Gene silencing of sugar-dependent 1 in
The present invention relates to the field of plant molecular biology and gene silencing. More particularly, the present invention relates to gene silencing of Sugar-dependent 1 (JcSDP1) in Jatropha curcas . JcSDP1 encodes a patatin-domain triacylglyerol lipase. Silencing of JcSDP1 enhances seed oil accumulation in J. curcas.
1. A method of increasing seed oil accumulation in Jatropha comprising reducing the activity of a patatin-domain triacylglycerol lipase encoded by a sugar-dependent-1 (SDP1) gene in a Jatropha plant to cause increased seed oil accumulation, wherein the reduction in activity is achieved by an RNAi molecule comprising the nucleotide sequence set forth in SEQ ID NO:3.
2. The method of claim 1 , wherein the lipase has the amino acid sequence set forth in SEQ ID NO:2.
3. The method of claim 2 , wherein the lipase is encoded by a nucleic acid having a nucleotide sequence set forth in SEQ ID NO:1, or nucleotides 874-3450 of SEQ ID NO:1, or nucleotides 874-3447 of SEQ ID NO:1.
4. The method of claim 1 , wherein the activity of the lipase is reduced in seeds of the Jatropha plant.
5. The method of claim 4 , wherein the reduction in Jatropha seeds is accomplished using a Jatropha seed specific promoter.
6. The method of claim 5 , wherein the Jatropha seed specific promoter is a Jatropha curcas SDP1 promoter.
7. The method of claim 6 , wherein the Jatropha curcas SDP1 promoter has the nucleotide sequence set forth in nucleotides 1-722 of SEQ ID NO:1 or promoter functional fragment thereof.
8. The method of claim 1 , wherein the seed oil accumulation is increased by about 13% to about 30%.
9. An isolated nucleic acid hairpin construct comprising nucleotides 3161-3533 of SEQ ID NO:1, wherein the nucleic acid, when used in RNAi, reduces expression of the sugar-dependent 1 (SDP1) gene.
10. The isolated nucleic acid of claim 9 , wherein the nucleic acid comprises nucleotides 3161-3528 of SEQ ID NO:1.
11. The isolated nucleic acid of claim 9 which further comprises a plant operable promoter operably linked to the nucleic acid.
12. The isolated nucleic acid of claim 11 , wherein the promoter is a seed specific promoter.
13. The isolated nucleic acid of claim 12 , wherein the seed specific promoter is a Jatropha curcas SDP1 promoter.
14. The isolated nucleic acid of claim 13 , wherein the Jatropha curcas SDP1 promoter comprises nucleotides 1-722 of SEQ ID NO:1.
15. An isolated nucleic acid comprising the nucleotide sequence set forth in SEQ ID NO:3.
16. A nucleic acid construct or a vector comprising the hairpin construct of claim 9 .
17. A transgenic plant cell or transgenic plant having the hairpin construct of claim 9 stably integrated in its genome.
18. A method of making a transgenic plant cell comprising introducing the hairpin construct of claim 9 into a plant cell and selecting a transgenic plant cell in which the isolated nucleic acid is stably integrated in its genome.
19. A method of making a transgenic plant comprising introducing the hairpin construct of claim 9 into a plant cell, selecting a transgenic plant cell in which the isolated nucleic acid is stably integrated in its genome and regenerating a transgenic plant from the selected transgenic plant cell.
20. A nucleic acid molecule comprising a heterologous plant operable promoter operably linked to a nucleic acid encoding a protein comprising the amino acid sequence set forth in SEQ ID NO:2.
21. The isolated nucleic acid of claim 20 which comprises the nucleotide sequence set forth in nucleotides 874-3450 of SEQ ID NO:1 or nucleotides 874-3447 of SEQ ID NO:1.
22. The isolated nucleic acid of claim 20 , wherein the promoter is a heterologous seed specific promoter.