Methods in molecular biology (Clifton, N.J.)Jul 18, 2026

Producing Rice Plants with Only One Set of Genes by Removing a Protein Active in Egg Cells

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Jul 19, 2026

Abstract

In vivo haploid induction in rice can be achieved using egg cell-specific peptidase (ECS) knockout lines.

Doubled haploid technology provides a rapid method for crop breeding and genetic research.
Genotype dependence poses challenges in the anther culture method for rice haploid production.
CRISPR-Cas9 is employed to mutate the ECS for haploid induction.
Homozygous Osecs mutants are selected to facilitate haploid identification.
Flow cytometry is used to identify haploids in the offspring of Osecs mutants.

Simplified

Doubled haploid (DH) technology is a fast and convenient approach for crop breeding and genetic research. Currently, in vitro anther culture is the main method for rice haploid production. However, genotype dependence remains a major problem in the anther culture of most rice subspecies or cultivars for haploid induction. In this chapter, we describe a protocol for in vivo haploid induction in rice using egg cell-specific peptidase (ECS) knockout lines, including mutation of ECS using CRISPR-Cas9 system, selection of homozygous Osecs mutants, and identification of haploids in the offspring of Osecs by flow cytometry. The ECS mediated maternal in vivo haploid-induction system is a convenient, time-saving and labor-saving technique to produce rice DH lines.

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Producing Rice Plants with Only One Set of Genes by Removing a Protein Active in Egg Cells

Abstract

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