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 producing a transgenic plant comprising (a) transforming a plant cell with a construct that comprises (i) a functional Pinus radiata male cone 2 (PrMC2) promoter operably linked to (ii) a polynucleotide that comprises a sequence encoding a K27A barnase mutant; 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.
2. The method of claim 1 , wherein the sequence that encodes the K27A barnase mutant comprises the sequence of SEQ ID NO: 8.
3. The method of claim 1 , wherein the polynucleotide comprises a sequence that encodes the amino acid sequence of SEQ ID NO: 12.
4. The method of claim 1 , wherein the PrMC2 promoter comprises the sequence of SEQ ID NO: 16.
5. The method of claim 4 , wherein the sequence encoding the K27A barnase mutant comprises either (i) the sequence of SEQ ID NO: 8, or (ii) a polynucleotide that comprises a sequence that encodes the amino acid sequence of SEQ ID NO: 12.
6. The method of claim 1 , wherein the PrMC2 promoter/K27A barnase mutant construct comprises the sequence of SEQ ID NO: 14.
7. The method of claim 1 , further comprising obtaining wood from the transgenic plant.
8. The method of claim 1 , further comprising obtaining wood pulp from the transgenic plant.
9. The method of claim 8 , wherein the sequence encoding the K27A barnase mutant comprises either (i) the sequence of SEQ II) NO: 8, or (ii) a polynucleotide that comprises a sequence that encodes the amino acid sequence of SEQ ID NO: 12.
10. A construct comprising a Pinus radiata male cone 2 (PrMC2) promoter operably linked to a polynucleotide encoding a K27A barnase mutant, wherein expression of the K27A barnase mutant disrupts reproductive development of at least one of a male reproductive structure in a plant which expresses the construct.
11. The construct of claim 10 , wherein the sequence encoding the K27A barnase mutant comprises either (i) the sequence of SEQ ID NO: 8, or (ii) a polynucleotide that encodes SEQ ID NO: 12.
12. The construct of claim 11 , wherein the PrMC2 promoter comprises the sequence of SEQ ID NO: 16.
13. The construct of claim 10 , wherein the construct comprises the sequence of SEQ ID NO: 14.
14. A plant cell transformed with the construct of claim 10 .
15. A transgenic plant comprising the plant cell of claim 14 .
16. The transgenic plant of claim 15 , wherein the sequence encoding the K27A barnase mutant comprises either (i) the sequence of SEQ ID NO: 8, or (ii) a polynucleotide that encodes SEQ ID NO: 12.
17. The transgenic plant of claim 16 , wherein the PrMC2 promoter comprises the sequence of SEQ ID NO: 16.
18. The transgenic plant of claim 15 , wherein the construct comprises the sequence of SEQ ID NO: 14.
19. A transgenic progeny plant of the transgenic plant of claim 15 , wherein the transgenic progeny plant either (i) has a disrupted male reproductive structure, or (ii) expresses the construct.
20. The transgenic progeny plant of claim 19 , wherein the progeny plant is obtained from the cross of pitch pine Pinus rigida with loblolly pine Pinus taeda.