Method and apparatus for substantially isolating plant tissues
The present invention provides methods and devices for the rapid isolation of monocot plant embryos suitable for transformation or tissue culture. The invention includes mechanical devices for substantially isolating plant embryos for use as transformable explants. Media suitable for isolating plant embryos and methods for their preparation are also provided.
1. A method for obtaining embryos suitable for tissue culture and/or genetic transformation, comprising at least partially excising an embryo from a plant seed in a liquid medium consisting essentially of water and/or an osmotic agent with an osmolality of about 0 mOsm/kg to about 600 mOsm/kg, wherein the embryo remains viable for tissue culture and/or genetic transformation following excising the embryo from the plant seed; further comprising the step of genetically transforming the embryo, wherein the step of transforming comprises use of a co-culture medium comprising a bactericide, and wherein the co-culture medium comprises about 0.5-1.5 mg/L of 2,4-D.
2. The method of claim 1 , wherein the osmotic agent is selected from the group consisting of mannitol, sorbitol, glucose, and sucrose.
3. The method of claim 2 , wherein the medium consists essentially of water and mannitol in a concentration of from about 0.05 M to about 0.5 M.
4. The method of claim 2 , wherein the medium consists essentially of water and sucrose in a concentration of from about 0.05 M to about 0.5 M.
5. The method of claim 1 , wherein the bactericide is carbenicillin.
6. The method of claim 1 , further comprising regenerating a transgenic plant from the transformed embryo.
7. The method of claim 1 , further comprising at least partially excising a plurality of embryos from a population of plant seeds or ears in the liquid medium.
8. The method of claim 1 , wherein the embryo is a maize embryo, a millet embryo, a soybean embryo, or a cotton embryo.