Extending juvenility in grasses
The present invention relates to compositions and methods for modulating the juvenile to adult developmental growth transition in plants, such as grasses (e.g. maize). In particular, the invention provides methods for enhancing agronomic properties in plants by modulating expression of GRMZM2G362718, GRMZM2G096016, or homologs thereof. Modulation of expression of one or more additional genes which affect juvenile to adult developmental growth transition such as Glossy15 or Cg1, in conjunction with such modulation of expression is also contemplated. Nucleic acid constructs for down-regulation of GRMZM2G362718 and/or GRMZM2G096016 are also contemplated, as are transgenic plants and products produced there from, that demonstrate altered, such as extended juvenile growth, and display associated phenotypes such as enhanced yield, improved digestibility, and increased disease resistance. Plants described herein may be used, for example, as improved forage or feed crops or in biofuel production.
1. A polynucleotide molecule comprising a nucleotide sequence selected from the group consisting of:
(a) a nucleotide sequence encoding a polypeptide that comprises an amino acid sequence that is at least 96% identical over its full length to the amino acid sequence of SEQ ID NO: 17; wherein the polypeptide regulates juvenile to adult phase change in grass plant leaves;
(b) a nucleotide sequence comprising SEQ ID NO: 16;
(c) a nucleotide sequence comprising at least 95% sequence identity over its full length to the full length of SEQ ID NO:16, wherein the nucleotide sequence encodes a protein that regulates juvenile to adult phase change in grass plant leaves; and
(d) the nucleotide sequence complementary to (a), (b), or (c),
wherein the polynucleotide molecule further comprises a heterologous promoter functional in plants that is operably linked to the nucleotide sequence.
2. The polynucleotide molecule of claim 1 , wherein the nucleotide sequence comprises SEQ ID NO:16.
3. A recombinant vector comprising the polynucleotide molecule of claim 1 .
4. The recombinant vector of claim 3 , further comprising at least one additional sequence chosen from the group consisting of: a regulatory sequence, a selectable marker, a leader sequence and a terminator.
5. The recombinant vector of claim 4 , wherein the additional sequence is a heterologous sequence.
6. The recombinant vector of claim 3 , wherein the promoter is a tissue-specific promoter.
7. The recombinant vector of claim 3 , wherein the promoter directs expression in leaf tissue.
8. The recombinant vector of claim 3 , defined as an isolated expression cassette.
9. A recombinant vector construct comprising a polynucleotide sequence encoding a siRNA that targets a nucleotide sequence, wherein expression of the siRNA in a plant down regulates expression of the nucleotide sequence, wherein the nucleotide sequence is selected from the group consisting of:
(a) a nucleotide sequence encoding a polypeptide that comprises an amino acid sequence that is at least 95% identical over its full length to the amino acid sequence of SEQ ID NO: 17; wherein the polypeptide regulates juvenile to adult phase change in grass plant leaves;
(b) a nucleotide sequence comprising SEQ ID NO: 16;
(c) a nucleotide sequence comprising at least 95% sequence identity over its full length to the full length of SEQ ID NO:16, wherein the nucleotide sequence encodes a protein that regulates juvenile to adult phase change in grass plant leaves; and
(d) the nucleotide sequence complementary to (a), (b), or (c).
10. A transgenic plant comprising a recombinant vector, the recombinant vector comprising a heterologous promoter functional in plants that is operably linked to a nucleotide sequence selected from the group consisting of:
(a) a nucleotide sequence encoding a polypeptide that comprises an amino acid sequence that is at least 95% identical over its full length to the amino acid sequence of SEQ ID NO: 17; wherein the polypeptide regulates juvenile to adult phase change in grass plant leaves;
(b) a nucleotide sequence comprising SEQ ID NO: 16;
(c) a nucleotide sequence comprising at least 95% sequence identity over its full length to the full length of SEQ ID NO:16, wherein the nucleotide sequence encodes a protein that regulates juvenile to adult phase change in grass plant leaves; and
(d) the nucleotide sequence complementary to (a), (b), or (c).
11. A transgenic plant comprising the recombinant vector construct of claim 9 .
12. The transgenic plant of claim 10 , further defined as a monocotyledonous plant.
13. The transgenic plant of claim 10 , further defined as a member of the Poaceae, the Panicoideae or the Pooideae.
14. The transgenic plant of claim 10 , further defined as maize, rice, sorghum, or switchgrass.
15. A seed of the transgenic plant of claim 10 , wherein the seed comprises the recombinant vector.
16. The transgenic plant of claim 10 , wherein the last leaf with epicuticular wax is produced later during plant development relative to that found in an otherwise isogenic plant lacking the recombinant vector.
17. A cell transformed with the recombinant vector of claim 3 .
18. A method of altering the timing of juvenile to adult phase change in a plant, the method comprising expressing the construct of claim 3 in the plant or expressing the construct of claim 9 in the plant.
19. The method of claim 18 , wherein the timing of juvenile to adult phase change is calculated by a method comprising counting the last leaf displaying epicuticular wax.
20. The method of claim 18 , wherein the plant exhibits an altered trait selected from the group consisting of: an increase of at least one in the numbering of the last leaf which displays epicuticular wax or which does not contain abaxial trichomes; an altered proportion of juvenile, transitional, or adult leaves; enhanced yield of vegetative tissue; enhanced digestibility of vegetative tissue; enhanced resistance to a plant pest; and enhanced resistance to a plant disease, wherein the trait exhibited by the plant is altered relative to a wild type plant.
21. The method of claim 18 , wherein the plant has altered development or morphology when compared to a wild type plant, further wherein the plant displays an altered trait selected from the group consisting of: enhanced disease resistance, enhanced insect resistance, improved forage digestibility, enhanced abiotic stress tolerance, and improved utility for biofuel production, wherein the development, morphology, or trait is altered relative to a wild-type plant.
22. A method of producing plant biomass, the method comprising:
(a) obtaining a plant according to claim 10 ; and
(b) preparing biomass from said plant or a part thereof.
23. The method of claim 22 , further comprising producing biofuel, food or feed from the biomass.
24. The recombinant vector of claim 3 , further comprising an additional polynucleotide sequence that encodes all or part of a sequence of Glossy15 or Cg1.
25. The recombinant vector of claim 9 , further comprising an additional polynucleotide sequence which, after being transcribed, regulates the timing of the juvenile to adult phase change in a plant.
26. A transgenic plant comprising the recombinant vector of claim 3 .
27. A transgenic plant comprising the recombinant vector of claim 25 .
28. A seed of the transgenic plant of claim 26 wherein the seed comprises the recombinant vector.
29. The method of claim 18 , further comprising modulating the expression of at least a second gene which regulates the timing of the juvenile to adult phase change in a plant.
30. The method of claim 29 , wherein the second gene is selected from the group consisting of: Glossy15 and Cg1.
31. The method of claim 29 , wherein the timing of juvenile to adult phase in the plant is extended relative to a wild type plant.
32. The polynucleotide molecule of claim 1 , wherein the nucleotide sequence encodes a polypeptide that comprises an amino acid sequence that is at least 99% identical over its full length to the amino acid sequence of SEQ ID NO: 17.
33. The polynucleotide molecule of claim 1 , wherein the nucleotide sequence comprises at least 95% sequence identity over its full length to the full length of SEQ ID NO:16.