Terminal C-bands Research Articles

C-bands and chiasma distribution inScilla amoena, S. ingridae, andS. mischtschenkoana (Hyacinthaceae)

An effect of C-band pattern and polymorphism on chiasma distribution in pollen meiosis was recently demonstrated inScilla siberica. A further meiotic banding study has been performed in the alliesS. amoena, S. ingridae, andS. mischtschenkoana in order to analyze the effect, if any, of their specific C-band patterns and cytochemically different heterochromatin types on recombination. No clear evidence for a preferential formation of chiasmata adjacent to homozygous intercalary heterochromatin and no consistent reduction of chiasma frequency near strongly heterozygous intercalary heterochromatin blocks, as observed inS. siberica, could be found. Terminal C-band heteromorphism is suspected to cause distal chiasma defaults. The results suggest once more that there is no uniform effect of heterochromatin on crossover distribution.

Genome analysis of Thinopyrum junceiforme and T. sartorii

Genome constitutions of Thinopyrum junceiforme (A. Löve and D. Löve) A. Löve (2n = 4x = 28) and T. sartorii (Boiss. &Heldr.) A. Löve (2n = 4x = 28) were determined by studying (i) meiotic pairing patterns in hybrids involving the two species and other pertinent hybrids; (ii) mitotic chromosome karyotypes based on length, arm ratio, and satellites; and (iii) C-banding patterns. New hybrids synthesized and reported are T. sartorii × T. bessarabicum (2n = 3x = 21), T. sartorii × (T. bessarabicum × T. elongatum) amphidiploid (2n = 4x = 28), and the reciprocal of the latter. Mean meiotic pairing in the triploid hybrid and the two tetraploid hybrids were 3.83 I + 3.92 II + 3.11 III, 0.931 + 7.46 II + 0.62 III + 2.44 IV + 0.07 V + 0.03 VI, and 3.41 I + 9.39 II + 0.74 III + 0.88 IV, respectively. Based on the chromosome pairing data, it can be concluded that T. junceiforme and T. sartorii have two versions (Jb and Je) of the J genome and behave like true allotetraploids owing to bivalentization. Karyotype analyses of the species and their hybrids with T. bessarabicum revealed minor structural differentiations of the genomes in the two species. Thinopyrum sartorii has one genome basically unchanged from the Jb genome of T. bessarabicum while another is a modified Je genome of T. elongatum. One genome of T. junceiforme is modified from Jb and the other is modified from Je. There are two pairs of large and one pair of small satellites in T. sartorii, but there are only one pair of each in T. junceiforme. Thinopyrum junceiforme is rich and T. sartorii is poor in interstitial C-bands for both sets of genomes. Thinopyrum sartorii has larger terminal C-bands on the Jb genome chromosomes than those of T. junceiforme. These organizational changes of chromosomes could not be detected by studying chromosome pairing alone. However, this study demonstrates that meiotic pairing is the first criterion for determining basic genome symbols. Other techniques, such as those involving chromosome banding, isozymes, and DNA probes, may then be used to detect differences in chromosome (or DNA) organization and gene expression.Key words: genome, hybrid, meiosis, karyotype, chromosome banding, speciation.

DNA structure of the B chromosome of rye revealed by in situ hybridization using repetitive sequences.

To clarify the DNA structure of the B chromosome of rye, Secale cereale L., we carried out in situ hybridization using probes of two highly repetitive sequences (pSc74 and pSc119) of rye A chromosome, of total genomic DNAs from plants with (4 B) or without (0 B) B chromosomes, and probes of fractionated genomic DNA (1.14-kb and 1.28-kb sequences) from a 4 B plant. Clones pSc74 and pSc119 did not hybridize to the B chromosomes except at small interstitial sites. A probe consisting of the total DNA of a 0 B plant hybridized over the entire length of all the A and B chromosomes except for the telomeric C-band region of the long arm of the B chromosome. A probe of total DNA from a 4B plant produced signals only in the terminal C-band region after prehybridization with the 0 B plant DNA. A probe consisting of DraI-digested 1.14-kb fragments from a 4B plant hybridized to the telomeric region of the B chromosome, whereas one of 1.28-kb fragments did so to an interstitial region of the A chromosome. On the basis of the in situ hybridization, and the cytological and molecular biological results, we assume that the B chromosome in rye originated once in the distant past and was then perpetuated in the populations.

Mutability of constitutive heterochromatin (C-bands) during eukaryotic chromosomal evolution and their cytological meaning.

A quantitative analysis of the alterations of constitutive heterochromatin in eukaryotic chromosomal evolution was attempted using the accumulated C-banding data available for mammals, amphibians, fish, ants, grasshoppers, and plants. It was found that these eukaryotes could be classified into two types by their C-banding patterns: 1) Type I included mammals, fish, and ants, and 2) Type II included amphibians, grasshoppers, and plants. C-bands were rather scarce in Type I eukaryote chromosomes and were found around the pericentromeric region when present at all, whereas the predominance of interstitial or terminal C-bands was found in Type II eukaryote chromosomes. The Type I and II C-banding patterns can best be interpreted by assuming that in the former group of eukaryotes the saltatory increase in constitutive heterochromatin occurs preferentially at the pericentromeric regions of telocentric chromosomes induced by centric fission, with C-bands being eliminated almost completely by centric fusion and/or pericentric inversion. On the other hand, C-bands appear in the Type II eukaryotes both interstitially and in the telomeric regions of chromosomes, and there may be no effective mechanism to eliminate these bands once they are integrated.

Mapping seed storage protein loci Sec-1 and Sec-3 in relation to five chromosomal rearrangements in rye (Secale cereale L.)

Linkage relationships were established between the secalin loci, Sec 1 (40-K gamma and omega secalins, homologous to the wheat gliadins) and Sec 3 (HMW = high-molecular-weight secalins, homologous to the wheat HMW glutenin subunits), and five chromosomal rearrangements involving chromosome 1R of rye (Secale cereale L.). These were: interchanges T273W (1RL/5RS), T306W (1RS/5RL), and T850W (1RS/ 4RL), Robertsonian centromere split Rb1RW and the interchanged Robertsonian split Rb2R/248W. The analysis established the linkage relationships between the secalin loci and the breakpoints of the rearrangements, in addition to the quantitative effects of the rearrangements on the linkage. Sec-1 is located in the satellite at a position at least 2.5 cMorgan from the proximal border of the terminal C-band, and about 30 cMorgan from the nucleolar organizing region (NOR). The locus is also physically closer to the terminal C-band than to the NOR, but not as much as corresponds with the map distances. Similarly, the physical distance between Sec-3 and the centromere is greater than corresponds with the recombination frequency (0%-9%). Although overall recombination in 1RL remains the same, recombination between the centromere and Sec-3 is greatly reduced in the Robertsonian split combined with the interchange. This is not the case with the single Robertsonian split.

Relationship of mouse minor satellite DNA to centromere activity

Chromosomes from a female mouse cell line were identified by Q-banding prior to in situ hybridization with 3H-labeled mouse minor satellite (satellite II) DNA. No cell was found in which every chromosome was labeled, but grain counts showed that every active centromeric region had minor satellite sequences. In the mouse T (10;13)199H translocation, the breakpoint was within the minor satellite array, leaving clusters of minor satellite at the C-bandless active centromere of the 13(10) chromosome and at the interstitial C-band of the 10(13) chromosome, which is not associated with centromeric activity. In a mouse A9 (L-cell derived) marker chromosome with one terminal and two interstitial C-bands, only the terminal C-band was adjacent to an active centromere, but minor satellite DNA was present at all three sites. Minor satellite DNA was not detected on the Y chromosome, although the presence of a small amount of divergent satellite sequences on this chromosome could not be ruled out.

Synaptonemal complex formation in rye (Secale cereale) heterozygous for telomeric C-bands

Zygotene–pachytene nuclei from a rye line heterozygous for most terminal C-bands were found by electron microscopic spreading analysis to have unequal lateral elements at most synaptonemal complex telomeres. An unpaired lateral element protruded beyond the end of the synaptonemal complex at each such telomere. Another rye line with only one C-band heterozygous telomere had only one uneven synaptonemal complex telomere. The length differences between paired lateral elements in heterozygotes (both total complement and individual synaptonemal complexes) was considerably less than the difference in DNA content or somatic metaphase chromosome size between C-band positive and C-band negative lines. There was no evidence of a synaptic adjustment effect reducing the telomere length differences in later pachytene–diplotene stage nuclei. Heterozygosity for the 1RL terminal C-band resulted in a slight reduction in chiasma frequency in that arm but no shift in chiasma position.Key words: rye, C-bands, telomeres, chromosome pairing, synaptonemal complex.

Genetic variability in Muscari comosum L. (Liliaceae). II. Characterization and effects of the polymorphic variants of chromosome 2 on chiasmata formation

Muscari comosum L. (Liliaceae) displays a striking chromosomal polymorphism in the second largest chromosome. This polymorphism involves four cosmopolitan types. Two of these are shorter than the other two homologues. One of these is submetacentric (SSM) and the other is subtelocentric (SST). The two longer types also include a submetacentric (LSM) and a subtelocentric (LST) morph. Each of the two submetacentric chromosomes has one interstitial C-band in the short arm and each of the two subtelocentric morphs has an interstitial C-band in the long arm. The change of position of this interstitial C-band is most easily explained by a pericentric inversion. Furthermore, all four types of chromosome 2 have a centromeric C-band, while the two subtelocentrics have an additional terminal C-band in the long arm. The variability in the size of the second chromosome is most likely the consequence of an unequal interchange or an insertional translocation. The meiotic behaviour of the chromosome 2 bivalents in individuals heterozygous for the pericentric inversion is characterized by normal pairing between homologues with no inversion loops, though asynapsis was present in some meiocytes. Chiasmata are absent in two regions of chromosome 2 bivalents in these heterozygotes in which they regularly form in both classes of homozygotes. In individuals heterozygous for the long morphs of chromosome 2 the bivalents again showed normal pairing at pachytene, with chiasmata again absent in some regions in which they normally form. The net result is that homozygotes have significantly higher chiasmata frequencies than hterozygotes. Key words: genetic variability, chiasma formation, Muscari.

The heterochromatin distribution and genome evolution in diploid species of Elymus and Agropyron

The acetocarmine–Giemsa C-banding technique was used to study heterochromatin distribution in somatic chromosomes of diploid Elymus junceus (= Psathyrostachys juncea) (2n = 14) (genome designation Ju = N) and nine diploid Agropyron species (2n = 14): A. cristatum (C = P), A. imbricatum (C = P), A. elongatum (= Elytrigia elongata = Thinopyrum elongatum) (E = J), A. junceum (= E. bessarabicum = T. bessarabicum) (J = E), A. spicatum (= Pseudoroegneria spicata) (S), A. libanoticum (= P. libanotica) (S), A. ferganense (S), A. stipifolium (= P. stipifolia) (S), and A. velutinum (V). With the exception of A. elongatum and A. velutinum, which were self-fertile, all species were cross-pollinating and self-sterile. The cross-pollinating species showed large terminal C-bands and a high level of C-band polymorphism. Agropyron elongatum, moderately self-fertile, showed small terminal and interstitial bands and a minimal C-band polymorphism. Agropyron velutinum, fully self-fertile, almost totally lacked C-bands. The Ju, C, E, and J genomes appeared to be distinctive and the equivalence of the E and J genomes was not supported from their C-banding patterns. Four species sharing the S genome, A. spicatum, A. libanoticum, A. ferganense, and A. stipifolium had C-band patterns similar to one another, although C-bands were less prominent in A. stipifolium than others.Key words: C-banding, karyotype, wheatgrass, cytology.

Meiotic behaviour, stability, transmission and recombination in four balanced tertiary trisomics of rye (Secale cereale L.)

The F3 and F4 progeny of four balanced tertiary trisomics (BTTs) of rye carrying the short translocation chromosomes of their corresponding interchanges were karyotyped and scored for the monogenic recessively inherited tigrina marker (ti) which is located terminally on the short arm of chromosome 5R. The segment carrying the dominant allele (Ti) was included in the extra translocation chromosome of each BTT, while the normal 5R pair carried the recessive allele. Recombination fractions between ti and the interchange breakpoint varied between 0.005±0.003 and 0.032±0.005 for the different BTTs. Interstitial chiasma formation could be demonstrated in only one BTT, in 1% of the first metaphase configurations without, however, resulting in recombination. Chiasma localization just proximal to the terminal C-band of 5RS was concluded to cause the low recombination observed. Less than 0.5% of the progeny consisted of plants with a deviant karyotype able to reach maturity, indicating a high level of meiotic stability of the trisomics. The results further suggest that selection towards low male and high female transmission rates of the extra chromosome is possible. The suitability of the BTTs investigated for use in hybrid breeding programs is evaluated.

Production of trigeneric (barley � wheat) � rye hybrids

Rye (Secale cereale cv. Prolific 2n=14 and 2n =14 + 2B was crossed onto hybrids between barley (Hordeum vulgare 2n = 14) and wheat (Triticum aestivum 2n= 42). Pollinated florets were injected with GA3 to promote fertilization and hybrid embryo development. At 16 days after pollination the watery caryopses were removed, embryos dissected and cultured on a modified B5 medium. Approximately 20% of the cultured embryos produced both roots and coleoptile and developed into viable seedlings. Viable seeds were also obtained at a low frequency from the same cross combinations. The hybrids were wheat-like except for the hairy neck characteristic of rye. There were 35 chromosomes in somatic tissue; 21 wheat, 7 barley and 7 rye. The rye chromosomes were distinguishable by their larger size and terminal C-bands. A lower seed set was obtained using pollen from rye plants with 2n=14 + 2B chromosomes than from plants without B chromosomes.

Regularities and restrictions governing C-band variation in acridoid grasshoppers

C-band patterns have been analysed in embryonic neuroblast chromosomes of 23 Australian species of acridoids. All of them showed paracentromeric C-bands but these varied considerably in size both within and between species. Many of them also showed interstitial C-bands in from 1–5 members of the haploid complement and in two cases (Atractomorpha similis and Genus nov. 95 ochracea) larger numbers of interstitial bands were present. Terminal C-bands were the least common though again when present they could be found in 1–6 members of the complement except in the cases of A. similis and Genus nov. 95 ochracea where still larger numbers occur. In 5 of the 23 species the megameric chromosome pair was distinctively C-banded. The B-chromosomes found in 3 of the species were also strikingly different in C-band characteristics compared to the standard A-chromosomes. Differences in the number of very small chromosomes present in different species clearly cannot be explained in terms of differences in their C-band content. Neither are differences in genome size simply related to differences in the total amount of C-band material indicating that changes in the size of the genome in this group have involved alterations in both eu and heterochromatin content. Finally similar amounts of C-band material may be distributed throughout the complement in very different ways in different species.

Conservation and chromosomal localization of DNA satellites in balenopterid whales.

DNA satellites were isolated from three balenopterid species, viz. the minke, sei, and fine whales. In each of them at least two DNA satellites were recognizable with buoyant densities in neutral CsCl of rho = 1.702/1.703 and rho = 1.710/1;711, respectively. cRNAs from each satellite group were used for filter and in situ hybridisations. Homo-and heterologous DNA-cRNA hybrids within each satellite group yielded virtually identical melting curve profiles showing conservation of at least a considerable part of the DNA satellite sequences. There was no evident sequence homology between the rho = 1.702/1.703 and the rho = 1.710/1;711 satellites by filter hybridisation.--The in situ hybridisation showed that in each species the rho = 1.702/1.703 satellite was located in centromeric-paracentromeric C-bands in a few pairs, whereas the rho = 1.710/1.711 satellite was located in terminal C-bands throughout the karyotypes.--The data on the whale DNA satellites indicate that the quantitative evolution of the sateliite DNA sequences preceded species divergence of the balenopterids and that the satellite sequences have remained relatively unaltered since the divergence took place. The function of satellite DNA is considered to imply the introduction of both chromosomal and genic polymorphisms and thus being of great importance in speciation, Based upon these concepts a model is postulated for the function of satellite DNA. According to this model at meiotic pairing euchromatinheterochromatin overlapping between homologous chromosomes is considered to be of a general occurrence. This overlapping is presumed to be accentuated by the size heteromorphism frequently observed between homologous heterochromatic segments (C-bands). In the region of such euchromatinheterochromatin overlapping, cross-over would be excluded. The overlapping is suggested to be rectified progresssively in the chromosome arms, leaving unaffected crossing-over distant to the euchromatin-heterochromatin junctions. The consequence of this will be that genes in the proximity of the junctions are collectively inherited and selected, whereas genes distant to the the heterochromatin will be independently assorted and selected.

Giemsa C-banding patterns in Aedes (Stegomyia) mosquitoes

Chromosomes from gonads of 14–24 h old pupae of nine species of Stegomyia mosquitoes have been examined using the Giemsa C-banding technique. The species studied were Aedes albopictus, A. polynesienis, A. scutellaris, A. alcasidi, A. seatoi, A. pseudalbopictus, A. melallicus, A. annandalei and A. vittatus. The diploid chromosome number of all species is six. All species possess C-bands in the centromeric regions of each of the three pairs of chromosomes. Besides, an intercalary C-band is present on the female determining (=m) chromosome but absent from the male—determining (= M) chromosome of all species except A. vittatus. In A. vittatus, the m and M chromosomes possess a terminal C-band. Thus, the nine species of Aedes analysed showed two distinct patterns of C-banding. —The evolution of heterochromatin patterns in various species is also discussed. The Giemsa C-banding technique should prove useful in studies of chromosomal speciation in culicine mosquitoes.

Population cytogenetics of the genus Caledia (Orthoptera: Acridinae). II. Variation in the pattern of C-banding.

The distribution of constitutive heterochromatin has been investigated in four chromosomal races of the grasshopper Caledia captiva (2n=23 male/24 female) by the C-banding technique. Each of the four races was found to have a distinctive banding pattern which is associated with the inter-racial differences in chromosomal rearrangements. The "Ancestral" race has a telocentric chromosome complements with large procentric C-bands which are structurally double on six pairs of chromosomes. The centromeres are unstained. The "General Purpose" race has a C-banding pattern very similar to that seen in other Acridine grasshoppers with the majority of its chromosomes showing a centromeric localisation of the bands. The two southern races, which show a complex polymorphism for presumed pericentric inversions on all twelve chromosomes, also show an unusually high level of interstitial and terminal C-bands. The different locations and numbers of these bands allow unambiguous identification of all the chromosome pairs within the complement. In two cases, there is good evidence to indicate that a C-band redistribution between acrocentric and metacentric chromosomes has occurred by pericentric inversion. Furthermore, C-band variation on the long arm of the metacentric X-chromosome indicates the presence of a large paracentric inversion. This double inversion system has involved over 95% of the X-chromosome. The interstitial and terminal C-bands probably have not resulted from heterochromatin movement within the complement but, more likely, have arisen by saltatory duplication of pre-existing sequences on the chromosome. A new nomenclature system for banded chromosomes is proposed which allows most kinds of chromosomal restructuring and rearrangement to be adequately enumerated.

A comparative banding analysis of chromosomes in three species of lemurs (Primates,Lemuridae)

Banded karyotypes were compared in 3 species of lemurs,Lemur catta (2n=56),L. f. fulvus (2n=60) andL. mongoz (2n=60). The karyotypes of the latter 2 species were indistinguishable from each other in both Q-band and G-band patterns, except thatL. mongoz had an unusually large Y chromosome. The karyotypic differences betweenL. catta and the other 2 species were mainly explained by centric fusions (or fissions) and pericentric inversions. Contrary to the general similarity in Q-band and G-band patterns, the C-band patterns were highly variable among the 3 species. All chromosomes ofL. fulvus had a distinctive C-band in the centromeric region, whileL. mongoz had only a few chromosomes with an apparent C-band.L. catta was remarkable by showing interstitial and terminal C-bands in some elements, in addition to the centromeric band which was observed in about 20 pairs.