Method and apparatus for substantially isolating plant tissues

Abstract

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 - 56 . (canceled) 
     
     
         57 . A method for preparation of plant embryos suitable for tissue culture and/or genetic transformation, wherein an apparatus for holding a kernel or other tissue is centrifuged in a container to apply force to the kernel or other tissue. 
     
     
         58 . An apparatus for obtaining plant embryos suitable for tissue culture and/or genetic transformation comprising
 (a) at least a first fluid jet for directing a medium onto a corn kernel or other tissue comprising a plant embryo; and   (b) an apparatus for holding the kernel or other tissue in the path of the medium.   
     
     
         59 . The apparatus of  claim 58 , wherein the embryo is comprised within a corn kernel. 
     
     
         60 . The apparatus of  claim 59 , wherein the corn kernel is comprised on an ear of corn. 
     
     
         61 . The apparatus of  claim 58 , wherein the apparatus is further defined as comprising a first and a second fluid jet. 
     
     
         62 . The apparatus of  claim 60 , wherein the apparatus for holding the kernel comprises means for moving the ear of corn relative to the first and second fluid streams to control the angle of contact between the first and second fluid streams and the kernel. 
     
     
         63 . The apparatus of  claim 58 , further comprising a detector to identify excised endosperm tissue and embryos. 
     
     
         64 . The apparatus of  claim 58 , further comprising at least a first separator to isolate embryos from non-embryo tissue. 
     
     
         65 . The apparatus of  claim 58 , wherein the apparatus for holding the kernel or other tissue comprises a sheet or a cylindrical sheet?. 
     
     
         66 . The apparatus of  claim 58 , wherein the apparatus for holding the kernel or other tissue comprises a mesh, a plurality of slots; or a pressure cam or screw that applies force to the tissue. 
     
     
         67 . The apparatus of  claim 58 , wherein seed or fruit tissue is held onto the apparatus for holding the kernel or other tissue by a mechanical force, friction, centrifugal force, or a suction force. 
     
     
         68 . The apparatus of  claim 58 , wherein the apparatus for holding the kernel or other tissue is suspended in a gaseous phase, a liquid phase, or is partially suspended in gaseous and liquid phases. 
     
     
         69 . The apparatus of  claim 58 , wherein the apparatus for holding the kernel or other tissue is fixed relative to a fluid force. 
     
     
         70 . The apparatus of  claim 58 , wherein the apparatus for holding the kernel or other tissue is movable relative to a fluid force. 
     
     
         71 . The apparatus of  claim 64 , wherein the separator separates embryos suitable for tissue culture from non-embryo tissue by a method selected from the group consisting of size exclusion, differential density and differential hydrophobicity. 
     
     
         72 . The apparatus of  claim 71 , further defined as comprising a sieve for separating embryo from non-embryo tissue based on size. 
     
     
         73 . The apparatus of  claim 60 , wherein the apparatus for holding the kernel comprises at least a first motor for moving the ear of corn relative to the fluid jet. 
     
     
         74 . The method of  claim 57 , wherein the kernel is comprised on an ear of corn. 
     
     
         75 . The method of  claim 74 , wherein the ear of corn is rotated about its longitudinal axis. 
     
     
         76 . The method of  claim 57 , wherein the kernel or other tissue is held onto the apparatus for holding the kernel or other tissue by a mechanical force, a friction force, a centrifugal force, or a suction force. 
     
     
         77 . The method of  claim 57 , wherein the apparatus for holding a kernel or other tissue comprises a sheet or a cylinder. 
     
     
         78 . The method of  claim 77 , wherein the apparatus for holding a kernel or other tissue comprises plastic or metal. 
     
     
         79 . The method of  claim 77 , wherein the apparatus for holding a kernel or other tissue comprises holes or slots. 
     
     
         80 . The method of  claim 57 , wherein the plant embryo is a maize embryo, a millet embryo, a soybean embryo, a canola embryo, or a cotton embryo. 
     
     
         81 . The method of  claim 57 , wherein the container comprises multiple plant embryos. 
     
     
         82 . The method of  claim 57 , wherein the force is centrifugal force. 
     
     
         83 . The method of  claim 57 , wherein the plant embryo is separated from other tissue by differential density. 
     
     
         84 . The method of  claim 57  wherein the top of the kernel is removed prior to applying the force. 
     
     
         85 . The method of  claim 57  wherein the plant embryo is ejected from the kernel.