Chromosome Banding and DNA Methylation Patterns, Chromatin Organisation and Nuclear DNA Content in Zingeria biebersteiniana

Chromosome structure and chromatin organisation of a two-chromosome model cereal Zingeria biebersteiniana (Claus) P. Smirnov were studied: nuclear DNA content was determined by microdensitometric analysis after Feulgen staining; Feulgen absorption at different thresholds of absorbance in interphase nuclei also provided evidence on the organisation of chromatin, allowing quantitative estimation of condensed chromatin within interphasic nucleus. The DNA methylation pattern of Z. biebersteiniana metaphase chromosomes was examined with a specific monoclonal antibody. 5-methyl-cytosine residues are present in several chromosome sites and differences may be present between corresponding regions of homologues. Chromosome banding pattern reveals large bands in the centromeric regions of each chromosome, showing constitutive heterochromatin; by fluorochromes staining pericentromeric blocks are evidenced. After the cold and 9-aminoacridine pre-treatments and after aceto-carmine and aceto-orceine staining, respectively, the metaphase chromosomes were analysed by image analysis system revealing a segmentation of the chromosome body that resembles Giemsa/Reverse banding in animal chromosomes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Production and cytogenetics of hybrids of Triticum aestivum x Leymus innovatus

Summary Hybrid plants were obtained between Triticum aestivum (2n=6x=42, AABBDD) and Leymus innovatus (2n=4x=28, JJNN) at a frequency varying from 0.4% to 1.2% of the pollinated florets. Improvement of the embryo culture medium resulted in a higher frequency of embryo rescue. Eight of ten hybrids had the expected chromosome number of 35 (ABDJN). Meiotic analysis indicated that there was no homology between the genomes of the two species. Two hybrids had only 28 chromosomes. Comparison of chromosome pairing between the two types of hybrids suggested that Leymus innovatus carries genes that affect chromosome pairing and behavior. The relatively high occurrence of spontaneous doubling in the meiocytes of these hybrids may indicate that backcrossing of the hybrids to wheat should be possible, although frequent chromosome irregularities observed in the meiocytes of the hybrids may decrease the probability of success of this step, which is essential to the process of gene transfer from L. innovatus to wheat.Contrib. no. 366     

New insights on the origin of B chromosomes in <Emphasis Type="Italic">Astyanax scabripinnis</Emphasis> obtained by chromosome painting and FISH

Chromosome painting (CP) with a probe of B chromosome obtained by microdissection and fluorescence in situ hybridization (FISH) with probes of As51 satellite DNA, C _o t−1 DNA, and 18S and 5S rDNA confirmed sharing of some repetitive DNA but not rDNA between A and B chromosomes in the fish Astyanax scabripinnis. Meiotic analysis revealed a pachytene B chromosome bivalent nearly half the size of its mitotic configuration, suggesting a self-pairing of B chromosome arms. Such an isochromosome nature of somatic B chromosome was further evidenced by CP and FISH. All the findings obtained suggest (i) intraspecific origin of B chromosome, and (ii) evolutionary enrichment of repetitive DNA classes, especially those contained in the C _o t−1 and the As51 probes, in B chromosome. However, the precise origin of B chromosome in the present species remains to be elucidated by further molecular cytogenetic analysis because of painting of some A chromosome regions with the B chromosome-derived probe.     

The eects of a ring chromosome on the meiotic segregation of other chromosomes in Saccharomyces cerevisiae

Meiotic chromosome segregation must occur with high fidelity in order to prevent the generation of aneuploid cells. We have previously described the identification and genetic characterization of a yeast mutant with defects in meiotic sister-chromatid segregation. We attributed the phenotype in this mutant to a dominant allele, which we referred to as SID1-1. These mutants appeared to exhibit high levels of nondisjunction and precocious separation of sister-chromatids of chromosome III, as well as precocious separation of sister chromatids of chromosome VIII and a univalent artificial chromosome. We show here that the unusual meiotic behavior of chromosome III in these strains is due to the presence of a ring III chromosome, rather than a mutant gene. Additional experiments demonstrate that a ring III/rod III pair alters the meiotic segregation of a univalent artificial chromosome.     

CHROMOSOME PULVERIZATION IN MICRONUCLEI INDUCED BY TRITIATED THYMIDINE

Cultures of a pseudodiploid cell line (Don) of Chinese hamster origin were exposed to varying doses of tritiated thymidine (TdR-³H) for relatively long periods of time. In addition to previously observed chromosomal aberrations) such as breaks and reunions, a substantial number of interphasic cells with micronuclei and of metaphases associated with pulverized chromosomes was found; both phenomena were dependent on exposure time to and concentration of TdR-³H. The former phenomenon appeared to result from the effects of the β-emissions originating in the TdR-³H. A possible interpretation for chromosome pulverization induction is presented, emphasizing the derivation of the pulverized material from micronuclei in a common cytoplasm with a metaphase nucleus. These observations further substantiate our previously advanced hypothesis regarding the essential role played by substances present in a mitotic cell in the induction of chromosome pulverization and nuclear membrane dissolution.     

Ploidy Manipulation and Polyploid Detection in the White Shrimp <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> (Boone 1931) (Decapoda,Penaeidae)

Ploidy manipulation has been rarely used in the genetic improvement of cultured marine shrimps. Although polyploid induction has been proven to be successful in Penaeids, including the species Litopenaeus vannamei, the methodology still requires some improvements. In the present work, different thermal shock treatments on ploidy manipulation were tested and a protocol for detecting polyploid individuals was also established. Fertilized eggs were treated by cold (10°C) and heat (38°C) thermal shocks for 8, 12, 15, 18, 20, and 22 min to induce polyploidy. Nuclear measurements within distinct treatments revealed a significant deviation in relation to the mean diameter of nuclei in the control individuals. Triploid and tetraploid metaphases were observed within treated individuals, confirming the increase of interphasic nuclear diameter. The cold thermal shock was more efficient than the hot ones, besides leading to a higher and more homogeneous hatchery rate. A mean number of three nucleoli per nucleus were observed in diploid individuals, while treated samples usually presented up to five nucleoli per nucleus. The standardization of protocols to obtain and detect polyploid products allows further utilization of such methods on a commercial scale in order to evaluate the performance of polyploid individuals in the genetic improvement of L. vannamei.     

Meiotic confirmation of the identification of chromosomes 9 and 13 in T(9; 19)163H,T(5; 13)264 Ca,and T(1; 13)70H stocks in Mus musculus

Quinacrine fluorescent banding patterns of chromosomes 9 and 13 are very similar in mitotic preparations of Mus musculus. Meiotic studies were carried out in male and female mice heterozygous for two translocations involving these chromosomes to determine whether the translocations have a common chromosome. The results indicate that chromosome 9 is involved in the T163H translocation but not in either the T70H or T264Ca translocations. The T70H and T264Ca translocations, but not the T163H, have chromosome 13 in common. These results support the interpretations based on mitotic studies.     

MEIOTIC CHROMOSOME BEHAVIOR IN SOME INTERGENERIC HYBRIDS OF THE VANDA ALLIANCE

Tanaka , R., and H. Kamemoto . (U. Hawaii, Honolulu.) Meiotic chromosome behavior in some intergeneric hybrids of the Vanda alliance. Amer. Jour. Bot. 48(7): 573–582. Illus. 1961.—Meiotic chromosome behavior of 11 groups of diploid intergeneric hybrids (2n=38) of the Vanda alliance was investigated. Vanda Miss Joaquim × Luisia teretifolia and Ascocentrum curvifolium × Vanda lamellata usually produced 19 bivalents at metaphase I, indicating good homology of the parental chromosomes. Vanda tricolor var. sanderae × Vandopsis lissochiloides, Vandopsis lissochiloides × Vanda sanderiana, Vandopsis lissochiloides × Vanda Tatzeri and Arachnis flos-aeris × Vandopsis lissochiloides showed fair homology of parental genomes with formation of 12–15 bivalents at metaphase I. Trichoglottis brachiata × Vanda sanderiana, Vanda tricolor var. purpurea × Phalaenopsis denevei, Vanda Tatzeri × Aerides lawrenceae, Renanthera monachica × Vanda luzonica, Renanthera storiei × Vanda Clara Shipman Fisher and Aerides lawrenceae × Saccolabium giganteum, formed an average of 2–7 bivalents, thereby indicating poor homology of parental genomes. The hybrids, Renanthera monachica × Phalaenopsis sanderiana and Arachnis hookeriana × Vanda suavis, lacked chromosome pairing. On the basis of chromosome affinity at meiosis, a cytotaxonomic scheme was developed for the several genera of the Vanda alliance studied. Cytotaxonomy, evolution and breeding behavior in the Vanda tribe were briefly discussed.     

Deoxyribonucleic acid of the crab Cancer pagurus : III. Intracellular localization of poly d(A---T) of Cancer pagurus by hybridization in situ

The satellite DNA poly [d(A---T) · d(T---A)] of the crab Cancer pagurus has been localized in situ by DNA-DNA hybridization in the nuclei of various spermatogenetic, midgut gland, intestinal and tegument cells. The specificity of hybridization was checked by various tests before, during and after hybridization. The nuclear sites revealed by this method were compared with those shown by quinacrine mustard or Giemsa staining. The A---T-rich satellite DNA appears to be highly dispersed and does not seem to have any preferential localization inside the crab interphasic nucleus. This situation was compared with that presented by mouse nuclei using similar methods.     

Identification of candidate genes for nicotine withdrawal in C57BL/6J × DBA/2J recombinant inbred mice

Nicotine is the reinforcing ingredient in tobacco. Following chronic exposure, sudden cessation of nicotine use produces negative symptoms of withdrawal that contribute to dependence. The molecular mechanisms underlying nicotine withdrawal behaviors, however, are poorly understood. Using recombinant inbred mice, chronic nicotine was delivered by minipump and withdrawal induced using mecamylamine. Somatic signs of withdrawal, and anxiety-like behavior using elevated plus maze, were then assessed. Interval mapping was used to identify associations between genetic variation and withdrawal behaviors, and with basal gene expression. Differential gene expression following nicotine exposure and withdrawal was also assessed in progenitor mice using microarrays. Quantitative trait loci mapping identified chromosome intervals with significant genetic associations to somatic signs of withdrawal or withdrawal-induced anxiety-like behavior. Using bioinformatics, and association with basal gene expression in nucleus accumbens, we implicated Rb1, Bnip3l, Pnma2, Itm2b, and Kif13b as candidate genes for somatic signs of withdrawal, and Galr1, which showed trans-regulation from a region of chromosome 14 that was associated with somatic signs of withdrawal. Candidate genes within the chromosome 9 region associated with anxiety-like withdrawal behavior included Dixdc1, Ncam1, and Sorl1. Bioinformatics identified six genes that were also significantly associated with nicotine or alcohol traits in recent human genome-wide association studies. Withdrawal-associated somatic signs and anxiety-like behavior had strong non-overlapping genetic associations, respectively, with regions of chromosome 14 and chromosome 9. Genetic, behavioral and gene expression correlations, and bioinformatics analysis identified several candidate genes that may represent novel molecular targets for modulating nicotine withdrawal symptoms.     

CYTOLOGY AND REPRODUCTIVE CHARACTERISTICS IN PENNISETUM SETACEUM

A green ecotype of fountaingrass, Pennisetum setaceum (Forsk.) Chiov., was found to be triploid (3x = 27) with 9 II + 9 I at metaphase I. Meiotic behavior of the bivalents was normal, but the univalents lagged and divided precociously at anaphase I. The univalent halves lagged at anaphase II and were distributed at random. Micronuclei were common, and stainable pollen was approximately 40%. The green type produced four progeny with 54 chromosomes. A purple strain of P. setaceum was found to be hexaploid (6x = 54) with varying chromosomal associations. Meiotic behavior was highly irregular and stainable pollen was almost zero. Megasporogenesis was initiated in both types, but the megaspores degenerated and nucellar and integument cells developed into four-nucleate, aposporous embryo sacs. Following self-pollination, the green type set approximately 10% seed and the purple type set 0.05% seed. Purple fountaingrass set as high as 18% seed following the application of pollen from Pennisetum ciliare (L.) Link (4x = 36), exemplifying a rare form of pseudogamy between two species. Improvement of P. setaceum through a standard breeding program appears unlikely.     

Chromosome differentiation in diploid species of Lotus (Leguminosae)

Summary Meiotic chromosome behavior was studied in seven diploid species of Lotus (L. alpinus Schleich., L. japonicus (Regel) Larsen, L. filicaulis Dur., L. schoelleri Schweinf., L. krylovii Schischk. and Serg., L. tenuis Waldst. et Kit., L. corniculatus var. minor Baker) and in 51 interspecific hybrids from 16 different crosses. Meiosis in the diploid species was quite regular. In a high proportion of the PMC's of the hybrids there was close chromosome homology with a normal association of 6 II's. However, meiotic irregularities including bridges, lagging chromosomes, univalents, and quadrivalents, occurred in a small percentage of the cells. The late separation of bivalents, the presence of quadrivalents, and inversion bridges with fragments, would indicate for some hybrids that certain chromosomes were structurally differentiated. The large number of rod bivalents observed at diakinesis was also highly suggestive that genetic nonhomology in one chromosome arm could contribute to the frequency of this type of bivalent. Therefore, the maximum number of 6 II's which occurred in a high percentage of cells may be misleading in that cryptic structural differences between chromosome arms, or segments, are not revealed. Pollen fertility in the species and hybrids was not correlated with meiotic irregularities suggesting that pollen fertility is genotypically controlled.     

A simple and rapid method for visualization of male meiotic chromosomes in Arabidopsis thaliana

Meiotic chromosomes are of basic interest to the geneticist and cell biologist who study their behavior. A rapid and highly repeatable method for visualization of meiotic chromosomes is useful. Here we describe a fast staining protocol for Arabidopsis male meiotic chromosomes. Meiocytes were squashed into a labeling buffer, the chromosome morphology could be analyzed using fluorescence without any additional treatment.     

Chromosome numbers of miscellaneous Malagasy Acanthaceae

Meiotic chromosome numbers are reported for 12 species in eight genera of Acanthaceae from Madagascar. Chromosome numbers of 11 species are reported for the first time. Counts inMendoncia (n=19) andNeuracanthus (n=20) are the first for these genera. A new chromosome number (n=30) is reported inJusticia. Systematic implications of the chromosome counts are addressed and basic chromosome numbers for these eight genera of Malagasy Acanthaceae are discussed.     

Aberrant meiosis and spindle abnormalities in Paspalum paspaloides (Michx.) Scribn (Gramineae)

Meiotic chromosome behaviour was studied in two collections of Paspalum paspaloides (Michx.) Scribn. In one of these chromosome mosaicism, restitution nuclei, multinucleate microsporocytes and multipolar and divergent spindles were observed. Chromosome mosaicism resulted from abnormal cell cleavage. In microsporocytes exhibiting chromosome mosaicism, the number of univalents per cell decreased as the chromosome number per cell increased. The increase in chromosome number brought it close to the euploid number (2n=60). Chromosome mosaicism never involved any increase above the cuploid number. In the other collection, meiosis was relatively normal.The work has been carried out at the Department of Botany, Gorakhpur University.     

Meiotic behaviour of tetraploid wheats (<Emphasis Type="Italic">Triticum turgidum</Emphasis> L.) and their synthetic hexaploid wheat derivates influenced by meiotic restitution and heat stress

Meiotic restitution is considered to be a common mechanism of polyploidization in plants and hence is one of the most important processes in plant speciation. Meiotic behaviour of plant chromosomes is influenced by both genetic and environmental factors. In this study, the meiotic behaviour of cereal crops was investigated, which includes tetraploid wheat genotypes (with and without the meiotic restitution trait) and their derivates (synthetic hexaploid wheats and a doubled haploid (DH) line), grown at two planting dates in the field. In addition, two local landraces of emmer wheat (Triticum turgidum ssp. dicoccum), one wheat cultivar (Chinese spring), one DH triticale cultivar (Eleanor) and one rye accession were included. Immature spikes of mid-autumn and end-winter sowing plants were collected in April and May 2008, respectively, fixed in Carnoy’s solution and stained with hematoxylin. Pollen mother cells (PMCs) from anthers at different stages of meiotic process were analysed for their chromosomal behaviour and irregularities. Meiotic aberrations such as laggards, chromosome bridges, micronuclei, abnormal cytokines, chromatin pulling and meiotic restitution were observed and the studied genotypes were accordingly ranked as follows: triticale > synthetic hexaploid wheats > tetraploid wheats possessing meiotic restitution > tetraploid wheats lacking meiotic restitution > rye. The results indicated that the samples that had been planted in the autumn, thus experiencing an optimum temperature level at the flowering stage, exhibited less meiotic irregularities than winter planting samples that encountered heat stress at the flowering period.     

Localization of chromosome regions in potoroo nuclei (<Emphasis Type="Italic"> Potorous tridactylus </Emphasis>Marsupialia: Potoroinae)

Chromosome paints of the rat kangaroo ( Aepyprymnus rufuscens, 2n =32) were used to define chromosome regions in the long nosed potoroo ( Potorous tridactylus, 2n =12 female, 13 male) karyotype and localize these regions in three-dimensionally preserved nuclei of the potoroo to test the hypothesis that marsupial chromosomes have a radial distribution. In human nuclei chromosomes are distributed in a proposed radial fashion. Gene-rich chromosomes in the human interphase nucleus are preferentially located in the central area while gene-poor chromosomes are found more at the periphery of the nucleus; this feature is conserved in primates and chicken. Chromosome ordering in nuclei of P. tridactylus is related to their size and centromere position. Its relationship with replication patterns in interphase nuclei and metaphase was studied. In addition it was observed that the nucleus was not a smooth entity but had projections occupied by specific chromosome regions. Edited by: R. Allshire     

Visualization of ribosomal DNA loci in spore interphasic nuclei of glomalean fungi by fluorescence in situ hybridization

 Fluorescence in situ hybridization (FISH) was applied to interphasic nuclei isolated from spores of four species of AM fungi : Scutellospora castanea, Glomus mosseae, Glomus intraradices and Gigaspora rosea. Ribosomal DNA loci were visualized using digoxigenin-labeled 25 S rDNA probes obtained by nested PCR. Several hybridization sites were detected per nucleus and an internuclear variability was observed in the number of loci. This is the first report of successful application of FISH to analyse the genomes of glomalean fungi. Accepted: 16 September 1998     

RAD‐seq linkage mapping and patterns of segregation distortion in sedges: meiosis as a driver of karyotypic evolution in organisms with holocentric chromosomes

Meiotic drive, the class of meiotic mechanisms that drive unequal segregation of alleles among gametes, may be an important force in karyotype evolution. Its role in holocentric organisms, whose chromosomes lack localized centromeres, is poorly understood. We crossed two individuals of Carex scoparia (Cyperaceae) with different chromosome numbers (2n = 33^II = 66 × 2n = 32^II = 64) to obtain F1 individuals, which we then self‐pollinated to obtain second‐generation (F2) crosses. RAD‐seq was performed for 191 individuals (including the parents, five F1 individuals and 184 F2 individuals). Our F2 linkage map based on stringent editing of the RAD‐seq data set yielded 32 linkage groups. In the final map, 865 loci were located on a linkage map of 3966.99 cM (linkage groups ranged from 24.39 to 193.31 cM in length and contained 5–51 loci each). Three linkage groups exhibit more loci under segregation distortion than expected by chance; within linkage groups, loci exhibiting segregation distortion are clustered. This finding implicates meiotic drive in the segregation of chromosome variants, suggesting that selection of chromosome variants in meiosis may contribute to the establishment and fixation of chromosome variants in Carex, which is renowned for high chromosomal and species diversity. This is an important finding as previous studies demonstrate that chromosome divergence may play a key role in differentiation and speciation in Carex.     

STAG3‐mediated stabilization of REC8 cohesin complexes promotes chromosome synapsis during meiosis

Cohesion between sister chromatids in mitotic and meiotic cells is promoted by a ring‐shaped protein structure, the cohesin complex. The cohesin core complex is composed of four subunits, including two structural maintenance of chromosome (SMC) proteins, one α‐kleisin protein, and one SA protein. Meiotic cells express both mitotic and meiosis‐specific cohesin core subunits, generating cohesin complexes with different subunit composition and possibly separate meiotic functions. Here, we have analyzed the in vivo function of STAG3, a vertebrate meiosis‐specific SA protein. Mice with a hypomorphic allele of Stag3, which display a severely reduced level of STAG3, are viable but infertile. We show that meiocytes in homozygous mutant Stag3 mice display chromosome axis compaction, aberrant synapsis, impaired recombination and developmental arrest. We find that the three different α‐kleisins present in meiotic cells show different dosage‐dependent requirements for STAG3 and that STAG3‐REC8 cohesin complexes have a critical role in supporting meiotic chromosome structure and functions.