RESUMO
Phosphinothricin (PPT) resistant hybrid plants between Brassica napus L. cv. Kalinovsky and Orychophragmus violaceus (L.) O.E. Shulz. were obtained as a result of somatic hybridization experiments. The hybrids inherited PPT resistance from O. violaceus plants which were previously transformed by the vector containing Spm/dSpm Zea mays transposon system with bar gene located within the nonautonomous transposon. The obtained plants had intermediate morphology. Their hybrid nature has been confirmed by isozyme (esterase and amilase activity) and PCR (bar, gus, Spm/dSpm integration) analyses. The hybrids combined B. napus plastom and O. violaceus mithochondrion that was revealed by PCR-RFLP. The hybrid plants might be included to rapeseed breeding programme after examination of their oil quality as well as to chloroplast transformation experiments that is still urgent for B. napus.
Assuntos
Aminobutiratos/farmacologia , Brassica napus/genética , Brassicaceae/genética , Herbicidas/farmacologia , Hibridização Genética , Plantas Geneticamente Modificadas , Brassica napus/efeitos dos fármacos , Brassica napus/crescimento & desenvolvimento , Brassicaceae/efeitos dos fármacos , Brassicaceae/crescimento & desenvolvimento , Cloroplastos , Elementos de DNA Transponíveis/genética , DNA de Plantas/genética , Resistência a Herbicidas/genética , Zea mays/genéticaRESUMO
Plastid genetic transformation has been performed using both the PEG-treatment of protoplasts of somatic hybrids of B. oleracea carrying A. thaliana chloroplasts and the particle bombardment of regenerable calluses of B. napus sv. Westar. The chloroplast transformation vector pCB040 carried resistance (aadA) gene flanked by rapeseed plastid DNA sequences to target its insertion between the trnV-rps7 fragments. Selection of transplastomic cell lines has been performed according to their ability to grow on the medium supplied with spectinomycin and streptomycin in high concentrations. Antibiotic resistant cell lines have been obtained using the both transformation methods. The presence of the aadA gene in the A. thaliana and B. napus plastomes was confirmed by PCR analysis for two cell lines of B. oleracea (+ A. thaliana) and three lines of B. napus.
Assuntos
Arabidopsis/genética , Brassica/genética , Plantas Geneticamente Modificadas , Espectinomicina/farmacologia , Estreptomicina/farmacologia , Transformação Genética , Arabidopsis/citologia , Arabidopsis/efeitos dos fármacos , Brassica/citologia , Brassica/efeitos dos fármacos , Brassica napus/citologia , Brassica napus/efeitos dos fármacos , Brassica napus/genética , Cloroplastos/efeitos dos fármacos , DNA de Plantas/genética , Genes de Plantas , Engenharia Genética , Plasmídeos , Reação em Cadeia da Polimerase , Protoplastos/efeitos dos fármacos , Seleção GenéticaRESUMO
Functionally asymmetric somatic hybrids possessing heterologous transposable element Spm/dSpm were obtained following intertribal somatic hybridization between Brassica napus and transgenic Arabidopsis thaliana. Mobile genetic elements actively transposed in the hybrid genomes. Complete elimination of A. thaliana genome was not observed.
Assuntos
Arabidopsis/genética , Brassica napus/genética , Elementos de DNA Transponíveis/genética , DNA de Plantas/genética , Hibridização Genética/genética , Zea mays/genética , Genes de Plantas , Plantas Geneticamente Modificadas , Transcrição GênicaRESUMO
Intertribal somatic hybridization between wild type Brassica juncea (L.) Czern. & Coss. and transgenic A. thaliana L. has been carried out. Genome of A. thaliana plants contained heterologous transposable element Spm/dSpm, reporter GUS gene, selective genes for kanamycin- (npt II) and phosphinothricin (bar) resistance. Hybrid nature of obtained plants was confirmed with their morphology, GUS hystochemical assay, PCR-RFLP, RAPD and isozyme analyses. It was determined that heterologous transposable element Spm/dSpm is able to function in hybrid plants. There was no complete elimination of A. thaliana genetic material in the hybrids and the transgenes were stably maintained.
Assuntos
Arabidopsis/genética , Elementos de DNA Transponíveis/genética , DNA de Plantas/genética , Hibridização Genética/genética , Mostardeira/genética , Plantas Geneticamente Modificadas , Amilases/metabolismo , Arabidopsis/enzimologia , Arabidopsis/crescimento & desenvolvimento , Primers do DNA , Eletroforese em Gel de Ágar , Esterases/metabolismo , Genes de Plantas/genética , Genes Reporter/genética , Vetores Genéticos , Mostardeira/enzimologia , Mostardeira/crescimento & desenvolvimento , Brotos de Planta/enzimologia , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Protoplastos/enzimologia , Seleção GenéticaRESUMO
A method of Agrobacterium-mediated genetic transformation of sugarbeet (Beta vulgaris L.) by vacuum infiltration has been developed. Transgenic sugarbeet plants of Ukraine breeding were selected for their resistance either to the antibiotic kanamycin or to the herbicide glufosinate ammonium. Integration of transgenes was confirmed by PCR and GUS-assay.
Assuntos
Agrobacterium tumefaciens/genética , Beta vulgaris/genética , Plantas Geneticamente Modificadas/genética , Transformação Genética , Beta vulgaris/crescimento & desenvolvimento , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plasmídeos/genéticaRESUMO
A phylogenetic cladogram for 23 species of genera Solanum and Lycopersicon was constructed based on restriction analysis of 10 amplified regions of chloroplast DNA. Three major clusters were detected. All Lycopersicon species and S. lycopersicoides formed the first group. Second cluster was occupied by S. verrucosum, polyploid species of series Longipedicellata, and all dyploid and polyploid species native to South America. Two diploid series Pinnatisecta and Bulbocastana were grouped together in the last claster. Previous morphological and molecular markers strongly support this type of phylogenetic analysis. Based on presented results it is evident that combination of chloroplast DNA region amplification and restriction could be a valuable alternative to a standard-RFLP analysis.
Assuntos
DNA de Cloroplastos/genética , Variação Genética/genética , Solanum lycopersicum/genética , Solanum/genética , Análise por Conglomerados , Mutação , Filogenia , Polimorfismo de Fragmento de RestriçãoRESUMO
Restriction analysis of amplificated chloroplast DNA fragments has allowed us to determine the source of cytoplasm for several natural diploids and amphidiploids of the genus Nicotiana. N. corymbosa or species of Trygonophyllae section donated cytoplasm to N. bigelovii and N. clevelandii, while N. arentsii inherited cytoplasm from N. undulata during the evolution of the genus. Plastomes of Repandae and Nudicaules species orginated from now extinct species of Trygonophyllae section.
Assuntos
DNA de Cloroplastos/genética , Variação Genética , Nicotiana/genética , Sequência de Bases , Dados de Sequência Molecular , Polimorfismo de Fragmento de Restrição , Técnica de Amplificação ao Acaso de DNA PolimórficoRESUMO
Chloroplast DNA of 18 Solanaceae species classified into 9 different tribes was analyzed using combined amplified fragment length polymorphism (AFLP) analysis. UPGMA phylogenetic tree was constructed based on the observed data set. In general, the conclusions of this study confirmed the morphology-based classification of these species. Cluster analysis has placed Mandragora into a monophiletic branch and grouped Schizantus and Capsicum into the independent clusters.
Assuntos
DNA de Cloroplastos/genética , Filogenia , Polimorfismo de Fragmento de Restrição , Solanaceae/genética , Sequência de Bases , Dados de Sequência Molecular , Solanaceae/classificação , Especificidade da EspécieRESUMO
Phylogenetic relationships of 46 Nicotiana species were estimated from the sequences of internal transcribed spacer region of nuclear ribosomal DNA. Phylogenetic tree, built by parsimony method, was highly concordant with the conclusions of morphological and cytogenetical investigations. The reliability of dendrogram obtained was assessed by both bootstrap and jackknife analyses.
