Methods of seed breeding using high throughput nondestructive seed sampling

Patent metadata

Field	Value
Publication number	US-9986699-B2
Application number	US-201514629235-A
Country	US
Kind code	B2
Filing date	Feb 23, 2015
Priority date	Aug 26, 2004
Publication date	Jun 5, 2018
Grant date	Jun 5, 2018

Abstract

Novel methods are provided to facilitate germplasm improvement activities through the use of high throughput, nondestructive sampling of seeds. A method for introgressive hybridization, for example, generally includes removing tissue samples from individual seeds using an automated seed sampler without affecting germination viability of the seeds, and analyzing nucleic acids extracted from the tissue samples for at least one genetic marker. The method then further includes selecting the sampled seeds that possess the at least one genetic marker, cultivating fertile plants from the selected seeds, and crossing the fertile plants with other plants.

First claim

What is claimed is:

1. A method for introgressive hybridization, the method comprising:
removing tissue samples from individual seeds using an automated seed sampler without affecting germination viability of the seeds;
analyzing nucleic acids extracted from the tissue samples for at least one genetic marker;
selecting the sampled seeds that possess the at least one genetic marker;
cultivating a fertile plant from a selected one of the sampled seeds; and
crossing the fertile plant with another plant.

2. The method of claim 1 , wherein crossing the fertile plant with another plant occurs at an F 1 stage of a breeding program.

3. The method of claim 1 , wherein crossing the fertile plant with another plant occurs at an F 2 stage of a breeding program.

4. The method of claim 1 , wherein crossing the fertile plant with another plant occurs at an F 3 stage of a breeding program.

5. The method of claim 1 , wherein cultivating a fertile plant from a selected one of the sampled seeds comprises cultivating no more than 50 fertile plants from selected ones of the sampled seeds in a single round of breeding.

6. The method of claim 1 , wherein cultivating a fertile plant from a selected one of the sampled seeds comprises cultivating the fertile plant in a first geographic location; and further comprising:
cultivating a fertile plant from another selected one of the sampled seeds in a second geographic location that is geographically distinct from the first geographic location.

7. The method of claim 6 , wherein the at least two geographically distinct locations are sufficiently distant from each other so that adverse weather in one does not affect plant welfare in another.

8. The method of claim 1 , wherein the at least one genetic marker includes at least two genetic markers.

9. The method of claim 1 , wherein the at least one genetic marker is linked with a sequence feature selected from the group consisting of a single nucleotide polymorphism, a simple sequence repeat, a restriction fragment length polymorphism, a haplotype, a tag SNP, an alleles of a genetic marker, a gene, a DNA-derived sequence, an RNA-derived sequence, a promoter, a 5′ untranslated region of a gene, a 3′ untranslated region of a gene, microRNA, siRNA, a QTL, a satellite marker, a transgene, mRNA, ds mRNA, a transcriptional profile, and a methylation pattern.

10. The method of claim 1 , wherein the at least one genetic marker is linked with a haplotype associated with a QTL selected from the group consisting of herbicide tolerance, disease resistance, insect or pest resistance, altered fatty acid, protein or carbohydrate metabolism, increased grain yield, increased oil, increased nutritional content, increased growth rates, enhanced stress tolerance, preferred maturity, enhanced organoleptic properties, altered morphological characteristics, other agronomic traits, traits for industrial uses, traits for improved consumer appeal, and a combination of traits as a multiple trait index.

11. The method of claim 1 , wherein the at least one genetic marker is linked with a QTL selected from the group consisting of herbicide tolerance, disease resistance, insect or pest resistance, altered fatty acid, protein or carbohydrate metabolism, increased grain yield, increased oil, increased nutritional content, increased growth rates, enhanced stress tolerance, preferred maturity, enhanced organoleptic properties, altered morphological characteristics, other agronomic traits, traits for industrial uses, traits for improved consumer appeal, and a combination of traits as a multiple trait index.

12. The method of claim 1 , further comprising coating the selected seeds with a polymer and/or a fungicide after removing the tissue samples therefrom to further preserve germination viability.

13. The method of claim 1 wherein the nucleic acids are analyzed by polymerase chain reaction.

14. The method of claim 1 wherein the nucleic acids are analyzed by microarray.

15. The method of claim 1 wherein the nucleic acids are analyzed by DNA sequencing.

16. The method of claim 1 , further comprising extracting the nucleic acids from the tissue samples.

17. The method of claim 1 , wherein removing tissue samples from individual seeds using an automated seed sampler without affecting germination viability of the seeds includes removing the tissue samples from the individual seeds using the automated seed sampler while maintaining the germination viability of greater than fifty percent of the seeds.

18. The method of claim 1 , further comprising preserving single seed identity of the selected seeds.

19. The method of claim 1 , wherein removing tissue samples from individual seeds using an automated seed sampler without affecting germination viability of the seeds includes removing the tissue samples from endosperm portions of the seeds.

20. The method of claim 1 , further comprising:
collecting seeds from the another plant, after crossing the fertile plant with the another plant;
removing tissue samples from the collected seeds using the automated seed sampler without affecting germination viability of the seeds;
analyzing nucleic acids extracted from the tissue samples for at least one genetic marker; and
selecting the sampled seeds that possess the at least one genetic marker.

Assignees

• Monsanto Technology Llc

Inventors

• Deppermann Kevin L
• Petersen Michael W
• Butruille David
• Dotson Stanton
• Eathington Sam
• Forbes Heather
• Schnicker Bruce
• Tamulonis John

Classifications

• A01H1/02 Primary

  Methods or apparatus for hybridisation; Artificial pollination {; Fertility} · CPC title

• A01H1/04

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

• C12Q1/6895 Primary

  for plants, fungi or algae · CPC title

• C12Q2600/13

  Plant traits · CPC title

• C12Q2600/156

  Polymorphic or mutational markers · CPC title