Terpene synthases from ylang ylang (
The present invention relates to the field of plant molecular biology. More particularly, the present invention relates to the isolation of nucleic acids encoding terpene synthases (TPSs), including a novel, multifunctional TPS identified herein as CoTPS2.
1. A cDNA encoding a Cananga odorata terpene synthase 2 protein (CoTPS2 protein) having the amino acid sequence set forth in SEQ ID NO:10 or a modified CoTPS2 protein, wherein the modified CoTPS2 protein has the amino acid sequence set forth in SEQ ID NO:10 with 1-10 conservative amino acid substitutions, wherein the modified CoTPS2 protein has the motifs R(R)X 8 W SEQ ID NO:1), DDXXD (SEQ ID NO:2) and NSE/DTE (SEQ ID NO:3/SEQ ID NO:4) and wherein the modified CoTPS2 protein has terpene synthase activity.
2. The cDNA of claim 1 , wherein the modified CoTPS2 protein has 1-6 conservative amino acid substitutions.
3. The cDNA of claim 1 , wherein the cDNA has the nucleotide sequence set forth in SEQ ID NO:9.
4. A nucleic construct comprising the cDNA of claim 1 operably linked to a plant operable promoter.
5. An expression vector comprising the cDNA of claim 1 .
6. A transgenic plant cell, plant or plant seed comprising cDNA of claim 1 stably integrated into its genome.
7. The transgenic plant cell, plant or plant seed of claim 6 , wherein the plant is a member of the Cananga genus.
8. The transgenic plant cell, plant or plant seed of claim 7 , wherein the plant is a ylang ylang ( Cananga odorata ) plant.
9. A method for producing a transgenic plant which comprises introducing the cDNA of claim 1 or an expression vector comprising the cDNA into a plant, wherein the transgenic plant has the cDNA stably integrated in its genome.
10. A method for producing a transgenic plant which comprises transfecting the cDNA of claim 1 or an expression vector comprising the cDNA into a plant cell or plant cells and regenerating a transgenic plant from the transfected plant cell or transfected plant cells, wherein the transgenic plant has the cDNA stably integrated in its genome.
11. The method of claim 9 , wherein the plant is a member of the Cananga genus.
12. The method of claim 11 , wherein the transgenic plant is a ylang ylang ( Cananga odorata ) plant.
13. A method of synthesizing at least one terpene or sesquiterpene in a plant which comprises introducing the cDNA of claim 1 or an expression vector comprising the cDNA into a plant, wherein the cDNA is expressed in the plant thereby synthesizing the at least one terpene or sesquiterpene.
14. A method of synthesizing at least one terpene or sesquiterpene in a plant comprising transfecting the cDNA of claim 1 or an expression vector comprising the cDNA into a plant cell or plant cells and growing a plant from the transfected plant cell or transfected plant cells, wherein the cDNA is expressed in the plant thereby synthesizing the at least one terpene or sesquiterpene.
15. The method of claim 13 , wherein the at least one sesquiterpene is β-ylangene, β-copaene or β-cubebene.
16. The method of claim 13 , wherein the plant is a member of the Cananga genus.
17. The method of claim 16 , wherein the transgenic plant is a ylang ylang ( Canangga odorata ) plant.
18. The method of claim 13 , wherein the cDNA is overexpressed.
19. A method of producing at least one terpene or sesquiterpene in a yeast cell, the method comprising exogenously expressing within the yeast cell the cDNA of claim 1 , thereby producing the at least one sesquiterpene in the yeast cell.
20. The method of claim 19 , wherein the at least one sesquiterpene is β-ylangene, β-copaene or β-cubebene.
21. The cDNA of claim 1 , wherein the modified CoTPS2 protein has 1 or 2 conservative amino acid substitutions.
22. An expression vector comprising the nucleic acid construct of claim 4 .