Doubled haploid inducer
AssigneePioneer Hi-Bred International, Inc.
InventorsJON AARON TUCKER REINDERS, MARK E. WILLIAMS, HUAXUN YE
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Abstract
Methods of creating doubled haploid inducers are provided. These doubled haploid inducers are used in the production of maternal doubled haploids.
Claims60
A method of producing a doubled haploid plant, comprising:
- a) providing a first plant, wherein the first plant comprises at least one introduced genetic chromosome doubling agent;
- b) crossing the first plant with a second plant, wherein the at least one introduced genetic chromosome doubling agent of the first plant induces chromosome doubling of a fertilized egg cell of the second plant in the absence of an exogenously applied chemical or biochemical chromosome doubling agent;
- c) obtaining a diploidized embryo comprising a pair of chromosomes inherited from the second plant and not comprising the introduced genetic chromosome doubling agent;
- and
- d) regenerating a diploid plant from the diploidized embryo.
A method of producing a doubled haploid inducer (DHI) plant, the method comprising:
- a) providing a transformable cell or tissue from a non-transgenic haploid inducer plant;
- b) transforming the cell or tissue with a heterologous DNA construct comprising a genetic chromosome doubling agent;
- and
- c) obtaining a double haploid inducer (DHI) line comprising the introduced genetic chromosome doubling agent.
A method of producing a doubled haploid inducer (DHI) line, the method comprising:
- a) providing a transformed first haploid inducer line comprising a heterologous DNA construct comprising a genetic chromosome doubling agent;
- b) providing a second haploid inducer line containing a marker;
- c) crossing the first haploid inducer line with the second haploid inducer line to produce a combination of maternal and diploid embryos;
- d) selecting the maternal embryos based on the absence of the selectable marker from the second haploid inducer line;
- and
- e) obtaining a double haploid inducer (DHI) line comprising the introduced genetic chromosome doubling agent, thereby producing the DHI line.
A method of eliminating or reducing the step of exogenously inducing chromosome doubling after embryo formation in the production of an inbred population, the method comprising crossing a doubled haploid inducer (DHI) line comprising an introduced genetic chromosome doubling agent with a second line and obtaining a doubled haploid progeny of the second line without a separate step of chromosome doubling due to exogenous treatment of a chromosome doubling agent, wherein the double haploid progeny does not comprise the introduced genetic chromosome doubling agent.
A method of maintaining a doubled haploid inducer (DHI) line comprising a genetic chromosome doubling agent, the method comprising:
- a) providing a maintainer line comprising a heterologous maintainer construct, the maintainer construct comprising (i) a repressor or an inhibitor component for the genetic chromosome doubling agent, (ii) a pollen transmission prevention component, and (iii) a selectable marker component to identify seeds that contain the maintainer construct;
- b) self-fertilizing the maintainer line having the maintainer construct and the genetic chromosome doubling agent;
- c) identifying seeds containing the maintainer construct from the self-fertilization, wherein the maintainer construct is present in about 50% of the F1 seeds;
- and
- d) growing the seeds containing the maintainer construct, thereby maintaining the DHI line in a diploid state for further seed increases or haploid induction crosses.
A doubled haploid inducer (DHI) maintainer line comprising a genetic chromosome doubling agent and a heterologous maintainer construct, the maintainer construct comprising (i) a repressor or an inhibitor component for the genetic chromosome doubling agent, (ii) a pollen transmission prevention component, and (iii) a selectable marker component to identify seeds that contain the maintainer construct.
Dependencies are inferred from the claim text.