Improvement of <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation in Hi-II maize (<Emphasis Type="Italic">Zea mays</Emphasis>) using standard binary vectors |
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Authors: | Juan M Vega Weichang Yu Angela R Kennon Xinlu Chen Zhanyuan J Zhang |
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Institution: | (1) Departamento de Genetica, Facultad de Biologia, Universidad Complutense, 28040 Madrid, Spain;(2) Division of Biological Science, College of Art and Science, University of Missouri, Columbia, MO 65211-7400, USA;(3) Plant Transformation Core Facility, Division of Plant Sciences, College of Agriculture, Food and Natural Resources, University of Missouri, Columbia, MO 65211-7140, USA |
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Abstract: | High-frequency transformation of maize (Zea mays L.) using standard binary vectors is advantageous for functional genomics and other genetic engineering studies. Recent advances
in Agrobacterium tumefaciens-mediated transformation of maize have made it possible for the public to transform maize using standard binary vectors without
a need of the superbinary vector. While maize Hi-II has been a preferred maize genotype to use in various maize transformation
efforts, there is still potential and need in further improving its transformation frequency. Here we report the enhanced
Agrobacterium-mediated transformation of immature zygotic embryos of maize Hi-II using standard binary vectors. This improved transformation
process employs low-salt media in combined use with antioxidant l-cysteine alone or l-cysteine and dithiothreitol (DTT) during the Agrobacterium infection stage. Three levels of N6 medium salts, 10, 50, and 100%, were tested. Both 10 and 50% salts were found to enhance
the T-DNA transfer in Hi-II. Addition of DTT to the cocultivation medium also improves the T-DNA transformation. About 12%
overall and the highest average of 18% transformation frequencies were achieved from a large number of experiments using immature
embryos grown in various seasons. The enhanced transformation protocol established here will be advantageous for maize genetic
engineering studies including transformation-based functional genomics. |
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Keywords: | Agrobacterium tumefaciens Binary vector Hi-II maize Zea mays Transformation |
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