Evaluation of a morphological marker selection and excision system to generate marker-free transgenic cassava plants |
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Authors: | Laddawan Saelim Salak Phansiri Malinee Suksangpanomrung Supatcharee Netrphan Jarunya Narangajavana |
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Institution: | (1) Department of Biotechnology, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok, 10400, Thailand;(2) Scientific Equipment Center, KURDI, Kasetsart University, Bangkok, Thailand;(3) National Center for Genetic Engineering and Biotechnology (BIOTEC), Pathumthani, Thailand;(4) Center for Cassava Molecular Biotechnology (CASS-MOL-BIOTECH), Faculty of Science, Mahidol University, Bangkok, Thailand |
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Abstract: | The efficacy of the ipt-type Multi-Auto-Transformation (MAT) vector system to transform the extensively grown cassava cultivar “KU50” was evaluated.
This system utilizes the isopentenyltransferase (ipt) gene as morphological marker for visual selection of transgenic lines. The extreme shooty phenotype (ESP) of transgenic
lines is lost due to the removal of ipt gene mediated by the yeast Rint/RS system. As a result, phenotypically normal shoots, considered marker-free transgenic plants,
could be obtained. When transforming KU50 cassava cultivar with two different ipt-type MAT vectors, transformation frequency at 19–21% was observed. Among the total number of ESP explants, 32–38% regained
normal extended shoot phenotype and 88–96% of which were confirmed to represent the marker-free transgenic plants. This is
the first demonstration of the efficacy of Rint/RS system in promoting excision of ipt marker gene in cassava specie, with the consequent rapid production of marker-free transgenic plants. The high efficiency
of this system should facilitate pyramiding a number of transgenes by repeated transformation without having to undergo through
laborious, expensive and time-consuming processes of sexual crossing and seed production. The generation of marker-free, thus
environmentally safe, genetically modified cassava clones should also ease the public concerns regarding the use of transgenic
cassava in both food and nonfood industries. |
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Keywords: | Cassava Marker-free transgenic plant Morphological marker Excision system |
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