Composition and method for producing tobacco plant and product having reduced or eliminated sucker

Abstract

To provide methods and compositions for controlling sucker growth in tobacco by altering the expression of different target genes.SOLUTION: A modified tobacco plant comprises a mutation in an endogenous gene encoding polypeptide selected from the group consisting of cyclin, cyclin-dependent kinase (CDK), a CDK inhibitor, MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and wherein the modified tobacco plant comprises no suckers or reduced suckers after topping as compared to the control tobacco plant when grown under comparable growth conditions.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 62 / 854,139, filed May 29, 2019, the entire contents of which are incorporated herein by reference.

[0002] Field The present disclosure provides methods and compositions for improving the expression of nucleic acids and proteins useful for reducing or eliminating suckers in plants.

[0003] Incorporating a sequence listing This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy, created on May 21, 2020, is titled P34703WO00_SL.txt and is 380,869 bytes in size. [Background technology]

[0004] background Tobacco is a plant species that exhibits very strong apical dominance. Molecular signals from the shoot apical meristem (SAM) mediate hormonal signals that effectively inhibit axillary bud growth. Removal of the SAM (also known as "topping") induces physiological and molecular changes that allow the growth of new shoots (or "suckers") from the axillary meristem (bud). Sucker growth results in reduced yield and leaf quality. Suckers have been removed by manual removal and chemical application. Maleic hydrazide and flumetralin are routinely used on topped plants to inhibit axillary bud growth ("sucker development"). However, the labor and chemical costs of removing suckers are very high. Suppressing sucker development in tobacco through conventional breeding, mutation breeding, and transgenic approaches has been a major goal for decades, but to date, these approaches have not successfully inhibited or eliminated sucker development. Although recent molecular studies have produced transgenic plants with reduced or eliminated suckers, leaky expression of axillary bud decomposition genes can result in seed and embryo death, preventing the production of successive generations of transgenic plants. Thus, the development of methods and compositions for preventing the expression of axillary bud decomposition genes in undesired tissues and / or organs would result in reduced use of chemical agents and reduce the costs and labor associated with tobacco production. Summary of the Invention

[0005] overview In one aspect, the present disclosure provides a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to the control tobacco plant.

[0006] In one aspect, the present disclosure provides a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety.

[0007] In one aspect, the disclosure provides a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor operably linked to a heterologous promoter, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions.

[0008] In one aspect, the present disclosure provides a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter.

[0009] In one aspect, the present disclosure provides a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions.

[0010] In one aspect, the present disclosure provides a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor.

[0011] In one aspect, the present disclosure provides a tobacco product comprising cured tobacco material from a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to the control tobacco plant.

[0012] In one aspect, the present disclosure provides a tobacco product comprising cured tobacco material from a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety.

[0013] In one aspect, the present disclosure provides a tobacco product comprising cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter, wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant.

[0014] In one aspect, the present disclosure provides a tobacco product comprising cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter.

[0015] In one aspect, the present disclosure provides a tobacco product comprising cured tobacco material from a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant.

[0016] In one aspect, the present disclosure provides a tobacco product comprising cured tobacco material from a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor.

[0017] In one aspect, the present disclosure provides a method for producing a modified tobacco plant, comprising: (a) inducing a mutation in at least one tobacco cell at a genomic locus encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; (b) selecting at least one tobacco cell containing the mutation from step (a); and (c) regenerating a modified tobacco plant from the at least one tobacco cell selected in step (b), wherein the modified tobacco plant contains no or reduced suckers after topping when grown under equivalent growing conditions compared to a control tobacco plant lacking the mutation. The present invention provides a method comprising:

[0018] In one aspect, the present disclosure provides a method for producing a modified tobacco plant, comprising: (a) introducing a recombinant DNA construct into at least one tobacco cell, wherein the recombinant DNA construct comprises a heterologous promoter operably linked to a nucleic acid encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; (b) selecting at least one tobacco cell comprising the recombinant DNA construct; and (c) regenerating a modified tobacco plant from the at least one tobacco cell selected in step (b), wherein the modified tobacco plant, when grown under equivalent growing conditions, comprises no or reduced suckers after topping compared to a control tobacco plant lacking the recombinant DNA construct. The present invention provides a method comprising:

[0019] In one aspect, the present disclosure provides a method for producing a modified tobacco plant, comprising: (a) introducing a recombinant DNA construct into at least one tobacco cell, wherein the recombinant DNA construct comprises a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; (b) selecting at least one tobacco cell comprising the recombinant DNA construct; and (c) regenerating a modified tobacco plant from the at least one tobacco cell selected in step (b), wherein the modified tobacco plant, when grown under equivalent growing conditions, comprises no or reduced suckers after topping compared to a control tobacco plant lacking the recombinant DNA construct. The present invention provides a method comprising:

[0020] In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and wherein the modified tobacco plant, when grown under equivalent growing conditions, comprises no or reduced suckers after topping compared to the control tobacco plant.

[0021] In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter, wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant.

[0022] In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter, wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant.

[0023] In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter, wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant.

[0024] In one aspect, the present disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter.

[0025] In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant.

[0026] In one aspect, the present disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor.

[0027] In one aspect, the disclosure provides a method comprising transforming a tobacco cell with a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor.

[0028] In one aspect, the disclosure provides a method comprising transforming a tobacco cell with a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor.

[0029] In one aspect, the present disclosure provides a method for producing a modified tobacco plant, comprising: (a) crossing at least one tobacco plant of a first tobacco variety with at least one tobacco plant of a second tobacco variety, wherein the at least one tobacco plant of the first tobacco variety comprises a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and wherein the at least one tobacco plant, when grown under equivalent growing conditions, produces no or reduced suckers after topping compared to the control tobacco plant; and (b) selecting progeny tobacco plants that produce no or reduced suckers after topping compared to the control tobacco plant of the same hybrid grown under equivalent growing conditions. The present invention provides a method comprising:

[0030] In one aspect, the present disclosure provides a method for producing a modified tobacco plant, comprising: (a) crossing at least one tobacco plant of a first tobacco variety with at least one tobacco plant of a second tobacco variety, wherein the at least one tobacco plant of the first tobacco variety comprises a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, and wherein the at least one tobacco plant, when grown under equivalent growing conditions, exhibits no or reduced suckers after topping compared to a control tobacco plant; and (b) selecting progeny tobacco plants that exhibit no or reduced suckers after topping compared to a control tobacco plant of the same hybrid grown under equivalent growing conditions. The present invention provides a method comprising:

[0031] In one aspect, the present disclosure provides a method for producing a modified tobacco plant, comprising: (a) crossing at least one tobacco plant of a first tobacco variety with at least one tobacco plant of a second tobacco variety, wherein the at least one tobacco plant of the first tobacco variety comprises a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous genomic locus encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, and wherein the at least one tobacco plant, when grown under equivalent growing conditions, produces no or reduced suckers after topping compared to a control tobacco plant; and (b) selecting progeny tobacco plants that produce no or reduced suckers after topping compared to a control tobacco plant of the same hybrid grown under equivalent growing conditions. The present invention provides a method comprising: [The present invention 1001] 1. A modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, A modified tobacco plant, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and wherein the modified tobacco plant, when grown under equivalent cultivation conditions, has no or reduced suckers after topping compared to the control tobacco plant. [The present invention 1002] 1. A modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, A modified tobacco plant, wherein said mutation is not present in said endogenous gene in a control tobacco plant of the same variety. [The present invention 1003] 1. A modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter, A modified tobacco plant, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions. [The present invention 1004] A modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter. [The present invention 1005] 1. A modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; A modified tobacco plant, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions. [The present invention 1006] A recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing the expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor. A modified tobacco plant, comprising: [The present invention 1007] The modified tobacco plant of the present invention 1001 or 1002, wherein the endogenous gene comprises a polynucleotide sequence at least 80% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1 to 44 and 113. [The present invention 1008] The modified tobacco plant of the present invention 1001 or 1002, wherein the endogenous gene encodes a polypeptide comprising an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45 to 88 and 114. [The present invention 1009] The modified tobacco plant of any of claims 1003 to 1006, wherein the heterologous promoter comprises a polynucleotide sequence that is at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 89 to 109. [The present invention 1010] The modified tobacco plant of claim 1005 or 1006, wherein the small RNA molecule comprises a polynucleotide sequence having at least 90% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. [The present invention 1011] A tobacco leaf of the modified tobacco plant of any one of 1001 to 1006 of the present invention. [The present invention 1012] A tobacco seed of the modified tobacco plant of any one of 1001 to 1006 of the present invention. [The present invention 1013] The tobacco leaf of the present invention 1054, wherein the tobacco leaf is a dried tobacco leaf. [The present invention 1014] A tobacco product comprising an alkaloid extracted from the modified tobacco plant of any one of claims 1001 to 1006 or a part thereof. [The present invention 1015] 1. A tobacco product comprising cured tobacco material from a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, A tobacco product wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to the control tobacco plant. [The present invention 1016] 1. A tobacco product comprising cured tobacco material from a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, A tobacco product, wherein said mutation is not present in said endogenous gene in a control tobacco plant of the same variety. [The present invention 1017] 1. A tobacco product comprising cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter, A tobacco product in which the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions. [The present invention 1018] A tobacco product comprising cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter. [The present invention 1019] 1. A tobacco product comprising cured tobacco material from a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, A tobacco product in which the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions. [The present invention 1020] A tobacco product comprising cured tobacco material from a modified tobacco plant containing a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of cyclin, cyclin-dependent kinase (CDK), CDK inhibitor, MYB, and WRKY transcription factor. [The present invention 1021] 1. A method for producing a modified tobacco plant, comprising: (a) inducing a mutation in at least one tobacco cell in a genomic locus encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; (b) selecting at least one tobacco cell containing the mutation from step (a); (c) regenerating a modified tobacco plant from the at least one tobacco cell selected in step (b), wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant lacking the mutation. [The present invention 1022] 1. A method for producing a modified tobacco plant, comprising: (a) introducing a recombinant DNA construct into at least one tobacco cell, wherein the recombinant DNA construct comprises a heterologous promoter operably linked to a nucleic acid encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; (b) selecting at least one tobacco cell containing the recombinant DNA construct; (c) regenerating a modified tobacco plant from the at least one tobacco cell selected in step (b), wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant lacking the recombinant DNA construct. [The present invention 1023] 1. A method for producing a modified tobacco plant, comprising: (a) introducing a recombinant DNA construct into at least one tobacco cell, wherein the recombinant DNA construct comprises a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; (b) selecting at least one tobacco cell containing the recombinant DNA construct; (c) regenerating a modified tobacco plant from the at least one tobacco cell selected in step (b), wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant lacking the recombinant DNA construct. [The present invention 1024] Preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor. A method comprising: wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to the control tobacco plant. [The present invention 1025] preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter; A method comprising: The method, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions. [The present invention 1026] preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter; A method comprising: The method, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions. [The present invention 1027] preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter; A method comprising: [The present invention 1028] preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; A method comprising: The method, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions. [The present invention 1029] preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; A method comprising: [The present invention 1030] transforming tobacco cells with a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; A method comprising: [The present invention 1031] Transforming tobacco cells with a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of cyclin, cyclin-dependent kinase (CDK), CDK inhibitor, MYB, and WRKY transcription factor. A method comprising: [The present invention 1032] 1. A method for producing a modified tobacco plant, comprising: (a) crossing at least one tobacco plant of a first tobacco variety with at least one tobacco plant of a second tobacco variety, wherein said at least one tobacco plant of said first tobacco variety comprises a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, said mutation being absent in said endogenous gene in a control tobacco plant of the same variety, and wherein said at least one tobacco plant, when grown under equivalent growing conditions, comprises no or reduced suckers after topping compared to said control tobacco plant; (b) selecting progeny tobacco plants that exhibit no or reduced suckers after topping compared to control tobacco plants of the same hybrid grown under comparable growing conditions; [The present invention 1033] 1. A method for producing a modified tobacco plant, comprising: (a) crossing at least one tobacco plant of a first tobacco variety with at least one tobacco plant of a second tobacco variety, wherein said at least one tobacco plant of said first tobacco variety comprises a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, and wherein said at least one tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant; (b) selecting progeny tobacco plants that exhibit no or reduced suckers after topping compared to control tobacco plants of the same hybrid grown under comparable growing conditions; [The present invention 1034] 1. A method for producing a modified tobacco plant, comprising: (a) crossing at least one tobacco plant of a first tobacco variety with at least one tobacco plant of a second tobacco variety, wherein said at least one tobacco plant of said first tobacco variety comprises a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous genomic locus encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, and wherein said at least one tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant; (b) selecting progeny tobacco plants that exhibit no or reduced suckers after topping compared to control tobacco plants of the same hybrid grown under comparable growing conditions; [This invention 1035] 1003. The modified tobacco plant of the present invention, wherein the polynucleotide sequence comprises a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113.

[0032] A brief description of arrays SEQ ID NOs: 1 to 44, 89 to 113 and 118 to 126 are nucleic acid sequences.

[0033] SEQ ID NOs: 45 to 88 and 114 to 117 are amino acid sequences.

[0034] Further description of the SEQ ID NOs provided herein can be found in Table 1 below.

[0035] Table 1: Sequence description TIFF2025134829000001.tif177153TIFF2025134829000002.tif243153TIFF2025134829000003.tif242153 [Brief explanation of the drawings]

[0036] [Figure 1] Venn diagram showing overlap of differentially expressed genes in axillary buds between pairwise comparisons of water-treated and maleic hydrazide-treated tobacco plants 4 hours, 24 hours, and 72 hours after topping. MH4 refers to 4 hours after maleic hydrazide treatment, W4 refers to 4 hours after water treatment, MH24 refers to 24 hours after maleic hydrazide treatment, W24 refers to 24 hours after water treatment, MH72 refers to 72 hours after maleic hydrazide treatment, and W72 refers to 72 hours after water treatment. [Figure 2] Shown is the total mass of suckers from individual P1_2.4::MYB plants one week after topping. [Figure 3] Shown is the total mass of suckers from individual P1_2.4::CDKI plants one week after topping. [Figure 4]The total mass of suckers from individual plants containing the amiRNA-1 construct is shown one week after topping. [Figure 5] The total mass of suckers from individual plants containing the amiRNA-2 construct is shown one week after topping. [Figure 6] The total mass of suckers from individual plants containing the amiRNA-3 construct is shown one week after topping. DETAILED DESCRIPTION OF THE INVENTION

[0037] Detailed Description Unless otherwise defined, all technical and scientific terms used have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Where a term is provided in the singular, the inventors also contemplate aspects of this disclosure described by the plural of that term. In the event of a discrepancy in terms and definitions used in references incorporated by reference, the terms used in this application shall have the definitions provided herein. Other technical terms used have their ordinary meaning in the art in which they are used, as exemplified by various art-specific dictionaries, such as "The American Heritage® Science Dictionary" (Editors of the American Heritage Dictionaries, 2011, Houghton Mifflin Harcourt, Boston and New York), "McGraw-Hill Dictionary of Scientific and Technical Terms" (6th edition, 2002, McGraw-Hill, New York), or "Oxford Dictionary of Biology" (6th edition, 2008, Oxford University Press, Oxford and New York).

[0038] Any references cited herein, including, for example, all patents, published patent applications, and non-patent publications, are hereby incorporated by reference in their entirety.

[0039] When a group of alternatives is presented, all combinations of the members comprising that group of alternatives are specifically contemplated. For example, if an item is selected from the group consisting of A, B, C, and D, the inventors specifically contemplate each alternative individually (e.g., A alone, B alone, etc.), as well as combinations such as A, B, and D; A and C; B and C. The term "and / or," when used in a list of two or more items, refers to any one of the listed items alone or in combination with any one or more of the other listed items. For example, the phrase "A and / or B" is intended to mean either or both A and B, i.e., A alone, B alone, or a combination of A and B. The phrase "A, B, and / or C" is intended to mean A alone, B alone, C alone, A and B in combination, A and C in combination, B and C in combination, or A, B, and C in combination.

[0040] When a numerical range is provided herein, the range is understood to include the endpoints of the range, as well as any number between the defined endpoints of the range. For example, "1 to 10" includes any number between 1 and 10, as well as the numbers 1 and 10.

[0041] As used herein, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" can include a plurality of compounds, including mixtures thereof.

[0042] In one aspect, the modified tobacco plants provided herein include reduced or no sucker development after topping compared to control tobacco plants when grown under comparable conditions.

[0043] As used herein, "sucker development" refers to the development and / or growth of an axillary (or lateral) bud ("sucker") from an axillary meristem that grows between the leaf and the stalk. An axillary bud is an embryonic shoot that includes the axillary meristem, surrounding leaf tissue, and surrounding stem tissue. See, e.g., U.S. Patent Application Publication Nos. 2016 / 0281100; and 2017 / 0260535, which are incorporated by reference in their entireties.

[0044] As used herein, "topping" refers to the removal of the stem tip, including the shoot apical meristem, flowers, and up to some adjacent leaves, when the plant is near maturity. Tipping of tobacco plants results in the loss of apical dominance. Prior to topping, sucker development is kept largely dormant by hormonal signals emanating from the shoot apical meristem; topping can remove the hormonal signals and allow sucker elongation ("topping-induced sucker development"). If sucker development is sufficiently controlled, topping can increase yield, increase value per acre, and result in desirable modifications to the physical and chemical properties of tobacco leaves.

[0045] As used herein, "equivalent growing conditions" refers to similar environmental conditions and / or agronomic practices for growing and making meaningful comparisons between two or more plant genotypes, such that none of the environmental conditions and agronomic practices contribute to or explain any differences observed between the two or more plant genotypes. Environmental conditions include, for example, light, temperature, water, humidity, and nutrients (e.g., nitrogen and phosphorus). Agronomic practices include, for example, sowing, clipping, undercutting, transplanting, topping, and sucker development. See Chapters 4B and 4C of "Tobacco, Production, Chemistry and Technology," Davis & Nielsen eds., Blackwell Publishing, Oxford (1999), pp. 70-103. It is recognized in the art that "equivalent growing conditions" do not require identical growing conditions.

[0046] As used herein, "modified" refers to plants, seeds, plant parts, plant cells and plant genomes that have been subjected to mutagenesis, genome editing, genetic transformation, or a combination thereof.

[0047] In one aspect, the present disclosure provides modified tobacco plants that, when grown under comparable conditions, have no or reduced suckers after topping compared to control tobacco plants. As used herein, a "reduction" in the number of suckers, sucker size, and / or the impact of suckers on agronomic performance refers to a statistically significant reduction. As used herein, "statistically significant" refers to a p-value of less than 0.05 when using an appropriate measure of statistical significance (e.g., a one-tailed, two-sample t-test).

[0048] In one aspect, the modified plants or methods provided herein require reduced management to control sucker development compared to control plants grown under comparable conditions. As used herein, "management" refers to manual removal of suckers, application of chemicals (e.g., maleic hydrazide, flumetralin) to inhibit or remove suckers, or both. In one aspect, the modified plants or methods provided herein require reduced frequency of manual sucker removal, reduced frequency of chemical application, reduced amount of chemical application, or a combination thereof, compared to control plants grown under comparable conditions. See, e.g., Fisher et al., "Topping, Managing Suckers, and Using Ethephon," pages 96-117 In:2016 Flue-Cured Tobacco Information, North Carolina State University, incorporated herein by reference in its entirety.

[0049] In one aspect, the modified plants or methods provided herein require a reduced frequency of maleic hydrazide application to reduce or eliminate suckers. In one aspect, the modified plants or methods provided herein require a reduced amount of maleic hydrazide application to reduce or eliminate suckers. In one aspect, the modified plants or methods provided herein require a reduced frequency of flumetralin application to reduce or eliminate suckers. In one aspect, the modified plants or methods provided herein require a reduced amount of flumetralin application to reduce or eliminate suckers.

[0050] In one aspect, a modified plant or method provided herein requires manual removal of suckers 1% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers 5% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers 10% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers 20% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers 30% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers 40% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers 50% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers 60% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers 70% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers 80% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers 90% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers 95% as frequently as a control plant when grown under equivalent conditions.

[0051] In one aspect, a modified plant or method provided herein requires manual removal of suckers less than 1% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers less than 5% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers less than 10% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers less than 20% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers less than 30% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers less than 40% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers less than 50% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers less than 60% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers less than 70% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers less than 80% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers less than 90% as frequently as a control plant when grown under equivalent conditions. In one aspect, a modified plant or method provided herein requires manual removal of suckers less than 95% as frequently as a control plant when grown under equivalent conditions.

[0052] In one aspect, the modified plants provided herein require manual removal of suckers at 10% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers at 20% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers at 30% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers at 40% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers at 50% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers at 60% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers at 70% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers at 80% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers at 90% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers at 10% to 75% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers at 10% to 50% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers at 10% to 25% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers 50% to 75% as frequently as control plants when grown under comparable conditions.In one aspect, the modified plants provided herein require manual removal of suckers 25% to 75% as frequently as a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers 25% to 50% as frequently as a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers 1% to 50% as frequently as a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers 1% to 25% as frequently as a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require manual removal of suckers 1% to 10% as frequently as a control plant when grown under equivalent conditions.

[0053] In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 1% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 5% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 10% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 20% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 30% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 40% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 50% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 60% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 70% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 80% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 90% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 95% the frequency of a control plant when grown under equivalent conditions.

[0054] In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at less than 1% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at less than 5% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at less than 10% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at less than 20% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at less than 30% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at less than 40% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at less than 50% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at less than 60% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at less than 70% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at less than 80% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at less than 90% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at less than 95% the frequency of a control plant when grown under equivalent conditions.

[0055] In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 10% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 20% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 30% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 40% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 50% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 60% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 70% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 80% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 90% to 95% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 10% to 75% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 10% to 50% the frequency of a control plant when grown under comparable conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 10% to 25% the frequency of a control plant when grown under comparable conditions.In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 50% to 75% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 25% to 50% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 25% to 75% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 1% to 50% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical application to inhibit sucker development at 1% to 25% the frequency of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require chemical applications to inhibit sucker development at 1% to 10% the frequency of control plants when grown under comparable conditions.

[0056] In one aspect, the modified plants provided herein require a chemical spray volume that is 1% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is 5% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is 10% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is 20% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is 30% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is 40% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is 50% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is 60% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is 70% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is 80% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is 90% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions.In one aspect, the modified plants provided herein require 95% of the chemical spray volume used to suppress sucker development in control plants when grown under equivalent conditions.

[0057] In one aspect, the modified plants provided herein, when grown under equivalent conditions, require a chemical spray volume that is less than 1% of the volume used to suppress sucker development of a control plant. In one aspect, the modified plants provided herein, when grown under equivalent conditions, require a chemical spray volume that is less than 5% of the volume used to suppress sucker development of a control plant. In one aspect, the modified plants provided herein, when grown under equivalent conditions, require a chemical spray volume that is less than 10% of the volume used to suppress sucker development of a control plant. In one aspect, the modified plants provided herein, when grown under equivalent conditions, require a chemical spray volume that is less than 20% of the volume used to suppress sucker development of a control plant. In one aspect, the modified plants provided herein, when grown under equivalent conditions, require a chemical spray volume that is less than 30% of the volume used to suppress sucker development of a control plant. In one aspect, the modified plants provided herein, when grown under equivalent conditions, require a chemical spray volume that is less than 40% of the volume used to suppress sucker development of a control plant. In one aspect, the modified plants provided herein require a chemical spray volume that is less than 50% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is less than 60% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is less than 70% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is less than 80% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require a chemical spray volume that is less than 90% of the volume used to suppress sucker development of a control plant when grown under equivalent conditions.In one aspect, the modified plants provided herein require a chemical spray volume that is 95% less than the volume used to suppress sucker development in control plants when grown under equivalent conditions.

[0058] In one aspect, the modified plants provided herein require 10% to 95% less of the chemical spray volume of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 20% to 95% less of the chemical spray volume of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 30% to 95% less of the chemical spray volume of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 40% to 95% less of the chemical spray volume of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 50% to 95% less of the chemical spray volume of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 60% to 95% less of the chemical spray volume of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 70% to 95% less of the chemical spray volume of a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 80% to 95% less chemical spray volume than a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 90% to 95% less chemical spray volume than a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 10% to 75% less chemical spray volume than a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 10% to 50% less chemical spray volume than a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 10% to 25% less chemical spray volume than a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 25% to 50% less chemical spray volume than a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 25% to 75% less chemical spray volume than a control plant when grown under equivalent conditions.In one aspect, the modified plants provided herein require 50% to 75% less chemical spray volume than a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 1% to 50% less chemical spray volume than a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 1% to 25% less chemical spray volume than a control plant when grown under equivalent conditions. In one aspect, the modified plants provided herein require 1% to 10% less chemical spray volume than a control plant when grown under equivalent conditions.

[0059] In one aspect, reduced suckers include fewer total suckers, a smaller average sucker size, or both, compared to a control tobacco plant when grown under comparable conditions. In another aspect, the smaller average sucker size includes a measurement selected from the group consisting of a reduced average mass, a reduced average length, a reduced average diameter, or any combination thereof, compared to the suckers of a control tobacco plant when grown under comparable growing conditions. In another aspect, the smaller average sucker size includes a reduced average mass of the suckers. In a further aspect, the smaller average sucker size includes a reduced average length of the suckers. In a further aspect, the smaller average sucker size includes a reduced average diameter of the suckers.

[0060] In one aspect, the modified tobacco plants comprise fewer total suckers compared to control tobacco plants when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise a smaller average sucker size compared to control tobacco plants when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise a shorter average sucker length compared to control tobacco plants when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise a lower average sucker mass compared to control tobacco plants when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise a shorter average sucker diameter compared to control tobacco plants when grown under equivalent conditions.

[0061] In one aspect, the modified tobacco plant contains at least one less sucker than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least two fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least three fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least four fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least five fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least seven fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 10 fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 15 fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 20 fewer suckers compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 25 fewer suckers compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 30 fewer suckers compared to a control tobacco plant when grown under equivalent conditions.

[0062] In one aspect, the modified tobacco plant contains at least 5% fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 10% fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 15% fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 20% fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 25% fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 30% fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 40% fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 50% fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 60% fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 70% fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 80% fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 90% fewer suckers than a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant contains at least 95% fewer suckers than a control tobacco plant when grown under equivalent conditions.

[0063] In one aspect, the average sucker mass is measured using fresh sucker weight. In another aspect, the average sucker mass is measured using dry sucker weight.

[0064] In one aspect, the modified tobacco plant comprises an average sucker mass that is at least 5% lower than the average sucker mass of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker mass that is at least 10% lower than the average sucker mass of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker mass that is at least 15% lower than the average sucker mass of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker mass that is at least 20% lower than the average sucker mass of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker mass that is at least 25% lower than the average sucker mass of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker mass that is at least 30% lower than the average sucker mass of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker mass that is at least 40% lower than the average sucker mass of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker mass that is at least 50% lower than the average sucker mass of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker mass that is at least 60% lower than the average sucker mass of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker mass that is at least 70% lower than the average sucker mass of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker mass that is at least 80% lower than the average sucker mass of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker mass that is at least 90% lower than the average sucker mass of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise an average sucker mass that is at least 95% lower compared to the average sucker mass of a control tobacco plant when grown under equivalent conditions.

[0065] In one aspect, the average sucker length is measured from the base of the sucker, where the sucker joins the main tobacco stem, to the distal tip of the sucker stem.

[0066] In one aspect, the modified tobacco plant comprises an average sucker length that is at least 1 centimeter shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 2 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 3 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 4 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 5 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 10 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise an average sucker length that is at least 15 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions, hi one aspect, the modified tobacco plants comprise an average sucker length that is at least 20 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions.

[0067] In one aspect, the modified tobacco plants comprise an average sucker length that is 0.5 centimeters to 30 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise an average sucker length that is 0.5 centimeters to 25 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise an average sucker length that is 0.5 centimeters to 20 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise an average sucker length that is 0.5 centimeters to 15 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise an average sucker length that is 0.5 centimeters to 10 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise an average sucker length that is 0.5 centimeters to 5 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise an average sucker length that is 1 centimeter to 30 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise an average sucker length that is 1 centimeter to 20 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise an average sucker length that is 1 centimeter to 10 centimeters shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions.

[0068] In one aspect, the modified tobacco plant comprises an average sucker length that is at least 5% shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 10% shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 15% shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 20% shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 25% shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 30% shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 40% shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 50% shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 60% shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 70% shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 80% shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker length that is at least 90% shorter than the average sucker length of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plants comprise an average sucker length that is at least 95% shorter compared to the average sucker length of a control tobacco plant when grown under equivalent conditions.

[0069] In one aspect, the sucker diameter is measured at the base of the sucker, where the sucker joins the main stem of the plant.

[0070] In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 5% shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 10% shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 15% shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 20% shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 25% shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 30% shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 40% shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 50% shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 60% shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 70% shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 80% shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 90% shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 95% shorter compared to the average sucker diameter of a control tobacco plant when grown under equivalent conditions.

[0071] In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 1 millimeter shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 2 millimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 3 millimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 4 millimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 5 millimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 6 millimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 7 millimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 8 millimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 9 millimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 1 centimeter shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 1.5 centimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 2 centimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions. In one aspect, the modified tobacco plant comprises an average sucker diameter that is at least 3 centimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions.In one aspect, the modified tobacco plants comprise an average sucker diameter that is at least 4 centimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions, hi one aspect, the modified tobacco plants comprise an average sucker diameter that is at least 5 centimeters shorter than the average sucker diameter of a control tobacco plant when grown under equivalent conditions.

[0072] Any nucleic acid molecule, protein, or polypeptide provided herein is contemplated for use with any method provided herein. Any nucleic acid molecule, protein, or polypeptide provided herein is contemplated for use with any plant, plant part, cell, or seed provided herein. Any nucleic acid molecule, protein, or polypeptide provided herein is contemplated for use with any tobacco plant, tobacco plant part, tobacco cell, or tobacco seed provided herein.

[0073] The use of the terms "polynucleotide" or "nucleic acid molecule" is not intended to limit the present disclosure to polynucleotides containing deoxyribonucleic acid (DNA). For example, ribonucleic acid (RNA) molecules are also contemplated. Those skilled in the art will recognize that polynucleotides and nucleic acid molecules can contain ribonucleotides and combinations of ribonucleotides and deoxyribonucleotides. Such deoxyribonucleotides and ribonucleotides include both naturally occurring molecules and synthetic analogs. Polynucleotides of the present disclosure encompass all forms of sequence, including, but not limited to, single-stranded forms, double-stranded forms, hairpins, stem-loop structures, and the like. In one aspect, the nucleic acid molecules provided herein are DNA molecules. In another aspect, the nucleic acid molecules provided herein are RNA molecules. In one aspect, the nucleic acid molecules provided herein are single-stranded. In another aspect, the nucleic acid molecules provided herein are double-stranded. The nucleic acid molecules can encode polypeptides or small RNAs.

[0074] Nucleic acids can be isolated using techniques common in the art. For example, nucleic acids can be isolated using any method, including, but not limited to, recombinant nucleic acid technology and / or polymerase chain reaction (PCR). General PCR techniques are described, for example, in PCR Primer: A Laboratory Manual, Dieffenbach & Dveksler, Eds., Cold Spring Harbor Laboratory Press, 1995. Recombinant nucleic acid technology includes, for example, restriction enzyme digestion and ligation, which can be used to isolate nucleic acids. Isolated nucleic acids can also be chemically synthesized as single nucleic acid molecules or as a series of oligonucleotides. Polypeptides can be purified from natural sources (e.g., biological samples) by known methods, such as DEAE ion exchange, gel filtration, and hydroxyapatite chromatography. Polypeptides can also be purified, for example, by expressing nucleic acids in expression vectors. In addition, purified polypeptides can be obtained by chemical synthesis. The purity of a polypeptide can be measured using any appropriate method, such as column chromatography, polyacrylamide gel electrophoresis, or HPLC analysis.

[0075] In one aspect, the present disclosure provides a method for detecting recombinant nucleic acids and polypeptides in plant cells.Nucleic acids can also be detected using hybridization, but this is not limited to this.Nucleic acid hybridization is discussed in detail in Sambrook et al. (1989, Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY).

[0076] In one aspect, the endogenous genes provided herein comprise a polynucleotide sequence at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the endogenous genes provided herein comprise a polynucleotide sequence at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the endogenous genes provided herein comprise a polynucleotide sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the endogenous genes provided herein comprise a polynucleotide sequence at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the endogenous genes provided herein comprise a polynucleotide sequence at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the endogenous genes provided herein comprise a polynucleotide sequence at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the endogenous genes provided herein comprise a polynucleotide sequence at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the endogenous genes provided herein comprise a polynucleotide sequence at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the endogenous genes provided herein comprise a polynucleotide sequence 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113.

[0077] As used herein, the term "polypeptide" refers to a chain of at least two covalently linked amino acids. A polypeptide can be encoded by a polynucleotide provided herein. A protein provided herein can be encoded by a nucleic acid molecule provided herein. A protein can include a polypeptide provided herein. As used herein, "protein" refers to a chain of amino acid residues that can provide structure or enzymatic activity to a cell.

[0078] Polypeptides can be detected using antibodies. Techniques for detecting polypeptides using antibodies include enzyme-linked immunosorbent assay (ELISA), Western blot, immunoprecipitation, and immunofluorescence. The antibodies provided herein can be polyclonal or monoclonal. Antibodies with specific binding affinity to the polypeptides provided herein can be generated using methods well known in the art. The antibodies provided herein can be attached to a solid support, such as a microtiter plate, using methods well known in the art.

[0079] Detection (e.g., of amplification products, of hybridization complexes, of polypeptides) can be achieved using a detectable label. The term "label" is intended to encompass the use of direct as well as indirect labeling. Detectable labels include enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials.

[0080] In one aspect, the endogenous genes provided herein encode a polypeptide comprising an amino acid sequence at least 70% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114. In one aspect, the endogenous genes provided herein encode a polypeptide comprising an amino acid sequence at least 75% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114. In one aspect, the endogenous genes provided herein encode a polypeptide comprising an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114. In one aspect, the endogenous genes provided herein encode a polypeptide comprising an amino acid sequence at least 85% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114. In one aspect, the endogenous genes provided herein encode a polypeptide comprising an amino acid sequence at least 90% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114. In one aspect, the endogenous genes provided herein encode a polypeptide comprising an amino acid sequence at least 95% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114. In one aspect, the endogenous genes provided herein encode a polypeptide comprising an amino acid sequence at least 96% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114. In one aspect, the endogenous genes provided herein encode a polypeptide comprising an amino acid sequence at least 97% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114. In one aspect, the endogenous genes provided herein encode a polypeptide comprising an amino acid sequence at least 98% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114. In one aspect, the endogenous genes provided herein encode a polypeptide comprising an amino acid sequence at least 99% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114.In one aspect, the endogenous gene provided herein encodes a polypeptide comprising an amino acid sequence 100% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114.

[0081] The term "percent identity" or "percent identical" as used herein with respect to two or more nucleotide or amino acid sequences is calculated by (i) comparing two optimally aligned sequences (nucleotides or amino acids) over a comparison window (one or more "alignable" regions), (ii) determining the number of positions where the same nucleic acid base (for nucleotide sequences) or the same amino acid residue (for proteins and polypeptides) occurs in both sequences to obtain the number of matched positions, (iii) dividing the number of matched positions by the total number of positions within the comparison window, and then (iv) multiplying this quotient by 100% to obtain the percent identity. When "percent identity" is calculated with respect to a reference sequence without specifying a specific comparison window, the percent identity is determined by dividing the number of matched positions over the region of alignment by the total length of the reference sequence. Thus, in this application, when two sequences (query and subject) are optimally aligned (allowing for gaps in their alignment), the "percent identity" of the query sequence is equal to the number of identical positions between the two sequences divided by the total number of positions in the query sequence over the length of the query sequence (or comparison window), then multiplied by 100%.

[0082] When the percentage of sequence identity is used in relation to amino acids, it is recognized that non-identical residue positions often differ by conservative amino acid substitutions, in which an amino acid residue is replaced with another amino acid residue having similar chemical properties (e.g., charge or hydrophobicity), thus not changing the functional properties of the molecule. When sequences differ by conservative substitutions, the percent sequence identity can be adjusted upward to correct for the conservative nature of the substitution. Sequences that differ by such conservative substitutions are said to have "sequence similarity" or "similarity."

[0083] Various pairwise or multiple sequence alignment algorithms and programs are known in the art, such as ClustalW or Basic Local Alignment Search Tool® (BLAST™), which can be used to compare sequence identity or similarity between two or more nucleotide or amino acid sequences to optimally align the sequences and calculate their percent identity. Although other alignment and comparison methods are known in the art, the alignment and percent identity between two sequences (including the above-mentioned percent identity ranges) can be determined by the ClustalW algorithm. See, for example, Chenna et al., "Multiple sequence alignment with the Clustal series of programs," Nucleic Acids Research 31:3497-3500 (2003); Thompson et al., "Clustal W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice," Nucleic Acids Research 22:4673-4680 (1994); Larkin MA et al., "Clustal W and Clustal X version 2.0," Bioinformatics 23:2947-48 (2007); and Altschul et al., "Basic local alignment search tool," J. Mol. Biol. 215:403-410 (1990), the entire contents and disclosures of which are incorporated herein by reference.

[0084] The term "percent complementarity" or "percent complementary," as used herein with respect to two nucleotide sequences, is similar to the concept of percent identity, but refers to the percentage of nucleotides in a query sequence that optimally base-pair or hybridize with nucleotides in a subject sequence when the query and subject sequences are aligned linearly and optimally base-pair without secondary folding structures such as loops, stems, or hairpins. Such percent complementarity can be between two DNA strands, two RNA strands, or a DNA strand and an RNA strand. "Percent complementarity" can be calculated by (i) optimally base-pairing or hybridizing two nucleotide sequences in a linear and fully extended configuration (i.e., without folding or secondary structures) over a window of comparison, (ii) determining the number of base-pairing positions between the two sequences over the window of comparison to obtain the number of complementary positions, (iii) dividing the number of complementary positions by the total number of positions in the window of comparison, and (iv) multiplying this quotient by 100% to obtain the percent complementarity of the two sequences. The optimal base pairing of two sequences can be determined based on the known pairing of nucleotide bases such as GC, AT and AU through hydrogen bonds.When " percent complementarity " is calculated relative to a reference sequence without specifying a specific comparison window, percent identity is determined by dividing the number of complementary positions between two linear sequences by the total length of the reference sequence.Therefore, in this application, when two sequences (query and subject) are optimally base-paired (allowing for mismatched or non-base-paired nucleotides), the " percent complementarity " of the query sequence is equal to the number of base-paired positions between two sequences divided by the total number of positions in the query sequence over the length of the query sequence, and then multiplied by 100%.

[0085] In one aspect, the present disclosure provides a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to the control tobacco plant.

[0086] In another aspect, the present disclosure provides a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety.

[0087] In a further aspect, the present disclosure provides a method for producing a modified tobacco plant, comprising: (a) inducing a mutation in at least one tobacco cell at a genomic locus encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; (b) selecting at least one tobacco cell containing the mutation from step (a); and (c) regenerating a modified tobacco plant from the at least one tobacco cell selected in step (b), wherein the modified tobacco plant contains no or reduced suckers after topping when grown under equivalent growing conditions compared to a control tobacco plant lacking the mutation. In one aspect, the method further comprises (d) cultivating the modified tobacco plant regenerated in step (c). In another aspect, the method further comprises (e) crossing the modified tobacco plant cultivated in step (d) with a second tobacco plant, and (f) obtaining at least one seed from the cross in step (e).

[0088] In one aspect, the modified tobacco plants, tobacco plant parts, tobacco seeds, tobacco cells, or tobacco genomes provided herein comprise one or more mutations in an endogenous gene encoding a cyclin. In one aspect, the modified tobacco plants, tobacco plant parts, tobacco seeds, tobacco cells, or tobacco genomes provided herein comprise one or more mutations in an endogenous gene encoding a cyclin-dependent kinase. In one aspect, the modified tobacco plants, tobacco plant parts, tobacco seeds, tobacco cells, or tobacco genomes provided herein comprise one or more mutations in an endogenous gene encoding a cyclin-dependent kinase inhibitor. In one aspect, the modified tobacco plants, tobacco plant parts, tobacco seeds, tobacco cells, or tobacco genomes provided herein comprise one or more mutations in an endogenous gene encoding a MYB. In one aspect, the modified tobacco plants, tobacco plant parts, tobacco seeds, tobacco cells, or tobacco genomes provided herein comprise one or more mutations in an endogenous gene encoding a WRKY transcription factor.

[0089] Cyclins regulate the progression of cells through the cell cycle. Cyclins often play a role in the activation of CDKs. Cyclins are characterized by a conserved cyclin box domain. Cyclin box domains typically contain approximately 150 amino acids, which are organized into five alpha helical regions. The cyclin box domain is important for binding other proteins, such as CDKs. See, for example, Noble et al., "The cyclin box fold: protein recognition in cell-cycle and transcription control," Trends Biochem. Sci. 22:482-487 (1997); and Menges et al., "Genomic Organization and Evolutionary Conservation of Plant D-Type Cyclins," Plant Physiology, 145:1558-1576 (2007), both of which are incorporated herein by reference in their entireties.

[0090] G1 (Gap 1 phase) is the first part of interphase during the cell cycle in eukaryotic cells. Cells synthesize mRNA and proteins during G1 phase, and G1 phase ends with transition to S phase. S phase (synthesis phase) is the second part of interphase during the cell cycle, during which DNA is replicated. S phase ends with transition to G2 phase. G2 (Gap 2 phase) is typically accompanied by rapid cell growth as cells prepare to transition to M phase (mitosis). Cyclins and CDKs are important regulators during the G1 to S and G2 to M phase transitions and at checkpoints during G1, S, G2, and M phases.

[0091] There are two main groups of cyclins: G1 / S cyclins, which are required for the regulation of cell cycle through G1 / S transition, and G2 / M cyclins, which are required for the regulation of cell cycle through G2 / M transition.If the required cyclin or CDK cannot function properly, the cell cycle may stop at G1 / S transition or G2 / M transition, preventing the formation of daughter cells.In one aspect, the cyclin provided herein is G1 / S cyclin.In another aspect, the cyclin provided herein is G2 / M cyclin.

[0092] In one aspect, the present disclosure provides polynucleotides encoding cyclins. In another aspect, the present disclosure provides cyclins.

[0093] In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene, wherein the mutated polynucleotide encodes a cyclin that is mutated compared to a cyclin encoded by the endogenous gene lacking the mutation. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene, wherein the mutated polynucleotide encodes a cyclin that is truncated compared to a cyclin encoded by the endogenous gene lacking the mutation. In another aspect, the present disclosure provides a non-naturally occurring truncated cyclin. In one aspect, the present disclosure provides a polynucleotide encoding a mutated mRNA of an endogenous gene that comprises a premature stop codon, wherein the mutated mRNA encodes a cyclin that is truncated compared to the mRNA of the endogenous gene lacking the premature stop codon.

[0094] In one aspect, the present disclosure provides a polynucleotide comprising a non-naturally occurring mutation in a sequence encoding a cyclin box domain. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a cyclin, wherein the mutated polynucleotide encodes a cyclin lacking a cyclin box domain. In another aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a cyclin, wherein the mutated polynucleotide encodes a cyclin comprising a truncated cyclin box domain compared to a cyclin encoded by the endogenous gene lacking the mutation. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a cyclin, wherein the mutated polynucleotide encodes a cyclin that is unable to bind a CDK. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a cyclin, wherein the mutated polynucleotide encodes a cyclin that exhibits reduced binding affinity for at least one CDK compared to a cyclin encoded by the endogenous gene lacking the mutation.

[0095] In one aspect, the mutated polynucleotide encodes a dominant negative allele of a cyclin. In another aspect, the mutated polynucleotide encodes a dominant positive allele of a cyclin. In a further aspect, the mutated polynucleotide encodes a constitutively active cyclin. In another aspect, the mutated polynucleotide encodes an inactive cyclin.

[0096] In one aspect, the endogenous gene encoding a cyclin comprises a polynucleotide sequence at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 6-32. In one aspect, the endogenous gene encoding a cyclin comprises a polynucleotide sequence at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 6-32. In one aspect, the endogenous gene encoding a cyclin comprises a polynucleotide sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 6-32. In one aspect, the endogenous gene encoding a cyclin comprises a polynucleotide sequence at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 6-32. In one aspect, the endogenous gene encoding a cyclin comprises a polynucleotide sequence at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 6-32. In one aspect, the endogenous gene encoding a cyclin comprises a polynucleotide sequence at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 6-32. In one aspect, the endogenous gene encoding the cyclin comprises a polynucleotide sequence at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 6 to 32. In one aspect, the endogenous gene encoding the cyclin comprises a polynucleotide sequence at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 6 to 32. In one aspect, the endogenous gene encoding the cyclin comprises a polynucleotide sequence 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 6 to 32.

[0097] In one aspect, the endogenous cyclin comprises an amino acid sequence at least 70% identical to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 75% identical to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the cyclin comprises an amino acid sequence 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 50-76.

[0098] In one aspect, the endogenous cyclin comprises an amino acid sequence at least 70% similar to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 75% similar to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 80% similar to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 85% similar to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 90% similar to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 95% similar to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 96% similar to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 97% similar to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 98% similar to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the endogenous cyclin comprises an amino acid sequence at least 99% similar to a sequence selected from the group consisting of SEQ ID NOs: 50-76. In one aspect, the cyclin comprises an amino acid sequence 100% similar to a sequence selected from the group consisting of SEQ ID NOs: 50-76.

[0099] Cyclin-dependent kinases (CDKs) are a family of protein kinases involved in the regulation of cell cycle, transcription, and mRNA processing. CDKs are recognized as belonging to Enzyme Commission class 2.7.11.22. CDKs contain a kinase domain and interact with or bind to cyclins. Because CDKs phosphorylate substrates on serine and threonine, they are also called serine-threonine kinases. CDKs, together with cyclins, are required for cells to progress through the cell cycle. See, for example, Mironov et al., "Cyclin-Dependent Kinases and Cell Division in Plants—The Nexus," Plant Cell, 11:509-521 (1999), the entire contents of which are incorporated herein by reference.

[0100] In one aspect, the present disclosure provides polynucleotides encoding CDKs. In another aspect, the present disclosure provides CDKs.

[0101] In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene, wherein the mutated polynucleotide encodes a CDK that is mutated compared to a CDK encoded by the endogenous gene lacking the mutation. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene, wherein the mutated polynucleotide encodes a CDK that is truncated compared to a CDK encoded by the endogenous gene lacking the mutation. In another aspect, the present disclosure provides a non-naturally occurring truncated CDK. In one aspect, the present disclosure provides a polynucleotide encoding a mutated mRNA of an endogenous gene that comprises a premature stop codon, wherein the mutated mRNA encodes a CDK that is truncated compared to the mRNA of the endogenous gene lacking the premature stop codon.

[0102] In one aspect, the present disclosure provides a polynucleotide comprising a non-naturally occurring mutation in a sequence encoding a kinase domain. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a CDK, wherein the mutated polynucleotide encodes a CDK lacking a kinase domain. In another aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a CDK, wherein the mutated polynucleotide encodes a CDK comprising a truncated kinase domain compared to a CDK encoded by the endogenous gene lacking the mutation. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a CDK, wherein the mutated polynucleotide encodes a CDK that is unable to bind a cyclin. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a CDK that exhibits reduced binding affinity for at least one cyclin compared to a CDK encoded by an endogenous gene lacking the mutation.

[0103] In one aspect, the mutated polynucleotide encodes a dominant negative allele of a CDK. In another aspect, the mutated polynucleotide encodes a dominant positive allele of a CDK. In a further aspect, the mutated polynucleotide encodes a constitutively active CDK. In another aspect, the mutated polynucleotide encodes an inactive CDK.

[0104] In one aspect, the endogenous gene encoding the CDK comprises a polynucleotide sequence at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-4. In one aspect, the endogenous gene encoding the CDK comprises a polynucleotide sequence at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-4. In one aspect, the endogenous gene encoding the CDK comprises a polynucleotide sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-4. In one aspect, the endogenous gene encoding the CDK comprises a polynucleotide sequence at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-4. In one aspect, the endogenous gene encoding the CDK comprises a polynucleotide sequence at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-4. In one aspect, the endogenous gene encoding the CDK comprises a polynucleotide sequence at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-4. In one aspect, the endogenous gene encoding the CDK comprises a polynucleotide sequence at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-4. In one aspect, the endogenous gene encoding the CDK comprises a polynucleotide sequence that is at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 1 to 4. In one aspect, the endogenous gene encoding the CDK comprises a polynucleotide sequence that is 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 1 to 4.

[0105] In one aspect, the endogenous CDK comprises an amino acid sequence at least 70% identical to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 75% identical to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 45 to 48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 45 to 48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 45 to 48. In one aspect, the CDK comprises an amino acid sequence 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 45 to 48.

[0106] In one aspect, the endogenous CDK comprises an amino acid sequence at least 70% similar to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 75% similar to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 80% similar to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 85% similar to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 90% similar to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 95% similar to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 96% similar to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 97% similar to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 98% similar to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the endogenous CDK comprises an amino acid sequence at least 99% similar to a sequence selected from the group consisting of SEQ ID NOs: 45-48. In one aspect, the CDK comprises an amino acid sequence 100% similar to a sequence selected from the group consisting of SEQ ID NOs: 45-48.

[0107] Cyclin-dependent kinase inhibitors (CDKIs) are proteins that inhibit CDKs. CDKIs bind to CDKs and prevent them from binding to cyclins, thereby negatively controlling the cell cycle in eukaryotic cells. In plants, CDKIs are classified into two families: KIP-related proteins (KRPs) and SIAMESE-related proteins (SMRs). See, for example, Kumar and Larkin, "Why do plants need so many cyclin-dependent kinase inhibitors?" Plant Signaling & Behavior, 12: el282021 (2017), the entire contents of which are incorporated herein by reference.

[0108] In one aspect, the present disclosure provides a polynucleotide encoding a CDKI. In another aspect, the present disclosure provides a CDKI. In one aspect, the CDKI provided herein is KRP. In another aspect, the CDKI provided herein is SMR.

[0109] In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene, wherein the mutated polynucleotide encodes a CDKI that is mutated compared to a CDKI encoded by the endogenous gene lacking the mutation. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene, wherein the mutated polynucleotide encodes a CDKI that is truncated compared to a CDKI encoded by the endogenous gene lacking the mutation. In another aspect, the present disclosure provides a non-naturally occurring truncated CDKI. In one aspect, the present disclosure provides a polynucleotide encoding a mutated mRNA of an endogenous gene comprising a premature stop codon, wherein the mutated mRNA encodes a CDKI that is truncated compared to the mRNA of the endogenous gene lacking the premature stop codon.

[0110] In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a CDKI, wherein the mutated polynucleotide encodes a CDKI that is unable to bind a CDK. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a CDKI, wherein the mutated polynucleotide encodes a CDKI that exhibits reduced binding affinity for at least one CDK compared to a CDKI encoded by the endogenous gene lacking the mutation.

[0111] In one aspect, the mutated polynucleotide encodes a dominant negative allele of a CDKI. In another aspect, the mutated polynucleotide encodes a dominant positive allele of a CDKI. In a further aspect, the mutated polynucleotide encodes a constitutively active CDKI. In another aspect, the mutated polynucleotide encodes an inactive CDKI.

[0112] In one aspect, the endogenous gene encoding the CDKI comprises a polynucleotide sequence at least 80% identical to SEQ ID NO:5. In one aspect, the endogenous gene encoding the CDKI comprises a polynucleotide sequence at least 85% identical to SEQ ID NO:5. In one aspect, the endogenous gene encoding the CDKI comprises a polynucleotide sequence at least 90% identical to SEQ ID NO:5. In one aspect, the endogenous gene encoding the CDKI comprises a polynucleotide sequence at least 95% identical to SEQ ID NO:5. In one aspect, the endogenous gene encoding the CDKI comprises a polynucleotide sequence at least 96% identical to SEQ ID NO:5. In one aspect, the endogenous gene encoding the CDKI comprises a polynucleotide sequence at least 97% identical to SEQ ID NO:5. In one aspect, the endogenous gene encoding the CDKI comprises a polynucleotide sequence at least 98% identical to SEQ ID NO:5. In one aspect, the endogenous gene encoding the CDKI comprises a polynucleotide sequence at least 99% identical to SEQ ID NO:5. In one aspect, the endogenous gene encoding the CDKI comprises a polynucleotide sequence 100% identical to SEQ ID NO:5.

[0113] In one aspect, the endogenous CDKI comprises an amino acid sequence at least 70% identical to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 75% identical to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 80% identical to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 85% identical to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 90% identical to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 95% identical to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 96% identical to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 97% identical to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 98% identical to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 49. In one aspect, the CDKI comprises an amino acid sequence that is 100% identical to SEQ ID NO:49.

[0114] In one aspect, the endogenous CDKI comprises an amino acid sequence at least 70% similar to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 75% similar to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 80% similar to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 85% similar to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 90% identical to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 95% similar to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 96% similar to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 97% similar to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence at least 98% similar to SEQ ID NO: 49. In one aspect, the endogenous CDKI comprises an amino acid sequence that is at least 99% similar to SEQ ID NO: 49. In one aspect, the CDKI comprises an amino acid sequence that is 100% similar to SEQ ID NO:49.

[0115] MYB proteins (MYBs) are part of a family of transcription factors. In plants, MYBs contain a conserved MYB DNA-binding domain, which contains up to three imperfect repeats of approximately 55 amino acids each in a helix-turn-helix structure. These repeats are designated R1, R2, and R3, and the R2 / R3 repeat has been shown to bind directly to the major groove of DNA. A subfamily of plant-specific MYBs contains an R2R3-type MYB domain and is called R2R3-type MYBs. Other subfamilies of MYBs include R1-type, 3R-type, and 4R-type MYBs. See, for example, Ambawat et al., "MYB transcription factor genes as regulators for plant responses: an overview," Physiol. Mol. Biol. Plants, 19:307-321 (2013), incorporated herein by reference in its entirety.

[0116] In one aspect, the MYBs provided herein are R1-type MYBs. In another aspect, the MYBs provided herein are R2R3-type MYBs. In another aspect, the MYBs provided herein are 3R-type MYBs. In another aspect, the MYBs provided herein are 4R-type MYBs.

[0117] In one aspect, the disclosure provides polynucleotides encoding MYB. In another aspect, the disclosure provides MYB.

[0118] In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene, wherein the mutated polynucleotide encodes a mutated MYB compared to a MYB encoded by the endogenous gene lacking the mutation. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene, wherein the mutated polynucleotide encodes a truncated MYB compared to a MYB encoded by the endogenous gene lacking the mutation. In another aspect, the present disclosure provides a non-naturally occurring truncated MYB. In one aspect, the present disclosure provides a polynucleotide encoding a mutated mRNA of an endogenous gene comprising a premature stop codon, wherein the mutated mRNA encodes a truncated MYB compared to the mRNA of the endogenous gene lacking the premature stop codon.

[0119] In one aspect, the present disclosure provides a polynucleotide comprising a non-naturally occurring mutation in a sequence encoding a MYB DNA-binding domain. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a MYB, wherein the mutated polynucleotide encodes a MYB lacking the MYB DNA-binding domain. In another aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a MYB, wherein the mutated polynucleotide encodes a MYB comprising a truncated MYB DNA-binding domain compared to a MYB encoded by the endogenous gene lacking the mutation.

[0120] In one aspect, the disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a MYB, wherein the mutated polynucleotide encodes a MYB that is unable to bind DNA. In one aspect, the disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a MYB, wherein the mutated polynucleotide encodes a MYB that exhibits reduced binding affinity for DNA compared to a MYB encoded by the endogenous gene lacking the mutation.

[0121] In another aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a MYB, wherein the mutated polynucleotide encodes a MYB with a truncated R2R3-type MYB domain compared to a MYB encoded by the endogenous gene lacking the mutation. In another aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a MYB, wherein the mutated polynucleotide encodes a MYB with a truncated R1-type MYB domain compared to a MYB encoded by the endogenous gene lacking the mutation. In another aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a MYB, wherein the mutated polynucleotide encodes a MYB with a truncated 3R-type MYB domain compared to a MYB encoded by the endogenous gene lacking the mutation. In another aspect, the disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a MYB, wherein the mutated polynucleotide encodes a MYB that comprises a truncated 4R-type MYB domain compared to a MYB encoded by the endogenous gene lacking the mutation.

[0122] In one aspect, the mutated polynucleotide encodes a dominant negative allele of MYB. In another aspect, the mutated polynucleotide encodes a dominant positive allele of MYB. In a further aspect, the mutated polynucleotide encodes a constitutively active MYB. In another aspect, the mutated polynucleotide encodes an inactive MYB.

[0123] In one aspect, the endogenous gene encoding MYB comprises a polynucleotide sequence at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 33-44. In one aspect, the endogenous gene encoding MYB comprises a polynucleotide sequence at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 33-44. In one aspect, the endogenous gene encoding MYB comprises a polynucleotide sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 33-44. In one aspect, the endogenous gene encoding MYB comprises a polynucleotide sequence at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 33-44. In one aspect, the endogenous gene encoding MYB comprises a polynucleotide sequence at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 33-44. In one aspect, the endogenous gene encoding MYB comprises a polynucleotide sequence at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 33-44. In one aspect, the endogenous gene encoding MYB comprises a polynucleotide sequence that is at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 33 to 44. In one aspect, the endogenous gene encoding MYB comprises a polynucleotide sequence that is at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 33 to 44. In one aspect, the endogenous gene encoding MYB comprises a polynucleotide sequence that is 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 33 to 44.

[0124] In one aspect, the endogenous MYB comprises an amino acid sequence at least 70% identical to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 75% identical to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the MYB comprises an amino acid sequence 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 77-88.

[0125] In one aspect, the endogenous MYB comprises an amino acid sequence at least 70% similar to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 75% similar to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 80% similar to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 85% similar to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 90% similar to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 95% similar to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 96% similar to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 97% similar to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 98% similar to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the endogenous MYB comprises an amino acid sequence at least 99% similar to a sequence selected from the group consisting of SEQ ID NOs: 77-88. In one aspect, the MYB comprises an amino acid sequence 100% similar to a sequence selected from the group consisting of SEQ ID NOs: 77-88.

[0126] WRKY transcription factors are one of the largest families of transcriptional regulators known in plants. WRKY transcription factors are proteins containing a "WRKY domain." WRKY domains contain approximately 60-70 amino acids and are involved in DNA binding. WRKY domains contain a highly conserved "WRKY core domain" with the amino acid motif WRKYGQK (SEQ ID NO: 110). The conserved core of WRKY domains can vary, with the amino acid motif WRKYGKK (SEQ ID NO: 111) being a commonly observed "WRKY core variant domain." WRKY domains contain a globular shape composed of five antiparallel β-strands, with the conserved core found on the second β-strand. The third β-strand also contains the highly conserved amino acid motif PRSYY (SEQ ID NO: 112). In many WRKY transcription factors, the PR and SYY amino acids of the "PRSYY motif" (SEQ ID NO: 112) are encoded by different exons, and the location of the intervening intron is highly conserved among WRKY transcription factors. The WRKY domain also contains a "zinc finger region" comprising the amino acid motif CX4-5CX22-23HXH (SEQ ID NO: 115) or CX7CX23HXC (SEQ ID NO: 116), where X can be any amino acid, C is cysteine, and H is histidine. The WRKY domain also contains a "DWK salt bridge" motif, in which the conserved tryptophan (W) in the WRKY core (SEQ ID NO: 117) forms a triad with an aspartic acid (D) located 4 amino acids upstream of the W and a lysine (K) located 29 amino acids downstream of the W. The DWK salt bridge is thought to be important for stabilizing the WRKY domain. See, e.g., Rushton et al., "WRKY transcription factors," Trends in Plant Science, 15:247-258 (2010), incorporated herein by reference in its entirety.

[0127] In one aspect, the present disclosure provides a polynucleotide encoding a WRKY transcription factor. In another aspect, the present disclosure provides a WRKY transcription factor.

[0128] In one aspect, the present disclosure provides a polynucleotide comprising a non-naturally occurring mutation, wherein the polynucleotide encodes a WRKY transcription factor. In one aspect, the present disclosure provides a polynucleotide encoding a truncated WRKY transcription factor. In another aspect, the present disclosure provides a non-naturally occurring truncated WRKY transcription factor. In one aspect, the present disclosure provides a polynucleotide encoding an mRNA comprising a premature stop codon, wherein the mRNA encodes a truncated WRKY transcription factor.

[0129] In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene, wherein the mutated polynucleotide encodes a WRKY transcription factor that is mutated compared to a WRKY transcription factor encoded by the endogenous gene lacking the mutation. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene, wherein the mutated polynucleotide encodes a WRKY transcription factor that is truncated compared to a WRKY transcription factor encoded by the endogenous gene lacking the mutation. In another aspect, the present disclosure provides a non-naturally occurring truncated WRKY transcription factor. In one aspect, the present disclosure provides a polynucleotide encoding a mutated mRNA of an endogenous gene that comprises a premature stop codon, wherein the mutated mRNA encodes a WRKY transcription factor that is truncated compared to the mRNA of the endogenous gene lacking the premature stop codon.

[0130] In one aspect, the present disclosure provides a polynucleotide comprising a non-naturally occurring mutation in a sequence encoding a WRKY domain. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor lacking a WRKY domain. In another aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor comprising a truncated WRKY domain compared to a WRKY transcription factor encoded by the endogenous gene lacking the mutation.

[0131] In one aspect, the present disclosure provides a polynucleotide comprising a non-naturally occurring mutation in a sequence encoding a WRKY core domain. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor lacking a WRKY core domain. In another aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor comprising a truncated WRKY core domain compared to a WRKY transcription factor encoded by the endogenous gene lacking the mutation.

[0132] In one aspect, the present disclosure provides a polynucleotide comprising a non-naturally occurring mutation in a sequence encoding a WRKY core variant domain. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor lacking a WRKY core variant domain. In another aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor comprising a truncated WRKY core variant domain compared to a WRKY transcription factor encoded by the endogenous gene lacking the mutation.

[0133] In one aspect, the disclosure provides a polynucleotide comprising a non-naturally occurring mutation in a sequence encoding a PRSYY motif (SEQ ID NO: 112). In one aspect, the disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor lacking a PRSYY motif (SEQ ID NO: 112). In another aspect, the disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor comprising a truncated PRSYY motif (SEQ ID NO: 112) compared to a WRKY transcription factor encoded by the endogenous gene lacking the mutation.

[0134] In one aspect, the present disclosure provides a polynucleotide comprising a non-naturally occurring mutation in a sequence encoding a zinc finger region. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor lacking a zinc finger region. In another aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor comprising a truncated zinc finger region compared to a WRKY transcription factor encoded by the endogenous gene lacking the mutation.

[0135] In one aspect, the present disclosure provides a polynucleotide comprising a non-naturally occurring mutation in a sequence encoding a DWK salt bridge motif. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor lacking a DWK salt bridge motif. In another aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor comprising a truncated DWK salt bridge motif compared to a WRKY transcription factor encoded by the endogenous gene lacking the mutation.

[0136] In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor that is unable to bind DNA. In one aspect, the present disclosure provides a mutated polynucleotide comprising a mutation in an endogenous gene encoding a WRKY transcription factor, wherein the mutated polynucleotide encodes a WRKY transcription factor that exhibits reduced binding affinity for DNA compared to a WRKY transcription factor encoded by the endogenous gene lacking the mutation.

[0137] In one aspect, the mutated polynucleotide encodes a dominant negative allele of a WRKY transcription factor. In another aspect, the mutated polynucleotide encodes a dominant positive allele of a WRKY transcription factor. In a further aspect, the mutated polynucleotide encodes a constitutively active WRKY transcription factor. In another aspect, the mutated polynucleotide encodes an inactive WRKY transcription factor.

[0138] In one aspect, the endogenous gene encoding the WRKY transcription factor comprises a polynucleotide sequence at least 80% identical to SEQ ID NO: 113. In one aspect, the endogenous gene encoding the WRKY transcription factor comprises a polynucleotide sequence at least 85% identical to SEQ ID NO: 113. In one aspect, the endogenous gene encoding the WRKY transcription factor comprises a polynucleotide sequence at least 90% identical to SEQ ID NO: 113. In one aspect, the endogenous gene encoding the WRKY transcription factor comprises a polynucleotide sequence at least 95% identical to SEQ ID NO: 113. In one aspect, the endogenous gene encoding the WRKY transcription factor comprises a polynucleotide sequence at least 96% identical to SEQ ID NO: 113. In one aspect, the endogenous gene encoding the WRKY transcription factor comprises a polynucleotide sequence at least 97% identical to SEQ ID NO: 113. In one aspect, the endogenous gene encoding the WRKY transcription factor comprises a polynucleotide sequence at least 98% identical to SEQ ID NO: 113. In one aspect, the endogenous gene encoding the WRKY transcription factor comprises a polynucleotide sequence at least 99% identical to SEQ ID NO: 113. In one aspect, the endogenous gene encoding the WRKY transcription factor comprises a polynucleotide sequence 100% identical to SEQ ID NO: 113.

[0139] In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 70% identical to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 75% identical to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 80% identical to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 85% identical to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 90% identical to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 95% identical to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 96% identical to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 97% identical to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 98% identical to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 99% identical to SEQ ID NO: 114. In one aspect, the WRKY transcription factor comprises an amino acid sequence 100% identical to SEQ ID NO: 114.

[0140] In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 70% similar to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 75% similar to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 80% similar to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 85% similar to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 90% similar to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 95% similar to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 96% similar to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 97% similar to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 98% similar to SEQ ID NO: 114. In one aspect, the endogenous WRKY transcription factor comprises an amino acid sequence at least 99% similar to SEQ ID NO: 114. In one aspect, the WRKY transcription factor comprises an amino acid sequence 100% similar to SEQ ID NO: 114.

[0141] In one aspect, the modified tobacco plants, tobacco plant parts, tobacco seeds, tobacco cells, or tobacco genomes provided herein contain one or more mutations. As used herein, "mutation" refers to a non-naturally occurring change to DNA compared to an endogenous reference DNA sequence. When identifying a mutation, it is understood that the endogenous reference DNA sequence should be derived from the same tobacco variety. For example, if the modified tobacco plant containing the mutation is derived from the TN90 variety, the endogenous reference sequence must be the endogenous TN90 sequence, rather than a homologous sequence derived from a different tobacco variety (e.g., K326). Similarly, if the modified tobacco cell containing the mutation is a TN90 cell, the endogenous reference sequence must be the endogenous TN90 sequence, rather than a homologous sequence derived from a tobacco cell from a different tobacco variety (e.g., K326).

[0142] In one aspect, the mutations provided herein create a dominant allele at the mutated locus. A dominant allele is an allele that masks the contribution of a second allele at the same locus. A dominant allele can be a "dominant-negative allele" or a "dominant-positive allele." A dominant-negative allele or antimorph is an allele that acts against the function of a normal allele. A dominant-negative allele typically does not function normally, directly inhibiting the activity of a wild-type protein (e.g., through dimerization) or inhibiting the activity of a second protein required for the normal function of the wild-type protein (e.g., an activator or downstream component of a pathway). For example, a dominant-negative allele suppresses or reduces the normal function of an allele in a heterozygous or homozygous state. A dominant-positive allele can increase normal gene function (e.g., a hypermorph) or provide a new function to a gene (e.g., a neomorph). A semi-dominant allele occurs when the penetrance of the associated phenotype in an individual heterozygous for the allele is less than the penetrance observed in an individual homozygous for the allele.

[0143] In one aspect, the mutations provided herein create a dominant negative allele of the mutated locus. In another aspect, the mutations provided herein create a dominant positive allele of the mutated locus.

[0144] As used herein, "inducing" a mutation refers to generating a mutation in a polynucleotide sequence through human intervention. Many suitable methods for inducing mutations in tobacco are known in the art. Non-limiting examples of such methods include the use of chemical mutagens, radiation, and nucleases. In one aspect, inducing a mutation includes the use of an agent selected from the group consisting of chemical mutagens, radiation, transposons, Agrobacterium, and nucleases.

[0145] In one aspect, inducing mutations comprises the use of a chemical mutagen. In one aspect, the chemical mutagen comprises ethyl methanesulfonate (EMS).

[0146] In another aspect, inducing mutations involves the use of irradiation. In one aspect, irradiation involves gamma rays, X-rays, or ionizing radiation. In another aspect, irradiation involves the use of fast neutrons.

[0147] In one aspect, inducing mutations involves the use of transposons, hi another aspect, inducing mutations involves the use of Agrobacterium.

[0148] In a further aspect, inducing mutations involves the use of a nuclease. In one aspect, the nuclease is selected from the group consisting of meganucleases, zinc finger nucleases, transcription activator-like effector nucleases, CRISPR / Cas9 nucleases, CRISPR / Cpf1 nucleases, CRISPR / CasX nucleases, CRISPR / CasY nucleases, and Csm1 nucleases. In one aspect, inducing mutations involves the use of CRISPR / Cas9 nucleases. In one aspect, inducing mutations involves the use of CRISPR / Cpf1 nucleases. In one aspect, inducing mutations involves the use of CRISPR / CasX nucleases. In one aspect, inducing mutations involves the use of CRISPR / CasY nucleases. In one aspect, inducing mutations involves the use of Csm1 nucleases.

[0149] Several types of mutations are known in the art. In one aspect, a mutation includes an insertion. "Insertion" refers to the addition of one or more nucleotides or amino acids to a given polynucleotide or amino acid sequence, respectively, compared to an endogenous reference polynucleotide or amino acid sequence. In another aspect, a mutation includes a deletion. "Deletion" refers to the removal of one or more nucleotides or amino acids from a given polynucleotide or amino acid sequence, respectively, compared to an endogenous reference polynucleotide or amino acid sequence. In another aspect, a mutation includes a substitution. "Substitution" refers to the replacement of one or more nucleotides or amino acids from a given polynucleotide or amino acid sequence, respectively, compared to an endogenous reference polynucleotide or amino acid sequence. In another aspect, a mutation includes an inversion. "Inversion" refers to when a segment of a polynucleotide or amino acid sequence is inverted end-to-end. In one aspect, the mutations provided herein include a mutation selected from the group consisting of an insertion, a deletion, a substitution, and an inversion.

[0150] Mutations in the coding region of a gene (e.g., mutations in an exon) can result in a truncated protein or polypeptide when the mutated messenger RNA (mRNA) is translated into a protein or polypeptide. In one aspect, the present disclosure provides mutations that result in truncation of a protein or polypeptide. As used herein, a "truncated" protein or polypeptide contains at least one fewer amino acid than an endogenous control protein or polypeptide. For example, if endogenous protein A contains 100 amino acids, a truncated form of protein A can contain 1 to 99 amino acids.

[0151] Without being limited by any scientific theory, one way to cause protein or polypeptide truncation is by introducing a premature stop codon into the mRNA transcript of an endogenous gene. In one aspect, the present disclosure provides a mutation that results in a premature stop codon in the mRNA transcript of an endogenous gene. As used herein, a "stop codon" refers to a nucleotide triplet within an mRNA transcript that signals the termination of protein translation. A "premature stop codon" refers to a stop codon that is located earlier (e.g., 5') than the normal stop codon position in the endogenous mRNA transcript. Several stop codons are known in the art, including, but not limited to, "UAG," "UAA," "UGA," "TAG," "TAA," and "TGA."

[0152] In one aspect, the mutation provided herein includes a null mutation. As used herein, "null mutation" refers to a mutation that confers complete loss of function to the protein encoded by the gene containing the mutation, or a mutation that confers complete loss of function to the small RNA encoded by the genomic locus. A null mutation can cause a lack of mRNA transcript production, a lack of small RNA transcript production, a lack of protein function, or a combination thereof.

[0153] The mutations provided herein can be located in any part of the endogenous gene. In one aspect, the mutations provided herein are located in the exons of the endogenous gene. In another aspect, the mutations provided herein are located in the introns of the endogenous gene. In a further aspect, the mutations provided herein are located in the 5'-untranslated region of the endogenous gene. In yet another aspect, the mutations provided herein are located in the 3'-untranslated region of the endogenous gene. In yet another aspect, the mutations provided herein are located in the promoter of the endogenous gene.

[0154] Screening and selection of mutagenized tobacco plants can be performed by any method known to those skilled in the art. Examples of screening and selection methods include, but are not limited to, Southern analysis, PCR amplification for detecting polynucleotides, Northern blot, RNase protection, primer extension, RT-PCR amplification for detecting RNA transcripts, Sanger sequencing, next-generation sequencing technology (e.g., Illumina, PacBio, Ion Torrent, 454) enzyme assay for detecting enzyme or ribozyme activity of polypeptides and polynucleotides, and protein gel electrophoresis, Western blot, immunoprecipitation, and enzyme-linked immunoassay for detecting polypeptides. Other techniques, such as in situ hybridization, enzyme staining, and immunostaining, can also be used to detect the presence or expression of polypeptides and / or polynucleotides. Methods for performing all of the cited techniques are known.

[0155] In one aspect, a mutation in an endogenous gene results in a reduced level of expression compared to an endogenous gene lacking the mutation. In another aspect, a mutation in an endogenous gene results in an increased level of expression compared to an endogenous gene lacking the mutation. In a further aspect, a mutation in an endogenous gene results in a reduced level of activity by a protein or polypeptide encoded by an endogenous gene having the mutation compared to a protein or polypeptide encoded by an endogenous gene lacking the mutation. In a further aspect, a mutation in an endogenous gene results in an increased level of activity by a protein or polypeptide encoded by an endogenous gene having the mutation compared to a protein or polypeptide encoded by an endogenous gene lacking the mutation.

[0156] In one aspect, a mutation in a genomic locus results in a reduced level of expression compared to a genomic locus lacking the mutation. In another aspect, a mutation in a genomic locus results in an increased level of expression compared to a genomic locus lacking the mutation. In a further aspect, a mutation in a genomic locus results in a reduced level of activity by a protein or polypeptide encoded by a genomic locus having the mutation compared to a protein or polypeptide encoded by a genomic locus lacking the mutation. In a further aspect, a mutation in a genomic locus results in an increased level of activity by a protein or polypeptide encoded by a genomic locus having the mutation compared to a protein or polypeptide encoded by a genomic locus lacking the mutation.

[0157] The level of gene expression is routinely investigated in the art.As a non-limiting example, gene expression can be measured using quantitative reverse transcriptase PCR (qRT-PCR), RNA sequencing or Northern blot.In one aspect, gene expression is measured using qRT-PCR.In another aspect, gene expression is measured using Northern blot.In another aspect, gene expression is measured using RNA sequencing.

[0158] The level of protein activity is also routinely investigated in the art, for example, CDK activity can be measured using phosphorylation assays.

[0159] As used herein, the term "heterologous" refers to a combination of two or more DNA molecules or sequences, such as a promoter and associated transcribable DNA sequence, coding sequence or gene, where such combination is artificial and not normally found in nature.

[0160] In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by at least 1% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by at least 5% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by at least 10% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by at least 15% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by at least 20% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by at least 25% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by at least 50% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by at least 75% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by at least 90% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by at least 95% compared to the expression level of the endogenous gene lacking the mutation.

[0161] In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 1% to 99% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 1% to 90% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 1% to 75% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 1% to 50% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 1% to 25% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 1% to 10% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 75% to 99% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 50% to 99% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 25% to 99% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 10% to 99% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 50% to 75% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 25% to 75% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is reduced by 10% to 50% compared to the expression level of the endogenous gene lacking the mutation.

[0162] In one aspect, the level of expression of the endogenous gene comprising the mutation is increased by at least 1% compared to the level of expression of the endogenous gene lacking the mutation. In one aspect, the level of expression of the endogenous gene comprising the mutation is increased by at least 5% compared to the level of expression of the endogenous gene lacking the mutation. In one aspect, the level of expression of the endogenous gene comprising the mutation is increased by at least 10% compared to the level of expression of the endogenous gene lacking the mutation. In one aspect, the level of expression of the endogenous gene comprising the mutation is increased by at least 15% compared to the level of expression of the endogenous gene lacking the mutation. In one aspect, the level of expression of the endogenous gene comprising the mutation is increased by at least 20% compared to the level of expression of the endogenous gene lacking the mutation. In one aspect, the level of expression of the endogenous gene comprising the mutation is increased by at least 25% compared to the level of expression of the endogenous gene lacking the mutation. In one aspect, the level of expression of the endogenous gene comprising the mutation is increased by at least 50% compared to the level of expression of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by at least 75% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by at least 90% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by at least 95% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by at least 100% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by at least 150% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by at least 200% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by at least 250% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the level of expression of the endogenous gene containing the mutation is increased by at least 300% compared to the level of expression of the endogenous gene lacking the mutation.In one aspect, the level of expression of the endogenous gene comprising the mutation is increased by at least 400% compared to the level of expression of the endogenous gene lacking the mutation. In one aspect, the level of expression of the endogenous gene comprising the mutation is increased by at least 500% compared to the level of expression of the endogenous gene lacking the mutation. In one aspect, the level of expression of the endogenous gene comprising the mutation is increased by at least 750% compared to the level of expression of the endogenous gene lacking the mutation. In one aspect, the level of expression of the endogenous gene comprising the mutation is increased by at least 1000% compared to the level of expression of the endogenous gene lacking the mutation.

[0163] In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 1% to 1000% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 1% to 750% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 1% to 500% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 1% to 400% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 1% to 300% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 1% to 200% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 1% to 100% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 1% to 75% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 1% to 50% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 1% to 25% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 1% to 10% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 100% to 1000% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 100% to 750% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the level of expression of the endogenous gene containing the mutation is increased by 100% to 500% compared to the level of expression of the endogenous gene lacking the mutation, hi one aspect, the level of expression of the endogenous gene containing the mutation is increased by 100% to 250% compared to the level of expression of the endogenous gene lacking the mutation.In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 750% to 1000% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 500% to 1000% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 250% to 1000% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 1% to 100% compared to the expression level of the endogenous gene lacking the mutation. In one aspect, the expression level of the endogenous gene comprising the mutation is increased by 50% to 100% compared to the expression level of the endogenous gene lacking the mutation.

[0164] As used herein, the term "endogenous gene" or "native gene" refers to a gene that occurs within the tobacco genome. An "endogenous gene" is a gene that has not previously been modified by human action. In one aspect, the endogenous gene is a nuclear gene. In another aspect, the endogenous gene is a mitochondrial gene. In a further aspect, the endogenous gene is a chloroplast gene.

[0165] As used herein, "gene" refers to a polynucleotide capable of producing a functional unit (such as, but not limited to, a protein or a small RNA molecule). A gene can include a promoter, an enhancer sequence, a leader sequence, a transcription start site, a transcription stop site, a polyadenylation site, one or more exons, one or more introns, a 5'-untranslated region (UTR), a 3'-UTR, or any combination thereof. A "gene sequence" can include a polynucleotide sequence encoding a promoter, an enhancer sequence, a leader sequence, a transcription start site, a transcription stop site, a polyadenylation site, one or more exons, one or more introns, a 5'-UTR, a 3'-UTR, or any combination thereof. In one aspect, a gene encodes a small RNA molecule or a precursor thereof. In another aspect, a gene encodes a protein or polypeptide.

[0166] As used herein, "genomic locus" refers to a fixed position on a chromosome. In one aspect, a genomic locus comprises a polynucleotide encoding a gene. In one aspect, a genomic locus comprises a polynucleotide encoding an endogenous gene. In another aspect, a genomic locus comprises a polynucleotide encoding a transgene. In one aspect, a genomic locus is capable of transcribing DNA into RNA. In one aspect, a genomic locus encodes messenger RNA. In another aspect, a genomic locus encodes a small RNA molecule.

[0167] As commonly understood in the art, the term "promoter" refers to a DNA sequence that contains an RNA polymerase binding site, a transcription initiation site, and / or a TATA box and that supports or facilitates the transcription and expression of an associated transcribable polynucleotide sequence and / or gene (or transgene). Promoters can be synthetically produced, altered, or derived from known or naturally occurring or other promoter sequences. Promoters can also include chimeric promoters, which comprise a combination of two or more heterologous sequences. Thus, promoters of the present application can include variants of promoter sequences that are similar in composition to, but not identical to, other promoter sequence(s) known or provided herein.

[0168] A promoter that drives expression in all or most tissues of a plant is referred to as a "constitutive" promoter. A promoter that drives expression during a specific period or stage of development is called a "developmental" promoter. A promoter that drives enhanced expression in specific tissues of an organism relative to other tissues of the organism is called a "tissue-preferred" promoter. Thus, a "tissue-preferred" promoter causes relatively higher or preferential expression in specific tissues of a plant, but lower expression levels in other tissues of the plant. As a non-limiting example, a "axillary-preferred promoter" is a promoter that drives relatively higher expression in axillary tissues. A promoter that expresses in a specific tissue of an organism, with little or no expression in other tissues, may be called a "tissue-specific" promoter. As a non-limiting example, an "axillary bud-specific promoter" drives expression in axillary bud tissue, with little or no detectable expression in other plant tissue types. An "inducible" promoter is a promoter that initiates transcription in response to an environmental stimulus, such as heat, cold, drought, light, or other stimuli, such as wounding or chemical application.

[0169] In one aspect, the promoter provided herein is an axillary bud-specific promoter. In another aspect, the promoter provided herein is an axillary bud-preferred promoter. In one aspect, the promoter provided herein is a constitutive promoter. In another aspect, the promoter provided herein is an inducible promoter. In a further aspect, the promoter provided herein is a developmental promoter.

[0170] In one aspect, the present disclosure provides a heterologous promoter. In another aspect, the present disclosure provides a promoter operably linked to a heterologous polynucleotide. In another aspect, the present disclosure provides a polynucleotide sequence operably linked to a heterologous promoter.

[0171] In one aspect, the heterologous promoter comprises an axillary bud-specific promoter. In another aspect, the heterologous promoter comprises an axillary bud-preferred promoter. In one aspect, the heterologous promoter comprises a constitutive promoter. In one aspect, the heterologous promoter comprises an inducible promoter. In one aspect, the heterologous promoter comprises a developmental promoter.

[0172] As used herein, "operably linked" refers to a functional link between two or more elements. For example, an operable link between a polynucleotide of interest and a regulatory sequence (e.g., a promoter) is a functional link that allows the expression of the polynucleotide of interest. The operably linked elements can be contiguous or non-contiguous. In one aspect, the promoter provided herein is operably linked to a heterologous nucleic acid molecule.

[0173] In one aspect, the heterologous promoter comprises a polynucleotide sequence at least 90% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the heterologous promoter comprises a polynucleotide sequence at least 91% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the heterologous promoter comprises a polynucleotide sequence at least 92% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the heterologous promoter comprises a polynucleotide sequence at least 93% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the heterologous promoter comprises a polynucleotide sequence at least 94% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the heterologous promoter comprises a polynucleotide sequence at least 95% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the heterologous promoter comprises a polynucleotide sequence at least 96% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the heterologous promoter comprises a polynucleotide sequence at least 97% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the heterologous promoter comprises a polynucleotide sequence at least 98% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the heterologous promoter comprises a polynucleotide sequence at least 99% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the heterologous promoter comprises a polynucleotide sequence 100% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109.

[0174] In one aspect, the axillary bud-preferred promoter comprises a polynucleotide sequence at least 90% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the axillary bud-preferred promoter comprises a polynucleotide sequence at least 95% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the axillary bud-preferred promoter comprises a polynucleotide sequence at least 96% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the axillary bud-preferred promoter comprises a polynucleotide sequence at least 97% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the axillary bud-preferred promoter comprises a polynucleotide sequence at least 98% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the axillary bud-preferred promoter comprises a polynucleotide sequence at least 99% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the axillary bud-preferred promoter comprises a polynucleotide sequence that is 100% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109.

[0175] In one aspect, the axillary bud-specific promoter comprises a polynucleotide sequence at least 90% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the axillary bud-specific promoter comprises a polynucleotide sequence at least 95% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the axillary bud-specific promoter comprises a polynucleotide sequence at least 96% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the axillary bud-specific promoter comprises a polynucleotide sequence at least 97% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the axillary bud-specific promoter comprises a polynucleotide sequence at least 98% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the axillary bud-specific promoter comprises a polynucleotide sequence at least 99% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109. In one aspect, the axillary bud-specific promoter comprises a polynucleotide sequence that is 100% identical to a polynucleotide selected from the group consisting of SEQ ID NOs: 89-109.

[0176] In one aspect, the disclosure provides a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor operably linked to a heterologous promoter, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions.

[0177] In another aspect, the present disclosure provides a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter.

[0178] In a further aspect, the present disclosure provides a method for producing a modified tobacco plant, comprising: (a) introducing a recombinant DNA construct into at least one tobacco cell, wherein the recombinant DNA construct comprises a heterologous promoter operably linked to a nucleic acid encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; (b) selecting at least one tobacco cell comprising the recombinant DNA construct; and (c) regenerating a modified tobacco plant from the at least one tobacco cell selected in step (b), wherein the modified tobacco plant, when grown under equivalent growing conditions, comprises no or reduced suckers after topping compared to a control tobacco plant lacking the recombinant DNA construct. In one aspect, the method further comprises (d) cultivating the modified tobacco plant regenerated in step (c). In another aspect, the method further comprises (e) crossing the modified tobacco plant cultivated in step (d) with a second tobacco plant, and (f) obtaining at least one seed from the cross in step (e).

[0179] In one aspect, the present disclosure provides a method comprising transforming a tobacco cell with a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, hi one aspect, the method further comprises regenerating a modified tobacco plant from the tobacco cell, wherein the modified tobacco plant comprises the recombinant DNA construct.

[0180] In one aspect, the present disclosure provides a recombinant DNA construct comprising a polynucleotide provided herein. As used herein, the term "recombinant DNA construct" refers to a construct formed by laboratory methods of genetic recombination, such as molecular cloning. In one aspect, the recombinant DNA construct is produced synthetically. In another aspect, the recombinant DNA construct comprises a promoter operably linked to a heterologous polynucleotide sequence. In one aspect, the recombinant DNA construct comprises a polynucleotide sequence encoding an amino acid sequence. In another aspect, the recombinant DNA construct comprises a polynucleotide sequence encoding a small RNA or a precursor thereof. In one aspect, the recombinant DNA construct comprises a plasmid. In another aspect, the recombinant DNA construct comprises a vector.

[0181] As used herein, the terms "vector" and "plasmid" are used interchangeably and refer to a circular double-stranded DNA molecule that is physically separated from chromosomal DNA. In one aspect, a plasmid or vector used herein can replicate in vivo. A "transformation vector" used herein is a plasmid that can transform plant cells. In one aspect, the plasmids provided herein are bacterial plasmids. In another aspect, the plasmids provided herein are Agrobacterium Ti plasmids or are derived from Agrobacterium Ti plasmids.

[0182] In one aspect, the vectors provided herein comprise a promoter. In one aspect, the vectors provided herein comprise an axillary bud-specific promoter. In one aspect, the vectors provided herein comprise an axillary bud-preferred promoter. In another aspect, the vectors provided herein comprise a small RNA. In another aspect, the vectors provided herein comprise a small RNA precursor. In one aspect, the vectors provided herein comprise an artificial miRNA. In another aspect, the vectors provided herein comprise an artificial miRNA precursor. In another aspect, the vectors provided herein comprise a sequence at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 89-113, or a fragment thereof. In another aspect, the vectors provided herein comprise a sequence at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 89-113, or a fragment thereof. In another aspect, the vectors provided herein comprise a sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 89-113, or a fragment thereof. In another aspect, the vectors provided herein comprise a sequence at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 89-113, or a fragment thereof. In another aspect, the vectors provided herein comprise a sequence at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 89-113, or a fragment thereof. In another aspect, the vectors provided herein comprise a sequence at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 89-113, or a fragment thereof. In another aspect, the vectors provided herein comprise a sequence at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 89-113, or a fragment thereof. In another aspect, the vectors provided herein comprise a sequence at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 89-113, or a fragment thereof. In another aspect, the vectors provided herein comprise a sequence 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 1-44 and 89-113, or a fragment thereof.

[0183] As used herein, a "fragment" of a nucleic acid or amino acid sequence includes a contiguous segment of the sequence comprising 1% to 99.99% of the total length of the reference sequence. For example, if a nucleic acid sequence comprises 1000 nucleotides, a fragment of the nucleic acid sequence can include any number of contiguous nucleotides of the reference sequence, from 10 to 999. The present disclosure explicitly provides fragments of each of SEQ ID NOs: 1-114 whenever SEQ ID NOs: 1-114 are referenced.

[0184] Numerous methods for introducing recombinant DNA constructs into plant cells are known in the art and can be used in accordance with the methods of the present application to generate transgenic plant cells and plants. Any suitable method or technique for transforming plant cells known in the art can be used in accordance with the methods of the present invention. Effective methods for plant transformation include bacterial-mediated transformation, such as Agrobacterium- or Rhizobium-mediated transformation, and biolistic transformation. Various methods are known in the art for transforming explants with transformation vectors via bacterial-mediated transformation or biolistic transformation, and then culturing the explants to regenerate or develop transgenic plants. Other methods for plant transformation, such as microinjection, electroporation, vacuum infiltration, pressure, sonication, silicon carbide fiber agitation, and polyethylene glycol (PEG)-mediated transformation, are also known in the art. Transgenic plants generated by these transformation methods can be chimeric or non-chimeric with respect to the transformation event, depending on the method and explant used.

[0185] The method of transforming plant cells is well known to those skilled in the art.For example, the specific description of transforming plant cells by particle bombardment with recombinant DNA coated particles (for example, biolistic transformation) can be found in United States Patent (USP) No. 5,550,318; United States Patent No. 5,538,880; United States Patent No. 6,160,208; United States Patent No. 6,399,861; United States Patent No. 6,153,812, and Agrobacterium-mediated transformation is described in United States Patent (USP) No. 5,159,135; United States Patent No. 5,824,877; United States Patent No. 5,591,616; United States Patent No. 6,384,301; United States Patent No. 5,750,871; United States Patent No. 5,463,174; and United States Patent No. 5,188,958, all of which are incorporated herein by reference. Further methods for transforming plants can be found, for example, in Compendium of Transgenic Crop Plants (2009) Blackwell Publishing. Any suitable method known to those of skill in the art can be used to transform tobacco cells with any of the nucleic acid molecules provided herein.

[0186] In one aspect, the method of providing a nucleic acid molecule to a tobacco cell comprises Agrobacterium-mediated transformation. In another aspect, the method of providing a nucleic acid molecule to a cell comprises PEG-mediated transformation. In another aspect, the method of providing a nucleic acid molecule to a cell comprises biolistic transformation. In another aspect, the method of providing a nucleic acid molecule to a cell comprises liposome-mediated transfection (lipofection). In another aspect, the method of providing a nucleic acid molecule to a cell comprises lentiviral transfection.

[0187] Lipofection is described, for example, in U.S. Patent Nos. 5,049,386, 4,946,787, and 4,897,355, and lipofection reagents are commercially available (e.g., Transfectam™ and Lipofectin™). Cationic and neutral lipids suitable for efficient receptor-recognition lipofection of polynucleotides include those described in WO 91 / 17424 and WO 91 / 16024. Delivery can be to cells (e.g., in vitro or ex vivo administration) or to target tissues (e.g., in vivo administration).

[0188] Any tobacco cell from which a fertile tobacco plant can be regenerated is contemplated as a useful recipient cell for carrying out the present disclosure. In one aspect, a recombinant DNA construct is introduced into a tobacco cell. In one aspect, a recombinant DNA construct is introduced into a tobacco protoplast cell. In another aspect, a recombinant DNA construct is introduced into a tobacco callus cell. In one aspect, a recombinant DNA construct is introduced into a tobacco cell selected from the group consisting of seed cells, fruit cells, leaf cells, cotyledon cells, hypocotyl cells, meristem cells, embryo cells, endosperm cells, root cells, shoot cells, stem cells, flower cells, inflorescence cells, stalk cells, pedicel cells, style cells, stigma cells, receptacle cells, petal cells, sepal cells, pollen cells, anther cells, filament cells, ovary cells, ovule cells, pericarp cells, and phloem cells.

[0189] Callus can be initiated from a variety of tissue sources, including, but not limited to, immature embryos or embryo parts, seedling apical meristems, microspores, etc. Cells that can grow as callus can serve as recipient cells for transformation. Practical transformation methods and materials for producing the transgenic plants of the present disclosure (e.g., various media and recipient target cells, transformation of immature embryos, and subsequent regeneration of fertile transgenic plants) are disclosed, for example, in U.S. Patent Nos. 6,194,636 and 6,232,526 and U.S. Patent Application Publication No. 2004 / 0216189, all of which are incorporated herein by reference.

[0190] In one aspect, the present disclosure provides a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions.

[0191] In another aspect, the present disclosure provides a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor.

[0192] In a further aspect, the present disclosure provides a method for producing a modified tobacco plant, comprising: (a) introducing a recombinant DNA construct into at least one tobacco cell, wherein the recombinant DNA construct comprises a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; (b) selecting at least one tobacco cell comprising the recombinant DNA construct; and (c) regenerating a modified tobacco plant from the at least one tobacco cell selected in step (b), wherein the modified tobacco plant, when grown under equivalent growing conditions, comprises no or reduced suckers after topping compared to a control tobacco plant lacking the recombinant DNA construct. In one aspect, the method further comprises (d) cultivating the modified tobacco plant regenerated in step (c). In another aspect, the method further comprises (e) crossing the modified tobacco plant cultivated in step (d) with a second tobacco plant, and (f) obtaining at least one seed from the cross in step (e).

[0193] In another aspect, the present disclosure provides a method comprising transforming a tobacco cell with a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor. In one aspect, the method further comprises regenerating a modified tobacco plant from the tobacco cell, wherein the modified tobacco plant comprises the recombinant DNA construct.

[0194] As used herein, the term "small RNA molecule" refers to any non-coding RNA molecule. In addition to providing small RNA molecules, the present disclosure also provides small RNA precursor molecules for each small RNA molecule.

[0195] In one aspect, the small RNA molecule comprises 18 to 30 nucleotides in length. In another aspect, the small RNA molecule comprises 18 to 24 nucleotides in length. In another aspect, the small RNA molecule comprises 18 to 22 nucleotides in length.

[0196] In another aspect, the small RNA molecule is 18 nucleotides in length. In another aspect, the small RNA molecule is 19 nucleotides in length. In another aspect, the small RNA molecule is 20 nucleotides in length. In another aspect, the small RNA molecule is 21 nucleotides in length. In another aspect, the small RNA molecule is 22 nucleotides in length. In another aspect, the small RNA molecule is 23 nucleotides in length. In another aspect, the small RNA molecule is 24 nucleotides in length. In another aspect, the small RNA molecule is 25 nucleotides in length. In another aspect, the small RNA molecule is 26 nucleotides in length. In another aspect, the small RNA molecule is 27 nucleotides in length. In another aspect, the small RNA molecule is 28 nucleotides in length.

[0197] In another aspect, the small RNA molecule is at least 18 nucleotides in length. In another aspect, the small RNA molecule is at least 19 nucleotides in length. In another aspect, the small RNA molecule is at least 20 nucleotides in length. In another aspect, the small RNA molecule is at least 21 nucleotides in length. In another aspect, the small RNA molecule is at least 22 nucleotides in length. In another aspect, the small RNA molecule is at least 23 nucleotides in length. In another aspect, the small RNA molecule is at least 24 nucleotides in length. In another aspect, the small RNA molecule is at least 25 nucleotides in length. In another aspect, the small RNA molecule is at least 26 nucleotides in length. In another aspect, the small RNA molecule is at least 27 nucleotides in length. In another aspect, the small RNA molecule is at least 28 nucleotides in length.

[0198] In one aspect, the small RNA molecule comprises 18 to 30 nucleotides. In another aspect, the small RNA molecule comprises 18 to 24 nucleotides. In another aspect, the small RNA molecule comprises 18 to 21 nucleotides. In another aspect, the small RNA molecule comprises 21 to 24 nucleotides. In another aspect, the small RNA molecule comprises 21 to 30 nucleotides.

[0199] In one aspect, the small RNA molecule provided herein is a microRNA (miRNA). In one aspect, the small RNA molecule provided herein is an artificial miRNA. In another aspect, the small RNA molecule provided herein is a small interfering RNA (siRNA). In another aspect, the small RNA molecule provided herein is a heterochromatin siRNA (hc-siRNA). In another aspect, the small RNA molecule provided herein is a Piwi-interacting RNA (piRNA). In one aspect, the small RNA molecule provided herein is a double-stranded RNA (dsRNA). In another aspect, the small RNA molecule provided herein is a hairpin double-stranded RNA (hp-dsRNA). In another aspect, the small RNA molecule provided herein is a trans-acting siRNA (ta-siRNA). In another aspect, the small RNA molecule provided herein is a naturally occurring antisense siRNA (nat-siRNA). In another aspect, the small RNA molecule provided herein is a Cas9-guide RNA (gRNA). In another aspect, the small RNA molecule provided herein is Cpf1-gRNA. In another aspect, the small RNA molecule provided herein is CasX-gRNA. In another aspect, the small RNA molecule provided herein is Csm1-gRNA.

[0200] In one aspect, the small RNA molecule is selected from the group consisting of dsRNA, siRNA, ta-siRNA, and miRNA. In another aspect, the small RNA molecule is selected from the group consisting of miRNA, siRNA, hc-siRNA, piRNA, dsRNA, hp-dsRNA, ta-siRNA, nat-siRNA, Cas9-gRNA, Cpf1-gRNA, CasX-gRNA, and Csm1-gRNA.

[0201] In one aspect, the small RNA molecules provided herein comprise a polynucleotide sequence having at least 90% identity or complementarity with an endogenous mRNA. In one aspect, the small RNA molecules provided herein comprise a polynucleotide sequence having at least 91% identity or complementarity with an endogenous mRNA. In one aspect, the small RNA molecules provided herein comprise a polynucleotide sequence having at least 92% identity or complementarity with an endogenous mRNA. In one aspect, the small RNA molecules provided herein comprise a polynucleotide sequence having at least 93% identity or complementarity with an endogenous mRNA. In one aspect, the small RNA molecules provided herein comprise a polynucleotide sequence having at least 94% identity or complementarity with an endogenous mRNA. In one aspect, the small RNA molecules provided herein comprise a polynucleotide sequence having at least 95% identity or complementarity with an endogenous mRNA. In one aspect, the small RNA molecules provided herein comprise a polynucleotide sequence having at least 96% identity or complementarity with an endogenous mRNA. In one aspect, the small RNA molecules provided herein comprise a polynucleotide sequence having at least 97% identity or complementarity with an endogenous mRNA. In one aspect, the small RNA molecules provided herein comprise a polynucleotide sequence having at least 98% identity or complementarity with an endogenous mRNA. In one aspect, the small RNA molecules provided herein comprise a polynucleotide sequence having at least 99% identity or complementarity with an endogenous mRNA. In one aspect, the small RNA molecules provided herein comprise a polynucleotide sequence having 100% identity or complementarity with an endogenous mRNA.

[0202] In one aspect, the small RNA molecule comprises a polynucleotide sequence having at least 85% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence having at least 90% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence having at least 91% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence having at least 92% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence having at least 93% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence having at least 94% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence having at least 95% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence having at least 96% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence having at least 97% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence having at least 98% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence having at least 99% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113.In one aspect, the small RNA molecule comprises a polynucleotide sequence having 100% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113.

[0203] In one aspect, the small RNA molecule comprises a polynucleotide sequence identical to or complementary to at least 16 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence identical to or complementary to at least 17 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence identical to or complementary to at least 18 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence identical to or complementary to at least 19 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence identical to or complementary to at least 20 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, a small RNA molecule comprises a polynucleotide sequence identical or complementary to at least 21 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, a small RNA molecule comprises a polynucleotide sequence identical or complementary to at least 22 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, a small RNA molecule comprises a polynucleotide sequence identical or complementary to at least 23 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, a small RNA molecule comprises a polynucleotide sequence identical or complementary to at least 24 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, a small RNA molecule comprises a polynucleotide sequence identical or complementary to at least 25 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113.In one aspect, the small RNA molecule comprises a polynucleotide sequence identical to or complementary to at least 26 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence identical to or complementary to at least 27 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence identical to or complementary to at least 28 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence identical to or complementary to at least 29 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the small RNA molecule comprises a polynucleotide sequence identical to or complementary to at least 30 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113.

[0204] In one aspect, the present disclosure provides a small RNA capable of reducing the expression of a polynucleotide encoding a cyclin. In one aspect, the present disclosure provides a small RNA capable of reducing the translation of a polynucleotide encoding a cyclin. In one aspect, the present disclosure provides a small RNA capable of reducing the expression of a polynucleotide encoding a CDK. In one aspect, the present disclosure provides a small RNA capable of reducing the translation of a polynucleotide encoding a CDK. In one aspect, the present disclosure provides a small RNA capable of reducing the expression of a polynucleotide encoding a CDKI. In one aspect, the present disclosure provides a small RNA capable of reducing the translation of a polynucleotide encoding a CDKI. In one aspect, the present disclosure provides a small RNA capable of reducing the expression of a polynucleotide encoding a MYB. In one aspect, the present disclosure provides a small RNA capable of reducing the translation of a polynucleotide encoding a MYB. In one aspect, the present disclosure provides a small RNA capable of reducing the expression of a polynucleotide encoding a WRKY transcription factor. In one aspect, the present disclosure provides a small RNA capable of reducing the translation of a polynucleotide encoding a WRKY transcription factor.

[0205] MicroRNAs (miRNAs) are non-protein-coding RNAs, typically about 19 to 25 nucleotides long (generally about 20 to 24 nucleotides in plants), that trans-guide the cleavage of target transcripts and negatively regulate the expression of genes involved in various regulatory and developmental pathways. In some cases, miRNAs also play a role in guiding in-phase processing of siRNA primary transcripts.

[0206] Many microRNA genes (MIR genes) have been identified and published in databases ("miRBase," available online at microrna(dot)sanger(dot)ac(dot)uk / sequences; see also Griffiths-Jones et al. (2003) Nucleic Acids Res., 31:439-441). MIR genes have been reported to occur in separate and clustered intergenic regions within the genome, but they can also be located completely or partially within introns of other genes (both protein-coding and non-protein-coding). Transcription of MIR genes may, at least in some cases, be under the facilitatory control of the MIR gene's own promoter. Primary transcripts, termed "pri-miRNAs," can be very large (several kilobases), may be polycistronic, and contain one or more pre-miRNAs (folded structures containing stem-loop arrangements that are processed into mature miRNAs) as well as the usual 5' "cap" and polyadenylated tail of mRNA.

[0207] Transgenic expression of miRNAs (whether naturally occurring or artificial) can be used to regulate the expression of one or more target genes of the miRNA. The inclusion of miRNA recognition sites in transgenically expressed transcripts is also useful for regulating the expression of those transcripts. Recognition sites for miRNAs have been identified in all regions of mRNAs, including the 5' untranslated, coding, and 3' untranslated regions, indicating that the location of a miRNA target site relative to the coding sequence may not necessarily affect repression.

[0208] Because miRNAs are important regulatory elements in eukaryotes, transgenic suppression of miRNAs is useful for manipulating biological pathways and responses. Various uses of miRNAs, miRNA precursors, miRNA recognition sites, and miRNA promoters are described in detail in U.S. Patent Application Publication No. 2006 / 0200878, which is incorporated herein by reference. Non-limiting examples of these uses include: (1) expression of natural miRNAs or miRNA precursor sequences to suppress target genes; (2) expression of artificial miRNAs or miRNA precursor sequences to suppress target genes; (3) expression of transgenes with miRNA recognition sites (transgenes are suppressed when mature miRNAs are expressed); and (4) expression of transgenes driven by miRNA promoters.

[0209] Designing an artificial miRNA sequence can be as simple as replacing nucleotides in the miRNA stem region of a miRNA precursor with a sequence complementary to the intended target, as demonstrated by Zeng et al. (2002) Mol. Cell, 9:1327-1333. One non-limiting example of a general method for determining nucleotide changes in a native miRNA sequence to generate an engineered miRNA precursor involves the following steps: (a) selecting a unique target sequence of at least 18 nucleotides specific to the target gene, for example, by using a sequence alignment tool such as BLAST (see, e.g., Altschul et al. (1990) J. Mol. Biol., 215:403-410; Altschul et al. (1997) Nucleic Acids Res., 25:3389-3402) of both tobacco cDNA and genomic DNA databases to identify any potential matches to orthologs of the target transcript and unrelated genes, thereby avoiding unintended silencing of non-target sequences; (b) analyzing the target gene for undesired sequences (e.g., matches to sequences from non-target species) and analysing the sequence by GC content, Reynolds score (Reynolds et al. (2004) Nature 106:101-102). Biotechnol., 22:326-330), and functional asymmetry characterized by a negative difference in free energy (".DELTA..DELTA.G" or "ΔΔG") (see Khvorova et al. (2003) Cell, 114:209-216). Preferably, 19mers are selected that have all or most of the following characteristics: (1) a Reynolds score >4, (2) about 40% to about 60% GC content, (3) a negative ΔΔG, (4) a terminal adenosine, (5) no stretch of four or more consecutive identical nucleotides, (6) location near the 3' end of the target gene, and (7) minimal differences from the miRNA precursor transcript.The position of every third nucleotide in an siRNA has been reported to be particularly important in influencing the efficacy of RNAi, and the algorithm "siExplorer" is publicly available at ma.chem.tu-tokyo.ac.jp / siexplorer.htm (see Katoh and Suzuki (2007) Nucleic Acids Res., 10.1093 / nar / gkl1120); (c) determining the reverse complement of the selected 19mer for use in generating a modified mature miRNA. The additional nucleotide at position 20 preferably matches the selected target sequence, and the nucleotide at position 21 is preferably selected to either not pair to prevent spread of silencing on the target transcript, or to pair with the target sequence to promote spread of silencing on the target transcript; and (d) transforming the artificial miRNA into a plant.

[0210] In one aspect, the artificial miRNAs provided herein are complementary to at least 18 contiguous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are complementary to at least 19 contiguous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are complementary to at least 20 contiguous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are complementary to at least 21 contiguous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are complementary to at least 22 contiguous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are complementary to at least 23 contiguous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are complementary to at least 24 contiguous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are complementary to at least 25 contiguous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are complementary to at least 26 contiguous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113.

[0211] In one aspect, the artificial miRNAs provided herein are at least 75% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are at least 80% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are at least 85% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are at least 90% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are at least 91% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are at least 92% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are at least 93% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are at least 94% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are at least 95% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are at least 96% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are at least 97% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are at least 98% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. In one aspect, the artificial miRNAs provided herein are at least 99% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113.In one aspect, the artificial miRNAs provided herein are 100% complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113.

[0212] In one aspect, the artificial miRNAs provided herein reduce or eliminate RNA transcription or protein translation of a target gene.

[0213] In one aspect, the artificial miRNA or its precursor is operably linked to an axillary bud-specific promoter. In another aspect, the artificial miRNA or its precursor is operably linked to an axillary bud-preferring promoter. In another aspect, the artificial miRNA or its precursor is operably linked to a promoter comprising a sequence selected from the group consisting of SEQ ID NOs: 89-109 and fragments thereof.

[0214] Tobacco is known in the art as a plant of the Solanaceae family. As used herein, tobacco plants include Nicotiana tabacum, Nicotiana amplexicaulis (PI 271989), Nicotiana benthamiana (PI 555478), Nicotiana bigelovii (PI 555485), Nicotiana debneyi (PI 555486), Nicotiana excelsior (PI 224063), Nicotiana glutinosa (PI 555507), Nicotiana goodspeedii (PI 241012), Nicotiana gossei (PI 230953), and Nicotiana hesperis (PI 230953). hesperis PI271991; Nicotiana knightiana PI555527, Nicotiana maritima PI555535; Nicotiana megalosiphon PI555536; Nicotiana nudicaulis PI555540; Nicotiana paniculata PI555545; Nicotiana plumbaginifolia PI555548; Nicotiana repanda PI555552; ​​Nicotiana rustica; Nicotiana suaveolens PI55555 suaveolens) PI230960; Nicotiana sylvestris (Nicotiana sylvestris) PI555569; Nicotiana tomentosa (Nicotiana tomentosa) PI266379, Nicotiana tomentosiformis (Nicotiana tomentosiformis);The tobacco plant may be from any plant in the genus Nicotiana, including, but not limited to, Nicotiana trigonophylla PI555572. In one aspect, the tobacco plant described herein is a Nicotiana tabacum plant.

[0215] In one aspect, the modified tobacco plants, seeds, cells, hybrids, varieties, or lines provided herein are essentially the same as or similar to BU64, CC101, CC200, CC13, CC27, CC33, CC35, CC37, CC65, CC67, CC301, CC400, CC500, CC600, CC700, CC800, CC900, CC1063, Coker176, Coker319, Coker371Gold, Coker48, CU263, DF911, Galpao, GL26H, GL338, GL350, GL395, GL600, GL73 7, GL939, GL973, GF157, GF318, RJR901, HB04P, K149, K326, K346, K358, K394, K399, K730, NC196, NC37NF, NC471, NC55, NC92, NC2326, NC95, NC925, PVH1118, PVH1452, PVH2110, PVH2254, PVH2275, VA116, VA119, KDH959, KT200, KT204LC, KY10, KY14, KY160, KY17, KY171, KY907, KY907LC, KTY14xL8 LC, Little Crittenden, McNair373, McNair944, Male Sterility KY14xL8, Narrow Leaf Madole, MS KY171, Narrow Leaf Madole(phph), MS Narrow Leaf Madole, MS TND950, PD7302LC, PD7305LC, PD7309LC, PD7312LC, PD7318LC, PD7319LC, MSTKS2002, TKF2002, TKF6400, TKF4028, TKF4024, KT206 LC, KT209LC, KT210LC, KT212LC, NC100, NC102, NC2000, NC291, NC297, NC299, NC3, NC4, NC5, NC6, NC7, NC606, NC71, NC72, NC810, NC BH 129, NC2002, Neal Smith Madole, OXFORD 207, "Perique", PVH03, PVH09, PVH19, PVH50, PVH51, R610, R630, R7-11, R7-12, RG17, RG81, RGH51, RGH4, RGH51, RS1410, Speight 168, Speight 172, Speight 179, Speight 210, Speight 220, Speight 225, Speight 227, Speight 234, Speight G-28, Speight G-70, Speight H-6, Speight H20, Speight NF3, TI1406, TI1269, TN86, TN86LC, TN90, TN90LC, TN97, TN97LC, TN D94, TN D950, TR (Tom Rosson) Madole, VA309, VA359 or any commercial tobacco variety bred by standard tobacco breeding techniques known in the art, or in these genetic backgrounds.

[0216] One-sided plant, modified plant, BU64 plant, CC101 plant, CC200 plant, CC13 plant, CC27 plant, CC33 plant, CC35 plant, CC37 plant Plants, CC65 plants, CC67 plants, CC301 plants, CC400 plants, CC500 plants, CC600 plants, CC700 plants, CC800 plants, CC900 plants, CC 1063 plant, Coker176 plant, Coker319 plant, Coker371Gold plant, Coker48 plant, CU263 plant, DF911 plant, Galpao plant, GL26H plant, GL338 plant, GL350 plant, GL395 plant, GL600 plant, GL737 plant, GL939 plant, GL973 plant, GF157 plants, GF318 plants, RJR901 plants, HB04P plants, K149 plants, K326 plants, K346 plants, K358 plants, K394 plants, K399 plants, K730 plants, NC196 plants, NC37NF plants, NC471 plants, NC55 plants, NC92 plants, NC2326 plants, NC95 plants, NC925 plants, PVH1118 plants, PVH1452 plants, PVH2110 plants, PVH2254 plants, PVH2275 plants, VA116 plants, VA119 plants, KDH959 plants, KT200 plants, KT204LC plants, KY10 plants, KY14 plants, KY160 plants, KY17 plants, KY171 plants, KY907 plants, KY907LC plants, KTY14xL8 LC plants, Little Crittenden plants, McNair373 plants, McNair944 plants, Male-Immature KY14xL8 plants, Narrow Leaf Madole plants, MS KY171 plants, Narrow Leaf Madole (phph) plants, MS Narrow Leaf Madole plants, MSSelected from the group consisting of TND950 plants, PD7302LC plants, PD7305LC plants, PD7309LC plants, PD7312LC plants, PD7318LC plants, PD7319LC plants, MSTKS2002 plants, TKF2002 plants, TKF6400 plants, TKF4028 plants, TKF4024 plants, KT206LC plants, KT209LC plants, KT210LC plants, KT212LC plants, NC100 plants, NC102 plants, NC2000 plants, NC29! plants, NC297 plants, NC299 plants, NC3 plants, NC4 plants, NC5 plants, NC6 plants, NC7 plants, NC606 plants, NC71 plants, NC72 plants, NC810 plants, NC BH 129 plants, NC2002 plants, Neal Smith Madole plants, OXFORD 207 plants, "Perique" plants, PVH03 plants, PVH09 plants, PVH19 plants, PVH50 plants, PVH51 plants, R610 plants, R630 plants, R7-11 plants, R7-12 plants, RG17 plants, RG81 plants, RG H51 plants, RGH4 plants, RGH51 plants, RS1410 plants, Speight168 plants, Speight172 plants, Speight179 plants, Speight210 plants, Speight220 plants, Speight225 plants, Speight227 plants, Speight234 plants, SpeightG-28 plants, SpeightG-70 plants, SpeightH-6 plants, SpeightH20 plants, SpeightNF3 plants, TI1406 plants, TI1269 plants, TN86 plants, TN86LC plants, TN90 plants, TN90LC plants, TN97 plants, TN97LC plants, TN D94 plants, TN D950 plants, TR(Tom Rosson)Madole plants, VA-309 plants and VA359 plants.

[0217] In another aspect, the modified tobacco cells are BU64 cells, CC101 cells, CC200 cells, CC13 cells, CC27 cells, CC33 cells, CC35 cells, CC37 cells, CC65 cells, CC67 cells, CC301 cells, CC400 cells, CC500 cells, CC600 cells, CC700 cells, CC800 cells, CC900 cells, CC1063 cells, Coker176 cells, Coker319 cells, Coker371Gold cells, Coker48 cells, CU263 cells, DF911 cells, Galpao cells, GL26H cells, GL338 cells, GL350 cells, GL395 cells, GL600 cells, GL737 cells, GL939 cells, GL973 cells, GF157 cells, GF318 cells, RJR901 cells, HB04P cells, K149 cells, K326 cells, K346 cells, K358 cells, K394 cells, K399 cells, K730 cells, NC196 cells, NC37NF cells, NC471 cells, NC55 cells, NC92 cells, NC2326 cells, NC95 cells, NC925 cells, PVH1118 cells, PVH1452 cells, PVH2110 cells, PVH2254 cells, PVH2275 cells, VA116 cells, VA119 cells, KDH959 cells, KT200 cells, KT204LC cells, KY10 cells, KY14 cells, KY160 cells, KY17 cells, KY171 cells, KY907 cells, KY907LC cells, KTY14xL8 LC cells, Little Crittenden cells, McNair373 cells, McNair944 cells, male sterile KY14xL8 cells, Narrow Leaf Madole cells, MS KY171 cells, Narrow Leaf Madole(phph) cells, MS Narrow Leaf Madole cells, MSTND950 cells, PD7302LC cells, PD7305LC cells, PD7309LC cells, PD7312LC cells, PD7318LC cells, PD7319LC cells, MSTKS2002 cells, TKF2002 cells, TKF6400 cells, TKF4028 cells, TKF4024 cells, KT206LC cells, KT209LC cells, KT210LC cells, KT212LC cells, NC100 cells, NC102 cells, NC2000 cells, NC291 cells, NC297 cells, NC299 cells, NC3 cells, NC4 cells, NC5 cells, NC6 cells, NC7 cells, NC606 cells, NC71 cells, NC72 cells, NC810 cells, NC BH 129 cells, NC2002 cells, Neal Smith Madole cells, OXFORD 207 cells, "Perique" cells, PVH03 cells, PVH09 cells, PVH19 cells, PVH50 cells, PVH51 cells, R610 cells, R630 cells, R7-11 cells, R7-12 cells, RG17 cells, RG81 cells, RG H51 cells, RGH4 cells, RGH51 cells, RS1410 cells, Speight168 cells, Speight172 cells, Speight179 cells, Speight210 cells, Speight220 cells, Speight225 cells, Speight227 cells, Speight234 cells, SpeightG-28 cells, SpeightG-70 cells, SpeightH-6 cells, SpeightH20 cells, SpeightNF3 cells, TI1406 cells, TI1269 cells, TN86 cells, TN86LC cells, TN90 cells, TN90LC cells, TN97 cells, TN97LC cells, TN D94 cells, TN D950 cells, TR (Tom) Rosson) Madole cells, VA309 cells and VA359 cells.

[0218] In another aspect, the modified tobacco seeds are BU64 seeds, CC101 seeds, CC200 seeds, CC13 seeds, CC27 seeds, CC33 seeds, CC35 seeds, CC37 seeds, CC65 seeds, CC67 seeds, CC301 seeds, CC400 seeds, CC500 seeds, CC600 seeds, CC700 seeds, CC800 seeds, CC900 seeds, CC1063 seeds, Coker176 seeds, Coker319 seeds, Coker371Gold seeds, Coker48 seeds, CU263 seeds, DF911 seeds, Galpao seeds, GL26H seeds, GL338 seeds, GL350 seeds, GL395 seeds, GL600 seeds, GL737 seeds, GL939 seeds, GL973 seeds, GF157 seeds, GF318 seeds, RJR901 seeds, HB04P seeds, K149 seeds, K326 seeds, K346 seeds, K358 seeds, K394 seeds, K399 seeds, K730 seeds, NC196 seeds, NC37NF seeds, NC471 seeds, NC55 seeds, NC92 seeds, NC2326 seeds, NC95 seeds, NC925 seeds, PVH1118 seeds, PVH1452 seeds, PVH2110 seeds, PVH2254 seeds, PVH2275 seeds, VA116 seeds, VA119 seeds, KDH959 seeds, KT200 seeds, KT204LC seeds, KY10 seeds, KY14 seeds, KY160 seeds, KY17 seeds, KY171 seeds, KY907 seeds, KY907LC seeds, KTY14xL8 LC seeds, Little Crittenden seeds, McNair373 seeds, McNair944 seeds, male sterile KY14xL8 seeds, Narrow Leaf Madole seeds, MS KY171 seeds, Narrow Leaf Madole(phph) seeds, MS Narrow Leaf Madole seeds, MSSelected from the group consisting of TND950 seeds, PD7302LC seeds, PD7305LC seeds, PD7309LC seeds, PD7312LC seeds, PD7318LC seeds, PD7319LC seeds, MSTKS2002 seeds, TKF2002 seeds, TKF6400 seeds, TKF4028 seeds, TKF4024 seeds, KT206LC seeds, KT209LC seeds, KT210LC seeds, KT212LC seeds, NC100 seeds, NC102 seeds, NC2000 seeds, NC291 seeds, NC297 seeds, NC299 seeds, NC3 seeds, NC4 seeds, NC5 seeds, NC6 seeds, NC7 seeds, NC606 seeds, NC71 seeds, NC72 seeds, NC810 seeds, NC BH 129 seeds, NC2002 seeds, Neal Smith Madole seeds, OXFORD 207 seeds, "Perique" seeds, PVH03 seeds, PVH09 seeds, PVH19 seeds, PVH50 seeds, PVH51 seeds, R610 seeds, R630 seeds, R7-11 seeds, R7-12 seeds, RG17 seeds, RG81 seeds, RG H51 seeds, RGH4 seeds, RGH51 seeds, RS1410 seeds, Speight168 seeds, Speight172 seeds, Speight179 seeds, Speight210 seeds, Speight220 seeds, Speight225 seeds, Speight227 seeds, Speight234 seeds, SpeightG-28 seeds, SpeightG-70 seeds, SpeightH-6 seeds, SpeightH20 seeds, SpeightNF3 seeds, TI1406 seeds, TI1269 seeds, TN86 seeds, TN86LC seeds, TN90 seeds, TN90LC seeds, TN97 seeds, TN97LC seeds, TN D94 seeds, TN D950 seeds, TR(Tom Rosson)Madole seeds, VA309 seeds and VA359 seeds.

[0219] As used herein, "tobacco plant" refers to the entire tobacco plant. Tobacco cell or tobacco tissue culture derived from a tobacco plant can include any tobacco plant part or organ (e.g., leaf, stem, root, etc.), tobacco plant tissue, tobacco seed, tobacco plant cell, and / or their progeny. The progeny plant can be derived from any hybrid generation, such as F1, F2, F3, F4, F5, F6, F7, etc. Tobacco plant cells are biological cells of a tobacco plant that are harvested from a tobacco plant or obtained through culture from cells harvested from a tobacco plant. As used herein, "seedling" refers to a tobacco plant that is equal to or less than 14 days after germination.

[0220] In one aspect, tobacco plant parts provided herein include, but are not limited to, leaves, stems, roots, seeds, flowers, pollen, anthers, ovules, pedicels, fruits, meristems, cotyledons, hypocotyls, pods, embryos, endosperm, explants, callus, tissue cultures, shoots, cells, and protoplasts. In a further aspect, the present disclosure provides tobacco plant cells, tissues, and organs that are not reproductive material and do not mediate the natural reproduction of plants. In another aspect, the present disclosure also provides tobacco plant cells, tissues, and organs that are reproductive material and mediate the natural reproduction of plants. In another aspect, the present disclosure provides tobacco plant cells, tissues, and organs that cannot sustain themselves through photosynthesis. In another aspect, the present disclosure provides somatic plant cells. Somatic cells, unlike germline cells, do not mediate the reproduction of plants.

[0221] The provided tobacco cells, tobacco tissues, and tobacco organs can be derived from seeds, fruits, leaves, cotyledons, hypocotyls, meristems, embryos, endosperms, roots, shoots, stems, sheaths, flowers, inflorescences, stalks, pedicels, styles, stigmas, receptacles, petals, sepals, pollen, anthers, filaments, ovaries, ovules, pericarp, phloem, and vascular tissues. In another aspect, the present disclosure provides tobacco plant chloroplasts. In a further aspect, the present disclosure provides epidermal cells, stomatal cells, trichome cells, root hairs, or storage roots.

[0222] In one aspect, the present disclosure provides a tobacco protoplast cell. In another aspect, the present disclosure provides a tobacco callus cell. In another aspect, the present disclosure provides a tobacco seed cell. In another aspect, the present disclosure provides a tobacco fruit cell. In another aspect, the present disclosure provides a tobacco leaf cell. In another aspect, the present disclosure provides a tobacco cotyledon cell. In another aspect, the present disclosure provides a tobacco hypocotyl cell. In another aspect, the present disclosure provides a tobacco meristem cell. In another aspect, the present disclosure provides a tobacco embryo cell. In another aspect, the present disclosure provides a tobacco root cell. In another aspect, the present disclosure provides a tobacco shoot cell. In another aspect, the present disclosure provides a tobacco stem cell. In another aspect, the present disclosure provides a tobacco flower cell. In another aspect, the present disclosure provides a tobacco inflorescence cell. In another aspect, the present disclosure provides a tobacco stalk cell. In another aspect, the present disclosure provides a tobacco pedicel cell. In another aspect, the present disclosure provides a tobacco style cell. In another aspect, the present disclosure provides a tobacco stigma cell. In another aspect, the present disclosure provides a tobacco receptacle cell. In another aspect, the present disclosure provides a tobacco petal cell. In another aspect, the present disclosure provides a tobacco sepal cell. In another aspect, the present disclosure provides a tobacco pollen cell. In another aspect, the present disclosure provides a tobacco anther cell. In another aspect, the present disclosure provides a tobacco filament cell. In another aspect, the present disclosure provides a tobacco ovary cell. In another aspect, the present disclosure provides a tobacco ovule cell. In another aspect, the present disclosure provides a tobacco pericarp cell. In another aspect, the present disclosure provides a tobacco phloem cell.

[0223] The present disclosure provides modified tobacco plants, modified tobacco plant parts, modified tobacco seeds, and modified tobacco cells, as well as methods for producing the same. In one aspect, the present disclosure provides tobacco leaves of modified tobacco plants. In another aspect, the present disclosure provides tobacco seeds of modified tobacco plants. In another aspect, the present disclosure provides tobacco stems of modified tobacco plants. In a further aspect, the present disclosure provides tobacco plant parts of modified tobacco plants. In another aspect, the present disclosure provides tobacco cells of modified tobacco plants. In a further aspect, the present disclosure provides dried tobacco leaves of modified tobacco plants. In yet a further aspect, the present disclosure provides cured tobacco leaves of modified tobacco plants. In yet another aspect, the present disclosure provides fermented tobacco leaves of modified tobacco plants.

[0224] In another aspect, the present disclosure provides alkaloids extracted from modified tobacco plants. In another aspect, the present disclosure provides nicotine extracted from modified tobacco plants. In another aspect, the present disclosure provides anatabine extracted from modified tobacco plants. In another aspect, the present disclosure provides anabasine extracted from modified tobacco plants. In another aspect, the present disclosure provides nornicotine extracted from modified tobacco plants.

[0225] In one aspect, the modified tobacco plant, plant part, seed, cell, or genome is cisgenic. As used herein, "cisgenic" refers to the genetic modification of a plant, plant cell, or plant genome in which all parts (e.g., promoters, donor nucleic acids, selection genes) have only plant origin (i.e., no parts of non-plant origin are used). The cisgenic plants, plant cells, and plant genomes provided herein can result in ready-to-use tobacco lines. In another aspect, the modified tobacco plants provided herein do not contain non-tobacco genetic material or sequences.

[0226] In one aspect, the modified plant comprises an increased leaf yield weight relative to a control tobacco plant when grown under equivalent conditions, hi one aspect, the leaf yield is selected from the group consisting of fresh leaf yield weight, dry leaf yield weight, and cured leaf yield weight.

[0227] In one aspect, the modified plant comprises at least 0.5% increased leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least 0.5% increased fresh leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least 0.5% increased dry leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least 0.5% increased dried leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least 1% increased leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least 1% increased fresh leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least 1% increased dry leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 1% increased dried leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 2% increased leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 2% increased fresh leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 2% increased dried leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 2% increased dried leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 3% increased leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 3% increased fresh leaf yield weight compared to a control tobacco plant when grown under equivalent conditions.In one aspect, the modified plant comprises an increased dry leaf yield weight of at least 3% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased dry leaf yield weight of at least 3% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased dry leaf yield weight of at least 4% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased fresh leaf yield weight of at least 4% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased dry leaf yield weight of at least 4% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased dry leaf yield weight of at least 4% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased dry leaf yield weight of at least 4% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased dry leaf yield weight of at least 5% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 5% increased fresh leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 5% increased dry leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 5% increased dry leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 250% increased leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 250% increased fresh leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 250% increased dry leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plants comprise an increased dry leaf yield mass of at least 250% compared to control tobacco plants when grown under equivalent conditions.In one aspect, the modified plant comprises at least a 25% increased leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 25% increased fresh leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 25% increased dry leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 25% increased dried leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 50% increased leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 50% increased fresh leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises at least a 50% increased dry leaf yield weight compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased dried leaf yield weight of at least 50% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased leaf yield weight of at least 75% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased fresh leaf yield weight of at least 75% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased dried leaf yield weight of at least 75% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased dried leaf yield weight of at least 75% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased dried leaf yield weight of at least 100% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plants comprise at least 100% increased fresh leaf yield mass compared to control tobacco plants when grown under equivalent conditions.In one aspect, the modified plant comprises an increased dry leaf yield mass of at least 100% compared to a control tobacco plant when grown under equivalent conditions. In one aspect, the modified plant comprises an increased dry leaf yield mass of at least 100% compared to a control tobacco plant when grown under equivalent conditions.

[0228] "Curing" is a ripening process that reduces moisture, results in the breakdown of chlorophyll, golden-colored tobacco leaves, and converts starch to sugars. Thus, cured tobacco has a higher reducing sugar content and a lower starch content compared to harvested green leaves. In one aspect, the tobacco plants or plant components provided herein can be dried using conventional means, such as hot air curing, barn-curing, flame-curing, air-drying, or sun-drying. For a description of different types of curing methods, see, for example, Chapter 1 of Tso (1999, Tobacco, Production, Chemistry and Technology, Davis & Nielsen, eds., Blackwell Publishing, Oxford). Cured tobacco is typically aged in a compressed state in wooden drums (e.g., vats) or cardboard boxes for several years (e.g., 2-5 years) at a moisture content ranging from 10% to about 25%. See U.S. Patent Nos. 4,516,590 and 5,372,149. The cured and aged tobacco can then be further processed, including conditioning the tobacco under vacuum with or without the introduction of steam at various temperatures, pasteurization, and fermentation.

[0229] In one aspect, the cured tobacco provided herein is selected from the group consisting of air-cured tobacco, flue-cured tobacco, sun-cured tobacco, and flue-cured tobacco. In another aspect, the cured tobacco material provided herein is selected from the group consisting of air-cured tobacco, flue-cured tobacco, sun-cured tobacco, and flue-cured tobacco. In one aspect, the cured tobacco is from a tobacco variety selected from the group consisting of flue-cured, bright, burley, Virginia, Maryland, dark, Oriental, and Turkish. In another aspect, the cured tobacco material is from a tobacco variety selected from the group consisting of flue-cured, bright, burley, Virginia, Maryland, dark, Oriental, and Turkish.

[0230] Fermentation is typically characterized by high initial moisture content, heat release, and a 10-20% loss in dry weight. See, e.g., U.S. Patent Nos. 4,528,993, 4,660,577, 4,848,373, and 5,372,149; U.S. Patent Application Publication No. 2005 / 0178398; and Tso (1999, Chapter 1 in Tobacco, Production, Chemistry and Technology, Davis & Nielsen, eds., Blackwell Publishing, Oxford). The dried, aged, and fermented tobacco can be further processed (e.g., cut, shredded, expanded, or blended). See, e.g., U.S. Patent Nos. 4,528,993, 4,660,577, and 4,987,907. In one aspect, the cured tobacco material of the present disclosure is hot-air cured, sun-cured, air-dried, or flame-dried.

[0231] In one aspect, the tobacco plants, seeds, plant parts, plant cells, and plant genomes provided herein are from a tobacco species selected from the group consisting of flue-cured, sun-cured, air-cured, dark air-cured, and dark flue-cured tobacco. In another aspect, the tobacco plants, seeds, plant parts, plant cells, and plant genomes provided herein are from a tobacco species selected from the group consisting of burley, Maryland, bright, Virginia, Oriental, Turkish, and Galpao tobacco.

[0232] In one aspect, the modified tobacco plants provided herein are of a tobacco variety selected from the group consisting of flue-cured, bright, burley, Virginia, Maryland, Galpao, dark, Oriental, and Turkish varieties.

[0233] In one aspect, the modified tobacco plants provided herein are selected from the group consisting of flue-cured tobacco plants, bright tobacco plants, burley tobacco plants, Virginia tobacco plants, Maryland tobacco plants, Galpao tobacco plants, dark tobacco plants, Oriental tobacco plants, and Turkish tobacco plants. In one aspect, the modified tobacco cells provided herein are selected from the group consisting of flue-cured tobacco cells, bright tobacco cells, burley tobacco cells, Virginia tobacco cells, Maryland tobacco cells, Galpao tobacco cells, dark tobacco cells, Oriental tobacco cells, and Turkish tobacco cells. In one aspect, the modified tobacco plant parts provided herein are selected from the group consisting of flue-cured tobacco plant parts, bright tobacco plant parts, burley tobacco plant parts, Virginia tobacco plant parts, Maryland tobacco plant parts, Galpao tobacco plant parts, dark tobacco plant parts, Oriental tobacco plant parts, and Turkish tobacco plant parts. In one aspect, the modified tobacco seeds provided herein are selected from the group consisting of flue-cured tobacco seeds, bright tobacco seeds, burley tobacco seeds, Virginia tobacco seeds, Maryland tobacco seeds, Galpao tobacco seeds, dark tobacco seeds, Oriental tobacco seeds, and Turkish tobacco seeds.

[0234] As used herein, a "hybrid" is produced by crossing two plants from different varieties or species so that the offspring contain genetic material from each parent. Those skilled in the art will recognize that higher-order hybrids can also be produced. For example, a first hybrid can be produced by crossing variety C with variety D to produce a C x D hybrid, and a second hybrid can be produced by crossing variety E with variety F to produce an E x F hybrid. The first and second hybrids can be further crossed to produce a higher-order hybrid (C x D) x (E x F) that contains genetic information from all four parent varieties. In one aspect, the modified tobacco plants provided herein are hybrid tobacco plants. In another aspect, the modified tobacco seeds provided herein are hybrid tobacco seeds.

[0235] As used herein, the term "crossing" refers to the intentional mating of two plants. In one aspect, crossing involves pollination and / or fertilization of a first tobacco plant with a second tobacco plant. The two tobacco plants being crossed can be distantly related, closely related, or identical. In one aspect, both of the two tobacco plants being crossed are modified tobacco plants. In one aspect, the two tobacco plants being crossed are of the same tobacco variety. In one aspect, the two tobacco plants being crossed are of two different tobacco varieties. In one aspect, one of the two tobacco plants being crossed is male sterile. In one aspect, one of the two tobacco plants being crossed is female sterile. In one aspect, at least one of the two tobacco plants being crossed is a hybrid tobacco plant. In one aspect, at least one of the two tobacco plants being crossed is a modified tobacco plant.

[0236] In one aspect, the present disclosure provides a method for producing a modified tobacco plant, comprising: (a) crossing at least one tobacco plant of a first tobacco variety with at least one tobacco plant of a second tobacco variety, wherein the at least one tobacco plant of the first tobacco variety comprises a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and wherein the at least one tobacco plant, when grown under equivalent growing conditions, exhibits no or reduced suckers after topping compared to the control tobacco plant; and (b) selecting progeny tobacco plants that exhibit no or reduced suckers after topping compared to the control tobacco plant of the same hybrid grown under equivalent growing conditions. The present invention provides a method comprising:

[0237] In another aspect, the present disclosure provides a method for producing a modified tobacco plant, comprising: (a) crossing at least one tobacco plant of a first tobacco variety with at least one tobacco plant of a second tobacco variety, wherein the at least one tobacco plant of the first tobacco variety comprises a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, and wherein the at least one tobacco plant, when grown under equivalent growing conditions, exhibits no or reduced suckers after topping compared to a control tobacco plant; and (b) selecting progeny tobacco plants that exhibit no or reduced suckers after topping compared to a control tobacco plant of the same hybrid grown under equivalent growing conditions. The present invention provides a method comprising:

[0238] In a further aspect, the present disclosure provides a method for producing a modified tobacco plant, comprising: (a) crossing at least one tobacco plant of a first tobacco variety with at least one tobacco plant of a second tobacco variety, wherein the at least one tobacco plant of the first tobacco variety comprises a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous genomic locus encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, and wherein the at least one tobacco plant, when grown under equivalent growing conditions, exhibits no or reduced suckers after topping compared to a control tobacco plant; and (b) selecting progeny tobacco plants that exhibit no or reduced suckers after topping compared to a control tobacco plant of the same hybrid grown under equivalent growing conditions. The present invention provides a method comprising:

[0239] In one aspect, the tobacco varieties provided herein are male sterile. In another aspect, the tobacco varieties provided herein are cytoplasmic male sterile (CMS). In one aspect, the modified tobacco plants provided herein are male sterile. In another aspect, the modified tobacco plants provided herein are cytoplasmic male sterile. Male sterile tobacco plants can be produced by any method known in the art. Methods for producing male sterile tobacco are described in Wernsman, EA, and Rufty, RC 1987. Chapter Seventeen. Tobacco. Pages 669-698 In: Cultivar Development. Crop Species. WH Fehr (ed.), MacMillan Publishing Go., Inc., New York, NY 761 pp.

[0240] In another aspect, the tobacco varieties provided herein are female sterile.For example, by mutating STIG1 gene, female sterile plants can be produced.See, for example, Goldman et al. 1994, EMBO Journal 13:2976-2984.In one aspect, the modified tobacco plants provided herein are female sterile.

[0241] Flue-cured tobacco (also known as "Virginia" or "bright" tobacco) accounts for approximately 40% of the world's tobacco production. Flue-cured tobacco is often called "bright tobacco" because of the light yellow to deep orange color it achieves during curing. Flue-cured tobacco has a light, vibrant aroma and taste. Flue-cured tobacco is generally high in sugar and low in oil. The main flue-cured tobacco-growing countries are Argentina, Brazil, China, India, Tanzania, and the United States. In one aspect, the modified tobacco plants or seeds provided herein are of a flue-cured tobacco variety selected from the group consisting of CC13, CC27, CC33, CC35, CC37, CC65, CC67, CC700, GF318, GL338, GL368, GL939, K346, K399, K326, NC102, NC196, NC291, NC297, NC299, NC471, NC55, NC606, NC71, NC72, NC92, PVH1118, PVH1452, PVH2110, SPEIGHT168, SPEIGHT220, SPEIGHT225, SPEIGHT227, SPEIGHT236, and any variety essentially derived from any one of the foregoing varieties. In another aspect, the modified tobacco plants or seeds provided herein are selected from the group consisting of Coker48, Coker176, Coker371-Gold, Coker319, Coker347, GL939, K149, K326, K340, K346, K358, K394, K399, K730, NC27NF, NC37NF, NC55, NC60, NC71, NC72, NC82, NC95, NC297, NC606, NC729, NC2326, McNair373, McNair944, Ox207, Ox414NF, Reams126, Reams713, Reams744, RG8, RG11, RG13, RG17, RG22, RG81, RG H4, RG H51, Speight H-20, Speight G-28, Speight G-58, Speight G-70, Speight G-108, Speight G-111, Speight G-117, Speight 168, Speight 179, Speight NF-3, Va116, Va182 and any variety essentially derived from any one of the foregoing varieties.See WO 2004 / 041006. In a further aspect, the modified tobacco plant or seed provided herein is a flue-colored variety selected from the group consisting of K326, K346, and NC196.

[0242] Air-cured tobaccos include "burley," "Maryland," and "dark" tobacco. The common factor that unites air-cured tobaccos is that the curing occurs primarily without artificial heat and moisture. Burley tobacco is light to dark brown in color, high in oil, and low in sugar. Burley tobacco is typically air-cured in sheds. Major burley-growing countries include Argentina, Brazil, Italy, Malawi, and the United States.

[0243] Maryland tobacco is very fluffy, has good burning characteristics, low nicotine and a neutral aroma. The main countries where Maryland is grown include the United States and Italy.

[0244] In one aspect, the modified tobacco plants or seeds provided herein are of a burley tobacco variety selected from the group consisting of Clay402, Clay403, Clay502, Ky14, Ky907, Ky910, Ky8959, NC2, NC3, NC4, NC5, NC2000, Tn86, Tn90, Tn97, R610, R630, R711, R712, NCBH129, HB4488PLC, PD7319LC, Bu21xKy10, HB04P, Ky14xL8, Kt200, Newton98, Pedigo561, Pf561, and Va509. In a further aspect, the modified tobacco plants or seeds provided herein are of a burley variety selected from the group consisting of TN90, KT209, KT206, KT212, and HB4488. In another aspect, the modified tobacco plants or seeds provided herein are of a Maryland tobacco variety selected from the group consisting of Md10, Md40, Md201, Md609, Md872, and Md341.

[0245] Dark air-cured tobacco is distinguished from other tobacco species primarily by its curing method, which gives it its medium to dark brown color and distinctive flavor. Dark air-cured tobacco is primarily used in the production of chewing tobacco and snuff. In one aspect, the modified tobacco plants or seeds provided herein are of a dark air-cured tobacco variety selected from the group consisting of Sumatra, Jatim, Dominican Cubano, Besuki, One Sucker, Green River, Virginia Sun-cured, and Paraguan Passado.

[0246] Dark flue-cured tobacco is generally cured using a low-burning wood fire on the floor of an enclosed curing shed. Dark flue-cured tobacco is typically used to make pipe blends, cigarettes, chewing tobacco, snuff, and strong cigars. The primary growing areas for dark flue-cured tobacco are Tennessee, Kentucky, and Virginia in the United States. In one aspect, the modified tobacco plants or seeds provided herein are of a dark flue-cured tobacco variety selected from the group consisting of Narrow Leaf Madole, Improved Madole, Tom Rosson Madole, Newton's VH Madole, Little Crittenden, Green Wood, Little Wood, Small Stalk Black Mammoth, DT508, DT518, DT592, KY171, DF911, DF485, TN D94, TN D950, VA309, and VA359.

[0247] Oriental tobacco is also called Greek, aromatic, and Turkish tobacco due to the fact that it is typically grown in Eastern Mediterranean regions such as Turkey, Greece, Bulgaria, Macedonia, Syria, Lebanon, Italy, and Romania. The small plant size, small leaf size, and unique aroma characteristics of Oriental tobacco varieties are the result of adaptation to the poor soil and stressful climatic conditions in which they have been grown. In one aspect, the modified tobacco plants or seeds provided herein are grown in the following regions: Izmir, Katerini, Samsun, Basma, Krumovgrad, Trabzon, Thesalian, Tasova, Sinop, Izmit, Hendek, Edirne, Şemdinli, Adıyaman The tobacco is of an oriental variety selected from the group consisting of Adiyanman, Yayladag, Iskenderun, Duzce, Macedonian, Mavra, Prilep, Bafra, Bursa, Bucak, Bitlis, Balikesir and any variety essentially derived from any one of the aforementioned varieties.

[0248] The tobacco plants, plant parts, and tobacco materials provided herein can be used in any tobacco product. As used herein, "tobacco product" is defined as any product made from or derived from tobacco, intended for human use or consumption. In one aspect, the tobacco products provided herein comprise dried ingredients from the tobacco plants provided herein. In another aspect, the tobacco products provided herein comprise cured tobacco leaves from the tobacco plants provided herein. In one aspect, the present disclosure provides a tobacco product comprising a cured tobacco material from any tobacco plant provided herein.

[0249] In one aspect, the present disclosure provides a tobacco product comprising cured tobacco material from a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and wherein the modified tobacco plant, when grown under equivalent growing conditions, produces no or reduced suckers after topping compared to the control tobacco plant. In another aspect, the disclosure provides a tobacco product comprising cured tobacco material from a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety.

[0250] In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and wherein the modified tobacco plant, when grown under equivalent growing conditions, produces no or reduced suckers after topping compared to the control tobacco plant. In another aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence operably linked to a heterologous promoter, encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the modified tobacco plant, when grown under equivalent growing conditions, produces no or reduced suckers after topping compared to the control tobacco plant.

[0251] In one aspect, the disclosure provides a tobacco product comprising cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence operably linked to a heterologous promoter, encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the modified tobacco plant, when grown under equivalent growing conditions, produces no or reduced suckers after topping compared to a control tobacco plant. In another aspect, the disclosure provides a tobacco product comprising cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence operably linked to a heterologous promoter, encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor.

[0252] In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter, wherein the modified tobacco plant, when grown under equivalent growing conditions, comprises no or reduced suckers after topping compared to a control tobacco plant. In another aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter.

[0253] In one aspect, the present disclosure provides a tobacco product comprising cured tobacco material from a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the modified tobacco plant, when grown under equivalent growing conditions, produces no or reduced suckers after topping compared to a control tobacco plant. In another aspect, the disclosure provides a tobacco product comprising cured tobacco material from a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor.

[0254] In one aspect, the present disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the modified tobacco plant, when grown under equivalent growing conditions, produces no or reduced suckers after topping compared to a control tobacco plant. In another aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor.

[0255] Alkaloid compounds can be extracted or isolated from any tobacco plant or plant part provided herein. In one aspect, the tobacco products provided herein comprise alkaloids extracted from modified tobacco plants or parts thereof. In another aspect, the tobacco products provided herein comprise nicotine extracted from tobacco plants or tobacco plant parts. In another aspect, the tobacco products provided herein comprise anatabine extracted from tobacco plants or tobacco plant parts. In another aspect, the tobacco products provided herein comprise anabasine extracted from tobacco plants or tobacco plant parts. In one aspect, the tobacco products provided herein comprise nornicotine extracted from tobacco plants or plant parts. In one aspect, the tobacco products provided herein comprise alkaloids extracted from modified tobacco plants, wherein the alkaloid is selected from the group consisting of nicotine, nornicotine, anatabine, and anabasine.

[0256] The alkaloid compounds extracted from tobacco plants or tobacco plant parts provided herein can be used to produce compositions suitable for use with non-combustion products. Exemplary non-combustion products include electronic cigarettes ("e-cigarettes"), electronic smoking articles, e-vapor products, aerosolized vapor products, and heated tobacco products. In one aspect, the non-combustion products provided herein comprise alkaloids extracted from tobacco plants or tobacco plant parts provided herein. In one aspect, the non-combustion products provided herein comprise nicotine extracted from tobacco plants or tobacco plant parts provided herein. In one aspect, the non-combustion products provided herein comprise anabasine extracted from tobacco plants or tobacco plant parts provided herein. In one aspect, the non-combustion products provided herein comprise anatabine extracted from tobacco plants or tobacco plant parts provided herein. In one aspect, the non-combustion products provided herein comprise nornicotine extracted from tobacco plants or tobacco plant parts provided herein.

[0257] In one aspect, the non-combustible product provided herein is an e-cigarette. In another aspect, the non-combustible product provided herein is an electronic smoking article. In another aspect, the non-combustible product provided herein is an aerosolized vapor product. In another aspect, the non-combustible product provided herein is a heated tobacco product. In another aspect, the non-combustible product provided herein is an e-vapor product.

[0258] Tobacco products provided herein include, but are not limited to, cigarette products (e.g., cigarettes, bidi cigarettes, kreteks), cigar products (e.g., cigars, cigar wrapping tobacco, cigarillos), pipe tobacco products, tobacco-derived products, tobacco-derived nicotine products, smokeless tobacco products (e.g., moist snuff, dry snuff, chewing tobacco, moist smokeless tobacco, fine cut chewing tobacco, long cut chewing tobacco, pouched chewing tobacco, snus), films, chewables (e.g., gums), lozenges, dissolving strips, tabs, tablets, shaped parts, gels, consumable units, insoluble matrices, hollow shapes, reconstituted tobacco, expanded tobacco, and the like. See, e.g., U.S. Patent Application Publication No. 2006 / 0191548.

[0259] As used herein, a "cigarette" refers to a tobacco product having a "rod" and a "filler." A cigarette "rod" includes cigarette paper, a filter, plug wrap (used to contain the filtration material), tipping paper that holds the cigarette paper (including the filler) to the filter, and any adhesives that hold these components together. "Filler" includes (1) all tobacco, including but not limited to reconstituted and expanded tobacco, (2) non-tobacco substitutes (including but not limited to herbs, non-tobacco plant materials, and other flavorings that may accompany tobacco wrapped in cigarette paper), (3) casing, (4) flavorings, and (5) all other additives (mixed with the tobacco and substitutes and wrapped into the cigarette).

[0260] In one aspect, the present disclosure provides nicotine from the modified tobacco plants provided herein and methods of producing nicotine from the modified tobacco plants provided herein for use in products.

[0261] In one aspect, the methods provided herein include preparing a tobacco product using cured tobacco leaves from the modified tobacco plants provided herein.

[0262] As used herein, "reconstituted tobacco" refers to a portion of tobacco filler made from tobacco dust and other tobacco scrap material, processed into sheet form, and cut into strips to resemble tobacco. In addition to cost savings, reconstituted tobacco is very important for contributing to the taste of cigarettes because it processes flavor development using a reaction between ammonia and sugars. In one aspect, the tobacco products provided herein include reconstituted tobacco derived from modified tobacco plants.

[0263] As used herein, "expanded tobacco" refers to a portion of tobacco filler that has been processed through expansion with a suitable gas so that the tobacco is "puffed" to reduce its density and provide greater filling capacity. Expanded tobacco reduces the weight of the tobacco used in a cigarette.

[0264] Tobacco products derived from plants of the present disclosure also include cigarettes and other smoking articles, particularly smoking articles comprising a filter element in which a rod of smokable material comprises tobacco cured in a tobacco blend. In one aspect, the tobacco product of the present disclosure is selected from the group consisting of cigarettes, kreteks, bidi cigarettes, cigars, cigarillos, non-ventilated cigarettes, vented recess filter cigarettes, pipe tobacco, snuff, snus, chewing tobacco, moist smokeless tobacco, fine-cut chewing tobacco, long-cut chewing tobacco, and packaged chewing tobacco products. In another aspect, the tobacco product of the present disclosure is selected from the group consisting of gums, tablets, lozenges, and dissolving strips.

[0265] In one aspect, the tobacco products provided herein comprise cured tobacco material from a modified tobacco plant. In another aspect, the tobacco products provided herein comprise cured tobacco leaf material from a modified tobacco plant. In another aspect, the tobacco products provided herein comprise cured tobacco stem material from a modified tobacco plant.

[0266] In another aspect, the tobacco product of the present disclosure is a smokeless tobacco product. Smokeless tobacco products are not combusted and include, but are not limited to, chewing tobacco, moist smokeless tobacco, snus, and dry snuff. Chewing tobacco is coarsely shredded tobacco typically packaged in a large pouch and used in plugs or twists. Moist smokeless tobacco is a more finely shredded moist tobacco provided in a loose or pouch form, typically packaged in a round can, and used as a pinch or in a pouch placed between the cheek and gum of an adult tobacco consumer. Snus is heat-treated smokeless tobacco. Dry snuff is finely ground tobacco that is held in the mouth or used nasally. In a further aspect, the tobacco product of the present disclosure is selected from the group consisting of loose-leaf chewing tobacco, stick chewing tobacco, moist snuff, and nasal snuff.

[0267] The present disclosure further provides methods for producing tobacco products containing tobacco material from the modified tobacco plants provided herein. In one aspect, the methods provided herein include conditioning aged tobacco material produced from the modified tobacco plants provided herein to increase its moisture content from about 12.5% ​​to about 13.5% to about 21%, and blending the conditioned tobacco material to produce a desired blend. In one aspect, the methods for producing tobacco products provided herein further include casing or flavoring the blend. Typically, during the casing process, casing or source materials are added to the blend to enhance the blend's quality by balancing its chemical composition and to produce certain desired flavor characteristics. Further details of the casing process can be found in Tobacco Production, Chemistry and Technology, Edited by L. Davis and M. Nielsen, Blackwell Science, 1999.

[0268] The present disclosure provides tobacco materials from modified tobacco plants or parts thereof. Tobacco materials obtained from the tobacco plants, cells, lines, varieties, or hybrids of the present disclosure can be used to manufacture tobacco products. In one aspect, the tobacco material includes leaf material. In one aspect, the tobacco material includes stem material. In one aspect, the tobacco material includes fresh tobacco material. In another aspect, the tobacco material includes dried tobacco material. In a further aspect, the tobacco material includes cured tobacco material. In yet another aspect, the tobacco material includes fermented tobacco material. In one aspect, the tobacco materials provided herein can be used in any tobacco product provided herein. In one aspect, the dried tobacco material provided herein includes air-cured tobacco material. In another aspect, the dried tobacco material provided herein includes flue-cured tobacco material. In another aspect, the dried tobacco material provided herein includes sun-cured tobacco material. In another aspect, the dried tobacco material provided herein includes flue-cured tobacco material. In another aspect, the cured tobacco material provided herein is selected from the group consisting of air-cured tobacco material, flue-cured tobacco material, sun-cured tobacco material, and flue-cured tobacco material.

[0269] The tobacco materials provided herein can also be processed using methods including, but not limited to, heat treatment (e.g., cooking, toasting), flavoring, enzyme treatment, expansion, and / or curing. Both fermented and non-fermented tobaccos can be processed using these techniques. Examples of suitable processed tobaccos include dark air-cured, dark flue-cured, burley, flue-cured, and cigar filler or wrapper, as well as products from whole leaf stemming operations. In one aspect, the tobacco fiber comprises up to 70% dark tobacco on a fresh weight basis. For example, the tobacco can be conditioned by heating, sweating, and / or pasteurization processes, as described in U.S. Patent Application Publication Nos. 2004 / 0118422 or 2005 / 0178398.

[0270] The tobacco materials provided herein can be subjected to fermentation. Fermentation is typically characterized by a high initial moisture content, heat generation, and a 10-20% reduction in dry weight. See, for example, U.S. Patent Nos. 4,528,993, 4,660,577, 4,848,373, and 5,372,149. In addition to modifying the aroma of the leaves, fermentation can also alter either or both the color and texture of the leaves. During the fermentation process, gases may be produced, oxygen may be taken up, pH may change, and the amount of water retained may change. See, for example, U.S. Patent Application Publication No. 2005 / 0178398 and Tso (1999, Chapter 1 in Tobacco, Production, Chemistry and Technology, Davis & Nielsen, eds., Blackwell Publishing, Oxford). The cured tobacco or the cured and fermented tobacco may be further processed (e.g., cut, expanded, blended, ground, or comminuted) before incorporation into an oral product. The tobacco is, in some cases, long-cut, fermented, dried, moist tobacco having an oven volatiles content of 48-50% by weight before being mixed with the copolymer and any flavorings and other additives that may be included as needed.

[0271] In one aspect, the tobacco materials provided herein can be processed to a desired size. In certain aspects, tobacco fibers can be processed to have an average fiber size of less than 200 micrometers. In one aspect, the tobacco fibers are 75 to 125 micrometers. In another aspect, the tobacco fibers are processed to have a size of 75 micrometers or less. In one aspect, the tobacco fibers include long cut tobacco that can be cut or shredded to widths of about 10 cuts per inch up to about 110 cuts per inch and lengths of about 0.1 inches up to about 1 inch. Double cut tobacco fibers can have a particle size range such that about 70% of the double cut tobacco fibers fall between -20 mesh and 80 mesh mesh sizes.

[0272] The tobacco materials provided herein can be processed to have a total oven volatiles content of about 10% by weight or more; about 20% by weight or more; about 40% by weight or more; about 15% to about 25% by weight; about 20% to about 30% by weight; about 30% to about 50% by weight; about 45% to about 65% by weight; or about 50% to about 60% by weight. Those skilled in the art will understand that "moist" tobacco typically refers to tobacco having an oven volatiles content of about 40% to about 60% by weight (e.g., about 45% to about 55% by weight, or about 50% by weight). As used herein, "oven volatiles" is determined by calculating the percentage weight loss of a sample after drying the sample for 3.25 hours in a preheated forced-air oven at 110°C. Oral products can have a total oven volatiles content that is different from the oven volatiles content of the tobacco fiber used to manufacture the oral product. The processing steps described herein can reduce or increase the oven volatile content.

[0273] The following exemplary, non-limiting embodiments are contemplated: 1. A modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to the control tobacco plant. 2. A modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein said mutation is not present in said endogenous gene in a control tobacco plant of the same variety. 3. A modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent cultivation conditions. 4. A modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter. 5. A modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing the expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent cultivation conditions. 6. A modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of cyclin, cyclin-dependent kinase (CDK), CDK inhibitor, MYB, and WRKY transcription factor. 7. The modified tobacco plant of embodiment 1 or 2, wherein the mutation comprises a mutation selected from the group consisting of an insertion, a deletion, a substitution, and an inversion. 8. The modified tobacco plant of any one of aspects 1, 2, or 7, wherein the mutation comprises a null mutation. 9. The modified tobacco plant of any one of aspects 1, 2, 7, or 8, wherein the mutation results in a premature stop codon in an mRNA transcript of the endogenous gene. 10. The modified tobacco plant of any one of aspects 1, 2, or 7-9, wherein the mutation results in a truncation of the polypeptide. 11. The modified tobacco plant of any one of aspects 1, 2, or 7-10, wherein the mutation is located within an exon of the endogenous gene. 12. The modified tobacco plant of any one of aspects 1, 2, or 7-10, wherein the mutation is located within an intron of the endogenous gene. 13. The modified tobacco plant of any one of aspects 1, 2, or 7, wherein the mutation is located within the 5'-untranslated region (UTR) or 3'-UTR of the endogenous gene. 14. The modified tobacco plant of any one of aspects 1, 2, or 7, wherein the mutation is located within a promoter of the endogenous gene. 15. The modified tobacco plant of any one of aspects 1, 2, or 7-14, wherein the endogenous gene comprises a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. 16. The modified tobacco plant of any one of aspects 1, 2, or 7-15, wherein the endogenous gene encodes a polypeptide comprising an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114. 17. The modified tobacco plant of any one of aspects 1, 2, or 7-16, wherein the mutation results in a reduced level of expression of the endogenous gene compared to the control tobacco plant. 18. The modified tobacco plant of any one of aspects 1, 2, or 7-16, wherein the mutation results in an increased level of expression of the endogenous gene compared to the control tobacco plant. 19. The modified tobacco plant of any one of aspects 1, 2, or 7-16, wherein the mutation results in a reduced level of activity of the polypeptide compared to the control tobacco plant. 20. The modified tobacco plant of any one of aspects 1, 2, or 7-16, wherein the mutation results in an increased level of activity of the polypeptide compared to the control tobacco plant. 21. The modified tobacco plant of any one of aspects 3 to 6, wherein the heterologous promoter comprises an axillary meristem-specific promoter. 22. The modified tobacco plant of any one of aspects 3 to 6, wherein the heterologous promoter comprises an axillary meristem-preferred promoter. 23. The modified tobacco plant of any one of aspects 3 to 6, wherein the heterologous promoter comprises a polynucleotide sequence that is at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 89 to 109. 24. The modified tobacco plant of any one of aspects 3 to 6, wherein the small RNA molecule is selected from the group consisting of double-stranded RNA, small interfering RNA (siRNA), trans-acting siRNA, and microRNA. 25. The modified tobacco plant of aspect 5 or 6, wherein the small RNA molecule comprises 18 to 30 nucleotides. 26. The modified tobacco plant of aspect 5 or 6, wherein the small RNA molecule comprises a polynucleotide sequence having at least 90% identity or complementarity to the endogenous mRNA. 27. The modified tobacco plant of aspect 5 or 6, wherein the small RNA molecule comprises 18 to 30 nucleotides. 28. The modified tobacco plant of aspect 5 or 6, wherein the small RNA molecule comprises a polynucleotide sequence having at least 90% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. 29. The modified tobacco plant of aspect 5 or 6, wherein the small RNA molecule comprises a polynucleotide sequence identical to or complementary to at least 18 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113. 30. The modified tobacco plant of any one of aspects 1 to 29, wherein the polypeptide is a cyclin. 31. The modified tobacco plant of any one of aspects 1 to 29, wherein the polypeptide is a CDK. 32. The modified tobacco plant of any one of aspects 1 to 29, wherein the polypeptide is a CDK inhibitor. 33. The modified tobacco plant of any one of aspects 1 to 29, wherein the polypeptide is MYB. 34. The modified tobacco plant of any one of aspects 1 to 29, wherein the polypeptide is a WRKY transcription factor. 35. The modified tobacco plant of aspect 30, wherein the cyclin is encoded by a polynucleotide sequence that is at least 80% identical to any of SEQ ID NOs: 6-32. 36. The modified tobacco plant of aspect 30, wherein the cyclin comprises an amino acid sequence that is at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 50 to 76. 37. The modified tobacco plant of aspect 31, wherein the CDK is encoded by a polynucleotide sequence that is at least 80% identical to SEQ ID NOs: 1-4. 38. The modified tobacco plant of aspect 31, wherein the CDK comprises an amino acid sequence at least 80% identical to or at least 80% similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45 to 48. 39. The modified tobacco plant of aspect 32, wherein the CDK inhibitor is encoded by a polynucleotide sequence that is at least 80% identical to SEQ ID NO:5. 40. The modified tobacco plant of aspect 32, wherein the CDK inhibitor comprises an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NO: 49. 41. The modified tobacco plant of aspect 33, wherein the MYB is encoded by a polynucleotide sequence that is at least 80% identical to any of SEQ ID NOs: 33-44. 42. The modified tobacco plant of aspect 33, wherein the MYB comprises an amino acid sequence at least 80% identical to or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 77 to 88. 43. The modified tobacco plant of embodiment 34, wherein the WRKY transcription factor is encoded by a polynucleotide sequence that is at least 80% identical to SEQ ID NO: 113. 44. The modified tobacco plant of embodiment 34, wherein the WRKY transcription factor comprises an amino acid sequence at least 80% identical or similar to an amino acid sequence at least 80% identical or similar to SEQ ID NO: 114. 45. The modified tobacco plant of any one of aspects 1 to 44, wherein the modified tobacco plant is of a tobacco variety selected from the group consisting of flue-cured, bright, burley, Virginia, Maryland, dark, Galpao, Oriental, and Turkish varieties. 46. Preface modified plant, BU64 plant, CC101 plant, CC200 plant, CC13 plant, CC27 plant, CC33 plant, CC35 plant, CC37 plant, CC65 Plants, CC67 plants, CC301 plants, CC400 plants, CC500 plants, CC600 plants, CC700 plants, CC800 plants, CC900 plants, CC1063 plants Plants, Coker176 plants, Coker319 plants, Coker371Gold plants, Coker48 plants, CU263 plants, DF911 plants, Galpao plants, GL26H plants, GL338 plants, GL350 plants, GL395 plants, GL600 plants, GL737 plants, GL939 plants, GL973 plants, GF157 plants, GF318 plants, RJR901 plants, HB04P plants, K149 plants, K326 plants, K346 plants, K358 plants, K394 plants, K399 plants, K730 plants, NC196 plants, NC37NF plants, NC471 plants, NC55 plants, NC92 plants, NC2326 plants, NC95 plants, NC925 plants, PVH1118 plants, PVH1452 plants, PVH2110 plants, PVH2254 plants, PVH2275 plants, VA116 plants, VA119 plants, KDH959 plants, KT200 plants, KT204LC plants, KY10 plants, KY14 plants, KY160 plants, KY17 plants, KY171 plants, KY907 plants, KY907LC plants, KTY14xL8 LC plants, Little Crittenden plants, McNair373 plants, McNair944 plants, Male-Immature KY14xL8 plants, Narrow Leaf Madole plants, MS KY171 plants, Narrow Leaf Madole (phph) plants, MS Narrow Leaf Madole plants, MSTND950 plant, PD7302LC plant, PD7305LC plant, PD7309LC plant, PD7312LC plant, PD7318LC plant, PD7319LC plant, MSTKS2002 plant, TKF2002 plant, TKF6400 plant, TKF4028 plant, TKF4024 plant, KT206LC plant, KT 209LC plant, KT210LC plant, KT212LC plant, NC100 plant, NC102 plant, NC2000 plant, NC291 plant, NC297 plant, NC299 plant, NC3 plant, NC4 plant, NC5 plant, NC6 plant, NC7 plant, NC606 plant, NC71 plant, NC72 plant, NC810 plant, NC BH 129 Plant, NC2002 Plant, Neal Smith Madole Plant, OXFORD 207 plants, "Perique" plants, PVH03 plants, PVH09 plants, PVH19 plants, PVH50 plants, PVH51 plants, R610 plants, R630 plants, R7-11 plants, R7-12 plants, RG17 plants, RG81 plants, RG 46. ​​The modified tobacco plant of any one of embodiments 1 to 45, wherein the modified tobacco plant is selected from the group consisting of an H51 plant, an RGH4 plant, an RGH51 plant, an RS1410 plant, a Speight168 plant, a Speight172 plant, a Speight179 plant, a Speight210 plant, a Speight220 plant, a Speight225 plant, a Speight227 plant, a Speight234 plant, a SpeightG-28 plant, a SpeightG-70 plant, a SpeightH-6 plant, a SpeightH20 plant, a SpeightNF3 plant, a TI1406 plant, a TI1269 plant, a TN86 plant, a TN86LC plant, a TN90 plant, a TN90LC plant, a TN97 plant, a TN97LC plant, a TN D94 plant, a TN D950 plant, a TR (Tom Rosson) Madole plant, a VA309 plant, and a VA359 plant. 47. The modified tobacco plant of any one of aspects 1-46, wherein the modified tobacco plant is a hybrid. 48. The modified tobacco plant of any one of aspects 1-47, wherein the modified tobacco plant is male sterile or cytoplasmic male sterile. 49. The modified tobacco plant of any one of aspects 1-47, wherein the modified tobacco plant is female sterile. 50. A modified tobacco plant described in any one of aspects 1, 3 or

[0009] , wherein the reduced suckers comprise fewer total suckers, a smaller average sucker size, or both, compared to the suckers of the control tobacco plant when grown under equivalent growing conditions. 51. The modified tobacco plant of embodiment 50, wherein the smaller average sucker size comprises a measurement selected from the group consisting of a reduced average mass, a reduced average length, a reduced average diameter, or any combination thereof, compared to suckers of the control tobacco plant when grown under equivalent cultivation conditions. 52. The modified tobacco plant of any one of aspects 1-51, wherein the modified plant has an increased leaf yield mass compared to the control tobacco plant when grown under equivalent growing conditions. 53. The modified tobacco plant of aspect 52, wherein the increased leaf yield mass comprises an increase of at least 0.5%. 54. A tobacco leaf of the modified tobacco plant of any one of aspects 1 to 53. 55. A tobacco seed of the modified tobacco plant of any one of aspects 1 to 53. 56. The tobacco leaf of aspect 54, wherein the tobacco leaf is a cured tobacco leaf. 57. The tobacco leaf of aspect 56, wherein the cured tobacco leaf is selected from the group consisting of air-cured tobacco leaf, flue-cured tobacco leaf, sun-cured tobacco leaf, and flue-cured tobacco leaf. 58. A tobacco product comprising an alkaloid extracted from the modified tobacco plant or part thereof of any one of aspects 1-53. 59. The tobacco product of aspect 58, wherein the alkaloid is selected from the group consisting of nicotine, nornicotine, anatabine, and anabasine. 60. The tobacco product of aspect 58 or 59, wherein the tobacco product is selected from t...

Claims

1. 1. A modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, A modified tobacco plant, wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and wherein the modified tobacco plant, when grown under equivalent cultivation conditions, has no or reduced suckers after topping compared to the control tobacco plant.

2. 1. A modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, A modified tobacco plant, wherein said mutation is not present in said endogenous gene in a control tobacco plant of the same variety.

3. 1. A modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter, A modified tobacco plant, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions.

4. A modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter.

5. 1. A modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; A modified tobacco plant, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions.

6. A recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing the expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor. A modified tobacco plant, comprising:

7. 3. The modified tobacco plant of claim 1, wherein the endogenous gene comprises a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113.

8. 3. The modified tobacco plant of claim 1 or 2, wherein the endogenous gene encodes a polypeptide comprising an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 45-88 and 114.

9. 7. The modified tobacco plant of claim 3, wherein the heterologous promoter comprises a polynucleotide sequence that is at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 89 to 109.

10. 7. The modified tobacco plant of claim 5 or 6, wherein the small RNA molecule comprises a polynucleotide sequence having at least 90% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113.

11. A tobacco leaf of the modified tobacco plant of any one of claims 1 to 6.

12. A tobacco seed of the modified tobacco plant of any one of claims 1 to 6.

13. 55. The tobacco leaf of claim 54, wherein the tobacco leaf is a cured tobacco leaf.

14. 10. A tobacco product comprising an alkaloid extracted from the modified tobacco plant or part thereof according to any one of claims 1 to 6.

15. 1. A tobacco product comprising cured tobacco material from a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, A tobacco product wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to the control tobacco plant.

16. 1. A tobacco product comprising cured tobacco material from a modified tobacco plant comprising a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, A tobacco product, wherein said mutation is not present in said endogenous gene in a control tobacco plant of the same variety.

17. 1. A tobacco product comprising: cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter; A tobacco product in which the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions.

18. A tobacco product comprising cured tobacco material from a modified tobacco plant comprising a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter.

19. 1. A tobacco product comprising cured tobacco material from a modified tobacco plant comprising a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, A tobacco product in which the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions.

20. A tobacco product comprising cured tobacco material from a modified tobacco plant containing a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of cyclin, cyclin-dependent kinase (CDK), CDK inhibitor, MYB, and WRKY transcription factor.

21. 1. A method for producing a modified tobacco plant, comprising: (a) inducing a mutation in at least one tobacco cell at a genomic locus encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; (b) selecting at least one tobacco cell containing the mutation from step (a); (c) regenerating a modified tobacco plant from the at least one tobacco cell selected in step (b), wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant lacking the mutation.

22. 1. A method for producing a modified tobacco plant, comprising: (a) introducing a recombinant DNA construct into at least one tobacco cell, wherein the recombinant DNA construct comprises a heterologous promoter operably linked to a nucleic acid encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; (b) selecting at least one tobacco cell containing the recombinant DNA construct; (c) regenerating a modified tobacco plant from the at least one tobacco cell selected in step (b), wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant lacking the recombinant DNA construct.

23. 1. A method for producing a modified tobacco plant, comprising: (a) introducing into at least one tobacco cell a recombinant DNA construct, wherein the recombinant DNA construct comprises a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; (b) selecting at least one tobacco cell containing the recombinant DNA construct; (c) regenerating a modified tobacco plant from the at least one tobacco cell selected in step (b), wherein the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant lacking the recombinant DNA construct.

24. preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; A method comprising: wherein the mutation is not present in the endogenous gene in a control tobacco plant of the same variety, and the modified tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to the control tobacco plant.

25. preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter; A method comprising: The method, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions.

26. preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter; A method comprising: The method, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions.

27. preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a polynucleotide sequence encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, operably linked to a heterologous promoter; A method comprising:

28. preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; A method comprising: The method, wherein the modified tobacco plant has no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions.

29. preparing a tobacco product using cured tobacco material from a modified tobacco plant containing a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; A method comprising:

30. transforming a tobacco cell with a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor; A method comprising:

31. Transforming tobacco cells with a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous mRNA encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor. A method comprising:

32. 1. A method for producing a modified tobacco plant, comprising: (a) crossing at least one tobacco plant of a first tobacco variety with at least one tobacco plant of a second tobacco variety, wherein said at least one tobacco plant of said first tobacco variety comprises a mutation in an endogenous gene encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, said mutation being absent in said endogenous gene in a control tobacco plant of the same variety, and wherein said at least one tobacco plant, when grown under equivalent growing conditions, comprises no or reduced suckers after topping compared to said control tobacco plant; (b) selecting progeny tobacco plants that exhibit no or reduced suckers after topping compared to control tobacco plants of the same hybrid grown under comparable growing conditions;

33. 1. A method for producing a modified tobacco plant, comprising: (a) crossing at least one tobacco plant of a first tobacco variety with at least one tobacco plant of a second tobacco variety, wherein said at least one tobacco plant of said first tobacco variety comprises a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, and wherein said at least one tobacco plant, when grown under equivalent growing conditions, has no or reduced suckers after topping compared to a control tobacco plant; (b) selecting progeny tobacco plants that exhibit no or reduced suckers after topping compared to control tobacco plants of the same hybrid grown under comparable growing conditions;

34. 1. A method for producing a modified tobacco plant, comprising: (a) crossing at least one tobacco plant of a first tobacco variety with at least one tobacco plant of a second tobacco variety, wherein said at least one tobacco plant of said first tobacco variety comprises a recombinant DNA construct comprising a heterologous promoter operably linked to a nucleic acid encoding at least one small RNA molecule capable of reducing expression of an endogenous genomic locus encoding a polypeptide selected from the group consisting of a cyclin, a cyclin-dependent kinase (CDK), a CDK inhibitor, a MYB, and a WRKY transcription factor, and wherein said at least one tobacco plant comprises no or reduced suckers after topping compared to a control tobacco plant when grown under equivalent growing conditions; (b) selecting progeny tobacco plants that exhibit no or reduced suckers after topping compared to control tobacco plants of the same hybrid grown under comparable growing conditions;

35. 4. The modified tobacco plant of claim 3, wherein the polynucleotide sequence comprises a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-44 and 113.

Images