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, comprising preparing the liquid medium in a media preparation system comprising:
(a) an inlet for water;
(b) an inlet for the osmotic agent; and
(c) a chamber for mixing the water and osmotic agent to produce the liquid medium, wherein the inlet for water and the inlet for osmotic agent are coupled to the chamber for mixing to allow delivery of the water and osmotic agent to the chamber for mixing.
2. The method of claim 1 , wherein the one or more of the inlet(s) receive water and/or the osmotic agent from a chamber for holding water or a chamber for holding the osmotic agent.
3. The method of claim 2 , further comprising measuring the fill level of one or more of the chamber for holding water, the chamber for holding the osmotic agent, or the chamber for mixing the water and the osmotic agent.
4. The method of claim 1 , further comprising coupling the chamber for mixing to a fluid jet apparatus.
5. The method of claim 1 , further comprising sterilizing the liquid medium with a sterilizer selected from the group consisting of a filter, a UV or gamma radiation source, and a sterilizing heat source.
6. The method of claim 5 , wherein the sterilizer sterilizes the water and/or osmotic agent prior to entering the mixing chamber.
7. The method of claim 5 , wherein the sterilizer sterilizes the water, osmotic agent, and/or liquid medium concurrently with and/or after entering the mixing chamber.
8. The method of claim 1 , wherein the liquid medium is sterilized after leaving the mixing chamber.
9. The method of claim 1 , further comprising controlling delivery of the water and the osmotic agent to the chamber for mixing the water and the osmotic agent.
10. The method of claim 9 , wherein controlling the delivery comprises electronically sensing the delivery of the water and/or the osmotic agent.