Genetic transformation in apple, with gene bvlI

O objetivo principal do presente trabalho foi introduzir o gene bvlI com potencial antif??ng??co confirmado da lectina de Bauhinia variegata, em macieira (Malus domestica Borkh), cv. Gala, para estabelecer resist??ncia ao fungo Coletotrichum gloeosporioides, causador da Mancha Foliar. Foram realizados experimentos de multiplica????o in vitro, regenera????o de explantes foliares e, dois protocolos de transforma????o via Agrobacterium tumefaciens visando a inser????o do gene bvll. Na multiplica????o de brota????es o meio que apresentou melhores resultados foi o MS suplementado com 0,1 mg L-1 ANA e 8,3 mg L-1 de 2iP e, para a regenera????o as melhores respostas foram obtidas com meio MS, suplementado com 30 g L-1 de sorbitol, 0,5 mg L-1 ANA e 5 mg L1 de TDZ, atingindo um percentual de 94,5% de regenera????es. Quanto aos protocolos de transforma????o, o que apresentou melhores resultados foi o protocolo P1, no qual atrav??s da an??lise de PCR foram obtidas 15% de brota????es contendo o gene bv/l. No entanto, a express??o da prote??na BVLI n??o foi detectada atrav??s de Dot Blot, indicando a necessidade de novos procedimentos, como refinamento do protocolo de extra????o de prote??nas, bem como utuliza????o de outras t??cnicas de an??lises, como Real Time.he main objective of this work was to introduce the gene bvlI with confirmed potential antifungal of lectin from Bauhinia variegata, Apple Tree (Malus domestica Borkh) cv. Gala, to establish resistance to fungus Coletotrichum gloeosporioides, causing Leaf Spot. Multiplication experiments were performed in vitro regeneration of leaf explant culture and two protocols via Agrobacterium tumefaciens transformation aimed at inserting the gene bvll. The multiplication of shoots the medium that has best results was MS supplemented with 0.1 mg L-1 ANA and 8.3 mg L-1 2iP and, for the regeneration of the best responses were obtained with using MS, supplemented with 30 g L-1 sorbitol, 0.5 mg L-1 ANA and 5 mg L-1 TDZ, reaching a rate of 94.5% of regenerations. As regards the processing protocols, which showed best results was the P1 Protocol, which through analysis of PCR were obtained 15% shoots containing the gene bv/l. However, the expression of BVLI protein was not detected by Dot Blot, indicating the need for further procedure, such as refinement of protein extraction protocol, as well as utilization other techniques of analyses, such as Real Time