scholarly journals Detecting Brachypodium distachyon Chromosomes Bd4 and Bd5 in MH- and X-Ray-Induced Micronuclei Using mcFISH

2019 ◽  
Vol 20 (11) ◽  
pp. 2848
Author(s):  
Arita Kus ◽  
Joanna Szymanowska-Pułka ◽  
Jolanta Kwasniewska ◽  
Robert Hasterok
Keyword(s):  
Brachypodium Distachyon ◽  
In Situ Hybridisation ◽  
5S Rdna ◽  
Artificial Chromosome ◽  
Proximal Region ◽  
Root Tip ◽  
Fluorescence In Situ Hybridisation ◽  
Dna Breaks ◽  
Submetacentric Chromosome ◽  
Chromosome Fragments

Micronuclei are biomarkers of genotoxic effects and chromosomal instability. They are formed when chromosome fragments or whole chromosomes fail to disjoin into daughter nuclei. We present qualitative and quantitative analyses of the involvement of specific chromosome regions of chromosomes Bd4 and Bd5 in the formation of micronuclei of Brachypodium distachyon root tip cells following maleic hydrazide (MH) treatment and X-radiation. This is visualised by cytomolecular approaches using bacterial artificial chromosome (BAC)-based multicolour fluorescence in situ hybridisation (mcFISH) in combination with 5S and 25S rDNA probes. The results showed that the long arm of submetacentric chromosome Bd4 forms micronuclei at twice the frequency of its short arm, suggesting that the former is more prone to double-strand breaks (DSBs). In contrast, no difference was observed in the frequency of micronuclei derived from the long and short arms of submetacentric chromosome Bd5. Interestingly, the proximal region of the short arm of Bd5 is more prone to DSBs than its distal part. This demonstrates that 5S rDNA and 35S rDNA loci are not “hot spots” for DNA breaks after the application of these mutagens.

Karyotyping of Brachypodium pinnatum (2n = 18) chromosomes using cross-species BAC–FISH

Genome ◽  
2013 ◽  
Vol 56 (4) ◽  
pp. 239-243 ◽  
Author(s):  
Elzbieta Wolny ◽  
Wojciech Fidyk ◽  
Robert Hasterok
Keyword(s):  
Brachypodium Distachyon ◽  
Genome Structure ◽  
In Situ Hybridisation ◽  
Artificial Chromosome ◽  
Fluorescence In Situ Hybridisation ◽  
Morphometric Features ◽  
Sequential Experiments ◽  
Brachypodium Pinnatum ◽  
Bac Fish

Identification of individual chromosomes in a complement is usually a difficult task in the case of most plant species, especially for those with small, numerous, and morphologically uniform chromosomes. In this paper, we demonstrate that the landmarks produced by cross-species fluorescence in situ hybridisation (FISH) of Brachypodium distachyon derived bacterial artificial chromosome (BAC) clones can be used for discrimination of Brachypodium pinnatum (2n = 18) chromosomes. Selected sets of clones were hybridised in several sequential experiments performed on exactly the same chromosome spreads, using reprobing of cytological preparations. Analysis of the morphometric features of B. pinnatum chromosomes was performed to establish their total length, the position of centromeres, and the position of BAC-based landmarks in relation to the centromere, thereby enabling their effective karyotyping, which is a prerequisite for more complex study of the grass genome structure and evolution at the cytomolecular level.

Effects of the diploidisation process upon the 5S and 35S rDNA sequences in the allopolyploid species of the Dilatata group of Paspalum (Poaceae, Paniceae)

2019 ◽  
Vol 67 (7) ◽  
pp. 521
Author(s):  
Magdalena Vaio ◽  
Cristina Mazzella ◽  
Marcelo Guerra ◽  
Pablo Speranza
Keyword(s):  
Evolutionary Dynamics ◽  
In Situ Hybridisation ◽  
5S Rdna ◽  
Its Region ◽  
Variable Number ◽  
Rrna Genes ◽  
Fluorescence In Situ Hybridisation ◽  
Rdna Sites ◽  
35S Rdna ◽  
Genome Restructuring

The Dilatata group of Paspalum includes species and biotypes native to temperate South America. Among them, five sexual allotetraploids (x = 10) share the same IIJJ genome formula: P. urvillei Steud, P. dasypleurum Kunze ex Desv., P. dilatatum subsp. flavescens Roseng., B.R. Arrill. & Izag., and two biotypes P. dilatatum Vacaria and P. dilatatum Virasoro. Previous studies suggested P. intermedium Munro ex Morong & Britton and P. juergensii Hack. or related species as their putative progenitors and donors of the I and J genome, respectively, and pointed to a narrow genetic base for their maternal origin. It has not yet been established whether the various members of the Dilatata group are the result of a single or of multiple allopolyploid formations. Here, we aimed to study the evolutionary dynamics of rRNA genes after allopolyploidisation in the Dilatata group of Paspalum and shed some light into the genome restructuring of the tetraploid taxa with the same genome formula. We used double target fluorescence in situ hybridisation of 35S and 5S rDNA probes and sequenced the nrDNA internal transcribed spacer (ITS) region. A variable number of loci at the chromosome ends were observed for the 35S rDNA, from 2 to 6, suggesting gain and loss of sites. For the 5S rDNA, only one centromeric pair of signals was observed, indicating a remarkable loss after polyploidisation. All ITS sequences generated were near identical to the one found for P. intermedium. Although sequences showed a directional homogeneisation towards the putative paternal progenitor in all tetraploid species, the observed differences in the number and loss of rDNA sites suggest independent ongoing diploidisation processes in all taxa and genome restructuring following polyploidy.

scholarly journals The role of chromosomal fusion in the karyotypic evolution of the genus Ageneiosus (Siluriformes: Auchenipteridae)

2013 ◽  
Vol 11 (2) ◽  
pp. 327-334 ◽  
Author(s):  
Roberto Laridondo Lui ◽  
Daniel Rodrigues Blanco ◽  
Juliana de Fatima Martinez ◽  
Vladimir Pavan Margarido ◽  
Paulo Cesar Venere ◽  
...  
Keyword(s):  
Chromosome Pair ◽  
5S Rdna ◽  
Proximal Region ◽  
Terminal Region ◽  
Metacentric Chromosome ◽  
South American ◽  
Submetacentric Chromosome ◽  
Araguaia River ◽  
The Family ◽  
Preferential Location

Ageneiosus is the most widely distributed genus of the family Auchenipteridae among South American river basins. Although chromosome studies in the family are scarce, this genus has the largest number of analyzed species, with 2n = 54 to 56 chromosomes, differing from the rest of the family (2n = 58). This study aimed to analyze Ageneiosus inermis from the Araguaia River basin. The diploid number found was of 56 chromosomes. Heterochromatin was allocated in terminal region of most chromosomes, plus a pericentromeric heterochromatic block in pair 1, a pair distinguished by size in relation to other chromosomes pairs. AgNORs were detected in only one submetacentric chromosome pair, which was confirmed by FISH. 5S rDNA was present in only one metacentric chromosome pair. Hybridization with [TTAGGG]n sequence marked the telomeres of all chromosomes, in addition to an ITS in the proximal region of the short arm of pair 1. The repetitive [GATA]n sequence was dispersed, with preferential location in terminal region of the chromosomes. Ageneiosus has a genomic organization somewhat different when compared to other Auchenipteridae species. Evidences indicate that a chromosomal fusion originated the first metacentric chromosome pair in A. inermis, rearrangement which may be a basal event for the genus

Karyotype analysis of Lilium longiflorum and Lilium rubellum by chromosome banding and fluorescence in situ hybridisation

Genome ◽  
2001 ◽  
Vol 44 (5) ◽  
pp. 911-918 ◽  
Author(s):  
Ki-Byung Lim ◽  
Jannie Wennekes ◽  
J Hans de Jong ◽  
Evert Jacobsen ◽  
Jaap M van Tuyl
Keyword(s):  
In Situ Hybridisation ◽  
5S Rdna ◽  
Lilium Longiflorum ◽  
Fluorescence In Situ Hybridisation ◽  
45S Rdna ◽  
C Banding ◽  
Rdna Sequences ◽  
Rdna Signal ◽  
5S And 45S Rdna

Detailed karyotypes of Lilium longiflorum and L. rubellum were constructed on the basis of chromosome arm lengths, C-banding, AgNO3 staining, and PI-DAPI banding, together with fluorescence in situ hybridisation (FISH) with the 5S and 45S rDNA sequences as probes. The C-banding patterns that were obtained with the standard BSG technique revealed only few minor bands on heterologous positions of the L. longiflorum and L. rubellum chromosomes. FISH of the 5S and 45S rDNA probes on L. longiflorum metaphase complements showed overlapping signals at proximal positions of the short arms of chromosomes 4 and 7, a single 5S rDNA signal on the secondary constriction of chromosome 3, and one 45S rDNA signal adjacent to the 5S rDNA signal on the subdistal part of the long arm of chromosome 3. In L. rubellum, we observed co-localisation of the 5S and 45S rDNA sequences on the short arm of chromosomes 2 and 4 and on the long arms of chromosomes 2 and 3, and two adjacent bands on chromosome 12. Silver staining (Ag-NOR) of the nucleoli and NORs in L. longiflorum and L. rubellum yielded a highly variable number of signals in interphase nuclei and only a few faint silver deposits on the NORs of mitotic metaphase chromosomes. In preparations stained with PI and DAPI, we observed both red- and blue-fluorescing bands at different positions on the L. longiflorum and L. rubellum chromosomes. The red-fluorescing or so-called reverse PI-DAPI bands always coincided with rDNA sites, whereas the blue-fluorescing DAPI bands corresponded to C-bands. Based on these techniques, we could identify most of chromosomes of the L. longiflorum and L. rubellum karyotypes.Key words: fluorescence in situ hybridisation, FISH, 5S rDNA, 45S rDNA, C-banding, reverse PI-DAPI banding.

Status and applications of genomic resources for the gray, short-tailed opossum, Monodelphis domestica, an American marsupial model for comparative biology

2006 ◽  
Vol 54 (3) ◽  
pp. 173 ◽  
Author(s):  
Paul B. Samollow
Keyword(s):  
In Situ Hybridisation ◽  
Artificial Chromosome ◽  
Monodelphis Domestica ◽  
Whole Genome Sequence ◽  
Whole Genome ◽  
Fluorescence In Situ Hybridisation ◽  
Resource Base ◽  
Bac Contig ◽  
Genomic Resources ◽  
Research Areas

Owing to its small size, favourable reproductive characteristics, and simple husbandry, the gray, short-tailed opossum, Monodelphis domestica, has become the most widely distributed and intensively utilised laboratory-bred research marsupial in the world today. This article provides an overview of the current state and future projections of genomic resources for this species and discusses the potential impact of this growing resource base on active research areas that use M. domestica as a model system. The resources discussed include: fully arrayed, bacterial artificial chromosome (BAC) libraries; an expanding linkage map; developing full-genome BAC-contig and chromosomal fluorescence in situ hybridisation maps; public websites providing access to the M. domestica whole-genome-shotgun sequence trace database and the whole-genome sequence assembly; and a new project underway to create an expressed-sequence database and microchip expression arrays for functional genomics applications. Major research areas discussed span a variety of genetic, evolutionary, physiologic, reproductive, developmental, and behavioural topics, including: comparative immunogenetics; genomic imprinting; reproductive biology; neurobiology; photobiology and carcinogenesis; genetics of lipoprotein metabolism; developmental and behavioural endocrinology; sexual differentiation and development; embryonic and fetal development; meiotic recombination; genome evolution; molecular evolution and phylogenetics; and more.

scholarly journals Evolution of the Human Chromosome 13 Synteny: Evolutionary Rearrangements, Plasticity, Human Disease Genes and Cancer Breakpoints

Genes ◽  
2020 ◽  
Vol 11 (4) ◽  
pp. 383 ◽  
Author(s):  
Rita Scardino ◽  
Vanessa Milioto ◽  
Anastasia A. Proskuryakova ◽  
Natalia A. Serdyukova ◽  
Polina L. Perelman ◽  
...  
Keyword(s):  
Human Chromosome ◽  
In Situ Hybridisation ◽  
Artificial Chromosome ◽  
Primate Species ◽  
Eutherian Mammal ◽  
Disease Genes ◽  
Fluorescence In Situ Hybridisation ◽  
Saguinus Oedipus ◽  
Chromosome 13 ◽  
High Level

The history of each human chromosome can be studied through comparative cytogenetic approaches in mammals which permit the identification of human chromosomal homologies and rearrangements between species. Comparative banding, chromosome painting, Bacterial Artificial Chromosome (BAC) mapping and genome data permit researchers to formulate hypotheses about ancestral chromosome forms. Human chromosome 13 has been previously shown to be conserved as a single syntenic element in the Ancestral Primate Karyotype; in this context, in order to study and verify the conservation of primate chromosomes homologous to human chromosome 13, we mapped a selected set of BAC probes in three platyrrhine species, characterised by a high level of rearrangements, using fluorescence in situ hybridisation (FISH). Our mapping data on Saguinus oedipus, Callithrix argentata and Alouatta belzebul provide insight into synteny of human chromosome 13 evolution in a comparative perspective among primate species, showing rearrangements across taxa. Furthermore, in a wider perspective, we have revised previous cytogenomic literature data on chromosome 13 evolution in eutherian mammals, showing a complex origin of the eutherian mammal ancestral karyotype which has still not been completely clarified. Moreover, we analysed biomedical aspects (the OMIM and Mitelman databases) regarding human chromosome 13, showing that this autosome is characterised by a certain level of plasticity that has been implicated in many human cancers and diseases.

scholarly journals Chromosomal Mapping of 18S-25S and 5S Ribosomal Genes on 15 Species of Fagaceae From Northern Thailand

2008 ◽  
Vol 57 (1-6) ◽  
pp. 5-13 ◽  
Author(s):  
P. Chokchaichamnankit ◽  
K. Anamthawat-Jónsson ◽  
W. Chulalaksananukul
Keyword(s):  
In Situ Hybridisation ◽  
5S Rdna ◽  
Ribosomal Genes ◽  
Chromosomal Mapping ◽  
Hybrid Origin ◽  
Fluorescence In Situ Hybridisation ◽  
Northern Thailand ◽  
Quercus Species ◽  
Rdna Sites ◽  
25S Rdna

Abstract Fifteen species of Fagaceae from Chiang Mai province, northern Thailand, were investigated: eight Castanopsis, four Lithocarpus and three Quercus species. The species were generally diploid with the chromosome number 2n = 24, and the basic number x =12 was confirmed in some species with meiosis. One tree belonging to Q. lenticellatus had 2n = 14. Chromosomal mapping of the highly repetitive 18S-25S and 5S ribosomal genes by fluorescence in situ hybridisation (FISH) was performed. Most species (from all three genera) showed four 18S-25S rDNA sites (two pairs: one subterminal major and one paracentromeric/intercalary minor loci) and two 5S rDNA sites (one pair: paracentromeric locus). Quercus kerrii also had two pairs of 18S-25S rDNA sites, but both were subterminal major loci. Two species, C. argentea and Q. brandisianus, only had one pair of 18S-25S rDNA sites. Two species, C. calathiformis and L. vestitus, showed an odd number of (unpaired) sites, and this indicated hybrid origin and/or polyploidy. Polyploid cells were detected in these species. The ribosomal gene maps based on both sequences together were genus-specific. In Castanopsis, the 18S-25S and the 5S genes were localized on three different chromosome pairs, and comprised species-specific maps. On the other hand, the ribosomal genes in Lithocarpus and Quercus were found only on two chromosome pairs, because one of the two 18S-25S rDNA loci was localized on the same chromosome as the 5S rDNA locus. The FISH markers may be used to clarify discrepancies arising from morphological assessments.

scholarly journals Chromosome Mapping of 5S Ribosomal Genes in Indo-Pacific and Atlantic Muraenidae: Comparative Analysis by Dual Colour Fluorescence In Situ Hybridisation

Genes ◽  
2020 ◽  
Vol 11 (11) ◽  
pp. 1319
Author(s):  
Elisabetta Coluccia ◽  
Federica Deidda ◽  
Cinzia Lobina ◽  
Riccardo Melis ◽  
Cristina Porcu ◽  
...  
Keyword(s):  
Gene Family ◽  
In Situ Hybridisation ◽  
5S Rdna ◽  
Chromosome Mapping ◽  
Ribosomal Gene ◽  
Ribosomal Genes ◽  
Chromosomal Mapping ◽  
Fluorescence In Situ Hybridisation ◽  
Species Specific

The Muraenidae is one of the largest and most complex anguilliform families. Despite their abundance and important ecological roles, morays are little studied, especially cytogenetically, and both their phylogenetic relationships and the taxonomy of their genera are controversial. With the aim of extending the karyology of this fish group, the chromosomal mapping of the 5S ribosomal gene family was performed on seven species belonging to the genera Muraena and Gymnothorax from both the Atlantic and Pacific oceans. Fluorescence in situ hybridisation (FISH) experiments were realized using species-specific 5S rDNA probes; in addition, two-colour FISH was performed to investigate the possible association with the 45S ribosomal gene family. Multiple 5S rDNA clusters, located either in species-specific or in possibly homoeologous chromosomes, were found. Either a syntenic or different chromosomal location of the two ribosomal genes was detected. Our results revealed variability in the number and location of 5S rDNA clusters and confirmed a substantial conservation of the number and location of the 45S rDNA.

scholarly journals Rapid Multi-Hybridisation FISH Screening for Balanced Porcine Reciprocal Translocations Suggests a Much Higher Abnormality Rate Than Previously Appreciated

Cells ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 250
Author(s):  
Rebecca E O’Connor ◽  
Lucas G Kiazim ◽  
Claudia C Rathje ◽  
Rebecca L Jennings ◽  
Darren K Griffin
Keyword(s):  
Semen Analysis ◽  
In Situ Hybridisation ◽  
Economic Loss ◽  
Environmental Damage ◽  
Fluorescence In Situ Hybridisation ◽  
Reciprocal Translocations ◽  
Chromosome Translocations ◽  
Farrowing Rate ◽  
Significant Economic Loss

With demand rising, pigs are the world’s leading source of meat protein; however significant economic loss and environmental damage can be incurred if boars used for artificial insemination (AI) are hypoprolific (sub-fertile). Growing evidence suggests that semen analysis is an unreliable tool for diagnosing hypoprolificacy, with litter size and farrowing rate being more applicable. Once such data are available, however, any affected boar will have been in service for some time, with significant financial and environmental losses incurred. Reciprocal translocations (RTs) are the leading cause of porcine hypoprolificacy, reportedly present in 0.47% of AI boars. Traditional standard karyotyping, however, relies on animal specific expertise and does not detect more subtle (cryptic) translocations. Previously, we reported development of a multiple hybridisation fluorescence in situ hybridisation (FISH) strategy; here, we report on its use in 1641 AI boars. A total of 15 different RTs were identified in 69 boars, with four further animals XX/XY chimeric. Therefore, 4.5% had a chromosome abnormality (4.2% with an RT), a 0.88% incidence. Revisiting cases with both karyotype and FISH information, we reanalysed captured images, asking whether the translocation was detectable by karyotyping alone. The results suggest that chromosome translocations in boars may be significantly under-reported, thereby highlighting the need for pre-emptive screening by this method before a boar enters a breeding programme.

scholarly journals Connecting structure to function with the recovery of over 1000 high-quality metagenome-assembled genomes from activated sludge using long-read sequencing

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Caitlin M. Singleton ◽  
Francesca Petriglieri ◽  
Jannie M. Kristensen ◽  
Rasmus H. Kirkegaard ◽  
Thomas Y. Michaelsen ◽  
...  
Keyword(s):  
16S Rrna ◽  
Wastewater Treatment Plants ◽  
In Situ Hybridisation ◽  
Amplicon Sequencing ◽  
Rrna Genes ◽  
Fluorescence In Situ Hybridisation ◽  
Sequencing Data ◽  
High Quality ◽  
16S Rrna Amplicon Sequencing ◽  
Long Read

AbstractMicroorganisms play crucial roles in water recycling, pollution removal and resource recovery in the wastewater industry. The structure of these microbial communities is increasingly understood based on 16S rRNA amplicon sequencing data. However, such data cannot be linked to functional potential in the absence of high-quality metagenome-assembled genomes (MAGs) for nearly all species. Here, we use long-read and short-read sequencing to recover 1083 high-quality MAGs, including 57 closed circular genomes, from 23 Danish full-scale wastewater treatment plants. The MAGs account for ~30% of the community based on relative abundance, and meet the stringent MIMAG high-quality draft requirements including full-length rRNA genes. We use the information provided by these MAGs in combination with >13 years of 16S rRNA amplicon sequencing data, as well as Raman microspectroscopy and fluorescence in situ hybridisation, to uncover abundant undescribed lineages belonging to important functional groups.

Sign in / Sign up

Export Citation Format

Share Document