Pachytene variations in Ricinus

The pachytene complement in microsporocytes was compared among ten races of Ricinus communis L., the castor plant (2n=20). Four kinds of pachytene variations were observed: (1) variation in degree of spreading of the chromosomes; (2) variation in degree of attenuation of heterochromatin; (3) variation in morphology, which appeared to be restricted to the two nucleolar organizing bivalents; (4) variation in frequency of association of each of these two bivalents with the nucleolus. It is suggested that variations in the pachytene chromosomes occur ubiquitously in this species.     

Ultrastructure and behavior of the achiasmatic,telosynaptic XY pair of the sand rat (Psammomys obesus)

The behavior of the X and Y chromosomes in somatic and testicular cells of the sand rat (P. obesus) has been investigated with light and electron-microscope procedures. The Y chromosome has been identified as the fourth longest of the complement, both by C-banding and by its meiotic behavior. The X chromosome is the longest of the complement and carries two major C-heterochromatic blocks, one in the distal part of the long arm and the other forming most of the short arm. During presynaptic stages in spermatocytes, separate C-heterochromatic blocks, representing the sex chromosomes, are observed in the nuclei. An XY body is regularly formed at pachytene. During first meiotic metaphase the X and Y chromosomes show variable associations, none of them chiasmatic. Second meiotic metaphases contain, as in other mammals, a single sex chromosome, suggesting normal segregation between the X and the Y. — Electron microscopic observations of the autosomal synaptonemal complexes (SCs) and the single axes of the X and Y chromosomes during pachytene permit accurate, statistically significant identification of each of the largest chromosomes of the complement and determination of the mean arm ratios of the X and Y axes. The X and Y axes always lie close to each other but do not form a SC. The ends of the X and Y axes are attached to the nuclear envelope and associate with each other in variable ways, both autologously (X with X or Y with Y) and heterologously (X with Y), with a tendency to form a maximum number (four) of associated ends. Analysis of 36 XY pairs showed no significant preference for any single specific attachment between arm ends. The eighth longest autosomal bivalent is frequently partially asynaptic during early pachytene, and only at that time is often near or touching one end of the X axis. — It is concluded that while axis formation and migration of the axes along the plane of the nuclear envelope proceed normally in the X and Y chromosomes, true synapsis (with SC formation) does not occur because the pairing region of the X chromosome has probably been relocated far from the chromosome termini by the insertion of distal C-heterochromatic blocks.     

Nucleolar cycle and localization of NORs in early embryos of Parascaris univalens

During early embryogenesis of the nematode Parascaris univalens (2n=2) the processes of chromatin diminution and segregation of the germ and somatic cell lineages take place simultaneously. In this study we analyzed the nucleolar cycle in early embryos, both in germinal and somatic blastomeres, by means of silver staining and antibodies against the nucleolar protein fibrillarin. We observed an identical nucleolar cycle in both types of blastomeres, hence, the chromatin diminution process has no effect on the nucleolar cycle of somatic blastomeres. We report the existence of outstanding differences between this cycle and those previously reported during early embryogenesis of other species. There is a true nucleolar cycle in early embryos that shows a peculiar nucleolar disorganization at prophase, and a preferential localization of prenucleolar bodies only on the euchromatic regions during nucleologenesis. Moreover, fibrillarin does not form a perichromosomal sheath in metaphase or anaphase holocentric chromosomes, probably owing to their special centromeric organization. The number and location of nucleolus organizer regions (NORs) in the chromosomal complement have been determined using silver impregnation, chromomycin A₃/distamycin A staining, and fluorescent in situ hybridization using an rDNA probe. There are only two NORs, one per chromosome, and these are lost in blastomeres after chromatin diminution. Moreover, the constant presence of two nucleoli in somatic blastomeres suggests that NORs are not affected during the fragmentation of euchromatic regions when this process occurs.     

Development of a quantitative pachytene chromosome map in Oryza sativa by imaging methods

A higher GC content region of an Oryza sativa chromosome can be specifically visualized by double staining with propidium iodide (PI) and 4, 6-diamidino-2-phenylindole (DAPI). This procedure allows identification of chromosome 9 from the other rice chromosomes at the pachytene stage. Using rice chromosome 9 as a model, an imaging method to construct a pachytene chromosomal map was developed by quantifying the fluorescence profile (FP) of each chromomere. The pachytene map of chromosome 9 consists of twenty-two chromomeres including four chromomeres within the nucleolar organizing region (NOR) and satellite region. The pachytene map was compared with the corresponding somatic prometaphase map and the linkage map. The differences among the three maps indicate that each map depicts specific biological information, which is difficult to be substituted by the other maps.     

Autosomal synaptonemal complexes and sex chromosomes without axes in Triatoma infestans (Reduviidae; Hemiptera)

Meiotic and somatic cells at interphase in Triatoma infestans are characterized by the formation of a large chromocenter, which was assumed to contain the whole of the three large pairs of autosomes and the sex chromosomes. Observations with C-banding techniques show that the chromocenter is formed only by the terminal and subterminal heterochromatic blocks of the three large pairs of autosomes and the sex chromosomes. During pachytene the two largest autosomal pairs loop on themselves and their condensed ends form the chromocenter, together with the single heterochromatic end of the third autosomal pair. The X and Y chromosomes seem to associate with these condensed ends by their affinity for C-heterochromatin. During a very short pachytene stage, bivalents and synaptonemal complexes (SCs) are observed. Pachytene is followed by a very long diffuse stage, during which SCs are disassembled, multiple complexes aggregate on the inner face of the chromocenter and finally all complexes disappear and a dense material is extruded to the cytoplasm through the annuli. The 3-dimensional reconstruction of early pachytene chromocenters show 3 SCs entering and tunnelling the chromocenter, while during mid-pachytene 4 SCs enter this mass and a 5th SC is in a separate small mass. The looping of a whole SC which has both ends in the chromocenter was shown by the reconstructions. These data are interpreted as the progressive looping of the two largest bivalents during pachytene, forming finally the association of 5 bivalent ends corresponding to the 5 C-banding blocks of the large autosomal pairs. No single axis or SC that could be ascribed to the sex chromosomes was found. This agrees with the pachytene microspreads, which show only 10 SCs corresponding to the autosomal bivalents. The X and Y chromosomes are enclosed in the chromocenter, as shown by the unravelling chromocenters at diplotene-diakinesis. Thus the sex chromosomes do not form axial condensations, and this fact may be related to the ability of the X and Y chromosomes to divide equationally at metaphase I. SCsThis paper is dedicated to the memory of the late Professor Francisco A. Saez     

Cytogenetics for the model system Arabidopsis thaliana

A detailed karyotype of Arabidopsis thaliana is presented using meiotic pachytene cells in combination with fluorescence in situ hybridization. The lengths of the five pachytene bivalents varied between 50 and 80 μm, which is 20–25 times longer than mitotic metaphase chromosomes. The analysis confirms that the two longest chromosomes (1 and 5) are metacentric and the two shortest chromosomes (2 and 4) are acrocentric and carry NORs subterminally in their short arms, while chromosome 3 is submetacentric and medium sized. Detailed mapping of the centromere position further revealed that the length variation between the pachytene bivalents comes from the short arms. Individual chromosomes were unambiguously identified by their combinations of relative lengths, arm-ratios, presence of NOR knobs and FISH signals with a 5S rDNA probe and chromosome specific DNA probes. Polymorphisms were found among six ecotypes with respect to the number and map positions of 5S rDNA loci. All ecotypes contain 5S rDNA in the short arms of chromosomes 4 and 5. Three different patterns were observed regarding the presence and position of a 5S rDNA locus on chromosome 3. Repetitive DNA clones enabled us to subdivide the pericentromeric heterochromatin into a central domain, characterized by pAL1 and 106B repeats, which accommodate the functional centromere and two flanking domains, characterized by the 17 A20 repeat sequences. The upper flanking domains of chromosomes 4 and 5, and in some ecotypes also chromosome 3, contain a 5S rDNA locus. The detection of unique cosmids and YAC sequences demonstrates that detailed physical mapping of Arabidopsis chromosomes by cytogenetic techniques is feasible. Together with the presented karyotype this makes Arabidopsis a model system for detailed cytogenetic mapping.     

Synaptonemal complex analysis of mouse chromosomal rearrangements

Synaptonemal complex (SC) analysis by electron microscopy of spermatocytes in surface microspreads was carried out in mice heterozygous for two paracentric inversions: either In(1)1RK or In(2)5Rk. Characteristic SC inversion loops are formed at synapsis in bivalents carrying the rearrangements. Although all loops were observed to be eliminated by late pachytene through synaptic adjustment, every spermatocyte at early pachytene contained a fully synapsed loop. Cells in the earliest stage of pachytene contained the longest loops and thus had undergone minimal adjustment. The SC estimates of inversion lengths and breakpoint positions in such cells corresponded well with those from mitotic chromosome banding and could be correlated with genetic maps of chromosomes # 1 and # 2, thus demonstrating the basis for the mapping of pachytene chromosomes. The regularity of loop formation and reproducibility of the SC analysis are reflected in the constant relative positions of the estimated breakpoints. The method is sensitive enough to reflect small, real, interstitial length differences between meiotic and mitotic chromosomes. The results demonstrate the feasibility and precision of detection and quantitative characterization of inversions at early meiotic prophase by SC analysis.This paper is warmly dedicated to Prof. Dr. Wolfgang Beermann, on the occasion of his 60th birthday     

Mapping of pachytene bivalents of female codling moth Cydia pomonella (Linnaeus) (Lep., Tortricidae)

Spread pachytene nuclei of codling moth Cydia pomonella (Linnaeus) (Lep., Tortricidae) females of a Syrian strain (SY) were used to investigate chromomere patterns of chromosome bivalents and determine their length. The karyotype of female codling moths consists of 28 chromosome bivalents, of which seven are clearly distinguishable using chromosome length and the number and size of the chromomeres in the pachytene stage. One autosome bivalent has two nucleolar organizing regions (NORs) that are located at the opposite ends of the chromosome and appear as distinct structural landmarks. In female codling moths, the WZ sex chromosome bivalent was easily identified in pachytene oocytes according to the heterochromatic thread of the W chromosome. This study contributed to the knowledge and identification of pachytene chromosomes of female codling moths.     

Morphology of the pachytene chromosomes ofPennisetum purpureum schumach

The morphology of the pachytene chromosomes ofPennisetum purpureum was studied. On the basis of relative lengths and arm ratios it was possible to identify them individually. They are numbered 1 to 14 in the order of their decreasing lengths and diagnostic characters are given for each chromosome. The first and the fourteenth chromosomes, the longest and the shortest of the complement respectively, are the nucleolus organizing chromosome. In all the fourteen chromosomes the centromeres are flanked by deep staining regions. This feature is common to the two species.P. typhoides andP. purpureum. Seven out of the fourteen chromosomes have terminal knobs in their long arms. This is a feature in which it differs from the other seven chromosome species.Pennisetum typhoides. These knobs, however, are not present uniformly in all populations; material obtained from Ghana did not show these knobs.     

Structure of and homology between pachytene and somatic metaphase chromosomes of the tomato

The pachytene and somatic metaphase chromosomes of tomato are structurally differentiated into proximal chromatic and distal achromatic parts. The pachytene chromosomes have very clear and characteristic chromosome markers, with the help of which all 12 bivalents can be clearly identified. Based on the size, the arm ratio, the ratio of chromatic parts and the presence and size of achromatic parts, all 12 pairs of somatic chromosomes can also be identified, and each pair be homologised with the corresponding pachytene bivalent. A comparison of the lengths of chromatic and achromatic parts of pachytene chromosomes with the chromatic and achromatic parts of the corresponding somatic chromosomes indicate, that, on an average, the chromatic parts are contracted by a factor of 4 to 5, whereas the achromatic parts are contracted by a factor of 30. The heteropycnosis near the centromere in tomato chromosomes therefore is not a special characteristic of meiotic chromosomes, but present in somatic metaphase chromosomes also.This study was part of a project resulting from a contract between the AssociationEuratom-I.T.A.L. and the Agricultural University of Wageningen.     

Giemsa C-banded karyotypes and taxonomic relationships of some North AmericanAllium species and their relationship to Old World species (Liliaceae)

The somatic karyotypes of six North AmericanAllium species and the EuropeanA. scorzonerifolium have been investigated using a Giemsa C-banding technique. All species have a chromosome number of 2n = 14. InA. scorzonerifolium and the three North American speciesA. dichlamydeum, A. fibrillum andA. unifolium C-bands are restricted to two pairs of nucleolar chromosomes. Each chromosome has a band proximal to the nucleolar constriction and a positively banded satellite. InA. acuminatum, in addition to the bands associated with the nucleolar constrictions, all chromosomes also have pericentromeric bands.A. cernuum exhibits a distinctive banding style: two chromosome pairs with bands adjacent to the nucleolar constrictions and four pairs with telomeric bands on their short arms. In the karyotype ofA. geyeri neither C-bands nor nucleolar chromosomes were found.—A comparison of the banding styles together with other cytological and morphological characters of these species with old world members ofAllium reveals:A. cernuum closely resembles species within subgenusRhizirideum, whereas the other species studies exhibit many similarities with subgenusMolium. Their sectional grouping and their relationships with Old World species are discussed.     

Heterochromosomes in Mytilicola intestinalis Steuer (Copepoda)

The male and female diploid complements in Mytilicola are formed both by 22 chromosomes. Two different somatic metaphase figures are found: with two minute-size chromosomes and with three minute-size chromosomes. The study of meiosis has shown that the female sex is heterogametic. The chromosome complement is 10_II+XX in the male and 10_II+XY in the female. A bivalent formed by two different-length homologues, is to be seen in the female meiosis at pachytene; it shows, sometimes, a light positive hetero-pycnosis.     

THE DISTRIBUTION OF HETEROCHROMATIN IN THE GENUS NICOTIANA (SOLANACEAE)

A survey of the species of the genus Nicotiana was carried out to determine the distribution and the cytological characteristics of heterochromatin in this genus. All examined species of the genus possess knob-type heterochromatin, which is defined as spherical, densely staining regions of the pachytene chromosomes. These knobs are most frequently located near the centromere, the nucleolar organizer, and the ends of the chromosomes. Block-type heterochromatin, defined as any longer-than-broad heterochromatic segment seen at pachytene, was found in three species of the section Paniculatae, three species of the section Tomentosae, and two species of the section Noctiflorae. Three categories of the block-type heterochromatin, corresponding to the three subgenera, were found to differ with respect to overall size, staining properties, and location of the blocks. The distribution of these three types of block heterochromatin is discussed in the light of the latest taxonomic treatments of the genus.     

Meiosis and pollen mitosis in diploid and triploid Colocasia antiquorum Schott.

The relationship between diploid and triploid forms of Colocasia antiquorum Schott. was assessed through comparative meiotic and pollen mitotic studies. Owing to poor spreading of the chromosomes of both materials, karyological observations on pachytene nuclei were limited to a few chromosomes. Among the two nucleolar chromosomes and a metacentric, telochromomere-bearing chromosome of the diploid, the latter and one of the nucleolar chromosomes characterized by a heteropycnotic short arm were identified in both bivalent and trivalent associations in the triploid. The homologues in these cases were homomorphic and intimately paired. Two types of heteromorphic bivalents exhibiting partial pairing of homomorphic segments were also recorded in the triploid. Among the 14 bivalents of the diploid at diakinesis, two were nucleolus-associated. In the triploid, chromosomal associations at diakinesis included trivalents (2 to 9), bivalents and univalents, and the chiasma frequency per paired chromosome was lower than in the diploids. In 21.6 percent of the PMCs at this stage intragenomic pairing of one or two chromosomes was observed. Post-diakinesis stages in the diploid were regular while in the triploid they were marked by various irregularities in a majority of the cells. However, fertility (stainability), size and divisional frequency of pollen in both materials were remarkably similar. Chromosome numbers in pollen nuclei in the triploid ranged from 8 to 25. Based on these data an autopolyploid origin for the triploid Colocasia and a lower base number than the gametic chromosome number for this genus are advanced.     

Variation of the chromosome phenotype in zea,solanum and salvia

Summary InSolanum lycopersicum pachytene chromosomes the gradient in chromomere size, originating on both sides of the kinetochore, reveals the following characteristics: 1. a relatively abrupt decrease in size of the large chromomeres, 2. the gradient is related to arm length in 9 of the 12 chromosomes, 3. the gradient is particularly irregular in the short arm of the nucleolar chromosome and in the long arm is not conspicuous, 4. chromosome 6 shows an abrupt interruption in the gradient close to the kinetochore. Salvia viridis andZea mays chromosomes represent intermediate conditions between species with well defined and species without gradients. InSalvia the intermediate condition is manifested by the presence of a very large chromomere on each side of the kinetochore followed by very small chromomeres. In two chromosomes the intermediate condition is particularly apparent. In these chromosomes two chromomeres of intermediate size are present in the proximal region of the long arm. The nucleolar organizing arm has also an irregular pattern in this species.Maize has a less distinct gradient than tomato in all its chromosomes. Chromosomes 3, 4, 5 and 8 are those where the gradient is the least sharp. The nucleolar organizing arm of chromosome 6 has also an irregular pattern.In a translocation between chromosomes 5 and 6 of maize, a segment composed of very small chromomeres from the distal region of 5 which was moved to the right of the kinetochore of chromosome 6, did not change appreciably its phenotype after ten years of cultivation. During the period of cultivation a selection was made for plants where the original phenotype was preserved so that this result cannot be considered as demonstrating an absence of change in chromomere phenotype with changed position.InDrosophila andChironomus salivary gland chromosomes where chromomeres are large, and no selection has been carried out with such a purpose, the pattern and nucleic acid content of the bands is known to change when rearrangements occur within the chromosome.Supported by a grant from the Swedish Natural Science Research Council toA. Lima-De-Faria. This work was partly carried out at the Department of Botany, University of Illinois, U.S.A. during a visit to this department byA. Lima-De-Faria.P. Sarvella's collaboration in this work was done during her stay at the University of Lund.     

Kinetochore and microtubules in two members of Chlorophyceae,Cladophora fracta and Spirogyra majuscula

Evidence is presented for the existence of a localised kinetochore with stratified fine structure in Cladophora and in Spirogyra. In the latter, there is the possibility of two kinetochores on the longer chromosomes. There is no evidence for a diffuse kinetochore. The nucleolus persists during mitosis in Cladophora on the nucleolar organising chromosomes, the granular material being lost from it very largely during metaphase and anaphase but the fibrillar material remaining. The persistent nucleolar material at metaphase and anaphase in Spirogyra is not attached to the nucleolar organising chromosomes but accumulates around all the chromosomes and chromatids, the microtubules of the spindle at anaphase passing through and possibly attaching to this nucleolar material and possibly assisting in the movement of the chromatids which are embedded within it.     

Non-relationship between nucleolus and sex chromosome system Xyp in Chelymorpha variabilis boheman (Coleoptera: Chrysomelidae)

Nucleolar-organizer region, nucleolus and mode of association of the sex bivalent were analyzed in spermatecytes of Chelymorpha variabilis Boheman. This species (2n=10II+Xy_p) shows the typical sex chromosome system of the group Polyphaga. The results of silver staining techniques showed the nucleolar organizer region localized in a subterminal position of an autosomal bivalent. During meiotic prophase the nucleolus was distinguished with the silver staining and acridine orange fluorescence technique up to diakinesis. The independence of nucleolus and sex bivalent Xy_p during meiosis is demonstrated. The positively silver staining but negatively orange-red material found within the parachute could be involved in the regular co-orientation of both sex chromosomes. After a longer hypotonic treatment, sex bivalents were observed elongated and paired only at one end during the pachytene stage. Along these sex chromosomes, C-bands showed positive blocks located in the pericentromeric and telomeric regions. Heterochromatic association of both sex chromosomes was suggested.