Method and apparatus for substantially isolating plant tissues

Patent metadata

Field	Value
Publication number	US-10433503-B2
Application number	US-201715452519-A
Country	US
Kind code	B2
Filing date	Mar 7, 2017
Priority date	Aug 5, 2003
Publication date	Oct 8, 2019
Grant date	Oct 8, 2019

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 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 the force is selected from the group consisting of: negative pressure, centrifugal force, linear acceleration, linear deceleration, bulk maceration, vibration, and ultrasound.

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 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 , wherein said tissue culture comprises regeneration.

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.

13. The method of claim 5 , wherein the corn ear is immature.

14. The method of claim 1 , wherein the pericarp is opened by using one or more of the following: a needle, an awl, a blade, an abrasive material, application of a chemical treatment, and application of an enzymatic treatment.

Assignees

• Monsanto Technology Llc

Inventors

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

Classifications

• C12N15/8205

  Agrobacterium mediated transformation · CPC title

• C12N15/8201

  Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation · CPC title

• A01G7/00

  Botany in general · CPC title

• A01H4/008

  Methods for regeneration to complete plants · CPC title

• A01H4/003 Primary

  Cutting apparatus specially adapted for tissue culture · CPC title