Methods for maize transformation coupled with adventitious regeneration utilizing nodal section explants and mature zygotic embryos


Ontology type: sgo:Patent     


Patent Info

DATE

2003-05-27T00:00

AUTHORS

Margaret M. Young , Nancy A. Reichert

ABSTRACT

Maize tissue may be regenerated from nodal extracts prepared from germinated mature seeds and germinated embryos grown in a medium containing plant growth regulators. Nodal section explants are secured from seedlings approximately 3-10 days old, preferably from 3-7 days old. The explants are grown on an induction medium until adventitious shoot formation is observed. The shoots are separated and elongated on an MS-based medium, and then rooted. Fast genotype-independent regeneration is obtained, in 12-14 weeks. These explants, as well as zygotic embryos, may be transformed with exogenous DNA using a biolistic approach, where DNA precipitated onto tungsten microprojectiles is accelerated at a minimum of 650 psi towards the explants at a distance of at least 7.5 cm. Improved frequency of transformation is obtained using microprojectiles which prior to DNA precipitation were frozen in glycerol, and suspending from a preparation of 2.5 M CaCl2. The combination of transformation process and regeneration can be used, independent of genotype, to provide new commercial crop organisms. More... »

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