Method and apparatus for substantially isolating plant tissues

Patent metadata

Field	Value
Publication number	US-11212972-B2
Application number	US-201916553911-A
Country	US
Kind code	B2
Filing date	Aug 28, 2019
Priority date	Aug 5, 2003
Publication date	Jan 4, 2022
Grant date	Jan 4, 2022

Abstract

The present invention discloses and claims methods and devices for the rapid mechanical isolation of monocot plant tissues suitable for transformation or tissue culture. The invention includes mechanical devices for substantially isolating target plant tissues for use as transformable explants, and propagation of transgenic plants and plant tissues.

First claim

What is claimed is:

1. A method of providing monocot embryos suitable for transformation or tissue culture, comprising:
(a) providing multiple monocot seeds containing immature embryos having an opening in the pericarp of said seeds; and
(b) applying force to said seeds sufficient to extract said immature embryos from said seeds, wherein said extracted immature embryos comprise embryos or pieces of embryos suitable for genetic transformation or tissue culture; wherein said force comprises mechanical positive pressure applied to the exterior of said seeds.

2. The method of claim 1 , further comprising separating said extracted immature embryos from associated non-embryo tissue, wherein said separated extracted immature embryos comprise embryos suitable for tissue culture.

3. The method of claim 1 , wherein said monocot is in the family Poaceae.

4. The method of claim 1 , wherein said monocot is a Zea species.

5. The method of claim 4 , wherein said multiple monocot seeds comprise multiple corn kernels on at least one corn ear.

6. The method of claim 1 , wherein said embryos suitable for tissue culture comprise intact embryos.

7. The method of claim 1 , wherein said embryos suitable for tissue culture comprise partial embryos.

8. The method of claim 1 , wherein said tissue culture comprises transformation.

9. The method of claim 1 , further comprising:
c) regenerating said extracted immature embryos.

10. The method of claim 9 , wherein said regeneration results in callus formation.

11. The method of claim 9 , wherein said regeneration results in at least one fertile plant.

12. The method of claim 2 , wherein said separating comprises use of at least one of size-exclusion, hydrophobic separation, and density differentials.

Assignees

• Monsanto Technology Llc

Inventors

• Adams Jr Whitney R
• Davis Brandon
• Kucher Lubomyr
• Lowe Brenda A
• Spencer Michael
• Mann Michael T

Classifications

• A01H4/001

  Culture apparatus for tissue culture · CPC title

• C12N15/82

  for plant cells {, e.g. plant artificial chromosomes (PACs)} · CPC title

• A01H4/003 Primary

  Cutting apparatus specially adapted for tissue culture · CPC title

• A01H4/008

  Methods for regeneration to complete plants · CPC title

• A01G7/00

  Botany in general · CPC title