Plant embryo storage and manipulation

Patent metadata

Field	Value
Publication number	US-10477859-B2
Application number	US-201515507032-A
Country	US
Kind code	B2
Filing date	Jun 4, 2015
Priority date	Aug 29, 2014
Publication date	Nov 19, 2019
Grant date	Nov 19, 2019

Abstract

Methods for obtaining genetic material from plant embryos while preserving their viability are provided. In the methods, preservation of viability may be maintained by suspending the embryos in an aqueous solution surrounded by an oil matrix. Genetic material may be obtained from an aliquot of the aqueous solution and may be used directly for molecular analysis, or whole genome amplification may be performed prior to molecular analysis.

First claim

We claim:

1. A method for performing molecular analysis of one or more plant embryos comprising agitating the plant embryos to remove cellular material for molecular analysis, storing the one or more plant embryos in an aqueous solution surrounded by a matrix of at least two oils, wherein one of the at least two oils is more dense than the aqueous solution and one of the at least two oils is less dense than the aqueous solution, obtaining genetic material from the cellular material removed from the plant embryos, and performing molecular analysis of the genetic material.

2. The method of claim 1 wherein the one or more plant embryos comprise one or more immature plant embryos.

3. The method of claim 1 wherein antimicrobial agents and/or minimal growth media are added to the aqueous solution.

4. The method of claim 1 wherein the one or more plant embryos are stored under dark conditions.

5. The method of claim 1 wherein the one or more plant embryos are stored at approximately 4 degrees Celsius.

6. The method of claim 1 further comprising transferring the one or more plant embryos for continued storage.

7. The method of claim 1 further comprising transferring the one or more plant embryos to a germination medium.

8. The method of claim 1 further comprising germinating at least one of the one or more plant embryos.

9. The method of claim 1 wherein the method of storing is automated.

10. The method of claim 1 wherein said molecular analysis is genotyping.

11. The method of claim 1 , further comprising the step of selecting or discarding the plant embryo based on the results of the molecular analysis.

12. The method of claim 1 , wherein the plant embryo is washed in the aqueous solution prior to agitation.

Assignees

• Pioneer Hi Bred Int

Inventors

• Arnold Randal
• Cope Matthew Paul
• Schares Justin Andrew
• Yun Yue

Classifications

• A01H4/00

  Plant reproduction by tissue culture techniques {; Tissue culture techniques therefor} · CPC title

• A01N3/00 Primary

  Preservation of plants or parts thereof, e.g. inhibiting evaporation, improvement of the appearance of leaves {or protection against physical influences such as UV radiation using chemical compositions} (preservation or chemical ripening of fruit or vegetables A23B7/00); Grafting wax · CPC title

• A01H1/04

  Processes of selection {involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection} · CPC title

• A01H4/001

  Culture apparatus for tissue culture · CPC title