scholarly journals Comparative chromosome studies in species of subtribe Orchidinae (Orchidaceae)

2021 ◽  
Vol 15 (4) ◽  
pp. 507-525
Author(s):  
Alessio Turco ◽  
Antonella Albano ◽  
Pietro Medagli ◽  
Robert P. Wagensommer ◽  
Saverio D'Emerico
Keyword(s):  
Silver Staining ◽  
5S Rdna ◽  
Giemsa Staining ◽  
Fish Mapping ◽  
Rdna Sequences ◽  
First Case ◽  
25S Rdna ◽  
First Time

In our study, FISH mapping using 18S-5.8S-25S rDNA and 5S rDNA sequences was performed for the first time on Ophrys tenthredinifera Willdenow, 1805, Serapias vomeracea (Burman f., 1770) Briquet, 1910 and Himantoglossum hircinum (Linnaeus, 1753) Sprengel, 1826. A detailed study was also performed on O. tenthredinifera using Giemsa-staining, silver-staining, CMA fluorescence banding and fluorescence in situ hybridisation (FISH) with rDNA probes. We analysed two subspecies, i.e. O. tenthredinifera subsp. neglecta (Parlatore, 1860) E.G. Camus, 1908 and O. tenthredinifera subsp. grandiflora (Tenore, 1819) Kreutz, 2004 by the traditional Feulgen method and constructed the karyotype. The cytotaxonomic implications for both taxa are also discussed. In Himantoglossum hircinum, FISH and silver staining highlighted differences in the number of two rDNA families (35S and 5S) with respect to Barlia robertiana (Loiseleur-Deslongchamps, 1807) Greuter, 1967. In addition, fluorescence in situ hybridisation was also applied to diploid (2n = 2x = 36) and triploid (2n = 3x = 54) Anacamptis morio (Linnaeus, 1753) R.M. Bateman, Pridgeon et M.W. Chase, 1997. As far as we are aware, this is the first case of autotriploidy observed in A. morio.

FISH-mapping of rDNAs and Arabidopsis BACs on pachytene complements of selected Brassicas

Genome ◽  
2002 ◽  
Vol 45 (1) ◽  
pp. 189-197 ◽  
Author(s):  
Piotr A Ziolkowski ◽  
Jan Sadowski
Keyword(s):  
In Situ Hybridization ◽  
Physical Mapping ◽  
5S Rdna ◽  
Artificial Chromosome ◽  
Rdna Locus ◽  
Fish Analysis ◽  
45S Rdna ◽  
Fish Mapping ◽  
Rdna Sequences

To improve resolution of physical mapping on Brassica chromosomes, we have chosen the pachytene stage of meiosis where incompletely condensed bivalents are much longer than their counterparts at mitotic metaphase. Mapping with 5S and 45S rDNA sequences demonstrated the advantage of pachytene chromosomes in efficient physical mapping and confirmed the presence of a novel 5S rDNA locus in Brassica oleracea, initially identified by genetic mapping using restriction fragment length polymorphism (RFLP). Fluorescence in situ hybridization (FISH) analysis visualized the presence of the third 5S rDNA locus on the long arm of chromosome C2 and confirmed the earlier reports of two 45S rDNA loci in the B. oleracea genome. FISH mapping of low-copy sequences from the Arabidopsis thaliana bacterial artificial chromosome (BAC) clones on the B. oleracea chromosomes confirmed the expectation of efficient and precise physical mapping of meiotic bivalents based on data available from A. thaliana and indicated conserved organization of these two BAC sequences on two B. oleracea chromosomes. Based on the heterologous in situ hybridization with BACs and their mapping applied to long pachytene bivalents, a new approach in comparative analysis of Brassica and A. thaliana genomes is discussed.Key words: Brassicaceae, pachytene chromosomes, FISH, rDNA, BACs.

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.

scholarly journals Karyotypic variation in the long-whiskered catfish Pimelodus blochii Valenciennes, 1840 (Siluriformes, Pimelodidae) from the lower Tapajós, Amazonas and Trombetas Rivers

2018 ◽  
Vol 12 (3) ◽  
pp. 285-298 ◽  
Author(s):  
Ivanny Coelho da Fonseca ◽  
Luan Aércio Melo Maciel ◽  
Frank Raynner Vasconcelos Ribeiro ◽  
Luís Reginaldo Ribeiro Rodrigues
Keyword(s):  
Silver Staining ◽  
Chromosome Pair ◽  
Fluorescent In Situ Hybridization ◽  
Species Complex ◽  
Diploid Number ◽  
Acrocentric Chromosome ◽  
5S Rdna ◽  
Karyotypic Formula ◽  
Karyotypic Variation

The genus Pimelodus LaCépède, 1803 comprises 35 formally recognized species distributed along the major neotropical river basins. Despite conservatism in diploid number with 2n=56, an intense variation of chromosomal morphology (karyotypic formula) has been documented in Pimelodus species. In the present study, we analyzed karyotypes of 20 specimens, identified as Pimelodusblochii Valenciennes, 1840 and collected from the lower courses of the Tapajós, Amazonas and Trombetas Rivers. The karyotypes were characterized by Giemsa conventional staining, C-banding, silver staining (Ag-NOR) and fluorescent in situ hybridization (FISH) with 5S and 18S rDNA probes. The karyotypes showed 2n=56 chromosomes in fish from the Tapajós River. In contrast, fish from the Amazonas and Trombetas Rivers had 2n=58. The nucleolus organizing regions were labeled on the short arm of an acrocentric chromosome as demonstrated by silver staining and FISH. Signals for 18S and 5S rDNA were co-localized on one chromosome pair. Our results demonstrate karyotypic divergence between Tapajós and Amazonas-Trombetas populations of P.blochii, interpreted as supporting the existence of a species complex in this taxon.

Distribution of FISH oligo-5S rDNA and oligo-(AGGGTTT)3 in Hibiscus mutabilis L.

Genome ◽  
2021 ◽  
pp. 1-10
Author(s):  
Xiaomei Luo ◽  
Zhoujian He
Keyword(s):  
Chromosome Number ◽  
Physical Map ◽  
Molecular Cytogenetics ◽  
5S Rdna ◽  
Chromosome Length ◽  
Chromosome Identification ◽  
Karyotype Asymmetry ◽  
First Time ◽  
Distribution Of Fish

Hibiscus exhibits high variation in chromosome number both within and among species. The Hibiscus mutabilis L. karyotype was analyzed in detail using fluorescence in situ hybridization (FISH) with oligonucleotide probes for (AG3T3)3 and 5S rDNA, which were tested here for the first time. In total, 90 chromosomes were counted in prometaphase and metaphase, and all exhibited similarly intense (AG3T3)3 signals at both ends. (AG3T3)3 showed little variation and thus did not allow discrimination among H. mutabilis chromosomes, but its location at both ends confirmed the integrity of each chromosome, thus contributing to accurate counting of the numerous, small chromosomes. Oligo-5S rDNA marked the proximal/distal regions of six chromosomes: weak signals on chromosomes 7 and 8, slightly stronger signals on chromosomes 15 and 16, and very strong signals on chromosomes 17 and 18. Therefore, 5S rDNA could assist in chromosome identification in H. mutabilis. Metaphase chromosome lengths ranged from 3.00 to 1.18 μm, indicating small chromosomes. The ratios of longest to shortest chromosome length in prometaphase and metaphase were 2.58 and 2.54, respectively, indicating karyotype asymmetry in H. mutabilis. These results provide an exact chromosome number and a physical map, which will be useful for genome assembly and contribute to molecular cytogenetics in the genus Hibiscus.

Evolutionary Dynamics of 5S rDNA and Recurrent Association of Transposable Elements in Electric Fish of the Family Gymnotidae (Gymnotiformes): The Case of Gymnotus mamiraua

2016 ◽  
Vol 149 (4) ◽  
pp. 297-303 ◽  
Author(s):  
Maelin da Silva ◽  
Patricia Barbosa ◽  
Roberto F. Artoni ◽  
Eliana Feldberg
Keyword(s):  
Electric Fish ◽  
Evolutionary Dynamics ◽  
Chromosomal Location ◽  
Molecular Mapping ◽  
5S Rdna ◽  
Mapping Data ◽  
Rdna Sequences ◽  
Rdna Sites ◽  
The Family

Gymnotidae is a family of electric fish endemic to the Neotropics consisting of 2 genera: Electrophorus and Gymnotus. The genus Gymnotus is widely distributed and is found in all of the major Brazilian river systems. Physical and molecular mapping data for the ribosomal DNA (rDNA) in this genus are still scarce, with its chromosomal location known in only 11 species. As other species of Gymnotus with 2n = 54 chromosomes from the Paraná-Paraguay basin, G. mamiraua was found to have a large number of 5S rDNA sites. Isolation and cloning of the 5S rDNA sequences from G. mamiraua identified a fragment of a transposable element similar to the Tc1/mariner transposon associated with a non-transcribed spacer. Double fluorescence in situ hybridization analysis of this element and the 5S rDNA showed that they were colocalized on several chromosomes, in addition to acting as nonsyntenic markers on others. Our data show the association between these sequences and suggest that the Tc1 retrotransposon may be the agent that drives the spread of these 5S rDNA-like sequences in the G. mamiraua genome.

Physical mapping of four sites of 5S rDNA sequences and one site of the α-amylase-2 gene in barley (Hordeum vulgare)

Genome ◽  
1993 ◽  
Vol 36 (3) ◽  
pp. 517-523 ◽  
Author(s):  
I. J. Leitch ◽  
J. S. Heslop-Harrison
Keyword(s):  
In Situ Hybridization ◽  
Hordeum Vulgare ◽  
Physical Mapping ◽  
5S Rdna ◽  
Chromosome 1 ◽  
Chromosome 2 ◽  
Hordeum Vulgare L ◽  
Rdna Sequences ◽  
5S Rdna Sequence

The 5S rDNA sequences have been mapped on four pairs of barley (Hordeum vulgare L.) chromosomes using in situ hybridization and barley monotelotrisomic lines. The 5S rDNA sequences are located, genetically and physically, on the short arm of chromosome 1 (7I) and the long arms of chromosomes 2 (2I) and 3 (3I). The 5S rDNA sequence is also located on the physically long arm of chromosome 4 (4I). Only one site on chromosome 2(2I) has previously been reported. The characteristic locations of the 5S rDNA sequences make them useful as molecular markers to identify each barley chromosome. The physical position of the low-copy α-amylase-2 gene was determined using in situ hybridization; the location of this gene on the long arm of chromosome 1 (7I) was confirmed by reprobing the same preparation with the 5S rDNA probe. The results show that there is a discrepancy between the physical and genetic position of the α-amylase-2 gene.Key words: genetic mapping, physical mapping, barley, mapping, 5S DNA, α-amylase, in situ hybridization.

FISH-aimed karyotyping and characterization of Renner complexes in permanent heterozygote Rhoeo spathacea

Genome ◽  
2005 ◽  
Vol 48 (1) ◽  
pp. 145-153 ◽  
Author(s):  
Hieronim Golczyk ◽  
Robert Hasterok ◽  
Andrzej J Joachimiak
Keyword(s):  
5S Rdna ◽  
Chromosome Identification ◽  
Rdna Sites ◽  
Rdna Loci ◽  
Target Fish ◽  
Telomere Association ◽  
25S Rdna ◽  
Dapi Fluorescence

Fluorescence in situ hybridization (FISH) using 25S rDNA, 5S rDNA, and telomere sequences as probes was carried out in the complex permanent heterozygote Rhoeo spathacea. Telomere sites were exclusively terminal. All 10 25S rDNA loci were located distally and appeared transcriptionally active after silver staining. Six distal and 2 interstitial 5S rDNA sites were detected; 2 of the distal sites strictly colocalized with 25S rDNA loci. The 2 intercalary 5S rDNA loci occurred in short arms of 2 chromosomes that conjoined at meiosis. Chromosomes differed as to the amount of AT-rich centric heterochromatin, suggesting involvement of pericentromeric regions in translocations. The possibility of Robertsonian-like rearrangements was discussed. Double target FISH with ribosomal probes along with DAPI fluorescence gave the basis for full chromosome identification in mitosis. The 2 Renner complexes are structurally balanced, both having 5 25S and 4 5S rDNA sites. Centromere clustering, telomere association, a high number of NOR sites, and a strong tendency for formation of joint nucleoli contribute to the preservation of highly polarized Rabl arrangement at interphase. These findings were discussed in relation to meiotic catenation in Rhoeo.Key words: chromosomes, complex heterozygotes, FISH, heterochromatin, interphase, meiotic multivalents, nucleolus, NOR, rDNA, Rhoeo, Renner complexes, translocations.

Distribution of highly repeated DNA sequences in species of the genus Lens Miller

Genome ◽  
2003 ◽  
Vol 46 (6) ◽  
pp. 1118-1124 ◽  
Author(s):  
Incoronata Galasso
Keyword(s):  
Dna Sequences ◽  
Lens Culinaris ◽  
Repetitive Sequences ◽  
5S Rdna ◽  
Chromosome Identification ◽  
Mitotic Chromosomes ◽  
Genomic Relationships ◽  
Lens Nigricans ◽  
25S Rdna

Multiple-target fluorescence in situ hybridization (FISH) was applied on mitotic chromosomes of seven Lens taxa using two highly repetitive sequences (pLc30 and pLc7) isolated from the cultivated lentil and the multigene families for the 18S–5.8S–25S (pTa71) and 5S rRNA (pTa794) from wheat simultaneously as probes. The number and location of pLc30 and pLc7 sites on chromosomes varied markedly among the species, whereas the hybridization pattern of 5S rDNA and 18S–5.8S–25S rDNA was less variable. In general, each species showed a typical FISH karyotype and few differences were observed among accessions belonging to the same species, except for the accessions of Lens odemensis. The most similar FISH karyotype to the cultivated lentil is that of Lens culinaris subsp. orientalis, whereas Lens nigricans and Lens tomentosus are the two species that showed the most divergent FISH patterns compared with all taxa for number and location of pLc30 and 18S–5.8S–25S rDNA sites.Key words: chromosome identification, comparative FISH karyotype, wild Lens species, genomic relationships.

Uniparental loss of ribosomal DNA in the allotetraploid grass Zingeria trichopoda (2n = 8)

Genome ◽  
2003 ◽  
Vol 46 (1) ◽  
pp. 156-163 ◽  
Author(s):  
Violetta Kotseruba ◽  
Dorota Gernand ◽  
Armin Meister ◽  
Andreas Houben
Keyword(s):  
Ribosomal Dna ◽  
5S Rdna ◽  
45S Rdna ◽  
Rdna Sequences ◽  
Genome Wide ◽  
Rdna Sites ◽  
Rdna Loci ◽  
Subsequent Elimination ◽  
Genome Level

Analysis of the grass Zingeria trichopoda (2n = 8, 2C = 5.3 pg) revealed a dynamic evolution with the following characteristics. (i) Genomic in situ hybridization (GISH) demonstrates that Z. trichopoda evolved from an interspecific hybrid involving a species like contemporary Zingeria biebersteiniana (2n = 4) and a second species with a similar low number of chromosomes. The nucleus of Z. trichopoda is spatially organized at the genome level and the two parental genomes occupy distinct and separate domains of lateral arrangements. (ii) The copy number of the Z. biebersteiniana specific pericentromeric tandem repeat family Zbcen1 is drastically reduced in Z. trichopoda. (iii) GISH in combination with labeled rDNA sequences simultaneously discriminated the two parental genomes and the corresponding 5S and 45S rDNA sites. Hence, following allopolyploidization of Z. trichopoda the Z. biebersteiniana like parental chromosomes probably underwent drastic loss of 45S rDNA. This could have arisen either through the loss ofZ. biebersteiniana derived 45S rDNA or through Z. trichopoda genome-wide homogenization of Z. biebersteiniana type 45S rDNA and subsequent elimination of 45S rDNA loci from Z. biebersteiniana derived chromosomes. Finally, 5S rDNA loci are present in both subgenomes of Z. trichopoda and the chromosomal position of these loci is similar for both Z. biebersteiniana and the Z. biebersteiniana like parental genome of Z. trichopoda.Key words: genome evolution, polyploidy, ribosomal DNA, Poaceae.

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