The barley chloroplast mutator (cpm) mutant: all roads lead to the Msh1 gene

The barley chloroplast mutator (cpm) is a nuclear gene mutant that induces a wide spectrum of cytoplasmically inherited chlorophyll deficiencies. Plastome instability of cpm seedlings was determined by identification of a particular landscape of polymorphisms that suggests failures in a plastome mismatch repair (MMR) protein. In Arabidopsis, MSH genes encode proteins that are in charge of mismatch repair and have anti-recombination activity. In this work, barley homologs of these genes were identified, and their sequences were analyzed in control and cpm mutant seedlings. A substitution, leading to a premature stop codon and a truncated MSH1 protein, was identified in the Msh1 gene of cpm plants. The relationship between this mutation and the presence of chlorophyll deficiencies was established in progenies from crosses and backcrosses. These results strongly suggest that the mutation identified in the Msh1 gene of the cpm mutant is responsible for the observed plastome instabilities. Interestingly, comparison of mutant phenotypes and molecular changes induced by the barley cpm mutant with those of Arabidopsis MSH1 mutants revealed marked differences.Instituto de GenéticaFil: Lencina, Franco Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; ArgentinaFil: Landau, Alejandra Mabel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; ArgentinaFil: Prina, Alberto Raul. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; 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

In vitro culture response of barley ( Hordeum vulgare ) ethylene synthesis mutant MC 169

Although it is generally accepted that plant in vitro culture response is influenced by the donor genotype, the genetic and molecular bases of this phenomenon are barely known. As a consequence, the optimization of tissue culture protocols is mainly empirically done. Researchers of the IGEAF studied the genetic basis of the in vitro regeneration of various plant species, including the tissue culture response of artificially induced barley mutants. One barley mutant, MC 169, carries a nuclear gene, recently described controlling the root growth in hydroponic cultivation. Under this condition, the roots of MC 169 mutant plants were longer than those of the original wild type line MC 182, a fact that was associated with a reduced ethylene biosynthesis. On the other hand, it is known that ethylene accumulation is inhibitory for in vitro regeneration of several plant species. In this study, we compared the in vitro culture response of mutant MC 169 with that of its mother line MC 182. The data about induction and regeneration of calli as well as those of habituated calli formation demonstrated that mutant MC 169 and its mother line MC 182 show a similar in vitro behaviour

In vitro culture response of barley ( Hordeum vulgare ) ethylene synthesis mutant MC 169

Abstract Although it is generally accepted that plant in vitro culture response is influenced by the donor genotype, the genetic and molecular bases of this phenomenon are barely known. As a consequence, the optimization of tissue culture protocols is mainly empirically done. Researchers of the IGEAF studied the genetic basis of the in vitro regeneration of various plant species, including the tissue culture response of artificially induced barley mutants. One barley mutant, MC 169, carries a nuclear gene, recently described controlling the root growth in hydroponic cultivation. Under this condition, the roots of MC 169 mutant plants were longer than those of the original wild type line MC 182, a fact that was associated with a reduced ethylene biosynthesis. On the other hand, it is known that ethylene accumulation is inhibitory for in vitro regeneration of several plant species. In this study, we compared the in vitro culture response of mutant MC 169 with that of its mother line MC 182. The data about induction and regeneration of calli as well as those of habituated calli formation demonstrated that mutant MC 169 and its mother line MC 182 show a similar in vitro behaviour

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

Data from: Plastome mutations and recombination events in barley chloroplast mutator seedlings

The barley chloroplast mutator (cpm) is an allele of a nuclear gene that when homozygous induces several types of cytoplasmically inherited chlorophyll deficiencies. In this work, a plastome TILLING (Targeting Induced Local Lesions in Genomes) strategy based on mismatch digestion was used on families that carried the cpm genotype through many generations. Extensive scanning of 33 plastome genes and a few intergenic regions was conducted. Numerous polymorphisms were detected on both genic and intergenic regions. The detected polymorphisms can be accounted for by at least 61 independent mutational events. The vast majority of the polymorphisms originated in substitutions and small indels (insertions/deletions) in microsatellites. The rpl23 and the rps16 genes were the most polymorphic. Interestingly, the variation observed in the rpl23 gene consisted of several combinations of five different one nucleotide polymorphisms. Besides, four large indels that have direct repeats at both ends were also observed, which appear to be originated from recombinational events. The cpm mutation spectrum suggests that the CPM gene product is probably involved in plastome mismatch repair. The numerous subtle molecular changes that were localized in a wide range of plastome sites show the cpm as a valuable source of plastome variability for plant research and/or plant breeding. Moreover, the cpm mutant appears to be as an interesting experimental material for investigating the mechanisms responsible for maintaining the stability of plant organelles DNA

In vitro culture response of barley ( Hordeum vulgare ) ethylene synthesis mutant MC 169

Abstract Although it is generally accepted that plant in vitro culture response is influenced by the donor genotype, the genetic and molecular bases of this phenomenon are barely known. As a consequence, the optimization of tissue culture protocols is mainly empirically done. Researchers of the IGEAF studied the genetic basis of the in vitro regeneration of various plant species, including the tissue culture response of artificially induced barley mutants. One barley mutant, MC 169, carries a nuclear gene, recently described controlling the root growth in hydroponic cultivation. Under this condition, the roots of MC 169 mutant plants were longer than those of the original wild type line MC 182, a fact that was associated with a reduced ethylene biosynthesis. On the other hand, it is known that ethylene accumulation is inhibitory for in vitro regeneration of several plant species. In this study, we compared the in vitro culture response of mutant MC 169 with that of its mother line MC 182. The data about induction and regeneration of calli as well as those of habituated calli formation demonstrated that mutant MC 169 and its mother line MC 182 show a similar in vitro behaviour

Plastome mutations and recombination events in barley chloroplast mutator seedlings

The barley chloroplast mutator (cpm) is an allele of a nuclear gene that when homozygous induces several types of cytoplasmically inherited chlorophyll deficiencies. In this work, a plastome Targeting Induced Local Lesions in Genomes (TILLING) strategy based on mismatch digestion was used on families that carried the cpm genotype through many generations. Extensive scanning of 33 plastome genes and a few intergenic regions was conducted. Numerous polymorphisms were detected on both genic and intergenic regions. The detected polymorphisms can be accounted for by at least 61 independent mutational events. The vast majority of the polymorphisms originated in substitutions and small indels (insertions/deletions) in microsatellites. The rpl23 and the rps16 genes were the most polymorphic. Interestingly, the variation observed in the rpl23 gene consisted of several combinations of 5 different one nucleotide polymorphisms. Besides, 4 large indels that have direct repeats at both ends were also observed, which appear to be originated from recombinational events. The cpm mutation spectrum suggests that the CPM gene product is probably involved in plastome mismatch repair. The numerous subtle molecular changes that were localized in a wide range of plastome sites show the cpm as a valuable source of plastome variability for plant research and/or plant breeding. Moreover, the cpm mutant appears to be an interesting experimental material for investigating the mechanisms responsible for maintaining the stability of plant organelle DNA.Inst. de Genética "Ewald A. Favret"- IGEAFFil: Landau, Alejandra Mabel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret"; ArgentinaFil: Lencina, Franco. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret"; ArgentinaFil: Pacheco, Maria Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret"; ArgentinaFil: Prina, Alberto Raul. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret"; Argentin