variation. By far the large majority of all plant breeding still depends on the generative cycle, although in some instances effective use has been made of the somatic phase at the plant level for the induction of variation by mutagenesis and polyploidization. The rapid and large-scale development of molecular and in-vitro cell techniques (see Bajaj 1986) is based primarily on the somatic phase, and application to the generative cycle lags far behind. It appears as if this will not change for a considerable period to come. It is not reasonable to expect that the new somatic approaches will ever be able to replace generative approaches in all or even most phases of plant breeding (Borlaug 1983), but in several instances it will soon be, or is already, necessary to make a choice. The increasing ease with which molecular and cell manipulations techniques can be applied will reduce the barrier against their application. Whereas a few years ago, shortage of capital and progressiveness were thought to be the main obstacles to the introduction of modern somatic techniques in practical breeding institutions, this is clearly changing. Some of these obstacles will remain, but the emphasis in the choice between molecular and cell biological somatic versus generative approaches will be placed more and more on the requirements of the breeding program and the real merits of the techniques.

The frequency of discolored sectors in the seedling leaves of Crotalaria intermedia is an index of two-hit radiation effects. Partitioning of an exposure to X-rays of repeatedly results in frequencies lower that the sum of the frequencies induced by two single exposures (“below additivity”). When the first dose is small relative to the second, and the interval is sufficiently, but not excessively, long, a total effect significantly lower than that produced by the second exposure alone is observed. In the set-up used, changes in the composition of the cell population are improbable. It is suggested that this “protection” is due to induced enzymic repair and/or protection systems which are liable to interfere with any interpretation of dose fractionation effects.

Intercrossing the Wageningen translocation tester set of rye and the series of ‘Imperial’ rye additions to ‘Chinese Spring’ wheat of Sears yielded 29 chromosome disomic translocation hybrids. Observation of trivalents led to the identification of the chromosomes of the tester set in terms of the terminology system used in the Triticinae. The analysis was complicated by very low chiasma frequencies in some short chromosome segments in the hybrids. Nevertheless, it could be safely concluded that 1R=VII; 2R=III; 3R=II; 4R=IV; 5R=VI; 6R=V; 7R=I, which deviates slightly from previous classifications based on other methods.

Relative multivalent and bivalent configuration frequencies at first meiotic metaphase of a translocation derived duplication heterozygote of rye have been used to study recombination (chiasma) patterns. After multivalent pairing chiasma frequency is greatly reduced in the segments proximal to the duplication even when no chiasma is formed. There is positive interference after multivalent pairing between the duplication and the two adjacent segments, possibly especially the interstitial segment of the donor chromosome. A variegated type of across-centromere interference is inferred for both chromosomes. The duplication can in principle be applied in a hybrid variety using chromosomal male sterility genes. The restriction of recombination is not as effective as claimed for some other systems working with excess chromosomal material.

The Giemsa banding pattern is given for eleven reciprocal translocations of rye, Secale cereale L., together involving all chromosomes at least once, and one telocentric substitution. It is possible to correlate the identification system based on the Giemsa pattern with that based on the translocation tester set. The location of the translocation break points could be determined very exactly for a number of translocations, somewhat less exactly for others. The variations in the banding pattern, resulting from genetic, environmental and technical variation, make definite identification with the nomenclature system of the different rye additions to wheat difficult. An attempt is made, but some caution is necessary.