Characterization of eight species of Aloe (Asphodelaceae) from the nucleolar organizing region

Nucleolar organizing region of eight species of Aloe was analyzed in somatic metaphases and interphase nuclei. All species showed a uniform 2n=14, with eight large chromosomes and six small chromosomes. Satellites were observed on the long arm of one or two pairs of large chromosomes and/or on the short arm of one of the small pairs. The silver-stained nucleolus organizing regions were located on the subtelomeric region of the long arm of one or two pairs of large chromosomes, except for Aloe dichotoma and Aloe maculata, which the AgNORs were located at a short arm of one of their small chromosomes. In most studied species, the active AgNOR number was four. However, this number changing from one to eight. For all species, the interphase number of nucleoli can be one or two, while, in Aloe excelsa, this number can be changing from one to eight. Polymorphism of active AgNORs and the number of interphase nucleoli were revealed, except for Aloe petricola, which active AgNORs were located only in the subtelomeric regions at the long arm of one of the L2 chromosomes, as well as in the L4 pair, which is agreement with the maximum number (three) of interphase nucleoli.Fil: Sánchez, Ysbelia. Universidad Nacional Experimental del Tachira; Venezuela. Universidad Pedagogica Experimental Libertador, Institute Pedagogico de Caracas; VenezuelaFil: Raymúndez, María B.. Universidad Central de Venezuela; VenezuelaFil: Imery, José. Universidad de Oriente - Venezuela; VenezuelaFil: Acosta, María Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Moscone, Eduardo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentin

Efficiency of cytogenetic methods in detecting a chromosome rearrangement induced by ionizing radiation in a cultivated chili pepper line (Capsicum baccatum var. pendulum – Solanaceae)

Purpose: To locate transient chromosome aberrations on a selected pepper cultivar and determine the tracing efficiency of different cytogenetic methods. Materials and methods: Seeds from Capsicum baccatum var. pendulum cultivar ‘Cayenne’ were treated with an acute dose of X-rays (300 Gy) and chromosome aberrations were analysed by different cytogenetic methods [Feulgen, silver staining for nucleolus organizer regions (silver positive nucleolus organizing regions or AgNOR), fluorescent banding, fluorescence in situ hybridization (FISH) and meiotic analysis]. Results: A rearranged chromosome carrying two nucleolus organizing regions (NOR) induced by ionizing radiation was detected in the cultivar, with the occurrence of a small reciprocal exchange between a chromosome of pair no. 1 and another chromosome of pair no. 3, both carrying active NOR in short arms and associated chromomycin A positive/diamidino-phenylindole negative (CMA+/DAPI−) heterochromatin. Meiotic analysis showed a quadrivalent configuration, confirming a reciprocal translocation between two chromosomes. Conclusions: The use of X-rays in Capsicum allowed us to develop and identify a pepper line with structural rearrangements between two NOR-carrying chromosomes. We postulate that all the cytological techniques employed in this research were efficient in the search for chromosome aberrations. Particularly, Feulgen and AgNOR were the most suitable in those cases of transient rearrangements, whereas fluorescent banding and FISH were appropriate for intransitive ones.Instituto de Patología VegetalInstituto de GenéticaFil: Scaldaferro, Marisel Analía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Grabiele, Mauro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Seijo, Jose Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste; ArgentinaFil: Debat, Humberto Julio. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; ArgentinaFil: Romero, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Ducasse, Daniel Adrian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; ArgentinaFil: Prina, Alberto Raul. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; ArgentinaFil: Moscone, Eduardo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal; Argentin

B Chromosomes in Nierembergia aristata (Solanaceae): Nucleolar Activity and Competition with the A Chromosomes

B chromosomes are additional dispensable chromosomes that may be present in some individuals, populations, or species, which have probably arisen from the A chromosomes but follow their own evolutionary pathway. Supposedly, B chromosomes do not contain major genes except for ribosomal DNA (rDNA) sequences that have been mapped on the supernumerary chromosomes of many plants and animals. This paper is a new report of B chromosome occurrence in plants. B chromosomes with nucleolar organizing regions (NORs) were found in a diploid sample of Nierembergiaaristata D. Don (sub nom. N. stricta Miers) (2n = 2x = 16). This is an extreme case in which B chromosomes possess not only strong nucleolar activity, as revealed by conventional staining methods, AgNOR and fluorescence banding, and fluorescent in situ hybridization (FISH), but also show nucleolar competition with the A chromosomes. The observed phenomenon could be analogous to the nucleolar dominance or ‘differential amphiplasty’ phenomenon that occurs in interspecific hybrids.Fil: Acosta, María Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); ArgentinaFil: Moscone, Eduardo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); Argentin

Karyological relationships among some South American species of Solanum (Solanaceae) based on fluorochrome banding and nuclear DNA amount

Fluorescent chromosome banding and measurements of nuclear DNA content by image cytometry of Feulgen-stained cells were performed in one sample each of eight diploid (2n = 24) species of Solanum: S. endoadenium, S. argentinum, S. pseudocapsicum, S. atropurpureum, S. elaeagnifolium, S. sisymbriifolium, S. chenopodioides, and S. palustre. The species studied could be distinguished by heterochromatin amount, banding patterns, and genome size. They exhibited only GC-rich heterochromatin and showed a comparatively low heterochromatin amount (expressed as percentage of haplotype karyotype length), ranging from 2.10 in S. argentinum to 8.37 in S. chenopodioides.Genome size displayed significant variation between species, with 1C-values ranging from 0.75 pg (735 Mbp) in S. palustre to 1.79 pg (1,754 Mbp) in S. sisymbriifolium. No significant correlation between genome size and heterochromatin amount was observed, butintrachromosomal asymmetry index (A1) was negative and significantly correlated with heterochromatin amount. DNA content was positively and significantly correlated withkaryotype length. DNA C-value distribution in the genus as well as karyotype affinities and relationships between species are discussed in relation to different infrageneric classifications of Solanum.Fil: Acosta, María Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); Argentina. Universidad Nacional de Córdoba; ArgentinaFil: Guerra, Marcelo. Universidad Federal de Pernambuco; BrasilFil: Moscone, Eduardo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); Argentina. Universidad Nacional de Córdoba; Argentin

Heterochromatin type, amount and distribution in wild species of chili peppers (Capsicum-Solanaceae)

Triple staining with the fluorochromes chromomycin A3, distamycin A and 4?-6-diamidino-2-phenylindole (CMA/DA/DAPI) was applied to somatic metaphases and interphase nuclei of 11 taxa of wild chili peppers (Capsicum), with 2n = 2x = 24 (C. annuum var. glabriusculum, C. cardenasii, C. chacoense, C. flexuosum, C. galapagoense, C. eximium, C. praetermissum and C. tovarii) and 2n = 2x = 26 (C. recurvatum, C. rhomboideum and C. villosum) to analyse heterochromatin type, amount and distribution in wild members of this genus. Heterochromatic banding patterns allowed the identification of all the taxa examined and contributed to their taxonomic grouping. GC-rich heterochromatin (CMA+/DAPI-) was typical in all taxa; only C. praetermissum possessed also AT-rich (CMA-/DAPI+) and mixed GC- and AT-rich (CMA+/DAPI+) bands. Heterochromatin amount (expressed as % of karyotype length) ranged between 1.72 (C. chacoense) and 16.82 (C. flexuosum) and was positively correlated with karyotype length in most of the taxa examined. Heterochromatin located mainly at terminal position of chromosomes but intercalary position prevailed in C. flexuosum. Nucleolus organizer regions (NOR)-associated GC-rich heterochromatin was exclusively terminal and included the distal macrosatellite and a small portion on the corresponding arm. In all the taxa analysed, an equilocal heterochromatin distribution in non-homologous chromosomes of karyotype was observed, suggesting concerted evolution of heterochromatin dispersion in Capsicum.Fil: Scaldaferro, Marisel Analía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); ArgentinaFil: Grabiele, Mauro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); ArgentinaFil: Moscone, Eduardo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); Argentin

Karyotype analysis in two species of Solanum (Solanaceae) sect. Cyphomandropsis based on chromosome banding

To contribute to the cytogenetic characterisation of the Cyphomandra clade of Solanum, fluorescence banding and silver staining were performed on Solanum fusiforme and Solanum stuckertii for the first time. Both species had 2n=24 chromosomes. Solanum fusiforme had a karyotype consisting of 9m+3sm chromosome pairs; one pair had active nucleolar organiser regions (NORs). Solanum stuckertii had 8m+4sm chromosome pairs with two pairs having active NORs.The two species exhibit similar amounts (11%) of constitutive CMA+/DAPI- heterochromatin, rich in GC base pairs. This heterochromatin was distributed in a complex pattern of mainly intercalary bands. Localisation of chromosome-specific markers was successful, allowing recognition of all chromosomes of the complements.With the aim of contributing to a better understanding of the interspecific relationships within the Cyphomandra clade, we discuss our results in conjunction with previous findings for species in this clade.Fil: Miguel, Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaFil: Acosta, María Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Moscone, Eduardo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentin

Using chromosomal data in the phylogenetic and molecular dating framework: Karyotype evolution and diversification in Nierembergia (Solanaceae) influenced by historical changes in sea level

Karyotype data within a phylogenetic framework and molecular dating were used to examine chromosome evolution in Nierembergia and to infer how geological or climatic processes have influenced in the diversification of this solanaceous genus native to South America and Mexico. Despite the numerous studies comparing karyotype features across species, including the use of molecular phylogenies, to date relatively few studies have used formal comparative methods to elucidate chromosomal evolution, especially to reconstruct the whole ancestral karyotypes. Here, we mapped on the Nierembergia phylogeny one complete set of chromosomal data obtained by conventional staining, AgNOR-, C- and fluorescent chromosome banding, and fluorescent in situ hybridisation. In addition, we used a Bayesian molecular relaxed clock to estimate divergence times between species. Nierembergia showed two major divergent clades: a mountainous species group with symmetrical karyotypes, large chromosomes, only one nucleolar organising region (NOR) and without centromeric heterochromatin, and a lowland species group with asymmetrical karyotypes, small chromosomes, two chromosomes pairs with NORs and centromeric heterochromatin bands. Molecular dating on the DNA phylogeny revealed that both groups diverged during Late Miocene, when Atlantic marine ingressions, called the 'Paranense Sea', probably forced the ancestors of these species to find refuge in unflooded areas for about 2 Myr. This split agrees with an increased asymmetry and heterochromatin amount, and decrease in karyotype length and chromosome size. Thus, when the two Nierembergia ancestral lineages were isolated, major divergences occurred in chromosomal evolution, and then each lineage underwent speciation separately, with relatively minor changes in chromosomal characteristics.Fil: Acosta, María Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Moscone, Eduardo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Cocucci, Andrea Aristides. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentin

Effects of ionizing radiation on Capsicum bacatum var. pendulum (Solanaceae)

Cytogenetic and somatic effects of various x-ray treatments were evaluated in pepper, Capsicum baccatum var. pendulum cv. “Cayenne”, with the aim to assess optimal conditions for obtaining viable lines. The cytogenetic effects were quantified by counting chromosome aberrations. The level of DNA fragmentation was estimated with TUNEL test (terminal transferase mediated dUTP-fluorescein nick end labeling). Irradiation to 20 Gy with 16-h presoaking can be a suitable treatment of the selected pepper cultivar for a mutagenesis program.Fil: Scaldaferro, Marisel Analía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); ArgentinaFil: Prina, A.. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Centro de Investigación de Ciencias Veterinarias y Agronómicas. Instituto de Genética; ArgentinaFil: Moscone, Eduardo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); ArgentinaFil: Kwasniewska, J.. University of Silesia; Poloni

25S–18S rDNA IGS of Capsicum: molecular structure and comparison

The primary and secondary structures of the intergenic spacer (IGS) between the 3′-end of 25S ribosomal RNA (rRNA) gene and the 5′-end of 18S rRNA gene are described for the cultivated chili pepper Capsicum pubescens. The recognized functional IGS is 2,078 bp in length. According to nucleotide base composition, regulatory elements, and conserved and repeated sequences the IGS can be divided into seven structural regions (SRI–VII). SRI comprises three copies of GAGGTTTTT-like motif, a probable transcription termination site in Solanaceae. At 3′-end, there are 21 bp matching the 18S rDNA. SRII is formed by 47 repeats of CACCATGG-like motif, the shortest repetitive region found in plant rDNA to date. SRIII is highly AT-rich, preceding SRIV, a highly conserved region in Solanaceae containing the transcription initiation site (TIS) TATATAAGGGGGG. The external transcribed spacer (ETS) is 966 bp in length. SRV-VII, downstream of the TIS, possesses eight inverted repeats, and three predicted stem-loops show pre-micro RNA (miRNA)-like structural features. Intragenomic variation is presented, and data are compared with characterized Solanaceae 25S–18S rDNA IGS.Instituto de Patología VegetalFil: Grabiele, Mauro. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); ArgentinaFil: Debat, Humberto Julio. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; ArgentinaFil: Moscone, Eduardo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); ArgentinaFil: Ducasse, Daniel Adrian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentin

Complex rearrangements are involved in Cephalanthera (Orchidaceae) chromosome evolution

The genus Cephalanthera is an excellent plant group for karyotype evolution studies because it exhibits a dysploid series and bimodal karyotypes. With the aim of understanding their chromosomal and phylogenetic relationships, rRNA genes and the Arabidopsis-type telomeric sequence were mapped by fluorescence in-situ hybridization (FISH), and the rDNA intergenic spacer (ITS) was sequenced for the first time in three European species: C. longifolia (2n = 4x = 32), C. damasonium (2n = 4x = 36) and C. rubra (2n = 4x = 44). One 45S and three 5S rDNA sites are observed in C. longifolia, one 45S and two 5S sites in C. damasonium, and two 45S and one 5S site in C. rubra. Telomeric signals were observed at every chromosome end in all three species and C. damasonium also displays interstitial signals on three chromosome pairs. In agreement with chromosome data, molecular analyses support C. longifolia and C. damasonium as closely related taxa, while C. rubra stands apart. Possible pathways of karyotype evolution are discussed in reference to a previous hypothesis. The results indicate that complex chromosomal rearrangements, possibly involving Robertsonian fusions and fissions, loss of telomeric repeats, gain or loss of rDNA sites and other heterochromatic sequences and inversions, may have contributed to generating the present-day karyotypes.Fil: Moscone, Eduardo Alberto. Universidad de Viena; Austria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Samuel, Rosabelle. Universidad de Viena; AustriaFil: Schwarzacher, Trude. University of Leicester; Reino UnidoFil: Schweizer, Dieter. Universidad de Viena; AustriaFil: Pedrosa-Harand, Andrea. Universidad de Viena; Austri