摘要:
The present invention provides methods and compositions which deliver Agrobacterium via microinjection directly into the embryo sac. At the time of injection, the embryo sac can comprise an egg cell, or alternatively, the embryo sac can be fertilized and comprise either a zygote or an embryo. Once inside the embryo sac, the Agrobacterium harboring a T-DNA having a polynucleotide of interest can express of the polynucleotide of interest in the plant. Further, the Agrobacterium can transfer the T-DNA having the polynucleotide of interest to the plant nucleus to produce a transformed plant. The polynucleotide of interest may be stably integrated into the genome of the egg cell, zygote, embryo, or endosperm, and any tissue, plant part, and/or plant generated therefrom.
摘要:
The invention provides improved plant transformation methods. In particular the method provides increased transformation frequency, especially in recalcitrant plants. The method includes various transformation protocols for monocots, such as maize and sorghum, using a combination of media and light conditions to achieve increased efficiency of monocot transformation and increased callus initiation frequencies.
摘要:
A method for increasing efficiency of germplasm screening for transformability may include providing a plurality of lines of plant target tissue to be transformed, characterizing each of the lines to provide characterization data, the characterization data comprises DNA or nucleic acid delivery technique response data and tissue culture response data, eliminating one or more of the plurality of lines based on the characterization data without performing transformation of the plurality of lines, such that a subset of the plurality of lines remains, and performing transformation experiments on the subset of the plurality of lines. The method may also include selecting a DNA or nucleic acid delivery technique protocol and a tissue culture protocol prior to the characterization.
摘要:
A method for regulating expression of a virulence gene of Agrobacterium is described. The method comprises the steps of stimulating cereal cells, such as sorghum, so as to produce an active, typically phenolic, compound and exposing the Agrobacterium to this compound. The compound induces expression of the virulence gene of the Agrobacterium, effecting T-DNA transfer from the Agrobacterium to the cereal cells, which are thereby transformed.