Assuntos
Genética/história , História do Século XX , História do Século XXI , Federação Russa , UcrâniaRESUMO
The review is dedicated to several aspects of sugar beet (Beta vulgaris L.) biotechnology: in vitro cultivation, callus induction, plant regeneration and genetic transformation. Media composition, methods of plant regeneration via somatic embryogenesis and protoplast culture are analysed. The use of Agrobacterium tumefaciens and gold particle bombardment is the base for modern genetic transformation methods.
Assuntos
Beta vulgaris/genética , Plantas Geneticamente Modificadas/genética , Transformação Genética , Beta vulgaris/crescimento & desenvolvimento , DNA de Plantas , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , ProtoplastosRESUMO
Ex situ germplasm bank of plant species of world flora has been created at the Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine as a result of investigations in the fields of botany and plant biotechnology. At present the bank contains more than 4000 samples in seed bank and more than 2000 cell lines in in vitro bank. The germplasm bank which has been put on the List of scientific objects of national property of Ukraine is the basis of elaboration ofbiotechnological methods of plant biodiversity preservation and of plant material screening for biologically active substances.
Assuntos
Agricultura/tendências , Biodiversidade , Biotecnologia/métodos , Conservação dos Recursos Naturais/métodos , Plantas , Desenvolvimento Vegetal , Plantas/genética , Sementes/genética , Sementes/crescimento & desenvolvimento , Especificidade da EspécieRESUMO
New model system of plastid transformation has been proposed using a wild representative of Solanaceae family--S. sinuata. Earlier obtained cybrid plants N. tabacum (+ S. sinuata) were used for transformation experiments by PEG treatment of protoplasts with aadA gene that confers resistance to spectinomycin. Transformed S. sinuata plastome was transferred from N. tabacum (+ S. sinuata) cybrid to S. sinuata wild type plants by somatic hybridization. Molecular analysis of nuclear and mitochondrial DNA has been performed.
Assuntos
DNA de Plantas/genética , Engenharia Genética , Nicotiana/genética , Plantas Geneticamente Modificadas/genética , Plastídeos/genética , Solanaceae/genética , DNA Mitocondrial/genética , DNA Espaçador Ribossômico/genética , Reação em Cadeia da Polimerase , Recombinação GenéticaRESUMO
The Orychophragmus violaceus chlorophylldefective line of "albino" type has been obtained by spectinomycin treatment. Somatic hybridization between Orychophragmus violaceus and Brassica napus was performed by fusion of green mesophyll protoplasts of rape and callus protoplasts of the O. violaceus "albino" line. Near two hundred of regenerant plants were selected according to the regeneration type and ability to become green, and were determined as hybrids. Chloroplast DNA in selected hybrids was identical to rape chlDNA, which was confirmed by the PCR-RFLP analysis of plastid DNA fragments. Fragments of hybrid mitochondrial DNA analyzed by the PCR-RFLP analysis were identical to fragments of O. violaceus. The nuclear genome of the majority of hybrids was represented by the O. violaceus genome, which was demonstrated by analyses of isoenzymes, DNA telomeric sequences, ribosomal and satellite DNAs, and the RAPD analysis. The cytogenetic analysis of a number of lines has shown variability in the number of chromosomes in the obtained lines.
Assuntos
Brassica napus/genética , Brassicaceae/genética , Cloroplastos/genética , Hibridização Genética , Brassica napus/enzimologia , Brassica napus/ultraestrutura , Brassicaceae/enzimologia , Brassicaceae/ultraestrutura , Cromossomos de Plantas/ultraestrutura , DNA Mitocondrial/genética , Proteínas de Plantas/genética , Protoplastos/metabolismo , Técnica de Amplificação ao Acaso de DNA Polimórfico , Seleção GenéticaRESUMO
The construct containing the structural gene of the type II DNA topoisomerase of Drosophila melanogaster fused to the sequence of the chloroplast transit peptide directed by the 35 S promoter was produced on the basis of the binary vector Bin 19. The construct was introduced into Nicotiana tabacum plants by Agrobacterium-mediated transformation. The integration of the construct into genomic DNA was demonstrated by PCR and Southern blot hybridization. Expression of the chimeric gene at the transcription level was studied by Northern blotting. The protein produce of the expression was identified in chloroplasts by Western blot hybridization with polyclonal anti-type II Drosophila topoisomerase antibodies.
Assuntos
Cloroplastos/genética , DNA Topoisomerases Tipo II/genética , Drosophila melanogaster/genética , Nicotiana/genética , Plantas Tóxicas , Animais , Cloroplastos/enzimologia , DNA/genética , Drosophila melanogaster/enzimologia , Regulação da Expressão Gênica de Plantas/genética , Vetores Genéticos/genética , Técnicas de Amplificação de Ácido Nucleico , Hibridização de Ácido Nucleico/genética , Plantas Geneticamente Modificadas , Plasmídeos/genética , Regiões Promotoras Genéticas/genética , Proteínas Recombinantes de Fusão/genética , Nicotiana/enzimologiaRESUMO
An electron microscope study of higher plant protoplasts and human cell fusion was made using polyethylene glycol. The successive stages of proteoplast cell fusion, and the formation of intraspecific, interfamiliar, or interkingdom "hybrid" cells, and some details of their submicroscopic changes at the early stages of culturing are shown.