DECREASING GENE EXPRESSION FOR INCREASED PROTEIN CONTENT IN PLANTS

1 . A plant or plant part comprising decreased activity of a protein-related polypeptide compared to a control plant or plant part, wherein said plant or plant part comprises a genetic mutation that decreases the activity of said protein-related polypeptide, and wherein said protein-related polypeptide is selected from the group consisting of cinnamyl-alcohol dehydrogenase 1 (CAD1), cinnamyl-alcohol dehydrogenase (CAD), stomatal cytokinesis defective 2 (SCD2), stomatal cytokinesis defective 2A (SCD2A), stomatal cytokinesis defective 2B (SCD2B), response to dehydration 22 (RD22), glucuronidase 3 (GUS3), glucuronidase 3A (GUS3A), glycosyl hydrolase family 10 protein B (GH10B), protein phosphatase 2A beta subunit (PP2AB), protein phosphatase 2A beta subunit A (PP2ABA), protein phosphatase 2A beta subunit B (PP2ABB), alpha/beta-hydrolases superfamily protein (ABH), alpha/beta-hydrolases superfamily protein A (ABHA), alpha/beta-hydrolases superfamily protein B (ABHB), calmodulin-binding transcription activator protein 2 (CAMTA2), calmodulin-binding transcription activator protein 2A (CAMTA2A), calmodulin-binding transcription activator protein 2B (CAMTA2B), beta-ketoacyl reductase 1 (KCR1), beta-ketoacyl reductase 1A (KCR1A), and beta-ketoacyl reductase 1B (KCR1B).

2 . (canceled)

3 . The plant or plant part of claim 1 , comprising increased protein content and/or white flake protein content compared to a control plant or plant part.

4 . The plant or plant part of claim 1 , wherein the mutation comprises one or more insertions, substitutions, or deletions in at least one native protein-related gene or homolog thereof or in a regulatory region thereof in said plant or plant part, wherein said at least one protein-related gene or homolog encodes said protein-related polypeptide, and wherein (a)_an expression level of said at least one protein-related gene or homolog thereof is reduced compared to an expression level the gene or homolog thereof in a plant or plant part without said mutation, and/or (b) said mutation reduces level or activity of said protein-related polypeptide compared to level or activity of a copy of said protein-related polypeptide in a plant or plant part without said mutation.

5 . (canceled)

6 . The plant or plant part of claim 4 , wherein:

(a) the mutation is located at least partially in the regulatory region, a promoter region, or 5′ untranslated region (5′UTR) of said at least one native protein-related gene or homolog thereof, wherein said at least one protein-related gene is at least one copy of CAD1, SCD2, SCD2A, SCD2B, RD22, GUS3, GUS3-A, GH10-B, PP2AB, PP2AB-A, PP2AB-B, A/BH, A/BH-A, A/BH-B, CAMTA2, CAMTA2-A, CAMTA2-B, KCR1, KCR1A, or KCR1B gene; and/or

(b) the mutation is located at least partially in a protein-related gene or regulatory region thereof, wherein:

(i) said protein-related gene comprises a nucleic acid sequence having at least 80% sequence identity to a nucleic acid sequence of any one of SEQ ID NOs: 1-15, wherein said nucleic acid sequence encodes a polypeptide that retains protein-related polypeptide activity;

(ii) said protein-related gene comprises the nucleic acid sequence of any one of SEQ ID NOs: 1-15;

(iii) said protein-related gene encodes a polypeptide comprising an amino acid sequence having at least 80% sequence identity to an amino acid sequence of any one of SEQ ID NOs: 16-30, wherein said polypeptide retains protein-related polypeptide activity;

(iv) said protein-related gene encodes a polypeptide comprising an amino acid sequence of any one of SEQ ID NOs: 16-30;

(v) said protein-related gene including said regulatory region thereof comprises a nucleic acid sequence having at least 80% sequence identity to a nucleic acid sequence of any one of SEQ ID NOs: 1-15, wherein said nucleic acid sequence encodes a polypeptide that retains protein-related polypeptide activity; and/or

(vi) said protein-related gene including said regulatory region thereof comprises the nucleic acid sequence of any one of SEQ ID NOs: 1-15.

7 - 9 . (canceled)

10 . The plant or plant part of claim 4 , comprising:

(i) a deletion of one or more nucleotides of SEQ ID NO: 1 in the Glycine max SCD2A gene;

(ii) a deletion of one or more nucleotides of SEQ ID NO: 2 in the Glycine max SCD2B gene;

(iii) a deletion of one or more nucleotides of SEQ ID NO: 3 in the Glycine max RD22 gene;

(iv) a deletion of one or more nucleotides of SEQ ID NO: 4 in the Glycine max GUS3-A gene;

(v) a deletion of one or more nucleotides of SEQ ID NO: 5 in the Glycine max GH10-B gene;

(vi) a deletion of one or more nucleotides of SEQ ID NO: 6 in the Glycine max PP2AB-A gene;

(vii) a deletion of one or more nucleotides of SEQ ID NO: 7 in the Glycine max PP2AB-B gene;

(viii) a deletion of one or more nucleotides of SEQ ID NO: 8 in the Glycine max A/BH-A gene;

(ix) a deletion of one or more nucleotides of SEQ ID NO: 9 in the Glycine max A/BH-B gene;

(x) a deletion of one or more nucleotides of SEQ ID NO: 10 in the Glycine max CAMTA2-A gene;

(xi) a deletion of one or more nucleotides of SEQ ID NO: 11 in the Glycine max CAMTA2-B gene;

(xii) a deletion of one or more nucleotides of SEQ ID NO: 12 in the Glycine max CAD1 gene;

(xiii) a deletion of one or more nucleotides of SEQ ID NO: 13 in the Glycine max CAD1 gene;

(xiv) a deletion of one or more nucleotides of SEQ ID NO: 14 in the Glycine max KCR1A gene;

(xv) a deletion of one or more nucleotides of SEQ ID NO: 15 in the Glycine max KCR1B gene;

(xvi) a polynucleotide comprising a nucleic acid sequence of SEQ ID: 60, or a deletion of nucleotides 428-431 of SEQ ID NO: 12 in the Glycine max CAD1 gene;

(xvii) a polynucleotide comprising a nucleic acid sequence of SEQ ID: 61, or a deletion of nucleotides 447-456 of SEQ ID NO: 13 in the Glycine max CAD1 gene;

(xviii) a polynucleotide comprising a nucleic acid sequence of SEQ ID: 62, or a deletion of nucleotides 416-429 of SEQ ID NO: 12 in the Glycine max CAD1 gene;

(xix) a polynucleotide comprising a nucleic acid sequence of SEQ ID: 63, or a deletion of nucleotides 452-458 of SEQ ID NO: 13 in the Glycine max CAD1 gene;

(xx) a polynucleotide comprising a nucleic acid sequence of SEQ ID: 60, or a deletion of nucleotides 428-431 of SEQ ID NO: 12 in the Glycine max CAD1 gene, and a polynucleotide comprising a nucleic acid sequence of SEQ ID: 61, or a deletion of nucleotides 447-456 of SEQ ID NO: 13 in the Glycine max CAD1 gene; and/or

(xxi) a polynucleotide comprising a nucleic acid sequence of SEQ ID: 62, or a deletion of nucleotides 416-429 of SEQ ID NO: 12 in the Glycine max CAD1 gene, and a polynucleotide comprising a nucleic acid sequence of SEQ ID: 63, or a deletion of nucleotides 452-458 of SEQ ID NO: 13 in the Glycine max CAD1 gene.

11 .- 21 . (canceled)

22 . A population of plants or plant parts comprising the plant or plant part of claim 1 , wherein the population comprises decreased activity of said protein-related polypeptide and/or increased protein content and/or white flake protein content compared to a control population.

23 . (canceled)

24 . A method for increasing protein content and/or white flake protein content in a plant or plant part, said method comprising reducing level or activity of at least one endogenous gene encoding a protein-related polypeptide in said plant or plant part, wherein said protein-related polypeptide is selected from the group consisting of cinnamyl-alcohol dehydrogenase 1 (CAD1), cinnamyl-alcohol dehydrogenase (CAD), stomatal cytokinesis defective 2 (SCD2), stomatal cytokinesis defective 2A (SCD2A), stomatal cytokinesis defective 2B (SCD2B), response to dehydration 22 (RD22), glucuronidase 3 (GUS3), glucuronidase 3A (GUS3A), glycosyl hydrolase family 10 protein B (GH10B), protein phosphatase 2A beta subunit (PP2AB), protein phosphatase 2A beta subunit A (PP2ABA), protein phosphatase 2A beta subunit B (PP2ABB), alpha/beta-hydrolases superfamily protein (ABH), alpha/beta-hydrolases superfamily protein A (ABHA), alpha/beta-hydrolases superfamily protein B (ABHB), calmodulin-binding transcription activator protein 2 (CAMTA2), calmodulin-binding transcription activator protein 2A (CAMTA2A), calmodulin-binding transcription activator protein 2B (CAMTA2B), beta-ketoacyl reductase 1 (KCR1), beta-ketoacyl reductase 1A (KCR1A), and beta-ketoacyl reductase 1B (KCR1B).

25 . A method for increasing protein content and/or white flake protein content in a plant or plant part, said method comprising introducing a genetic mutation that decreases activity of a protein-related polypeptide into said plant or plant part, wherein said protein-related polypeptide is selected from the group consisting of cinnamyl-alcohol dehydrogenase 1 (CAD1), cinnamyl-alcohol dehydrogenase (CAD), stomatal cytokinesis defective 2 (SCD2), stomatal cytokinesis defective 2A (SCD2A), stomatal cytokinesis defective 2B (SCD2B), response to dehydration 22 (RD22), glucuronidase 3 (GUS3), glucuronidase 3A (GUS3A), glycosyl hydrolase family 10 protein B (GH10B), protein phosphatase 2A beta subunit (PP2AB), protein phosphatase 2A beta subunit A (PP2ABA), protein phosphatase 2A beta subunit B (PP2ABB), alpha/beta-hydrolases superfamily protein (ABH), alpha/beta-hydrolases superfamily protein A (ABHA), alpha/beta-hydrolases superfamily protein B (ABHB), calmodulin-binding transcription activator protein 2 (CAMTA2), calmodulin-binding transcription activator protein 2A (CAMTA2A), calmodulin-binding transcription activator protein 2B (CAMTA2B), beta-ketoacyl reductase 1 (KCR1), beta-ketoacyl reductase 1A (KCR1A), and beta-ketoacyl reductase 1B (KCR1B).

26 . (canceled)

27 . The method of claim 25 , further comprising introducing the genetic mutation that decreases activity of said protein-related polypeptide into a plant cell, and regenerating said plant or plant part from said plant cell.

28 . (canceled)

29 . The method of claim 25 , wherein (a) the mutation is introduced to locate at least partially in the regulatory region, a promoter region, or 5′ untranslated region (5′UTR) of said at least one native protein-related gene or homolog thereof, wherein said at least one protein-related gene is at least one native CAD1, SCD2, SCD2A, SCD2B, RD22, GUS3, GUS3-A, GH10-B, PP2AB, PP2AB-A, PP2AB-B, A BH, A BH-A, A BH-B, CAMTA2, CAMTA2-A, CAMTA2-B, KCR1, KCR1A, or KCR1B gene; and/or

(b) the mutation is introduced at least partially into a protein-related gene or regulatory region thereof, wherein:

(i) said protein-related gene comprises a nucleic acid sequence having at least 80% sequence identity to a nucleic acid sequence of any one of SEQ ID NOs: 1-15, wherein said nucleic acid sequence encodes a polypeptide that retains protein-related polypeptide activity;

(ii) said protein-related gene comprises the nucleic acid sequence of any one of SEQ ID NOs: 1-15;

(iii) said protein-related gene encodes a polypeptide comprising an amino acid sequence having at least 80% sequence identity to an amino acid sequence of any one of SEQ ID NOs: 16-30, wherein said polypeptide retains protein-related polypeptide activity;

(iv) said protein-related gene encodes a polypeptide comprising an amino acid sequence of any one of SEQ ID NOs: 16-30;

(v) said protein-related gene including said regulatory region thereof comprises a nucleic acid sequence having at least 80% sequence identity to a nucleic acid sequence of any one of SEQ ID NOs: 1-15, wherein said nucleic acid sequence encodes a polypeptide that retains protein-related polypeptide activity; and/or

(vi) said protein-related gene including said regulatory region thereof comprises the nucleic acid sequence of any one of SEQ ID NOs: 1-15.

30 .- 32 . (canceled)

33 . The method of claim 25 , wherein:

(i) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 1 in the Glycine max SCD2A gene;

(ii) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 2 in the Glycine max SCD2B gene;

(iii) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 3 in the Glycine max RD22 gene;

(iv) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 4 in the Glycine max GUS3-A gene;

(v) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 5 in the Glycine max GH10-B gene;

(vi) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 6 in the Glycine max PP2AB-A gene;

(vii) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 7 in the Glycine max PP2AB-B gene;

(viii) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 8 in the Glycine max A/BH-A gene;

(ix) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 9 in the Glycine max A/BH-B gene;

(x) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 10 in the Glycine max CAMTA2-A gene;

(xi) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 11 in the Glycine max CAMTA2-B gene;

(xii) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 12 in the Glycine max CAD1 gene;

(xiii) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 13 in the Glycine max CAD1 gene;

(xiv) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 14 in the Glycine max KCR1A gene;

(xv) the mutation comprises a deletion of one or more nucleotides of SEQ ID NO: 15 in the Glycine max KCR1B gene;

(xvi) the mutation comprises a deletion of nucleotides 428-431 of SEQ ID NO: 12 in the Glycine max CAD1 gene, or said plant or plant part comprises SEQ ID NO: 60 when said mutation is introduced;

(xvii) the mutation comprises a deletion of nucleotides 447-456 of SEQ ID NO: 13 in the Glycine max CAD1 gene, or said plant or plant part comprises SEQ ID NO: 61 when said mutation is introduced;

(xviii) the mutation comprises a deletion of nucleotides 416-429 of SEQ ID NO: 12 in the Glycine max CAD1 gene, or said plant or plant part comprises SEQ ID NO: 62 when said mutation is introduced; and/or

(xix) the mutation comprises a deletion of nucleotides 452-458 of SEQ ID NO: 13 in the Glycine max CAD1 gene, or said plant or plant part comprises SEQ ID NO: 63 when said mutation is introduced;

(xx) the mutation comprises a deletion of nucleotides 428-431 of SEQ ID NO: 12 in the Glycine max CAD1 gene and a deletion of nucleotides 447-456 of SEQ ID NO: 13 in the Glycine max CAD1 gene, or said plant or plant part comprises SEQ ID NOs: 60 and 61 when said mutation is introduced; and/or

(xxi) the mutation comprises a deletion of nucleotides 416-429 of SEQ ID NO: 12 in the Glycine max CAD1 gene and a deletion of nucleotides 452-458 of SEQ ID NO: 13 in the Glycine max CAD1 gene, or said plant or plant part comprises SEQ ID NO: 62 and 63 when said mutation is introduced.

34 .- 37 . (canceled)

38 . The method of claim 28 , further comprising introducing editing reagents or a nucleic acid construct encoding said editing reagents into said plant, plant part, or plant cell.

39 . The method of claim 38 , wherein said editing reagents comprise at least one nuclease and/or one or more guide RNAs (gRNAs),

wherein the one or more gRNAs comprise a nucleic acid sequence complementary to a region of a genomic DNA sequence encoding said protein-related polypeptide or regulating transcription or translation of said protein-related polypeptide in said plant or plant part, and/or

wherein the at least one nuclease cleaves a target site in said at least one protein-related gene or homolog thereof or a regulatory region thereof in said plant, plant part, or plant cell, and said mutation is introduced at said cleaved target site.

40 .- 43 . (canceled)

44 . The method of claim 39 , wherein at least one of the one or more gRNAs comprises a nucleic acid sequence encoded by:

(i) a nucleic acid sequence that shares at least 80% sequence identity with a nucleic acid sequence of any one of SEQ ID NO: 1-15 and 57; or

(ii) the nucleic acid sequence of any one of SEQ ID NO: 1-15 and 57.

45 .- 48 . (canceled)

49 . A plant or plant part produced by the method of claim 25 , wherein said plant or plant part comprises (a)_reduced activity of said protein-related polypeptide compared to a control plant or plant part, and/or (b) increased protein content and/or white flake protein content compared to a control plant or plant part.

50 .- 51 . (canceled)

52 . A population of plants or plant parts produced by the method of claim 25 , wherein the population comprises (a) decreased activity of said protein-related polypeptide and/or (b) increased protein content and/or white flake protein content compared to a control population.

53 . (canceled)

54 . A composition produced from the plant, plant part, or population plants or plant parts of claim 1 , wherein the composition is a seed composition, a protein composition, of a food or beverage product.

55 .- 56 . (canceled)

57 . A nucleic acid molecule comprising a nucleic acid sequence of a mutated protein-related gene or coding sequence thereof, wherein said nucleic acid sequence comprises any one of SEQ ID NOs: 1-15 and 31-45 comprising one or more insertions, substitutions, or deletions therein, or any one of SEQ ID NOs: 60-63.

58 . (canceled)

59 . A DNA construct comprising, in operable linkage:

(i) a promoter that is functional in a plant cell; and

(ii) the nucleic acid molecule of claim 57 .

60 . A cell comprising the nucleic acid molecule of claim 57 , or the DNA construct comprising the nucleic acid molecule of claim 57 .

61 . (canceled)