Obtenção de plantas transgênicas de soja com a forma constitutiva do fator de transcrição AREB1
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Data
2012-05-04
Autores
Leite, Juliana Paula [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
No cenário atual de mudanças climáticas, com o aumento da população mundial e crescente demanda por alimentos, as ferramentas biotecnológicas veem sendo utilizadas na obtenção de plantas mais tolerantes a estresse ambientais como a seca, que provoca perdas financeiras e de produção significativas aos produtores. Os fatores de transcrição, são genes potenciais na estratégia de reduzir os prejuízos decorrentes de períodos de déficit hídrico, pois atuam controlando a expressão de genes estresse-induzidos e têm sido estudados na planta modelo Arabidopsis thaliana e em culturas de interesse agronômico. O fator de transcrição AtAREB1ΔQT, consiste na forma constitutivamente ativa de AREB1, (ABA Responsive Element Binding – elemento de ligação de resposta ao ABA) que está envolvido, na via de sinalização ABA (ácido abcísico) dependente de resposta ao estresse hídrico em plantas. O objetivo do presente trabalho foi introduzir em soja, via biobalística, a construção gênica pBI35SΩ:AtAREB1ΔQT e caracterizar molecularmente os eventos quanto ao número de cópias inseridas, e análise da expressão gênica do transgene. Para a caracterização molecular, foram utilizadas as metodologias de Southern blot e RT-qPCR. Um total de 12 linhagens independentes foram obtidas na geração T0, com uma eficiência de transformação de 0,59%. Somente três eventos (2651, 2639 e 2654) segregaram e passaram o gene para a geração T1. O número de cópias do transgene quantificado via qPCR, foi diferente, para cada linhagem geneticamente modificada (GM) analisada, variando de poucas cópias (1 a 2) a várias (17 cópias). Estes dados foram corroborados pelos resultados obtidos via Southern blot. O nível de expressão gênica relativa do transgene foi variável entre os eventos e a análise da segregação em plantas...
In the actual scenario of climatic changes, with world population increase and growing food demand, biotechnological tools are being used to develop plants more tolerant to environmental stresses such as drought, which causes to producers, significant financial losses and yield production. The transcription factors are potential genes to be strategically used to reduce damage due to water deficit conditions, because they acts controlling the expression of numerous stress-induced genes and has been studied in the model plant Arabidopsis thaliana as well as in agronomic crops. The transcription factor, AtAREB1ΔQT, consists in the constitutively active form of AREB1, (ABA Responsive Element Binding) that is involved, ABA signaling pathway (abscisic acid) dependent response pathway to drought in plants. The objective of this study was to insert the construction pBI35SΩ: AtAREB1ΔQT in soybean, via biolistics, and molecularly characterization of the events on the number of inserted copies, and analyze the relative expression level of transgene. Twelve independent lines were identified in T0 generation, with a transformation efficiency of 0.59%. Only three events (2651, 2639 and 2654) segregated and transmitted the gene for T1 generation. The number of copies quantified using qPCR was different for each modified genetically (GM) line, ranging from a few copies (1 to 2 copies) to many copies (17 copies). These data were corroborated by Southern blot results. The relative expression level of transgene was variable between events and segregation analysis in T2 generation showed that he transgene did not follow the Mendelian laws. Aiming to obtain more information on the effect of the transgene in response to water stress... (Complete abstract click electronic access below)
In the actual scenario of climatic changes, with world population increase and growing food demand, biotechnological tools are being used to develop plants more tolerant to environmental stresses such as drought, which causes to producers, significant financial losses and yield production. The transcription factors are potential genes to be strategically used to reduce damage due to water deficit conditions, because they acts controlling the expression of numerous stress-induced genes and has been studied in the model plant Arabidopsis thaliana as well as in agronomic crops. The transcription factor, AtAREB1ΔQT, consists in the constitutively active form of AREB1, (ABA Responsive Element Binding) that is involved, ABA signaling pathway (abscisic acid) dependent response pathway to drought in plants. The objective of this study was to insert the construction pBI35SΩ: AtAREB1ΔQT in soybean, via biolistics, and molecularly characterization of the events on the number of inserted copies, and analyze the relative expression level of transgene. Twelve independent lines were identified in T0 generation, with a transformation efficiency of 0.59%. Only three events (2651, 2639 and 2654) segregated and transmitted the gene for T1 generation. The number of copies quantified using qPCR was different for each modified genetically (GM) line, ranging from a few copies (1 to 2 copies) to many copies (17 copies). These data were corroborated by Southern blot results. The relative expression level of transgene was variable between events and segregation analysis in T2 generation showed that he transgene did not follow the Mendelian laws. Aiming to obtain more information on the effect of the transgene in response to water stress... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Soja - Transgene, Alimentos geneticamente modificados, Plantas - Efeito da seca, Desidratação (Hídrica), Genetica vegetal, Water stress, Soybean - Transgene, Southern blot
Como citar
LEITE, Juliana Paula. Obtenção de plantas transgênicas de soja com a forma constitutiva do fator de transcrição AREB1. 2012. xiv, 74 f. Dissertação (mestrado) - Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, 2012.