Generation of haploid plants and improved plant breeding

Patent metadata

Field	Value
Publication number	US-10912264-B2
Application number	US-201916385982-A
Country	US
Kind code	B2
Filing date	Apr 16, 2019
Priority date	Oct 6, 2009
Publication date	Feb 9, 2021
Grant date	Feb 9, 2021

Abstract

Methods and compositions for generating haploid organisms are described.

First claim

What is claimed is:

1. A transgenic plant comprising a heterologous transgene expression cassette, the expression cassette comprising a promoter operably linked to a polynucleotide encoding a polypeptide comprising a CENH3 tail domain linked to a heterologous CENH3 histone-fold domain, wherein the CENH3 tail domain and the CENH3 histone-fold domain are from different species.

2. The transgenic plant of claim 1 , wherein one or two alleles of the endogenous CENH3 genomic coding sequence of the plant is inactivated or knocked out.

3. The transgenic plant of claim 1 , wherein all alleles of the endogenous CENH3, genomic coding sequence of the plant are inactivated or knocked out.

4. The transgenic plant of claim 3 , wherein the plant, when crossed with a wildtype diploid plant, generates at least 0.1% haploid progeny.

5. The transgenic plant of claim 1 , wherein the polypeptide further comprises a further heterologous amino acid sequence of at least 5 amino acids at an amino terminus of the polypeptide.

6. The transgenic plant of claim 1 , wherein the CENH3 histone-fold domain is at least 95% identical to at least one of SEQ ID NOs: 49-94.

7. The transgenic plant of claim 1 , wherein the CENH3 histone-fold domain is from the same species as the transgenic plant.

8. An isolated polynucleotide encoding the polypeptide of claim 1 .

9. A method of generating an F1 progeny plant having half the ploidy of a parent plant expressing an endogenous CENH3 protein, the method comprising,
crossing the parent plant to the plant of claim 3 ; and
selecting F1 progeny generated from the crossing step having half the ploidy of the parent plant.

10. The method of claim 8 , wherein the parent plant is the pollen parent.

11. The method of claim 8 , wherein the parent plant is the ovule parent.

12. The method of claim 8 , wherein the parent plant is diploid and the selected F1 progeny is haploid and the method further comprises converting at least one selected haploid plant into a doubled haploid plant.

13. A method of making the transgenic plant of claim 1 , comprising, transforming plant cells with a nucleic acid comprising the expression cassette; and selecting transformants comprising the nucleic acid, thereby making the transgenic plant.

Assignees

• Univ California

Inventors

• Chan Simon
• Maruthachalam Ravi

Classifications

• C12N15/8241 Primary

  Phenotypically and genetically modified plants via recombinant DNA technology · CPC title

• C12N15/8287

  for fertility modification, e.g. apomixis · CPC title

• C12N15/8261

  with agronomic (input) traits, e.g. crop yield · CPC title

• C07K14/415

  from plants · CPC title

• Y02A40/146

  Genetically Modified [GMO] plants, e.g. transgenic plants · CPC title