Antisense gene systems of pollination control for hybrid seed production
AssigneePioneer Hi-Bred International, Inc.
InventorsSteven F. Fabijanski, Paul G. Arnison
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Abstract
A process is described for producing hybrid seed using male-sterile plants created by employing molecular techniques to manipulate antisense DNA and other genes that are capable of controlling the production of fertile pollen in plants. Transformation techniques are used to introduce constructs containing antisense DNA and other genes into plants. Said plants are functionally male-sterile and are useful for the production of hybrid seed by the crossing of said male-sterile plants with pollen from male-fertile plants. Hybrid seed production is simplified and improved by this invention and can be extended to plant crop species for which commercially acceptable hybrid seed production methods are not currently available.
Claims32
A method of producing hybrid seed from plants from a species of pollen-producing plants that is capable of being genetically transformed, comprising the steps of:
- (a) producing a genetically transformed plant by:
- (i) inserting into the genome of a plant cell of said species that can be regenerated into a differentiated, whole plant;
- I. a sense gene that confers on said plant resistance to a herbicide or an antibiotic
- and
- II. a recombinant DNA molecule comprising:
- (A) a DNA sequence that codes for RNA that is complementary to the RNA sequence encoded by said sense gene
- and
- (B) a pollen-specific Brassica promoter which functions in said plant cell to cause transcription of said DNA sequence into RNA,
- (ii) obtaining a transformed plant cell which has been transformed with said sense gene and said recombinant DNA and
- (iii) regenerating from said transformed plant cell a plant that is genetically transformed with said sense gene and said recombinant DNA molecule such that (A) said recombinant DNA molecule interferes with the expression of said sense gene and (B) said genetically transformed plant can be rendered male sterile by said herbicide or antibiotic;
- (b) increasing the number of genetically transformed plants by:
- (i) fertilizing said genetically transformed plant with pollen produced by a suitable male fertile plant and obtaining seed which, when germinated, yields a plurality of genetically transformed plants;
- or
- (ii) clonally propagating said genetically transformed plant to obtain a plurality of genetically transformed plants;
- and
- (c) effecting a hybrid cross by pollinating said genetically transformed plants with pollen from suitable male fertile plant donors.
A method of producing a plant which carries the male sterile trait, which method comprises the steps of:
- (a) transforming a cell of a pollen-producing plant which is from a species of pollen-producing plants that is capable of being regenerated into a differentiated whole plant, with a sense gene that confers on said plant resistance to a herbicide or an antibiotic;
- (b) regenerating from said transformed plant cell a genetically transformed plant which is resistant to the same herbicide or antibiotic;
- (c) inserting a recombinant DNA molecule into the genome of a plant cell of said herbicide- or antibiotic-resistant plant which is capable of being regenerated into a differentiated whole plant, wherein said recombinant DNA molecule comprises:
- (i) a DNA sequence that codes for RNA that is antisense relative to the RNA sequence encoded by said sense gene;
- and
- (ii) a pollen-specific Brassica promotor which functions in said plant cell to cause transcription of said DNA sequence into RNA;
- (d) obtaining a transformed plant cell;
- and
- (e) regenerating from said transformed plant cell a genetically transformed plant which has been transformed with the sense gene described in step (a) and the recombinant DNA molecule described in step (c) above such that said recombinant DNA interferes with the expression of said sense gene and said genetically transformed plant can be rendered male sterile by said herbicide or antibiotic.
A method of producing a hybrid seed with restored male fertility from plants of a species of pollen-producing plants that is capable of being genetically transformed, comprising the steps of:
- (a) producing a plant which carries a male sterile trait by:
- (i) inserting into the genome of a plant cell of said species that can be regenerated into a differentiated, whole plant:
- I. a sense gene which confers on said plant resistance to a herbicide or an antibiotic and
- II. a recombinant DNA molecule comprising
- (A) a DNA sequence that codes for RNA that is antisense relative to the RNA sequence encoded by said sense gene;
- and
- (B) a pollen-specific Brassica promoter which functions in said plant cell to cause transcription of said DNA sequence into RNA,
- (ii) obtaining a plant cell of a plant which has been transformed with the sense gene and recombinant DNA molecule described in step (i) above and
- (iii) regenerating from said transformed plant cell a genetically transformed plant which is transformed with the sense gene and recombinant DNA molecule described in step (i) above such that said recombinant DNA molecule interferes with the expression of said sense gene and said genetically transformed plant can be rendered male sterile by said herbicide or antibiotic;
- (b) increasing the number of genetically transformed plants by:
- (i) growing the genetically transformed plant described in step (a)(iii) above in isolation from the same herbicide or antibiotic to produce a self-fertile plant,
- (ii) permitting self-fertilization and
- (iii) growing seed of such self-fertile plant over a number of generations in isolation from the same herbicide or antibiotic to increase the number of genetically transformed plants;
- (c) exposing said genetically transformed plants to said herbicide or antibiotic to produce male sterile plants;
- and
- (d) effecting a hybrid cross by pollinating said male sterile plants with pollen derived from suitable male fertile donors.
A plant which has been transformed with:
- (a) a sense gene which confers on said plant resistance to a herbicide or an antibiotic;
- and
- (b) a recombinant, double-stranded DNA molecule comprising:
- (i) a DNA sequence that codes for RNA that is antisense relative to the RNA sequence encoded by said sense gene;
- (ii) a pollen-specific Brassica promotor which functions in said plant cell to cause transcription of said DNA sequence into RNA;
- and
- (iii) a terminator sequence which defines a termination signal during transcription of said DNA sequence.
Hybrid seed containing DNA comprising:
- (a) a sense gene which confers on a plant grown from said seed resistance to a herbicide or an antibiotic;
- and
- (b) a recombinant double stranded DNA molecule comprising:
- (i) a DNA sequence that codes for RNA that is antisense relative to the RNA sequence encoded by said sense gene;
- (ii) a pollen-specific Brassica promotor which functions in said plant cell to cause transcription of said DNA sequence into RNA;
- and
- (iii) a terminator sequence which defines a termination signal during transcription of said DNA sequence.
A cell line produced from genetically transformed cells that are the product of a process comprising the steps of (i) inserting into the genome of a plant cell of a pollen-producing plant a recombinant DNA sequence comprised of antisense DNA that renders developing pollen grains susceptible to a herbicide or to an antibiotic, wherein said plant is capable of regeneration into a differentiated, whole plant and wherein said antisense DNA is under the control of a pollen-specific Brassica promoter;
- and then (ii) obtaining a transformed plant cell of said plant.
Dependencies are inferred from the claim text.