Methods for improving competency of plant cells
The present invention provides methods for improving competency of plant cells for bacterial-mediated transformation comprising contacting the plant cells with an effective amount of polyethylene glycol (PEG) for a period of time prior to transformation. The ability to store and maintain competent plant cells for transformation and tissue culture allows more efficient planning and execution of large-scale experiments by providing flexibility of peak production hours, or during unplanned disruptions in the production process. These methods are useful in preserving the viability of plant cells in various storage conditions, thus improving their competency for transformation and tissue culture.
1. A method for improving competency of plant embryo cells for bacterial-mediated transformation comprising:
(a) contacting a mature plant embryo explant with an effective amount of a polyethylene glycol (PEG) containing composition having from about 10% to about 50% by volume of polyethylene glycol; and
(b) transforming at least one cell of the mature plant embryo explant with a heterologous DNA sequence via bacterial-mediated transformation,
wherein the transforming step (b) is performed after the contacting the step (a); wherein the PEG containing composition is removed prior to the transforming step (b); wherein the mature plant embryo explant is excised from a dry seed; and wherein the improved competency is measured as an increase in transformation frequency.
2. The method of claim 1 , wherein the at least one transformed cell of the mature plant embryo explant is a meristematic cell.
3. The method of claim 1 , wherein the mature plant embryo explant is from a soybean or cotton seed.
4. The method of claim 1 , wherein the PEG containing composition is removed by rinsing the mature plant embryo explant with a non-PEG containing composition prior to the transforming step (b).
5. The method of claim 1 , wherein the PEG containing composition in the contacting step (a) has from about 10% to about 25% by volume of polyethylene glycol.
6. The method of claim 1 , wherein the bacterial-mediated transformation is Agrobacterium -mediated, Rhizobium -mediated, Sinorhizobium -mediated, Mesorhizobium -mediated, or Bradyrhizobium -mediated transformation.
7. The method of claim 6 , wherein the bacterial-mediated transformation is Agrobacterium -mediated transformation.
8. The method of claim 1 , wherein the transforming step (b) is performed without any prior callus induction of the mature plant embryo explant.