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US 12,735,718 B2Granted
Parthenogenesis factors and methods of using same
AssigneePioneer Hi-Bred International, Inc.
InventorsShane E. Abbitt, Jon Aaron Tucker Reinders, Huaxun Ye
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Abstract
Parthenogenesis is a natural form of asexual reproduction wherein growth and development of embryos occur without fertilization by sperm. Peptides are used as parthenogenesis factors, specifically comprising polypeptides or polynucleotides encoding gene products for generating doubled haploids or haploid plants from female gametes.
Claims34
A method of producing a genome-edited, doubled haploid maize plantlet or plant, the method comprising:
- (a) providing to a maternal gametophyte:
- (i) a polynucleotide sequence encoding a parthenogenesis factor, wherein the polynucleotide sequence is operably linked to a regulatory element active in an egg cell, and wherein the parthenogenesis factor is an Ovule Development Protein 2 (ODP2) polypeptide having at least 95% sequence identity to SEQ ID NO:
- 20 and comprising a first motif having an amino acid sequence of SEQ ID NO:
- 36 and a second motif having an amino acid sequence of SEQ ID NO:
- 38;
- and
- (ii) a site-directed nuclease;
- (b) modifying a DNA target site in the maternal gametophyte via the site-directed nuclease, resulting in a modified DNA target site;
- (c) regenerating a T0 plant containing the polynucleotide sequence encoding the parthenogenesis factor and the modified DNA target site;
- (d) pollinating the T0 plant with pollen;
- (e) obtaining a genome-edited, doubled haploid embryo from a parthenogenic maternal gametophyte of the T0 plant;
- and
- (f) regenerating a genome-edited, doubled haploid maize plantlet or plant from the genome-edited, doubled haploid embryo.
10 more elements
A method of producing a doubled haploid maize plantlet or plant, the method comprising:
- (a) providing to a maternal gametophyte a polynucleotide sequence encoding a parthenogenesis factor, wherein the polynucleotide sequence encoding a parthenogenesis factor is operably linked to a regulatory element active in an egg cell, and wherein the parthenogenesis factor is an Ovule Development Protein 2 (ODP2) polypeptide having at least 95% sequence identity to SEQ ID NO:
- 20 and comprising a first motif having an amino acid sequence of SEQ ID NO:
- 36 and a second motif having an amino acid sequence of SEQ ID NO:
- 38;
- (b) regenerating a T0 plant containing the polynucleotide sequence encoding the parthenogenesis factor;
- (c) pollinating the T0 plant with pollen;
- (d) obtaining a doubled haploid embryo from a parthenogenic maternal gametophyte of the T0 plant;
- and
- (e) regenerating a doubled haploid maize plantlet or plant from the doubled haploid embryo.
6 more elements
Dependencies are inferred from the claim text.