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Efficient transformation method for sunflower and oil seeds based on positive selection

机译:基于正选择的向日葵和油料种子高效转化方法

摘要

The invention describes a highly improved, reproducible and a consistent method of transformation and regeneration that results in obtaining 12-15% transgenic plants. The present invention relates to a method of selecting genetically transformed Sunflower explants based on their ability to utilize Xylose as a sole carbohydrate source. Further disclosed is the nucleic acid sequence of the Xylose Isomerase gene, vector construction for incorporation of the selection marker gene and the process of Agrobacterium Mediated Transformation of target host plant with the vector comprising the gene encoding the enzyme Xylose isomerase under the functional combination of the heterologous regulatory sequences. Also disclosed is the method of selecting the putative transformants post transformation with the said vector that possess a metabolic advantage of utilizing Xylose as a sole carbon source. Increased efficiency of regeneration, better growth and survival has been observed on subjection to the described method of positive selection. The subject invention alleviates the disadvantages of negative selection methods such as the undesired elimination of the transformed cells and the potential environmental harm caused due to the dispersal of the antibiotic and the herbicide resistant genes.
机译:本发明描述了高度改良的,可再现的和一致的转化和再生方法,其导致获得12-15%的转基因植物。本发明涉及一种基于遗传转化的向日葵外植体利用木糖作为唯一碳水化合物源的能力的选择方法。进一步公开了木糖异构酶基因的核酸序列,掺入选择标记基因的载体构建以及农杆菌介导的用包含编码木糖酶基因的载体介导的靶宿主植物转化过程。异源调节序列功能组合下的异构酶。还公开了选择用所述载体转化后的推定转化体的方法,所述载体具有利用木糖作为唯一碳源的代谢优势。遵循所述的阳性选择方法,观察到再生效率提高,生长和存活更好。本发明减轻了负选择方法的缺点,例如不希望的转化的细胞的消除和由于抗生素和除草剂抗性基因的分散引起的潜在的环境危害。

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