Reproductive ablation constructs
View Patent ↗The present invention relates to the regulation of reproductive development, particularly to the genetic ablation of reproductive tissues in angiosperm and gymnosperm species. Reproductive-preferred promoters, regulatory elements, and cytotoxic nucleotide sequences are disclosed herein, as are constructs and methods for genetic ablation.
1. A method for obtaining wood pulp, comprising:
(a) introducing into a plant cell of a woody plant a construct comprising (i) a Pinus radiata AGAMOUS promoter operably linked to
(ii) a desired nucleic acid encoding an E73G barnase mutant that comprises the sequence of SEQ ID NO: 5 or which comprises a nucleotide sequence that encodes the amino acid sequence of SEQ ID NO: 9;
(b) culturing said transformed plant cell under conditions that promote growth of a plant; and
(c) obtaining wood pulp from said plant.
2. A construct, comprising (i) a Pinus radiata AGAMOUS promoter operably linked to the sequence of SEQ ID NO: 5, or (ii) the sequence depicted in SEQ ID NOS: 20 or 21.
3. The construct of claim 2 , further comprising in part (i) an Arabidopsis AGAMOUS floral-specific enhancer (AtAGenh) fused to the 5′ end of the promoter, wherein expression of the construct is capable of disrupting reproductive development of at least one reproductive structure in a plant.
4. A transgenic plant, comprising the construct of claim 2 or 3 .
5. A transgenic plant, comprising a construct that comprises a Pinus radiata AGAMOUS promoter operably linked to a desired nucleic acid that comprises the sequence of SEQ ID NO: 5 or which encodes the amino acid sequence of SEQ ID NO: 9.
6. A method for producing a transgenic plant, comprising:
(a) transforming a plant cell with a construct that comprises a Pinus radiata AGAMOUS promoter operably linked to a desired nucleic acid, wherein said desired nucleic acid has the sequence of SEQ ID NO:5 or encodes the amino acid of SEQ ID NO:9, wherein said promoter regulates the activity of said desired nucleic acid; and
(b) culturing said transformed plant cell under conditions that promote growth of a plant, wherein said plant is a transgenic plant that exhibits a phenotype that is different from a plant of the same species that does not contain said construct.
7. A construct comprising a Pinus radiata AGAMOUS promoter operably linked to a desired nucleic acid that comprises the sequence of SEQ ID NO: 5 or which encodes SEQ ID NO: 9, wherein said promoter regulates the expression of said desired nucleic acid in a plant cell transformed with the construct.
8. A hybrid progeny plant transformed with the construct of claim 2 or 3 .
9. The hybrid progeny plant of claim 8 , wherein the hybrid progeny plant is obtained from the cross of pitch pine ( Pinus rigida ) with loblolly pine ( Pinus taeda ).
10. A method for producing a transgenic plant, comprising:
(a) transforming a plant cell with a construct that comprises the sequence of SEQ ID NOS: 20 or 21; and
(b) culturing the transformed plant cell under conditions that promote growth of a plant, wherein said plant is a transgenic plant that exhibits a phenotype that is different from a plant of the same species that has not been transformed with the construct.
11. The method of claim 1 or 6 , wherein the construct further comprises an Arabidopsis AGAMOUS floral-specific enhancer (AtAGenh) fused to the 5′ end of the promoter, wherein expression of the construct is capable of disrupting reproductive development of at least one reproductive structure in a plant.
12. A nucleic acid encoding an E73G barnase mutant that comprises the sequence of SEQ ID NO: 5 or which comprises a nucleotide sequence that encodes the amino acid sequence of SEQ ID NO: 9.