REGULATORY POLYNUCLEOTIDES AND USES THEREOF
The present disclosure provides compositions and methods for regulating expression of transcribable polynucleotides in plant cells, plant tissues, and plants. Compositions include regulatory polynucleotide molecules capable of providing expression in plant tissues and plants. Methods for expressing polynucleotides in a plant cell, plant tissue, or plants using the regulatory polynucleotide molecules disclosed herein are also provided.
1 . An isolated regulatory polynucleotide comprising a polynucleotide molecule selected from the group consisting of
(a) a polynucleotide molecule comprising a nucleic acid molecule having a sequence selected from the group consisting of SEQ ID NOS: 1-105 that is capable of regulating transcription of an operably linked transcribable polynucleotide molecule;
(b) a polynucleotide molecule having at least about 70% sequence identity to a sequence selected from the group consisting of SEQ ID NOS:1-105 that is capable of regulating transcription of an operably linked transcribable polynucleotide molecule; and
(c) a fragment of the polynucleotide molecule of (a) or (b) capable of regulating transcription of an operably linked transcribable polynucleotide molecule.
2 . The isolated regulatory polynucleotide of claim 1 , wherein the molecule is (a) a polynucleotide molecule comprising a nucleic acid molecule having the sequence selected from the group consisting of SEQ ID NOS: 1-105 that is capable of regulating transcription of an operably linked transcribable polynucleotide molecule.
3 . The isolated regulatory polynucleotide of claim 1 , wherein the regulatory polynucleotide is capable of regulating tissue-specific transcription.
4 . The isolated regulatory polynucleotide of claim 1 , wherein the regulatory polynucleotide is capable of regulating root-specific transcription.
5 . (canceled)
6 . The isolated regulatory polynucleotide of claim 4 , wherein the regulatory polynucleotide is capable of regulating transcription in epidermal, vascular, cortex, phloem, pericycle, endodermal, meristem root cap tissues, or combinations thereof.
7 - 9 . (canceled)
10 . The isolated regulatory polynucleotide of claim 1 , wherein the molecule is (b) a polynucleotide molecule having at least about 70% sequence identity to a sequence selected from the group consisting of SEQ ID NOS:1-105 that is capable of regulating transcription of an operably linked transcribable polynucleotide molecule.
11 - 21 . (canceled)
22 . The isolated regulatory polynucleotide of claim 1 , wherein the polynucleotide molecule is (c) a fragment of the polynucleotide molecule of (a) or (b) capable of regulating transcription of an operably linked transcribable polynucleotide molecule.
23 - 28 . (canceled)
29 . A recombinant polynucleotide construct comprising the regulatory polynucleotide of claim 1 operably linked to a heterologous transcribable polynucleotide molecule.
30 . The recombinant polynucleotide construct of claim 29 , wherein the transcribable polynucleotide molecule encodes a protein of agronomic interest.
31 . The recombinant polynucleotide construct of claim 29 , wherein the transcribable polynucleotide molecule is operably linked to a 3′ transcription termination polynucleotide molecule.
32 . A chimeric polynucleotide molecule comprising:
(a) a first polynucleotide molecule selected from the group consisting of
(i) a polynucleotide molecule comprising a nucleic acid molecule having a sequence selected from the group consisting of SEQ ID NOS: 1-105 that is capable of regulating transcription of an operably linked transcribable polynucleotide molecule;
(ii) a polynucleotide molecule having at least about 70% sequence identity to a sequence selected from the group consisting of SEQ ID NOS:1-105 that is capable of regulating transcription of an operably linked transcribable polynucleotide molecule; and
(iii) a fragment of the polynucleotide molecule of (a) or (b) capable of regulating transcription of an operably linked transcribable polynucleotide molecule, and
(b) a second polynucleotide molecule capable of regulating transcription of an operably linked polynucleotide molecule, wherein the first polynucleotide molecule is operably linked to the second polynucleotide molecule.
33 . The chimeric polynucleotide of claim 32 , wherein the first polynucleotide molecule comprises a core promoter molecule and the second polynucleotide molecule is selected from the group consisting of a cis-element, an enhancer element, and an intron.
34 . The chimeric polynucleotide of claim 32 , wherein the first polynucleotide molecule is selected from the group consisting of a cis-element, an enhancer element, and an intron and the second polynucleotide molecule comprises a core promoter molecule.
35 . The chimeric polynucleotide of claim 32 , wherein the first polynucleotide molecule comprises an intron.
36 . The chimeric polynucleotide of claim 32 , wherein the second polynucleotide molecule is heterologous to the first polynucleotide molecule.
37 . The chimeric polynucleotide of claim 32 , wherein the first polynucleotide molecule is (iii) a fragment of the polynucleotide molecule of (i) or (ii) capable of regulating transcription of an operably linked transcribable polynucleotide molecule and the second polynucleotide molecule is a heterologous core promoter sequence.
38 . A transgenic host cell comprising the recombinant polynucleotide construct of claim 29 .
39 . The transgenic host cell of claim 38 , wherein the host cell is a plant cell.
40 . A transgenic plant stably transformed with the recombinant polynucleotide construct of claim 29 .
41 . The transgenic plant of claim 40 , wherein the plant is selected from the group consisting of a monocotyledonous and a dicotyledonous plant.
42 . The transgenic plant of claim 41 , wherein the plant is a monocotyledonous plant selected from the group consisting of wheat, corn, rice, turf grass, millet, sorghum, switchgrass, miscanthus, sugarcane, and Brachypodium.
43 . The transgenic plant of claim 41 , wherein the plant is a dicotyledonous plant selected from the group consisting of soybean, cotton, canola, and potato.
44 . Seed produced by the transgenic plant of claim 40 .
45 . An isolated polynucleotide molecule comprising a regulatory element derived from SEQ ID NOs: 1-105, wherein the regulatory element is capable of regulating transcription of an operably linked transcribable polynucleotide molecule.
46 . The isolated polynucleotide molecule of claim 45 , wherein the regulatory element is in operable linkage with a core promoter sequence.
47 . (canceled)
48 . The isolated polynucleotide molecule of claim 45 , wherein the regulatory element is selected from the group consisting of core promoter regions, cis-elements, introns, and leader sequences.
49 . The isolated polynucleotide molecule of claim 48 , wherein the regulatory element is an intron capable of enhancing the transcription of the operably linked transcribable polynucleotide molecule.
50 . A method of directing expression of a transcribable polynucleotide molecule in a host cell comprising:
(a) introducing the recombinant polynucleotide construct of claim 29 into a host cell to produce a transgenic host cell; and
(b) selecting a transgenic host cell exhibiting expression of the transcribable polynucleotide molecule.
51 . The method of claim 50 , wherein the transcribable polynucleotide molecule is selected from the group consisting of a coding sequence and a functional RNA.
52 . The method of claim 50 , wherein the host cell is a plant cell.
53 . The method of claim 52 , further comprising regenerating a plant comprising the introduced recombinant nucleic acid construct.
54 . A method of directing expression of a transcribable polynucleotide molecule in a plant comprising:
(a) introducing the recombinant polynucleotide construct of claim 29 into a plant cell;
(b) regenerating a plant from the plant cell; and
(c) selecting a transgenic plant exhibiting expression of the transcribable polynucleotide molecule.
55 . The method of claim 54 , wherein the transcribable polynucleotide molecule is selected from the group consisting of a coding sequence and a functional RNA.