Engineering single-gene-controlled staygreen potential into plants
View Patent ↗The enzymes of the ACC synthase family are used in producing ethylene. Nucleotide and polypeptide sequences of ACC synthases are provided along with knockout plant cells having inhibition in expression and/or activity in an ACC synthase and knockout plants displaying a staygreen phenotype, a male sterility phenotype, or an inhibition in ethylene production. Methods for modulating staygreen potential in plants, methods for modulating sterility in plants, and methods for inhibiting ethylene production in plants are also provided.
1. An isolated nucleic acid comprising a polynucleotide selected from the group consisting of:
a) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:56, or the complement thereof;
b) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:57, or the complement thereof;
c) a polynucleotide comprising the polynucleotide sequence of SEQ ID NOs:56 and 57, or the complement thereof;
d) a polynucleotide comprising nucleotides 781-1267 of the polynucleotide sequence of SEQ ID NO:5, or the complement thereof;
e) a polynucleotide comprising nucleotides 781-1267 of the polynucleotide sequence of SEQ ID NO:5 and the reverse complement thereof, or the complement thereof;
f) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:54, or the complement thereof;
g) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:55, or the complement thereof; and
h) a polynucleotide comprising the polynucleotide sequence of SEQ ID NOs:54 and 55, or the complement thereof.
2. The isolated nucleic acid of claim 1 , wherein the nucleic acid inhibits ethylene production when expressed in a plant.
3. An expression cassette comprising a promoter operably linked to the isolated nucleic acid of claim 1 .
4. The expression cassette of claim 3 , wherein the promoter is a constitutive promoter.
5. A plant cell transformed with a construct effective for inhibiting expression of at least one endogenous ACC synthase gene, wherein said construct comprises a promoter that functions in plants operably linked to a nucleic acid configured for RNA silencing or interference, wherein said nucleic acid comprises a polynucleotide selected from the group consisting of:
a) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:56, or the complement thereof;
b) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:57, or the complement thereof;
c) a polynucleotide comprising the polynucleotide sequence of SEQ ID NOs:56 and 57, or the complement thereof;
d) a polynucleotide comprising nucleotides 781-1267 of the polynucleotide sequence of SEQ ID NO:5, or the complement thereof;
e) a polynucleotide comprising nucleotides 781-1267 of the polynucleotide sequence of SEQ ID NO:5 and the reverse complement thereof, or the complement thereof;
f) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:54, or the complement thereof;
g) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:55, or the complement thereof; and
h) a polynucleotide comprising the polynucleotide sequence of SEQ ID NOs:54 and 55, or the complement thereof.
6. The plant cell of claim 5 , wherein the plant cell is from a monocot.
7. The plant cell of claim 5 , wherein the plant cell is from Zea mays.
8. The plant cell of claim 5 , wherein the promoter is a constitutive promoter.
9. A transgenic plant comprising a staygreen potential phenotype, said staygreen potential phenotype resulting from at least one introduced transgene which inhibits ethylene synthesis, wherein said transgene is in an RNA silencing or interference configuration, and wherein said transgene comprises a promoter operably linked to a nucleic acid comprising a polynucleotide selected from the group consisting of:
a) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:56, or the complement thereof;
b) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:57, or the complement thereof;
c) a polynucleotide comprising the polynucleotide sequence of SEQ ID NOs:56 and 57, or the complement thereof;
d) a polynucleotide comprising nucleotides 781-1267 of the polynucleotide sequence of SEQ ID NO:5, or the complement thereof;
e) a polynucleotide comprising nucleotides 781-1267 of the polynucleotide sequence of SEQ ID NO:5 and the reverse complement thereof, or the complement thereof;
f) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:54, or the complement thereof;
g) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:55, or the complement thereof; and
h) a polynucleotide comprising the polynucleotide sequence of SEQ ID NOs:54 and 55, or the complement thereof.
10. The plant of claim 9 , wherein the plant is a monocot.
11. The plant of claim 9 , wherein the plant is Zea mays.
12. The plant of claim 9 , wherein the promoter is a constitutive promoter.
13. An expression cassette comprising a promoter operably linked to a polynucleotide comprising the polynucleotide sequence of SEQ ID NOs:56 and 57, wherein transcription from the promoter produces a hairpin RNA.
14. A plant cell comprising the expression cassette of claim 13 .
15. A plant comprising the expression cassette of claim 13 .
16. The plant of claim 15 , wherein the promoter is a constitutive promoter that functions in the plant.
17. The plant of claim 15 , further comprising an expression cassette comprising a promoter operably linked to a polynucleotide comprising the polynucleotide sequence of SEQ ID NOs:54 and 55, wherein transcription from the promoter produces a hairpin RNA.
18. The plant of claim 15 , wherein the plant is a monocot.
19. The plant of claim 15 , wherein the plant is Zea mays.
20. An expression cassette comprising a promoter operably linked to a polynucleotide comprising the polynucleotide sequence of SEQ ID NOs:54 and 55, wherein transcription from the promoter produces a hairpin RNA.
21. A plant cell comprising the expression cassette of claim 20 .
22. A plant comprising the expression cassette of claim 20 .
23. The plant of claim 22 , wherein the promoter is a constitutive promoter that functions in the plant.
24. The plant of claim 22 , wherein the plant is a monocot.
25. The plant of claim 22 , wherein the plant is Zea mays.
26. The plant cell of claim 5 , wherein the plant cell is from Oryza sativa.
27. The plant of claim 9 , wherein the plant is Oryza sativa.
28. The plant of claim 15 , wherein the plant is Oryza sativa.
29. The plant of claim 22 , wherein the plant is Oryza sativa.