Methods for increasing trichome density and improving metabolite transport in plant trichomes - Patents.com

Abstract

The present disclosure relates to compositions and methods related to the modification of trichome density and metabolite transport, and their use in plants, including tobacco and cannabis. The provided transcription factors allow for increased trichome density in plants. TIFF2024505756000013.tif57170

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS AND INCORPORATION OF SEQUENCE LISTING This application claims the benefit of U.S. Provisional Patent Application No. 63 / 145,259, filed February 3, 2021; U.S. Provisional Patent Application No. 63 / 145,262, filed February 3, 2021; and U.S. Provisional Patent Application No. 63 / 145,263, filed February 3, 2021, all of which are incorporated by reference in their entireties. This application also includes a sequence listing, which has been submitted electronically in ASCII format and is incorporated by reference in its entirety. The ASCII copy, created on January 25, 2022, is named P335029WO00_SL.txt and is 141,791 bytes in size as measured on Microsoft Windows®.

[0002] Field The present disclosure relates to enhancing the formation and development of trichomes on the surface of plant tissues, as well as improving the transport of specialized metabolites to and exudation of specialized metabolites from the trichomes, and its application to alter the chemical composition of plants, including tobacco. [Background technology]

[0003] background Glandular trichomes are epidermal derivatives of plants that are the synthesis and storage part of metabolic compounds. Their presence on stem, leaf and flower bud tissues protects the plant from various biotic and abiotic stresses. Glandular trichomes also play a role in the biosynthesis, storage and secretion of specialized or secondary metabolites.

[0004] Metabolites produced and secreted by glandular trichomes are often hydrophobic (e.g., fatty acid derivatives, flavonoids, terpenoids). Terpenoids constitute the largest and most diverse class of plant metabolites. Their olefinic backbone is made of multiples of 5-carbon (C) isoprene units, with the major groups being monoterpenes (10C), sesquiterpenes (15C), and diterpenes (20C). These terpenoids are produced through condensation of 5-carbon isoprene units (dimethylallyl diphosphate [DMAPP] and isopentenyl diphosphate [IPP]) together, mostly by sequential head-to-tail addition of DMAPP to IPP.

[0005] The amount of secondary metabolites produced is often closely correlated with the density of glandular trichomes present on the plant epidermis (Chalvin et al., Cell, 25:477-487 (2020)). One way to increase the amount of secondary metabolite production in plants is to increase the density of trichomes present on the plant epidermis. Transcriptional regulation of trichome formation has been shown to involve members of the MYB and C2H2 zinc finger families of transcription factors. Transgenic overexpression of Artemisia annua MYB1 (AaMYB1) has been shown to increase trichome density and subsequently increase the production of artemisin (Matias-Hernandez et al., Plant Journal, 90:520-534 (2017)).

[0006] Lipid transfer protein (LTP) is important in the transport of specialized metabolites in glandular trichomes. Studies have shown that overexpression of LTP leads to increased exudation in plant glandular trichomes (Choi et al., Plant Journal, 70:480-491 (2012)).

[0007] Since glandular trichomes play an important role in the biosynthesis and secretion of terpenoids, there is a need to better understand the genes, regulators and signaling mechanisms involved in the control of trichome formation and development in plants.It is also important to understand the manner of secretion of these specialized metabolites into the epidermis of trichomes.In this disclosure, candidate genes are provided that can be used to modify trichome density in plants.Modifying trichome density will also improve the transport of specialized metabolites in glandular trichomes. [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] Chalvin et al., Cell, 25:477-487 (2020) [Non-Patent Document 2] Matias-Hernandez et al., Plant Journal, 90:520-534 (2017) [Non-Patent Document 3] Choi et al., Plant Journal, 70:480-491 (2012) Summary of the Invention

[0009] overview In one aspect, the disclosure provides a modified plant, seed, or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

[0010] In one aspect, the disclosure provides a cured tobacco material derived from a modified tobacco plant or tobacco plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

[0011] In one aspect, the disclosure provides a tobacco product comprising material from a modified tobacco plant, tobacco seed, or tobacco plant part that comprises a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

[0012] In one aspect, the disclosure provides a cannabis product comprising material from a modified cannabis plant, cannabis seed or cannabis plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein said nucleic acid sequence is operably linked to a heterologous promoter.

[0013] In one aspect, the present disclosure provides a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

[0014] In one aspect, the disclosure provides a method for producing a plant, comprising: (a) obtaining at least one plant comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter; (b) crossing the at least one plant with at least one plant of a second variety to produce at least one progeny seed; and (c) selecting the at least one progeny seed produced in step (b), or a plant germinated therefrom, wherein the at least one progeny seed or a plant germinated therefrom comprises the recombinant nucleic acid molecule.

[0015] In one aspect, the disclosure provides a method of producing a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a nucleic acid sequence at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b).

[0016] In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises in at least one plant cell a recombinant nucleic acid molecule comprising a nucleic acid sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

[0017] In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or part derived therefrom, wherein the modified cannabis plant or part derived therefrom comprises in at least one plant cell a recombinant nucleic acid molecule comprising a nucleic acid sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, said nucleic acid sequence operably linked to a heterologous promoter.

[0018] In one aspect, the disclosure provides a method comprising transforming a plant cell with at least one recombinant nucleic acid molecule for the plant cell, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs:13-17, 23-30 and 34-36, and the nucleic acid sequence is operably linked to a heterologous promoter.

[0019] In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions.

[0020] A brief description of the sequence SEQ ID NOs:1-12 are primers that can be used to clone the nucleic acid sequences described herein for expression in transgenic plants. SEQ ID NOs:13-17 are nucleic acid sequences that correspond to the coding sequences of the tobacco genes of interest described herein, specifically NtMYB86, NtGIS, NbGIS, and NtLTP1. SEQ ID NOs:18-22 are the amino acid sequences corresponding to the nucleic acid sequences of SEQ ID NOs:13-17. SEQ ID NOs:23-27 are nucleic acid sequences that correspond to the genomic sequences of the tobacco genes of interest described herein, specifically NtMYB86, NtGIS, NbGIS, and NtLTP1. SEQ ID NOs:28-30 are nucleic acid sequences corresponding to the coding sequences of the cannabis genes of interest described herein, specifically MYB61, GIS3, and non-specific lipid transfer protein 1-like. SEQ ID NOs:31-33 are the amino acid sequences corresponding to the nucleic acid sequences of SEQ ID NOs:28-30. SEQ ID NOs:34-36 are nucleic acid sequences corresponding to the genomic sequences of the cannabis genes of interest described herein, specifically MYB61, GIS3, and non-specific lipid transfer protein 1-like. SEQ ID NO:37 is a nucleic acid sequence that corresponds to the coding sequence of sweetwormwood (Artemisia annua), specifically the MYB transcription factor. SEQ ID NO:38 is the amino acid sequence corresponding to the nucleic acid sequence of SEQ ID NO:37. SEQ ID NOs:39-43 are nucleic acid sequences corresponding to the coding sequences of the tobacco genes of interest described herein, specifically MFS, NMD, CBTS2a (HM), GGPPS2 and cis-abienol synthase (ABS). SEQ ID NOs:44-48 are nucleic acid sequences that correspond to the genomic sequences of the tobacco genes of interest described herein, specifically MFS, NMD, CBTS2a (HM), GGPPS2 and cis-abienol synthase (ABS). SEQ ID NOs:49-53 are the amino acid sequences corresponding to the nucleic acid sequences of SEQ ID NOs:39-43. SEQ ID NOs:54-62 and 63-71 are nucleic acid sequences that correspond to the coding and genomic sequences of the tobacco LTP gene, respectively. SEQ ID NOs:72-80 are amino acid sequences corresponding to the nucleic acid sequences of SEQ ID NOs:54-62. SEQ ID NO:81 is the consensus LTP amino acid sequence. SEQ ID NOs:82-91 are partial sequences of SEQ ID NOs:21 and 72-80 as shown in Figure 9. SEQ ID NO:92 is the consensus amino acid sequence shown in Figure 9.

[0021] Table 1 provides the nucleic acid and amino acid sequences used in this disclosure.

[0022] Table 1: Sequences used in this disclosure TIFF2024505756000002.tif181146TIFF2024505756000003.tif244146TIFF2024505756000004.tif226146 [Brief description of the drawings]

[0023] [Figure 1] Includes panels A, B, and C. Panel A depicts glandular trichomes in Nicotiana benthamiana. Panel B depicts glandular trichomes in Nicotiana tabacum cultivar "TN90," and panel C depicts glandular trichomes in Nicotiana tabacum cultivar "Izmir Ego." [Diagram 2] 1 depicts expression vectors used for tobacco transformation. [Figure 3A]Figure 3 includes panels Figure 3A and Figure 3B. Figure 3A depicts a phylogenetic analysis of the DNA-binding domains of the MYB transcription factor family in tobacco plants (N=319). Figure 3B depicts a protein sequence pairwise alignment of the tobacco MYB86 gene (SEQ ID NO:18, XP_016450602.1) and sweetwormwood (Artemisia annua) MYB (SEQ ID NO:38, AGR40501.1), specifically highlighting the R2 and R3 DNA-binding domains (boxed). The black circle indicates MYB86 on the cladogram. The scale bar in Figure 3A represents the evolutionary distance used to infer the phylogenetic tree. [Figure 3B] Please see the legend to FIG. 3A. [Figure 4] Depicts the phylogenetic analysis of the Glabrous Inflorescence Stems (GIS) family of C2H2 transcription factors in tobacco plants (N=247). The black circle indicates the NtGIS gene (SEQ ID NO:19) on the cladogram. The scale bar represents the evolutionary distance used to infer the phylogenetic tree. [Diagram 5] Includes panels A, B, C, and D. Panel A depicts glandular trichome density in the abaxial leaf midvein of wild-type (WT) TN90. Panel B depicts glandular trichome density in the abaxial leaf midvein of TN90 overexpressing NtMYB86 (SEQ ID NO:13) under control of the CaMV 35S promoter, demonstrating increased glandular trichome density compared to wild-type TN90. An abaxial view of a pedicel of a wild-type (WT) N. benthamiana leaf is shown in panel C. Panel D depicts increased glandular trichome density in N. benthamiana overexpressing NtMYB86 under control of the CaMV 35S promoter compared to wild-type (panel C). Images are light micrographs shown at 10× magnification. [Figure 6]Includes Panel A and Panel B. Panel A depicts the glandular trichome density in the primary vegetative stem of wild-type (WT) Nicotiana benthamiana. Panel B depicts the glandular trichome density in the primary vegetative stem of N. benthamiana overexpressing NtMYB86 (SEQ ID NO:18). Images are light micrographs shown at 160x magnification. [Figure 7A] FIG. 7 includes panels FIG. 7A and FIG. 7B. FIG. 7A depicts light micrographs of glandular trichome density of tobacco leaves overexpressing NtMYB86 (SEQ ID NO:18) (b), NtGIS (SEQ ID NO:19) (c), and NbGIS (SEQ ID NO:20) (c) in the Izmir-Ego background compared to wild-type Nicotiana tabacum cultivar Izmir-Ego (a). Images are shown at 70× magnification and scale bars represent 1 mm. FIG. 7B depicts trichome numbers of leaves of Izmir-Ego WT, Izmir-Ego T0 transgenic lines overexpressing NtMYB86 (SEQ ID NO:18), NtGIS (SEQ ID NO:19), and NbGIS (SEQ ID NO:20). [Figure 7B] Please see the legend to FIG. 7A. [Figure 8A] Figure 8 includes panels 8A and 8B. Figure 8A depicts a phylogenetic tree analysis of the LTP gene family, with the location of the NtLTP1 gene marked by a black circle. Figure 8B depicts a sequence alignment and gene structure comparison in tobacco plants of two NtLTP1 isoforms (SEQ ID NOs: 21, 22, and 93, respectively, in order of appearance). [Figure 8B] Please see the legend to FIG. 8A. [Figure 9] Figure 1 depicts conserved motifs in 76 representative NtLTPs identified in the Nicotiana tabacum genome after the ClustalW multiple sequence alignment algorithm. Six conserved cysteine ​​residues (boxed) are particularly conserved in ≥90% of the identified NtLTPs. [Figure 10]Figure 1 depicts the relative gene expression of NtMYB86 (SEQ ID NO:14) in nine independent Izmir-Ego transgenic lines at the T0 generation. Relative gene expression was quantified following the 2(-ΔΔC(t)) method. For information regarding the 2(-ΔΔC(t)) method, see Livak and Schmittgen, Methods, 25:402-408 (2001). Expression of NtMYB86 in wild-type control Izmir-Ego plants was used as a baseline (e.g., wild-type expression set to 1) and is not shown. [Figure 11] Figure 1 depicts the relative gene expression of NtGIS (SEQ ID NO:16) in eight independent Izmir-Ego transgenic lines at the T0 generation. Relative gene expression was quantified following the 2-ΔΔCT method. Expression of NtGIS in wild-type control Izmir-Ego plants was used as a baseline (e.g., wild-type expression was set to 1) and is not shown. [Figure 12] Figure 1 depicts the relative abundance of duvatrienediols in leaves of T0 Izmir-Ego transgenic lines overexpressing NbGIS (SEQ ID NO:20), NtGIS (SEQ ID NO:19), NtMFS (SEQ ID NO:39), NtNMD (29837; SEQ ID NO:40), Nicotiana sylvestris cembratrienol synthase 2a (HM; SEQ ID NO:41), geranylgeranyl diphosphate synthase (GQ; SEQ ID NO:42), and cis-abienol synthase (AAB; SEQ ID NO:43). [Figure 13]Figure 1 depicts the relative abundance of neophytadiene in leaves of T0 Izmir-Ego transgenic lines overexpressing NbGIS (SEQ ID NO:20), NtGIS (SEQ ID NO:19), NtMFS (SEQ ID NO:39), NtNMD (29837; SEQ ID NO:40), Nicotiana sylvestris cembratrienol synthase 2a (HM; SEQ ID NO:41), geranylgeranyl diphosphate synthase (GQ; SEQ ID NO:42), and cis-abienol synthase (AAB; SEQ ID NO:43). [Figure 14] 1 depicts the relative abundance of 2,4-dimethyl-1-heptene in leaves of T0 Izmir-Ego transgenic lines overexpressing NbGIS (SEQ ID NO:20), NtGIS (SEQ ID NO:19), NtMFS (SEQ ID NO:39), NtNMD (29837; SEQ ID NO:40), Nicotiana sylvestris cembratrienol synthase 2a (HM; SEQ ID NO:41), geranylgeranyl diphosphate synthase (GQ; SEQ ID NO:42), and cis-abienol synthase (AAB; SEQ ID NO:43). [Figure 15] 1 depicts the relative abundance of cis-abienol in leaves of T0 Izmir-Ego transgenic lines overexpressing NbGIS (SEQ ID NO:20), NtGIS (SEQ ID NO:19), NtMFS (SEQ ID NO:39), NtNMD (29837; SEQ ID NO:40), Nicotiana sylvestris cembratrienol synthase 2a (HM; SEQ ID NO:41), geranylgeranyl diphosphate synthase (GQ; SEQ ID NO:42), and cis-abienol synthase (AAB; SEQ ID NO:43). [Figure 16] 1 depicts the relative gene expression levels of NtMYB86 (SEQ ID NO:13) under the control of CaMV 35S promoter in T1 transgenic tobacco lines of Izmir-Ego variety. [Figure 16]1 depicts the relative gene expression levels of NbGIS (SEQ ID NO:15) under the control of CaMV 35S promoter in T1 transgenic tobacco lines of Izmir-Ego variety. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] Detailed Description Unless otherwise defined, all technical and scientific terms used have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. Where a term is provided in the singular form, the inventors also contemplate aspects of this disclosure described by the plural form of that term. Where there is a difference 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 the common meaning in the art in which they are used, as exemplified in various technical 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).

[0025] All references cited herein, including, for example, all patents, published patent applications and non-patent publications, are hereby incorporated by reference in their entirety.

[0026] When a grouping of options is presented, any and all combinations of the members of that grouping of options 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 option individually (e.g., A alone, B alone, etc.), as well as combinations such as A and B and D; A and C; B and C, etc. The term "and / or," when used in a list of two or more items, means 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 combination of A and B, a combination of A and C, a combination of B and C, or a combination of A, B, and C.

[0027] When a range of numerical values ​​is provided herein, the range is understood to include the endpoints of the range as well as any number between the endpoints of the stated range. For example, "1-10" includes any number between 1 and 10, as well as the number 1 and the number 10.

[0028] When the term "about" is used in reference to numerical values, it is understood to mean + or - 10%. For example, "about 100" would include from 90 to 110.

[0029] As used herein, the singular forms "a," "an," and "the" include plural referents 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.

[0030] Any tobacco plant provided herein, or part thereof, is specifically contemplated for use with any method provided herein.Similarly, any modified tobacco plant, or part thereof, is specifically contemplated for use with any method provided herein.Any nucleic acid sequence, amino acid sequence, or other composition provided herein is specifically contemplated for use with any method provided herein.

[0031] Any cannabis plant provided herein, or part thereof, is specifically contemplated for use with any method provided herein.Similarly, any modified cannabis plant, or part thereof, is specifically contemplated for use with any method provided herein.Any nucleic acid sequence, amino acid sequence, or other composition provided herein is specifically contemplated for use with any method provided herein.

[0032] Trichomes are generally hair-like epidermal outgrowths that cover most above-ground plant tissues. Trichomes tend to be multicellular, although unicellular trichomes are also known. Multiple types of trichomes can be found in individual plants, and trichomes vary in shape, size, and cellular organization. Individual trichomes can be classified as glandular or non-glandular trichomes.

[0033] Glandular trichomes (see Figure 1) are characterized by the presence of a head consisting of cells capable of secreting or storing large amounts of specialized metabolites (e.g., terpenes). Within the group of glandular trichomes, trichomes can be further characterized as shield-like or capitate. Capitate glandular trichomes typically have a stalk that is more than twice the height of the head, and the number of cells in a trichome is highly variable. Shield-like trichomes are short-stalked trichomes with a large head consisting of 4 to 18 cells arranged in one or two concentric rings.

[0034] In some aspects, the trichomes are glandular trichomes. In some aspects, the glandular trichomes are capitate glandular trichomes. In some aspects, the glandular trichomes are shield-shaped glandular trichomes. In some aspects, the glandular trichomes are selected from the group consisting of capitate glandular trichomes and shield-shaped glandular trichomes.

[0035] In certain aspects, the formation and development of glandular trichomes is regulated by several transcription factors, including, but not limited to, genes belonging to the MYB and C2H2 transcription factor gene families.

[0036] In one aspect, the present disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide operably linked to a heterologous promoter for the formation and development of trichomes to improve their density.In another aspect, the present disclosure provides a modified plant, seed or plant part comprising a nucleic acid sequence encoding a polypeptide operably linked to a heterologous promoter for improving the secretion of metabolites into trichome epithelium.In a further aspect, the modified plant, seed or plant part is a plant, seed or plant part of the genus Nicotiana.In a further aspect, the modified plant, seed or plant part is a plant, seed or plant part of the genus Cannabis.

[0037] Transcription factor regulation of trichome formation Transcription factors are known regulators of various biological processes involved in plant growth and development. Several transcription factors have been identified as regulators of glandular trichome formation. These include, but are not limited to, the transcription factor gene families R2R3-MYB, HD-ZIP IV, MYC, and C2H2 (Chavlin et al., 2020). Overexpression of MYB1 (AaMYB1) in Artemisia annua significantly increases not only trichome density but also the production of artemisin, a sesquiterpene lactone (Matias-Hernandez et al., 2017). Another transcription factor that regulates trichome development as well as terpene biosynthesis is MYC1, a basic helix-loop-helix (bHLH). Knockdown of MYC1 (SlMYC1) in tomato led to a significant reduction in monoterpenes as well as a low density of ectopic development of smaller type VI glandular trichomes (Xu et al., Plant Cell, 30:2988-3005 (2018)). Transcription factors belonging to the C2H2 gene family are also involved in regulating trichome development in tobacco. Overexpression of Nicotiana benthamiana GIS (NbGIS) led to increased glandular trichome density in transgenic lines (Liu et al., Plant Molecular Biology, 98:153-167 (2018)).

[0038] Metabolite storage and transport in glandular trichomes Glandular trichomes produce large amounts of metabolites that can account for up to 20% of the leaf dry weight (Tissier et al., Trends in Plant Science, 22:930-938 (2017)). Trichomes express morphological features that allow the storage of secondary metabolites. The type, shape and size of these morphological features depend on the type of glandular trichome. Non-volatile compounds, including diterpenoids, are typically produced by capitate glandular trichomes and secreted directly from the tips of the trichomes, whereas glandular trichomes producing volatile compounds have dedicated structures for secretion and storage (Tissier et al., 2017). Transport of hydrophobic molecules in trichomes requires transporters and lipid transfer proteins (LTPs), which facilitate the movement of volatile organic compounds through the hydrophilic cell wall and prevent redistribution of VOCs to the plasma membrane (Tissier et al., 2017). LTPs are small (~10 kDa) soluble proteins characterized by a highly conserved cysteine-rich motif (Salminen et al., Planta, 244:971-997 (2016)). See Figure 10. LTPs are involved in various functions during plant growth and development, including but not limited to cuticular wax accumulation, pollen and seed development, and cell expansion (Salminen et al., 2016). Trichome-specific NtLTP1 was also reported to increase secretion in trichome exudates in tobacco plants and enhance pest defense when overexpressed (Choi et al., 2012).

[0039] plant In some aspects, the plants provided herein are modified plants. In some aspects, the seeds provided herein are modified seeds. In some aspects, the plant parts provided herein are modified plant parts. As used herein, "modified" in the context of a plant, seed or plant part refers to a plant, seed or plant part that is introduced for a specific purpose and contains genetic variations beyond natural polymorphisms. Without being limited thereto, modified plants, seeds or plant parts include recombinant nucleic acid molecules. In another aspect, modified plants, seeds or plant parts include genetic modifications. In some aspects, modified plants, seeds or plant parts are transgenic plants, seeds or plant parts.

[0040] In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 85% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 90% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 95% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed, or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 96% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter.In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 97% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 98% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 99% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 99.9% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed, or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence that is 100% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

[0041] In one aspect, the disclosure provides a modified plant, seed, or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.9%, or 100% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs:72-81, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

[0042] In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence at least 80% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence at least 85% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence at least 90% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence at least 95% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence at least 96% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence at least 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter.In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence at least 98% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence at least 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence at least 99.9% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter. In one aspect, the disclosure provides a modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence 100% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

[0043] In one aspect, the disclosure provides a modified plant, seed, or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99%, or 100% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs:54-71, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

[0044] In some aspects, at least one leaf of the modified plant comprises a greater average trichome density compared to the leaf of a control plant grown under comparable conditions.In some aspects, at least one leaf of the modified plant comprises a greater average trichome density on the abaxial surface of the leaf compared to the abaxial surface of the leaf of a control plant grown under comparable conditions.In some aspects, at least one leaf of the modified plant comprises a greater average trichome density on the adaxial surface of the leaf compared to the adaxial surface of the leaf of a control plant grown under comparable conditions.

[0045] In some aspects, at least two leaves of the modified plant contain a higher average trichome density compared to two leaves of a control plant grown under the same conditions.In some aspects, a majority of the leaves of the modified plant contain a higher average trichome density compared to the same number of leaves of a control plant grown under the same conditions.In some aspects, all of the leaves of the modified plant contain a higher average trichome density compared to all of the leaves of a control plant grown under the same conditions.

[0046] In some aspects, at least one stem of the modified plant comprises a higher average trichome density than the stem of a control plant grown under comparable conditions.In some aspects, at least one flower of the modified plant comprises a higher average trichome density than the flower of a control plant grown under comparable conditions.In some aspects, at least one root of the modified plant comprises a higher average trichome density than the root of a control plant grown under comparable conditions.

[0047] In some aspects, the modified plants contain a greater average density of glandular trichomes as compared to control plants, hi some aspects, the modified plants contain a greater average density of non-glandular trichomes as compared to control plants.

[0048] In some aspects, the modified tobacco plant leaves include at least 70 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include at least 75 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include at least 80 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include at least 85 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include at least 90 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include at least 95 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include at least 100 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include at least 105 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include at least 110 trichomes / square centimeter. In some aspects, the leaves of the modified tobacco plant comprise at least 115 trichomes / square centimeter. In some aspects, the leaves of the modified tobacco plant comprise at least 120 trichomes / square centimeter. In some aspects, the leaves of the modified tobacco plant comprise at least 125 trichomes / square centimeter. In some aspects, the leaves of the modified tobacco plant comprise at least 130 trichomes / square centimeter. In some aspects, the leaves of the modified tobacco plant comprise at least 140 trichomes / square centimeter. In some aspects, the leaves of the modified tobacco plant comprise at least 150 trichomes / square centimeter.

[0049] In some aspects, the modified tobacco plant leaves include 60-200 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include 60-190 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include 60-180 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include 60-170 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include 60-160 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include 60-150 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include 60-140 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include 60-130 trichomes / square centimeter. In some aspects, the leaves of the modified tobacco plant include between 60 and 120 trichomes per square centimeter. In some aspects, the leaves of the modified tobacco plant include between 60 and 110 trichomes per square centimeter. In some aspects, the leaves of the modified tobacco plant include between 60 and 100 trichomes per square centimeter. In some aspects, the leaves of the modified tobacco plant include between 60 and 90 trichomes per square centimeter. In some aspects, the leaves of the modified tobacco plant include between 60 and 80 trichomes per square centimeter. In some aspects, the leaves of the modified tobacco plant include between 60 and 70 trichomes per square centimeter.

[0050] In some aspects, the modified tobacco plant leaves include 80-130 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include 80-120 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include 80-100 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include 75-140 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include 75-130 trichomes / square centimeter. In some aspects, the modified tobacco plant leaves include 75-125 trichomes / square centimeter.

[0051] In some aspects, the modified tobacco plant leaves contain at least 1% more trichomes per square centimeter compared to a control leaf of the same age when grown under comparable conditions. In some aspects, the modified tobacco plant leaves contain at least 5% more trichomes per square centimeter compared to a control leaf of the same age when grown under comparable conditions. In some aspects, the modified tobacco plant leaves contain at least 10% more trichomes per square centimeter compared to a control leaf of the same age when grown under comparable conditions. In some aspects, the modified tobacco plant leaves contain at least 15% more trichomes per square centimeter compared to a control leaf of the same age when grown under comparable conditions. In some aspects, the modified tobacco plant leaves contain at least 20% more trichomes per square centimeter compared to a control leaf of the same age when grown under comparable conditions. In some aspects, the modified tobacco plant leaves contain at least 25% more trichomes per square centimeter compared to a control leaf of the same age when grown under comparable conditions. In some aspects, the modified tobacco plant leaves contain at least 30% more trichomes per square centimeter compared to a control leaf of the same age when grown under comparable conditions. In some aspects, the modified tobacco plant leaves contain at least 40% more trichomes per square centimeter compared to a control leaf of the same age when grown under comparable conditions. In some aspects, the modified tobacco plant leaves contain at least 50% more trichomes per square centimeter compared to a control leaf of the same age when grown under comparable conditions. In some aspects, the modified tobacco plant leaves contain at least 60% more trichomes per square centimeter compared to a control leaf of the same age when grown under comparable conditions. In some aspects, the modified tobacco plant leaves contain at least 70% more trichomes per square centimeter compared to a control leaf of the same age when grown under comparable conditions. In some aspects, the modified tobacco plant leaves contain at least 80% more trichomes per square centimeter compared to a control leaf of the same age when grown under comparable conditions. In one aspect, the leaves of the modified tobacco plants contain at least 90% more trichomes per square centimeter compared to control leaves of the same age when grown under comparable conditions.In some aspects, the leaves of the modified tobacco plants contain at least 100% more trichomes per square centimeter compared to control leaves of the same age when grown under comparable conditions. In some aspects, the leaves of the modified tobacco plants contain at least 110% more trichomes per square centimeter compared to control leaves of the same age when grown under comparable conditions. In some aspects, the leaves of the modified tobacco plants contain at least 125% more trichomes per square centimeter compared to control leaves of the same age when grown under comparable conditions. In some aspects, the leaves of the modified tobacco plants contain at least 150% more trichomes per square centimeter compared to control leaves of the same age when grown under comparable conditions. In some aspects, the leaves of the modified tobacco plants contain at least 200% more trichomes per square centimeter compared to control leaves of the same age when grown under comparable conditions.

[0052] Unless otherwise specified, it will be understood that comparisons between modified leaves and leaves from control plants grown under equivalent conditions should use the same number of leaves on the modified and control plants (e.g., a modified V5 leaf should be compared to a control V5 leaf).

[0053] In some aspects, the plant is a tobacco plant. In some aspects, the plant is a Nicotiana plant. In some aspects, the tobacco plant is a Nicotiana tabacum plant.

[0054] In one aspect, the Nicotiana plant, seed, or plant part is selected from the group consisting of Nicotiana tabacum, Nicotiana amplexicaulis PI 271989; Nicotiana benthamiana PI 555478; Nicotiana bigelovii PI 555485; Nicotiana debneyi; Nicotiana excelsior PI 224063; Nicotiana glutinosa PI 555507; Nicotiana goodspeedii PI 241012; Nicotiana gossei PI 241012; 230953;Nicotiana hesperis PI 271991;Nicotiana knightiana PI 555527;Nicotiana maritima PI 555535;Nicotiana megalosiphon PI 555536;Nicotiana nudicaulis PI 555540;Nicotiana paniculata PI 555545;Nicotiana plumbaginifolia PI 555548;Nicotiana repanda PI 555552;Nicotiana rustica rustica);Nicotiana suaveolens PI 230960;Nicotiana sylvestris PI 555569;Nicotiana tomentosa PI 266379;Nicotiana tomentosiformis;and Nicotiana trigonophylla PI 555572;

[0055] In some aspects, the seed is a tobacco seed. In some aspects, the seed is a Nicotiana seed. In some aspects, the tobacco seed is a Nicotiana tabacum or Nicotiana benthamiana seed.

[0056] In some aspects, the plant part is a tobacco plant part. In some aspects, the plant part is a Nicotiana plant part. In some aspects, the tobacco plant part is a Nicotiana tabacum plant part or a Nicotiana benthamiana plant part.

[0057] In some aspects, the plant is a cannabis plant. In some aspects, the plant is a Cannabis plant. In some aspects, the cannabis plant is a Cannabis sativa plant. In some aspects, the cannabis plant is a Cannabis indica plant. In some aspects, the cannabis plant is a Cannabis ruderalis plant. In some aspects, the cannabis plant is selected from the group consisting of Cannabis sativa, Cannabis indica, and Cannabis ruderalis.

[0058] In some aspects, the seed is a cannabis seed. In some aspects, the seed is a Cannabis seed. In some aspects, the cannabis seed is a Cannabis sativa seed. In some aspects, the cannabis seed is a Cannabis indica seed. In some aspects, the cannabis seed is a Cannabis ruderalis seed. In some aspects, the cannabis seed is selected from the group consisting of Cannabis sativa, Cannabis indica, and Cannabis ruderalis.

[0059] In some aspects, the plant part is a cannabis plant part. In some aspects, the plant part is a Cannabis plant part. In some aspects, the cannabis plant part is a Cannabis sativa plant part. In some aspects, the cannabis plant part is a Cannabis indica plant part. In some aspects, the cannabis plant part is a Cannabis ruderalis plant part. In some aspects, the cannabis plant part is selected from the group consisting of Cannabis sativa, Cannabis indica, and Cannabis ruderalis.

[0060] In some aspects, the plant parts provided include, but are not limited to, leaves, stems, roots, trichomes, seeds, flowers, pollen, anthers, ovules, pedicels, fruits, meristems, cotyledons, hypocotyls, pods, embryos, endosperms, explants, calli, tissue cultures, shoots, cells, and protoplasts. In some aspects, the plant parts do not include seeds. In some aspects, the present disclosure provides plant cells, tissues, and organs that are not reproductive material and do not mediate natural reproduction of plants. In another aspect, the present disclosure also provides plant cells, tissues, and organs that are reproductive material and mediate natural reproduction of plants. In another aspect, the present disclosure provides plant cells, tissues, and organs that cannot sustain themselves through photosynthesis. In another aspect, the present disclosure provides somatic plant cells. Somatic cells, in contrast to germ line cells, do not mediate plant reproduction.

[0061] The cells, tissues and organs can be from seeds, fruits, leaves, cotyledons, hypocotyls, meristems, embryos, endosperms, roots, shoots, stems, trichomes, pods, flowers, inflorescences, stalks, pedicels, styles, stigmas, receptacles, petals, calyxes, pollen, anthers, filaments, ovaries, ovules, pericarp, phloem, vascular tissues. In another aspect, the present disclosure provides plant chloroplasts. In a further aspect, the present disclosure provides epidermal cells, stoma cells, leaves or root hairs, storage roots, or tubers. In another aspect, the present disclosure provides tobacco protoplasts.

[0062] Those skilled in the art will appreciate that tobacco and cannabis plants reproduce naturally through seeds, rather than through asexual or vegetative reproduction. In one aspect, the present disclosure provides a plant endosperm.

[0063] The present disclosure provides cells derived from the plants provided herein.

[0064] As used herein, a "progeny plant" or "progeny seed" can be derived from any hybrid generation, e.g., F1, F2, F3, F4, F5, F6, F7, etc.

[0065] In one aspect, the tobacco plant, seed or plant part is of a tobacco variety selected from the group consisting of a flue cured variety, a bright variety, a Burley variety, a Virginia variety, a Maryland variety, a dark variety, a Galpao variety, an Oriental variety, and a Turkish variety.

[0066] In one aspect, the tobacco cells are of a tobacco variety selected from the group consisting of flue-cured, bright, burley, Virginia, Maryland, dark, Galpao, Oriental, and Turkish.

[0067] In one aspect, the tobacco leaf is of a tobacco variety selected from the group consisting of flue-cured, bright, burley, Virginia, Maryland, dark, Galpao, Oriental, and Turkish.

[0068] In one aspect, the cured tobacco leaves or plant parts are of a tobacco variety selected from the group consisting of flue-cured, bright, burley, Virginia, Maryland, dark, Galpao, Oriental, and Turkish. One of skill in the art will further appreciate that cured tobacco is not a component of a living organism and is incapable of growth or reproduction.

[0069] Flue-cured tobacco (also called "Virginia" or "Bright" tobacco) accounts for approximately 40% of the world's tobacco production. Flue-cured tobacco is often also referred to as "Bright tobacco" due to the golden yellow to deep orange color it attains during curing. Flue-cured tobacco has a light and elegant aroma and flavor. Flue-cured tobacco is generally high in sugar and low in oil. The major flue-cured tobacco growing countries are Argentina, Brazil, China, India, Tanzania and the United States. In one aspect, the tobacco plants, seeds or plant parts provided herein are of a flue-cured tobacco variety selected from the group consisting of the varieties listed in Table 2, and any variety essentially derived from any one of the aforementioned varieties. See WO 2004 / 041006 A1. In a further aspect, the tobacco plants, seeds or plant parts provided herein are of a flue-cured variety selected from the group consisting of K326, K346, and NC196.

[0070] Table 2. Flue-cured tobacco varieties TIFF2024505756000005.tif154145TIFF2024505756000006.tif158145

[0071] Air-cured tobaccos include "Burley," "Maryland," and "Dark" tobaccos. The common factor that unites air-cured tobaccos is that curing occurs primarily without artificial sources of heat and moisture. Burley tobacco is light to dark brown in color and is high in oils and low in sugars. Burley tobacco is typically air-cured in warehouses. Major burley-growing countries include Argentina, Brazil, Italy, Malawi, and the United States.

[0072] Maryland tobacco is very fluffy, has good burning characteristics, low nicotine and a neutral flavor. Major Maryland-growing countries include the United States and Italy.

[0073] In one aspect, the tobacco plants, seeds or plant parts provided herein are of a Burley tobacco variety selected from the group consisting of the tobacco varieties listed in Table 3, and any variety essentially derived from any one of the foregoing varieties. In a further aspect, the tobacco plants, seeds or plant parts provided herein are of a Burley variety selected from the group consisting of TN 90, KT 209, KT 206, KT212, and HB 4488.

[0074] (Table 3) Burley tobacco varieties TIFF2024505756000007.tif200162

[0075] In another aspect, the tobacco plant, seed or plant part provided herein is of a Maryland tobacco variety selected from the group consisting of the tobacco varieties listed in Table 4, and any variety essentially derived from any one of the foregoing varieties.

[0076] Table 4. Maryland Tobacco Varieties TIFF2024505756000008.tif72128

[0077] Dark air-cured tobacco is distinguished from other tobacco types primarily by its medium to dark brown color and its curing process, which gives it a distinct aroma. Dark air-cured tobacco is primarily used in the production of chewing tobacco and snuff. In one aspect, the tobacco plants, seeds, or plant parts 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, as well as any variety essentially derived from any one of the aforementioned varieties.

[0078] Dark flue-cured tobacco is generally dried in a low-burning wood fire on the floor of a closed drying warehouse. Dark flue-cured tobacco is typically used to produce pipe blends, cigarettes, chewing tobacco, snuff, and strong-tasting cigars. The main growing areas of dark flue-cured tobacco are Tennessee, Kentucky, and Virginia in the United States. In one aspect, the tobacco plant, seed, or plant part provided herein is of a dark flue-cured tobacco variety selected from the group consisting of the tobacco varieties listed in Table 5, and any variety essentially derived from any one of the aforementioned varieties.

[0079] Table 5. Dark, flue-cured tobacco varieties TIFF2024505756000009.tif158147

[0080] Oriental tobacco is also referred to as Greek, aroma and Turkish tobacco, as it is typically grown in the eastern Mediterranean region, 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 their adaptation to have evolved in poor soil and stressful climatic conditions. In one aspect, the tobacco plant, seed or plant part provided herein is of an Oriental tobacco variety selected from the group consisting of the tobacco varieties listed in Table 6, and any variety essentially derived from any one of the aforementioned varieties.

[0081] Table 6. Oriental tobacco varieties TIFF2024505756000010.tif107140

[0082] In one aspect, the tobacco plants, seeds or plant parts provided herein are of a cigar tobacco variety selected from the group consisting of the tobacco varieties listed in Table 7, and any variety essentially derived from any one of the foregoing varieties.

[0083] (Table 7) Cigar Tobacco Varieties TIFF2024505756000011.tif51157

[0084] In one aspect, the tobacco plants, seeds, or plant parts provided herein are of a tobacco variety selected from the group consisting of the tobacco varieties listed in Table 8, and any variety essentially derived from any one of the foregoing varieties.

[0085] (Table 8) Other tobacco varieties TIFF2024505756000012.tif38128

[0086] In one aspect, the tobacco plant or plant part is from a variety selected from the group consisting of tobacco varieties listed in Table 2, Table 3, Table 4, Table 5, Table 6, Table 7 and Table 8. In another aspect, the tobacco plant or plant part is from a variety listed in Table 2. In another aspect, the tobacco plant or plant part is from a variety listed in Table 3. In another aspect, the tobacco plant or plant part is from a variety listed in Table 4. In another aspect, the tobacco plant or plant part is from a variety listed in Table 5. In another aspect, the tobacco plant or plant part is from a variety listed in Table 6. In another aspect, the tobacco plant or plant part is from a variety listed in Table 7. In another aspect, the tobacco plant or plant part is from a variety listed in Table 8.

[0087] In one aspect, the tobacco seed is from a variety selected from the group consisting of tobacco varieties listed in Table 2, Table 3, Table 4, Table 5, Table 6, Table 7 and Table 8. In another aspect, the tobacco seed is from a variety listed in Table 2. In another aspect, the tobacco seed is from a variety listed in Table 3. In another aspect, the tobacco seed is from a variety listed in Table 4. In another aspect, the tobacco seed is from a variety listed in Table 5. In another aspect, the tobacco seed is from a variety listed in Table 6. In another aspect, the tobacco seed is from a variety listed in Table 7. In another aspect, the tobacco seed is from a variety listed in Table 8.

[0088] In one aspect, the tobacco cell is from a variety selected from the group consisting of tobacco varieties listed in Table 2, Table 3, Table 4, Table 5, Table 6, Table 7 and Table 8. In another aspect, the tobacco cell is from a variety listed in Table 2. In another aspect, the tobacco cell is from a variety listed in Table 3. In another aspect, the tobacco cell is from a variety listed in Table 4. In another aspect, the tobacco cell is from a variety listed in Table 5. In another aspect, the tobacco cell is from a variety listed in Table 6. In another aspect, the tobacco cell is from a variety listed in Table 7. In another aspect, the tobacco cell is from a variety listed in Table 8.

[0089] All of the above specific varieties of air-cured, dark air-cured, burley, Maryland, dark flame-cured, cigar, or Oriental type are listed for illustrative purposes only. Any additional air-cured, dark air-cured, burley, Maryland, dark flame-cured, cigar, or Oriental varieties are also contemplated in the present application.

[0090] In some aspects, the plants or varieties provided herein are inbred plants or varieties. As used herein, an "inbred" variety is a variety that has been bred for genetic homogeneity.

[0091] As used herein, a "hybrid" is produced by crossing two plants from different varieties or species, so that the offspring contains genetic material from each parent. Those skilled in the art will recognize that higher order hybrids can be produced as well. For example, a first hybrid can be produced by crossing variety C with variety D to produce a C×D hybrid, and a second hybrid can be produced by crossing variety E with variety F to produce an E×F hybrid. The first and second hybrids can be further crossed to produce a higher order hybrid (C×D)×(E×F) that contains all the genetic information of the four parent varieties. In some aspects, the plants or seeds provided herein are hybrid plants or seeds.

[0092] In some aspects, the tobacco plants provided herein are inbred tobacco plants. In some aspects, the tobacco seeds provided herein are inbred tobacco seeds. In some aspects, the tobacco plants provided herein are hybrid tobacco plants. In another aspect, the tobacco seeds provided herein are hybrid tobacco seeds.

[0093] In some aspects, the cannabis plant provided herein is an inbred cannabis plant. In some aspects, the cannabis seed provided herein is an inbred cannabis seed. In some aspects, the cannabis plant provided herein is a hybrid cannabis plant. In some aspects, the cannabis seed provided herein is a hybrid cannabis seed.

[0094] Unless otherwise specified, all comparisons with control plants require similar or equivalent growing conditions for the two plants to be compared. As used herein, "grown under equivalent conditions", "similar growing conditions" or "equivalent growing conditions" refers to similar environmental conditions and / or agricultural practices for growth and meaningful comparison between two or more plant genotypes, which do not contribute to or explain the differences observed between two or more plant genotypes. Environmental conditions include, for example, light, temperature, moisture (humidity), and nutrients (e.g., nitrogen and phosphorus). Agricultural practices include, for example, sowing, pruning, stubbling, planting, topping, and division. See Chapters 4B and 4C of Tobacco, Production, Chemistry and Technology, Davis & Nielsen, eds., Blackwell Publishing, Oxford (1999), pp 70-103.

[0095] As used herein, a "control plant" refers to a plant of identical or nearly identical genetic make-up to the modified plant being compared, except for the recombinant nucleic acid molecule provided herein that has been introduced into the modified plant.

[0096] In one aspect, the plant or variety provided herein is male sterile. In another aspect, the plant or variety provided herein is cytoplasmic male sterile (CMS). Male sterile 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 Co., Inc., New York, NY 761 pp.

[0097] In another aspect, the plant or variety provided herein is female-sterile.As a non-limiting example, female-sterile plants can be produced by mutating STIG1 gene.See, for example, Goldman et al. 1994, EMBO Journal 13:2976-2984.

[0098] In one aspect, the disclosure provides a method for producing a plant, comprising the steps of: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence being at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule. In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence being at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule.In one aspect, the disclosure provides a method for producing a plant, comprising the steps of: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence being at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule. In one aspect, the disclosure provides a method for producing a plant, comprising the steps of: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence being at least 92.5% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule.In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence being at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule. In one aspect, the disclosure provides a method for producing a plant, comprising the steps of: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence being at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule.In one aspect, the disclosure provides a method for producing a plant, comprising the steps of: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence being at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule. In one aspect, the disclosure provides a method for producing a plant, comprising the steps of: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence being at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule.In one aspect, the disclosure provides a method for producing a plant, comprising the steps of: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence being at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule. In one aspect, the disclosure provides a method for producing a plant, comprising the steps of: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence being at least 99.9% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule.In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence being selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule.

[0099] In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule. In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence encoding an amino acid sequence at least 85% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule.In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence encoding an amino acid sequence at least 90% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule. In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence encoding an amino acid sequence at least 95% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule.In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence encoding an amino acid sequence at least 96% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule. In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence encoding an amino acid sequence at least 97% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule.In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence encoding an amino acid sequence at least 98% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule. In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence encoding an amino acid sequence at least 99% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule.In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence encoding an amino acid sequence at least 99.9% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule. In one aspect, the disclosure provides a method for producing a plant, comprising: (a) crossing at least one plant of a first variety with at least one plant of a second variety to produce at least one progeny seed, wherein the at least one plant of the first variety comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence encoding an amino acid sequence 100% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; and (b) selecting at least one progeny seed produced in step (a), or a plant germinated therefrom, wherein the at least one progeny seed or plant germinated therefrom comprises the recombinant nucleic acid molecule.

[0100] In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide comprises a nucleic acid sequence that is at least 80% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide comprises a nucleic acid sequence that is at least 85% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide comprises a nucleic acid sequence that is at least 90% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions.In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide comprises a nucleic acid sequence that is at least 95% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide comprises a nucleic acid sequence that is at least 96% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide comprises a nucleic acid sequence that is at least 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions.In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide comprises a nucleic acid sequence that is at least 98% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide comprises a nucleic acid sequence that is at least 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide comprises a nucleic acid sequence that is at least 99.9% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions.In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide comprises a nucleic acid sequence that is 100% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions.

[0101] In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 85% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 90% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions.In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 92.5% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 95% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 96% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions.In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 97% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 98% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 99% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions.In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 99.9% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions. In one aspect, the disclosure provides a method for producing a plant with increased trichome density, comprising: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, wherein the polynucleotide encodes an amino acid sequence that is 100% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) regenerating a modified plant from the at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions.

[0102] In some aspects, the first plant variety and the second plant variety are the same variety. In some aspects, the first plant variety and the second plant variety are two different varieties. In some aspects, the second plant variety comprises the recombinant nucleic acid molecule.

[0103] In some aspects, the first plant variety is heterozygous for the recombinant nucleic acid molecule. In some aspects, the first plant variety is hemizygous for the recombinant nucleic acid molecule. In some aspects, the first plant variety is homozygous for the recombinant nucleic acid molecule. In some aspects, the second plant variety is heterozygous for the recombinant nucleic acid molecule. In some aspects, the second plant variety is hemizygous for the recombinant nucleic acid molecule. In some aspects, the second plant variety is homozygous for the recombinant nucleic acid molecule. In some aspects, the progeny seeds, or plants sprouted therefrom, are heterozygous for the recombinant nucleic acid molecule. In some aspects, the progeny seeds, or plants sprouted therefrom, are hemizygous for the recombinant nucleic acid molecule. In some aspects, the progeny seeds, or plants sprouted therefrom, are homozygous for the recombinant nucleic acid molecule.

[0104] In some aspects, the first plant variety is a tobacco plant variety. In some aspects, the second plant variety is a tobacco plant variety. In some aspects, the first plant variety is a cannabis plant variety. In some aspects, the second plant variety is a cannabis plant variety.

[0105] As used herein, the term "crossing" refers to the intentional mating of two plants. In some aspects, crossing includes pollination and / or fertilization of a first plant by a second plant. The two plants being crossed can be distantly related, closely related, or identical. In some aspects, both of the two plants being crossed are modified plants. In some aspects, the two plants being crossed are of the same variety. In some aspects, the two plants being crossed are of two different varieties. In some aspects, one of the two plants being crossed is male sterile. In some aspects, one of the two plants being crossed is female sterile. In some aspects, at least one of the two plants being crossed is a hybrid tobacco plant. In some aspects, at least one of the two plants being crossed is a modified plant.

[0106] In some aspects, a plant of a first variety is the male parent in a crossbreeding process. In some aspects, a plant of a first variety is the female parent in a crossbreeding process. In some aspects, a plant of a second variety is the male parent in a crossbreeding process. In some aspects, a plant of a second variety is the female parent in a crossbreeding process.

[0107] Nucleic acids and amino acids As used herein, "heterologous" refers to a sequence (nucleic acid or amino acid) that originates from a foreign species or, if derived from the same species, has been substantially modified in composition and / or genomic loci from its native form by deliberate human intervention. This term is also applicable to nucleic acid constructs, also referred to herein as "polynucleotide constructs". Thus, a "heterologous" nucleic acid construct is intended to mean a construct that originates from a foreign species or, if derived from the same species, has been substantially modified in composition and / or genomic loci from its native form by deliberate human intervention. Heterologous nucleic acid constructs include, but are not limited to, recombinant nucleotide constructs that are introduced into a plant or plant part thereof, for example, via transformation methods or subsequent breeding of a transgenic plant with another plant of interest. It will be understood that an endogenous promoter can be considered heterologous to the endogenous gene to which it is operably linked if the two are not naturally operably linked (e.g., human intervention is required to place them in operably linked state). As used herein, an "endogenous" nucleic acid sequence refers to a nucleic acid sequence that is naturally present in the genome of an organism.

[0108] In some aspects, the heterologous polynucleotide comprises a gene. In some aspects, the heterologous polynucleotide encodes a small RNA molecule or a precursor thereof. In some aspects, the heterologous polynucleotide encodes a polypeptide.

[0109] As used herein, "gene" refers to a polynucleotide capable of producing a functional unit (such as, but not limited to, a polypeptide 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'-UTR, a 3'-UTR, or any combination thereof. A "gene sequence" can include a polynucleotide sequence that encodes 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 polypeptide.

[0110] In one aspect, the gene comprises a nucleic acid sequence that is at least 80% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36. In one aspect, the gene comprises a nucleic acid sequence that is at least 85% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36. In one aspect, the gene comprises a nucleic acid sequence that is at least 90% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36. In one aspect, the gene comprises a nucleic acid sequence that is at least 92.5% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36. In one aspect, the gene comprises a nucleic acid sequence that is at least 95% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36. In one aspect, the gene comprises a nucleic acid sequence that is at least 96% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In one aspect, the gene comprises a nucleic acid sequence that is at least 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In one aspect, the gene comprises a nucleic acid sequence that is at least 98% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In one aspect, the gene comprises a nucleic acid sequence that is at least 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In one aspect, the gene comprises a nucleic acid sequence that is at least 99.5% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In one aspect, the gene comprises a nucleic acid sequence that is 100% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36.

[0111] In some aspects, the disclosure provides a nucleic acid sequence that is at least 80% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In some aspects, the disclosure provides a nucleic acid sequence that is at least 85% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In some aspects, the disclosure provides a nucleic acid sequence that is at least 90% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In some aspects, the disclosure provides a nucleic acid sequence that is at least 92.5% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In some aspects, the disclosure provides a nucleic acid sequence that is at least 95% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In some aspects, the disclosure provides a nucleic acid sequence that is at least 96% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In some aspects, the disclosure provides a nucleic acid sequence that is at least 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In some aspects, the disclosure provides a nucleic acid sequence that is at least 98% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In some aspects, the disclosure provides a nucleic acid sequence that is at least 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In some aspects, the disclosure provides a nucleic acid sequence that is at least 99.5% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. In one aspect, the disclosure provides a nucleic acid sequence that is 100% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36.

[0112] In one aspect, the gene comprises a nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the gene comprises a nucleic acid sequence encoding an amino acid sequence at least 85% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the gene comprises a nucleic acid sequence encoding an amino acid sequence at least 90% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the gene comprises a nucleic acid sequence encoding an amino acid sequence at least 92.5% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the gene comprises a nucleic acid sequence encoding an amino acid sequence at least 95% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the gene comprises a nucleic acid sequence encoding an amino acid sequence at least 96% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the gene comprises a nucleic acid sequence encoding an amino acid sequence at least 97% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the gene comprises a nucleic acid sequence encoding an amino acid sequence at least 98% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the gene comprises a nucleic acid sequence encoding an amino acid sequence at least 99% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the gene comprises a nucleic acid sequence encoding an amino acid sequence at least 99.9% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33.In one aspect, the gene comprises a nucleic acid sequence that encodes an amino acid sequence that is 100% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33.

[0113] In some aspects, the disclosure provides nucleic acid sequences that encode an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the disclosure provides nucleic acid sequences that encode an amino acid sequence at least 85% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the disclosure provides nucleic acid sequences that encode an amino acid sequence at least 90% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the disclosure provides nucleic acid sequences that encode an amino acid sequence at least 92.5% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the disclosure provides nucleic acid sequences that encode an amino acid sequence at least 95% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the disclosure provides nucleic acid sequences that encode an amino acid sequence at least 96% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the disclosure provides nucleic acid sequences that encode an amino acid sequence at least 97% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the disclosure provides nucleic acid sequences that encode an amino acid sequence at least 98% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the disclosure provides nucleic acid sequences that encode an amino acid sequence at least 99% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the disclosure provides nucleic acid sequences that encode an amino acid sequence at least 99.9% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33.In one aspect, the disclosure provides nucleic acid sequences that encode an amino acid sequence that is 100% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33.

[0114] In some aspects, the polypeptide comprises an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the polypeptide comprises an amino acid sequence at least 85% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the polypeptide comprises an amino acid sequence at least 90% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the polypeptide comprises an amino acid sequence at least 92.5% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the polypeptide comprises an amino acid sequence at least 95% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the polypeptide comprises an amino acid sequence at least 96% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the polypeptide comprises an amino acid sequence at least 97% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the polypeptide comprises an amino acid sequence at least 98% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the polypeptide comprises an amino acid sequence at least 99% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In some aspects, the polypeptide comprises an amino acid sequence at least 99.9% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the polypeptide comprises an amino acid sequence that is 100% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33.

[0115] The term "percent identity" or "percent identical" when used herein with respect to two or more nucleotide or amino acid sequences is calculated by (i) comparing two optimally aligned sequences (nucleotide or amino acid) over a comparison window (one or more regions that can be "aligned"); (ii) determining the number of positions in both sequences where the same nucleic acid base (for nucleotide sequences) or amino acid residue (for proteins and polypeptides) occurs to obtain the number of matched positions; (iii) dividing the number of matched positions by the total number of positions in 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, for the purposes of this application, when two sequences (query and subject) are optimally aligned (gaps allowed in their alignment), "percent identity" to the query sequence equals the number of identical positions between the two sequences divided by the total number of positions in the query sequence over its length (or comparison window) multiplied by 100%.

[0116] When percentages of sequence identity are used with respect 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), and therefore does not alter the functional properties of the molecule. When sequences differ by conservative substitutions, the percent sequence identity can be adjusted upwards to correct for the conservative nature of the substitution. Sequences that differ by such conservative substitutions are said to have "sequence similarity" or "similarity."

[0117] 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 for optimal alignment of sequences to calculate percent identity. Although other alignment and comparison methods are known in the art, alignment and percent identity between two sequences (including the percent identity ranges described above) can be determined by the ClustalW algorithm, see, e.g., 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.

[0118] 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 the query sequence that are optimally base-paired or hybridized to the nucleotides of the subject sequence when the query and subject sequences are aligned linearly and optimally base-paired, 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 alignment (i.e., without folding or secondary structures) over a comparison window, (ii) determining the number of base-paired positions between the two sequences over the comparison window to obtain the number of complementary positions, (iii) dividing the number of complementary positions by the total number of positions in the comparison window, 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 known pairing of nucleotide bases through hydrogen bonds (such as GC, AT and AU). When "percent complementarity" is calculated with respect to a reference sequence without a specific comparison window, the percent identity is determined by dividing the number of complementary positions between two linear sequences by the total length of the reference sequence. Therefore, for the purposes of this application, when two sequences (query and subject) are optimally base-paired (mismatched or non-base-paired nucleotides are allowed), the "percent complementarity" to a query sequence is equal to the number of base-paired positions between two sequences divided by the total number of positions over the length of the query sequence, multiplied by 100%.

[0119] The use of the term "polynucleotide" or "nucleic acid molecule" is not intended to limit the present disclosure to polynucleotides that include 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 include ribonucleotides and combinations of ribonucleotides and deoxyribonucleotides. Such deoxyribonucleotides and ribonucleotides include both naturally occurring molecules and synthetic analogs. Polynucleotides of the present disclosure also encompass sequences of any form, 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.

[0120] As used herein, a "recombinant nucleic acid molecule" refers to a nucleic acid molecule formed by experimental methods of genetic recombination, including but not limited to molecular cloning. Similarly, a "recombinant DNA construct" refers to a DNA molecule formed by experimental methods of genetic recombination.

[0121] In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 92.5% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs:13-17, 23-30 and 34-36, or a functional fragment thereof.In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 99.9% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, wherein the nucleic acid sequence is 100% identical to a sequence selected from the group consisting of SEQ ID NOs:13-17, 23-30 and 34-36, or a functional fragment thereof.

[0122] In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 85% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 90% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 95% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 96% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 97% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33.In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 98% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 99% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, wherein the polynucleotide encodes an amino acid sequence at least 99.9% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. In one aspect, the disclosure provides a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, wherein the polynucleotide encodes an amino acid sequence 100% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs:18-22 and 31-33.

[0123] Nucleic acids can be isolated using techniques conventional in the art. For example, nucleic acids can be isolated using any method, including but not limited to recombinant nucleic acid techniques 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 techniques include, for example, restriction enzyme digestion and ligation, which can be used to isolate nucleic acids. Isolated nucleic acids can also be chemically synthesized, either as a single nucleic acid molecule 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 degree of purity of a polypeptide can be measured using any suitable method, for example, column chromatography, polyacrylamide gel electrophoresis, or HPLC analysis.

[0124] In one aspect, the present disclosure provides a method for detecting recombinant nucleic acid and polypeptide in plant cells.Nucleic acid can also be detected using hybridization, but is not limited to this.Nucleic acid-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).

[0125] 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.

[0126] 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 for the polypeptides provided herein can be produced 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.

[0127] Detection (e.g., of an amplification product, of a hybridization complex, of a polypeptide) can be accomplished using a detectable label. The term "label" is intended to encompass the use of direct and indirect labels. Detectable labels include enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials and radioactive materials.

[0128] In one aspect, the disclosure provides a promoter. In one aspect, the promoter is heterologous to the nucleic acid sequence that is operably linked. As used herein, "operably linked" refers to the functional linkage between two or more elements. For example, the functional linkage between a polynucleotide of interest and a regulatory sequence (e.g., a promoter) is a functional relationship that allows the expression of the polynucleotide of interest. The elements that are operably linked can be contiguous or non-contiguous.

[0129] A promoter that drives enhanced expression in certain tissues of an organism compared to other tissues of the organism is referred to as a "tissue-preferred" promoter. Thus, a "tissue-preferred" promoter causes relatively high or preferential expression in certain tissues of a plant, but lower expression levels in other tissues of the plant. In some aspects, the promoter is a tissue-preferred promoter. In some aspects, the tissue-preferred promoter is a leaf tissue-preferred promoter. In some aspects, the tissue-preferred promoter is a flower tissue-preferred promoter, such as the promoter of the gene encoding anthocyanidin synthase (NtANS1) (Lim et al., Plant Cell Tissue Organ Cult 114(3):373-383 (2013)). In some aspects, the tissue-preferred promoter is a root tissue-preferred promoter, such as the promoter of extension-like protein (NtREL1) (Zhang et al. Plant Cell Rep 35,757-769 (2016)). In one aspect, the promoter is an epidermal tissue-preferred promoter, such as the promoter of the gene encoding the lipid transport protein in the root epidermis (Ntltp1) (Canevascini et al., Plant Physiol 112(2) 513-524 (1996)).

[0130] A promoter that drives expression in all or most tissues of a plant is called a "constitutive" promoter. In some aspects, the promoter is a constitutive promoter. In some aspects, the constitutive promoter is selected from the group consisting of the cauliflower mosaic virus 35S promoter, the ubiquitin promoter, the actin promoter, the opine promoter, and the alcohol dehydrogenase promoter.

[0131] An "inducible" promoter is a promoter that initiates transcription in response to an environmental stimulus, such as heat, cold, sunlight, light, or other stimulus, such as wounding or the application of a chemical. In one aspect, the promoter is an inducible promoter.

[0132] In one aspect, the promoter is capable of being expressed in a plant cell.

[0133] Terpenes Terpene is a class of aromatic organic compounds produced by plants and some insects.Terpene is a hydrocarbon molecule that plants often use to directly prevent feeding damage or to attract predators or parasites of plant herbivores.Non-limiting examples of terpene include citral, menthol, camphor, salvinorin A, cannabinoids, and curcuminoids.

[0134] In some aspects, terpenes are terpenoids.Terpenoids (also called isoprenoids) are modified terpenes that contain additional functional groups that may include oxygen.Terpenoids, which may be cyclic or acyclic, vary in size from 5-carbon hemiterpenes to long complex molecules that contain thousands of isoprene units.Terpenoids are produced through the condensation of 5-carbon isoprene units (e.g., dimethylallyl diphosphate (DMAPP) and isopentenyl diphosphate (IPP)) with each other, mostly by sequential head-to-tail addition of DMAPP to IPP.The initial cyclization process is catalyzed by different terpene synthases, and mutations in the enzymes lead to mutations in monoterpene structures.

[0135] Terpenoids are classified according to the number of isoprene units that comprise the parent terpene: Hemiterpenoids contain one isoprene unit; Monoterpenoids contain two isoprene units; Sesquiterpenoids contain three isoprene units; Diterpenoids contain four isoprene units; Sesterterpenoids contain five isoprene units; Triterpenoids contain six isoprene units; Tetraterpenoids contain eight isoprene units; Polyterpenoids contain more than eight isoprene units.

[0136] In some aspects, the terpene is menthol. In some aspects, the terpene is menthol or a related compound. In some aspects, the terpene is a labdanoid. In some aspects, the terpene is cembratrienediol. In some aspects, the terpene is levopimaric acid. In some aspects, the terpene is L-leucine. In some aspects, the terpene is neophytadiene. In some aspects, the labdanoid is cis-abienol. In some aspects, the terpene is selected from the group consisting of menthol or a related compound, labdanoid, cembratrienediol, levopimaric acid, and L-leucine. In some aspects, the terpene is selected from the group consisting of menthol or a related compound, labdanoid, cembratrienediol, levopimaric acid, L-leucine, and neophytadiene. In some aspects, the terpene is selected from the group consisting of menthol, labdanoids, cembratrienediol, levopimaric acid, and L-leucine.In some aspects, the terpene is selected from the group consisting of menthol, labdanoids, cembratrienediol, levopimaric acid, L-leucine, and neophytadiene.

[0137] As used herein, "menthol" has the chemical formula: 10 H 20Menthol refers to an organic compound having the formula: 5-methyl-2-(propan-2-yl)cyclohexan-1-ol and the International Union of Pure and Applied Chemistry (IUPAC) name: 5-methyl-2-(propan-2-yl)cyclohexan-1-ol. Menthol is also referred to as "(-)-menthol". Menthol-related compounds include, but are not limited to, (+)-menthol, (+)-isomenthol, (+)-neomenthol, (+)-neoisomenthol, (-)-isomenthol, (-)-neomenthol, and (-)-neoisomenthol. In one aspect, the menthol-related compound is selected from the group consisting of (+)-menthol, (+)-isomenthol, (+)-neomenthol, (+)-neoisomenthol, (-)-isomenthol, (-)-neomenthol, and (-)-neoisomenthol.

[0138] As used herein, "neophytadiene" has the chemical formula: 20 H 38 and the IUPAC name: 7,11,15-trimethyl-3-methylidenehexadec-1-ene.

[0139] As used herein, "cembratrienediol" refers to a compound having the chemical formula: 20 H 34 It refers to the organic compound with the O2 and IUPAC name: (1R,3R,4Z,8Z,12S,13Z)-1,5,9-trimethyl-12-propan-2-ylcyclotetradeca-4,8,13-triene-1,3-diol. Cembratrienediol is also known as "beta-cembrenediol".

[0140] As used herein, "levopimaric acid" refers to a compound having the chemical formula: 20 H 30 It refers to the organic compound with the O2 and IUPAC name: (1R,4aR,4bS,10aR)-1,4a-dimethyl-7-propan-2-yl-2,3,4,4b,5,9,10,10a-octahydrophenanthrene-1-carboxylic acid. Levopimaric acid is also known as "L-pimaric acid".

[0141] As used herein, "L-leucine" has the chemical formula: CH 12 It refers to the amino acid with NO2 and IUPAC name: (2S)-2-amino-4-methylpentanoic acid.

[0142] As used herein, "labdanoid" refers to a diterpene that is a terpenoid derivative of the basic parent labdane. Labdane has the chemical formula: 20 H 38 and has the IUPAC name: (1S,2S,4aS,8aR)-2,5,5,8a-tetramethyl-1-[(3R)-3-methylpentyl]-1,2,3,4,4a,6,7,8-octahydronaphthalene.

[0143] A non-limiting example of a labdanoid is cis-abienol. As used herein, "cis-abienol" has the chemical formula: 20 H 34 It refers to the organic compound having the following IUPAC name: (1R,2R,4aS,8aS)-2,5,5,8a-tetramethyl-1-[(2Z)-3-methylpenta-2,4-dienyl]-3,4,4a,6,7,8-hexahydro-1H-naphthalen-2-ol.

[0144] In some aspects, modified plants, seeds or plant parts comprising the recombinant nucleic acid provided herein comprise an increased amount of at least one terpene compared to a control plant, seed or plant part lacking the recombinant nucleic acid molecule when grown under equivalent conditions. In some aspects, modified tobacco plants, tobacco seeds or tobacco plant parts comprising the recombinant nucleic acid provided herein comprise an increased amount of at least one terpene compared to a control tobacco plant, tobacco seed or tobacco plant part lacking the recombinant nucleic acid molecule when grown under equivalent conditions. In some aspects, modified cannabis plants, cannabis seeds or cannabis plant parts comprising the recombinant nucleic acid provided herein comprise an increased amount of at least one terpene compared to a control cannabis plant, cannabis seed or cannabis plant part lacking the recombinant nucleic acid molecule when grown under equivalent conditions.

[0145] In some aspects, the increased amount of the at least one terpene comprises an increase of at least 0.5%. In some aspects, the increased amount of the at least one terpene comprises an increase of at least 1%. In some aspects, the increased amount of the at least one terpene comprises an increase of at least 2%. In some aspects, the increased amount of the at least one terpene comprises an increase of at least 3%. In some aspects, the increased amount of the at least one terpene comprises an increase of at least 4%. In some aspects, the increased amount of the at least one terpene comprises an increase of at least 5%. In some aspects, the increased amount of the at least one terpene comprises an increase of at least 10%. In some aspects, the increased amount of the at least one terpene comprises an increase of at least 12.5%. In some aspects, the increased amount of the at least one terpene comprises an increase of at least 15%. In some aspects, the increased amount of the at least one terpene comprises an increase of at least 17.5%. In some aspects, the increased amount of the at least one terpene comprises an increase of at least 20%. In some aspects, the increased amount of at least one terpene includes at least a 25% increase. In some aspects, the increased amount of at least one terpene includes at least a 30% increase. In some aspects, the increased amount of at least one terpene includes at least a 40% increase. In some aspects, the increased amount of at least one terpene includes at least a 50% increase. In some aspects, the increased amount of at least one terpene includes at least a 60% increase. In some aspects, the increased amount of at least one terpene includes at least a 70% increase. In some aspects, the increased amount of at least one terpene includes at least an 80% increase. In some aspects, the increased amount of at least one terpene includes at least a 90% increase. In some aspects, the increased amount of at least one terpene includes at least a 100% increase. In some aspects, the increased amount of at least one terpene includes at least a 150% increase. In some aspects, the increased amount of at least one terpene includes at least a 200% increase. In one aspect, the increased amount of at least one terpene comprises an increase of at least 250%.In one aspect, the increased amount of at least one terpene comprises an increase of at least 500%.

[0146] In some aspects, the increased amount of the at least one terpene includes an increase of 0.5% to 500%. In some aspects, the increased amount of the at least one terpene includes an increase of 0.5% to 250%. In some aspects, the increased amount of the at least one terpene includes an increase of 0.5% to 100%. In some aspects, the increased amount of the at least one terpene includes an increase of 0.5% to 75%. In some aspects, the increased amount of the at least one terpene includes an increase of 0.5% to 50%. In some aspects, the increased amount of the at least one terpene includes an increase of 0.5% to 25%. In some aspects, the increased amount of the at least one terpene includes an increase of 0.5% to 10%. In some aspects, the increased amount of the at least one terpene includes an increase of 0.5% to 5%. In some aspects, the increased amount of the at least one terpene includes an increase of 0.5% to 500%. In some aspects, the increased amount of the at least one terpene includes an increase of 5% to 250%. In some aspects, the increased amount of the at least one terpene includes an increase of 5% to 100%. In some aspects, the increased amount of the at least one terpene includes an increase of 5% to 50%. In some aspects, the increased amount of the at least one terpene includes an increase of 25% to 500%. In some aspects, the increased amount of the at least one terpene includes an increase of 25% to 250%. In some aspects, the increased amount of the at least one terpene includes an increase of 50% to 100%. In some aspects, the increased amount of the at least one terpene includes an increase of 100% to 500%.

[0147] The amount of terpene in plant can be measured by any method known in the art, including but not limited to gas chromatography-mass spectrometry (GC-MS), nuclear magnetic resonance spectroscopy and liquid chromatography-mass spectrometry.See The Handbook of Plant Metabolomics, edited by Weckwerth and Kahl, (Wiley-Blackwell) (May 28, 2013).In some aspects, the amount of at least one terpene refers to the concentration of at least one terpene in collected tissue.

[0148] product In some aspects, the present disclosure provides dried plant material from any plant or plant part provided herein. In some aspects, the present disclosure provides dried tobacco material from any tobacco plant or tobacco plant part provided herein.

[0149] In some aspects, the dried plant material is produced by a drying process selected from the group consisting of hot air drying, air drying, flame drying, and sun drying. In some aspects, the dried tobacco material is produced by a drying process selected from the group consisting of hot air drying, air drying, flame drying, and sun drying.

[0150] "Curing" is a aging process that reduces the moisture content and causes the destruction of chlorophyll, thereby giving tobacco leaves a golden color and converting starch to sugars. Cured tobacco therefore has a higher reducing sugar content and a lower starch content than harvested green leaves. In one aspect, the tobacco plants or plant components provided herein can be dried using conventional means (e.g., hot air drying, barn drying, flame drying, air drying, or sun drying). For a description of different types of curing methods, see, for example, Tso (1999, Chapter 1 in Tobacco, Production, Chemistry and Technology, Davis & Nielsen, eds., Blackwell Publishing, Oxford). Cured tobacco is typically aged in wooden drums (e.g., vats) or cardboard cartons under compressed conditions for several years (e.g., 2-5 years) at a moisture content ranging from 10% to about 25%. See U.S. Pat. Nos. 4,516,590 and 5,372,149. The cured and aged tobacco can then be further processed, which includes conditioning the tobacco under vacuum with or without steam injection at various temperatures, pasteurization, and fermentation.

[0151] Information regarding harvesting burley and dark tobacco varieties can be found in the 2019-2020 Burley and Dark Tobacco Production Guide (December 2018) published by the University of Kentucky, the University of Tennessee, Virginia Polytechnic Institute and University, and North Carolina State University, which is incorporated by reference in its entirety.

[0152] In some aspects, the dried tobacco material comprises a tobacco material selected from the group selected from dried leaf material, dried stem material, dried bud material, dried flower material, and dried root material. In some aspects, the dried tobacco material comprises dried leaf material, dried stem material, or both. In some aspects, the dried tobacco material comprises dried leaf material. In some aspects, the dried tobacco material comprises dried stem material.

[0153] In some aspects, the cured tobacco material comprises flue-cured tobacco material. In some aspects, the cured tobacco material comprises air-cured tobacco material. In some aspects, the cured tobacco material comprises flue-cured tobacco material. In some aspects, the cured tobacco material comprises sun-cured tobacco material. In some aspects, 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. In some aspects, the cured tobacco material is derived from a tobacco variety selected from the group consisting of flue-cured, bright, burley, Virginia, Maryland, dark, Oriental, and Turkish.

[0154] In some aspects, the cured tobacco provided herein is selected from the group consisting of air-cured, flue-cured, sun-cured, and flue-cured tobacco. In some aspects, the cured tobacco is from a tobacco variety selected from the group consisting of flue-cured, bright, burley, Virginia, Maryland, dark, Oriental, and Turkish.

[0155] Fermentation is typically characterized by high initial moisture content, heat generation, and a loss of 10-20% of dry weight. See, e.g., U.S. Pat. Nos. 4,528,993, 4,660,577, 4,848,373, 5,372,149; U.S. Patent Publication No. 2005 / 0178398; and Tso (1999, Chapter 1 in Tobacco, Production, Chemistry and Technology, Davis & Nielsen, eds., Blackwell Publishing, Oxford). The cured, aged and fermented tobacco can be further processed (e.g., cut, shredded, expanded, or blended). See, e.g., U.S. Pat. Nos. 4,528,993; 4,660,577; and 4,987,907. In one aspect, the present disclosure provides a fermented tobacco material from any of the tobacco plants or parts thereof provided herein. In another aspect, the present disclosure provides a fermented tobacco material derived from any of the modified tobacco plants or parts thereof provided herein.

[0156] Tobacco materials obtained from the tobacco lines, varieties or hybrids of the present disclosure can be used to manufacture tobacco products. As used herein, "tobacco products" are defined as any product made from or derived from tobacco intended for human use or consumption. In one aspect, the present disclosure provides tobacco products comprising plant material from the tobacco plants provided herein. In another aspect, the present disclosure provides tobacco products comprising plant material from the modified tobacco plants provided herein. In another aspect, the present disclosure provides tobacco products comprising dried tobacco materials. In another aspect, the present disclosure provides tobacco products comprising fermented tobacco materials. In another aspect, the present disclosure provides tobacco products comprising tobacco blends.

[0157] Tobacco products include, but are not limited to, cigarette products (e.g., cigarettes and bidi cigarettes), cigar products (e.g., cigar wrappers and cigarillos), pipe tobacco products, tobacco-derived products, tobacco-derived nicotine products, smokeless tobacco products (e.g., moist snuff, dry snuff, and chewing tobacco), films, chewables, tabs, shaped parts, gels, consumable units, insoluble matrices, hollow shapes, reconstituted tobacco, expanded tobacco, etc. See, e.g., U.S. Patent Publication No. US 2006 / 0191548.

[0158] As used herein, a "cigarette" refers to a tobacco product having a "rod" and a "filler." The "rod" of a cigarette includes the cigarette paper, the filter, the plug wrap (used to contain the filtration material), the tipping paper that secures the cigarette paper (including the filler) to the filter, and any adhesives that secure these components together. The "filler" includes (1) all tobacco, including but not limited to reconstituted tobacco and expanded tobacco, (2) non-tobacco substitutes (including but not limited to herbs, non-tobacco plant materials, and other spices that may accompany tobacco that is wrapped in cigarette paper), (3) casing, (4) flavorings, and (5) all other additives that are mixed with the tobacco and substitutes and wrapped into the cigarette.

[0159] In one aspect, the tobacco product comprises reconstituted tobacco. In another aspect, the present disclosure provides a reconstituted tobacco comprising dried tobacco material. As used herein, "reconstituted tobacco" refers to a member of tobacco filler made from tobacco dust and other tobacco scrap materials that are processed into sheets and cut into strips to resemble tobacco. In addition to cost savings, reconstituted tobacco is also highly valued for its contribution to cigarette flavor, since it uses a reaction between ammonia and sugars to drive flavor development.

[0160] In some aspects, the tobacco product includes expanded tobacco. As used herein, "expanded tobacco" refers to a member of tobacco filler that has been processed through a suitable gas expansion to "expand" the tobacco resulting in reduced density and greater filling capacity. Expanded tobacco reduces the weight of the tobacco used in the cigarette.

[0161] Tobacco products derived from the plants of the present disclosure also include cigarettes and other smoking articles, particularly smoking articles that include a filter element, where the rod of smokable material includes cured tobacco within a tobacco blend. In some aspects, the tobacco products of the present disclosure are selected from the group consisting of kretek, bidi cigarettes, cigarillos, non-ventilated recess filter cigarettes, vented recess filter cigarettes, cigars, snuff, pipe tobacco, cigar tobacco, cigarette tobacco, chewing tobacco, leaf tobacco, hookah, shredded tobacco, and cut tobacco.

[0162] In certain aspects, the tobacco product of the present disclosure is selected from the group consisting of cigarettes, heated tobacco products, kreteks, bidi cigarettes, cigars, cigarillos, non-ventilated cigarettes, ventilated recessed filter cigarettes, pipe tobacco, snuff, snus, chewing tobacco, moist smokeless tobacco, fine cut chewing tobacco, long cut chewing tobacco, pouched chewing tobacco products, gum, tablets, lozenges, and dissolving strips.

[0163] In some aspects, the tobacco product of the present disclosure is a smokeless tobacco product. In some aspects, the smokeless tobacco product is selected from the group consisting of loose leaf chewing tobacco, solid chewing tobacco, moist snuff, nasal snuff, dry snuff, and snus.

[0164] 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 divided tobacco leaves, typically packaged in large pouch-like packages and used in a solid or twisted form. Moist smokeless tobacco is moist, more finely divided tobacco, provided in loose form or in the form of a bag, typically packaged in a round tin and used as a pinch or in a bag between the cheek and gums of an adult tobacco consumer. Snus is smokeless tobacco that has been heat-treated. Dry snuff is finely powdered tobacco that is placed in the mouth or used intranasally.

[0165] In yet another aspect, the tobacco product of the present disclosure is selected from the group consisting of an electronically heated cigarette, an e-cigarette, and an electronic vaporization device.

[0166] In some aspects, the tobacco products of the present disclosure can be blended tobacco products.

[0167] In another aspect, the present disclosure provides a tobacco blend comprising a dried tobacco material. The tobacco blend may include any combination of dried tobacco, never-cured tobacco, fermented tobacco, unfermented tobacco, expanded tobacco, and reconstituted tobacco.

[0168] In some aspects, the tobacco blend comprises at least 5% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 10% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 15% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 20% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 25% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 30% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 35% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 40% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 45% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 50% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 55% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 60% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 65% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 70% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 75% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 80% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 85% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 90% by weight of cured tobacco. In some aspects, the tobacco blend comprises at least 95% by weight of cured tobacco.

[0169] In some aspects, the tobacco blend comprises at least 5% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 10% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 15% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 20% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 25% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 30% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 35% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 40% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 45% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 50% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 55% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 60% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 65% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 70% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 75% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 80% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 85% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 90% by volume of cured tobacco. In some aspects, the tobacco blend comprises at least 95% by volume of cured tobacco.

[0170] In some aspects, the disclosure provides a cannabis product comprising a material derived from a cannabis plant, cannabis seed or cannabis plant part provided herein. In some aspects, the cannabis product is a smokeless product. In further aspects, the smokeless cannabis product is an edible cannabis product. In further aspects, the smokeless cannabis product is a fiber-based product. In some aspects, the cannabis product is a smokable product. In some aspects, the cannabis product is derived from cannabis biomass. In some aspects, the cannabis product is a distillate derived from cannabis biomass.

[0171] In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 92.5% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof.In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof.In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 99.9% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product produced using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, the nucleic acid sequence being selected from the group consisting of SEQ ID NOs:13-17, 23-30 and 34-36, or a functional fragment thereof.

[0172] In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 80% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 85% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 90% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 92.5% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof.In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 95% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 96% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 97% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 98% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof.In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 99% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a portion derived therefrom, wherein the modified tobacco plant or a portion derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 99.9% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a tobacco product produced using cured tobacco material from a modified tobacco plant or portion thereof, wherein the modified tobacco plant or portion thereof comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, the nucleic acid sequence encoding an amino acid sequence selected from the group consisting of SEQ ID NOs:18-22 and 31-33, or a functional fragment thereof.

[0173] In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 92.5% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof.In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof.In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 99.9% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or part derived therefrom, wherein the modified cannabis plant or part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous polynucleotide, and wherein the nucleic acid sequence is 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof.

[0174] In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 80% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 85% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 90% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 92.5% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof.In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 95% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 96% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 97% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 98% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof.In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 99% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant or a part derived therefrom, wherein the modified cannabis plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 99.9% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the disclosure provides a method comprising preparing a cannabis product using material from a modified cannabis plant, or a part derived therefrom, wherein the modified cannabis plant, or a part derived therefrom, comprises a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof.

[0175] Transformation In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b).In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 92.5% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b).In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b).In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is at least 99.9% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b).

[0176] In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence is at least 85% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence is at least 92.5% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule includes a promoter operably linked to a heterologous nucleic acid sequence, and the nucleic acid sequence is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof.In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence is at least 96% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence is at least 97% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence is at least 98% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence is at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36, or a functional fragment thereof. In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule includes a promoter operably linked to a heterologous nucleic acid sequence, and the nucleic acid sequence is at least 99.9% identical to a sequence selected from the group consisting of SEQ ID NOs:13-17, 23-30 and 34-36, or a functional fragment thereof.In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous polynucleotide, and the nucleic acid sequence is 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, or a functional fragment thereof.

[0177] In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 80% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 85% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 90% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b).In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 92.5% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 95% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 96% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b).In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 97% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 98% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 99% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b).In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes an amino acid sequence at least 99.9% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b). In one aspect, the disclosure provides a method of making a modified plant, the method comprising: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a promoter operably linked to a heterologous nucleic acid sequence, wherein the nucleic acid sequence encodes a protein, or a functional fragment thereof, selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell comprises the recombinant nucleic acid molecule; and (c) regenerating a modified plant from the at least one plant cell selected in step (b).

[0178] In one aspect, the method provided herein comprises transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence encodes an amino acid sequence at least 80% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the method provided herein comprises transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence encodes an amino acid sequence at least 85% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, a method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence encodes an amino acid sequence at least 90% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, a method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence encodes an amino acid sequence at least 92.5% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and the nucleic acid sequence encodes an amino acid sequence at least 95% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof.In one aspect, a method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence encodes an amino acid sequence at least 96% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, a method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence encodes an amino acid sequence at least 97% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, a method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence encodes an amino acid sequence at least 98% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, a method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and wherein the nucleic acid sequence encodes an amino acid sequence at least 99% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof. In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and the nucleic acid sequence encodes an amino acid sequence at least 99.9% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, or a functional fragment thereof.In one aspect, the method provided herein includes transforming a plant cell with a recombinant nucleic acid molecule, wherein the recombinant nucleic acid molecule comprises a promoter operably linked to a heterologous nucleic acid sequence, and the nucleic acid sequence encodes an amino acid sequence selected from the group consisting of SEQ ID NOs:18-22 and 31-33, or a functional fragment thereof.

[0179] Numerous methods are known in the art for "introducing" recombinant nucleic acid molecules into plant cells, which can be used in accordance with the methods of the present application to produce modified plant cells, plants, seeds, or plant parts. As used herein, the terms "introducing" and "transforming" can be used interchangeably. Any suitable method or technique for the transformation of plant cells known in the art can be used in accordance with the present methods. Effective methods for plant transformation include bacterially mediated transformation, such as Agrobacterium- or Rhizobium-mediated transformation, and biolistic mediated transformation. A variety of methods are known in the art for transforming explants with transformation vectors via bacterially mediated transformation or biolistic bombardment, and then subsequently culturing or otherwise regenerating transgenic plants from the explants. Other methods for plant transformation, such as microinjection, electroporation, vacuum infiltration, pressurization, sonication, silicon carbide fiber agitation, polyethylene glycol (PEG)-mediated transformation, and the like, are also known in the art. The modified plants produced by these transformation methods can be chimeric or non-chimeric with respect to the transformation event depending on the method and explant used.

[0180] The method of transforming plant cells is well known to those skilled in the art.For example, the specific teaching of transforming plant cells by particle bombardment with recombinant DNA-coated particles (for example, biolistec transformation) can be found in U.S. Patent No. 5,550,318; No. 5,538,880, No. 6,160,208; No. 6,399,861; and No. 6,153,812, and Agrobacterium-mediated transformation is described in U.S. Patent No. 5,159,135; No. 5,824,877; No. 5,591,616; No. 6,384,301; No. 5,750,871; No. 5,463,174; and 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 skilled in the art can be used to transform plant cells (e.g., tobacco cells, cannabis cells) with any of the nucleic acid molecules provided herein.

[0181] In one aspect, the method of introducing a recombinant nucleic acid molecule into a plant cell comprises Agrobacterium-mediated transformation. In another aspect, the method of introducing a recombinant nucleic acid molecule into a plant cell comprises PEG-mediated transformation. In another aspect, the method of introducing a recombinant nucleic acid molecule into a plant cell comprises biolistec transformation. In another aspect, the method of introducing a recombinant nucleic acid molecule into a plant cell comprises liposome-mediated transfection (lipofection). In another aspect, the method of introducing a recombinant nucleic acid molecule into a plant cell comprises lentiviral transfection.

[0182] 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 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).

[0183] Any plant cell capable of regenerating fertile plants is contemplated as a useful recipient cell for the practice of the present disclosure.

[0184] In some aspects, the recombinant nucleic acid molecule is introduced into a tobacco cell. In some aspects, the recombinant nucleic acid molecule is introduced into a tobacco protoplast cell. In other aspects, the recombinant nucleic acid molecule is introduced into a tobacco callus cell. In some aspects, the recombinant nucleic acid molecule is introduced into a tobacco cell selected from the group consisting of a seed cell, a fruit cell, a leaf cell, a cotyledon cell, a hypocotyl cell, a meristem cell, an embryo cell, an endosperm cell, a root cell, a shoot cell, a stem cell, a flower cell, an inflorescence cell, a stalk cell, a pedicel cell, a style cell, a stigma cell, a receptacle cell, a petal cell, a calyx cell, a pollen cell, anther cell, a filament cell, an ovary cell, an ovule cell, a pericarp cell, and a phloem cell.

[0185] In some aspects, the recombinant nucleic acid molecule is introduced into a cannabis cell. In some aspects, the recombinant nucleic acid molecule is introduced into a cannabis protoplast cell. In another aspect, the recombinant nucleic acid molecule is introduced into a cannabis callus cell. In some aspects, the recombinant nucleic acid molecule is introduced into a cannabis cell selected from the group consisting of a seed cell, a fruit cell, a leaf cell, a cotyledon cell, a hypocotyl cell, a meristem cell, an embryo cell, an endosperm cell, a root cell, a shoot cell, a stem cell, a flower cell, an inflorescence cell, a stalk cell, a pedicel cell, a style cell, a stigma cell, a receptacle cell, a petal cell, a calyx cell, a pollen cell, anther cell, a filament cell, an ovary cell, an ovule cell, a pericarp cell, and a phloem cell.

[0186] Callus can be initiated from various tissue sources, including but not limited to immature embryos or embryo parts, seedling apical meristems, microspores, etc. Those cells that can grow as callus can serve as recipient cells for transformation. Practical transformation methods and materials for making 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. Patents 6,194,636 and 6,232,526 and U.S. Patent Application Publication 2004 / 0216189, all of which are incorporated herein by reference.

[0187] Aspects The following non-limiting example embodiments are contemplated. 1. A modified plant, seed or plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. 2. The modified plant, seed or plant part of embodiment 1, wherein the nucleic acid sequence comprises a sequence at least 90% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36. 3. The modified plant, seed or plant part of embodiment 1 or 2, wherein the nucleic acid sequence is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36. 4. The modified plant, seed or plant part of any one of aspects 1-3, wherein the nucleic acid sequence encodes a protein that is at least 90% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. 5. The modified plant, seed or plant part of any one of aspects 1-4, wherein the nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36. 6. The modified plant, seed or plant part of any one of aspects 1 to 5, wherein the amino acid sequence is selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. 7. The modified plant, seed, or part thereof of any one of aspects 1-6, wherein said modified plant, seed, or plant part is a tobacco plant, tobacco seed, or tobacco plant part. 8. The modified plant, seed, or part thereof of embodiment 7, wherein the tobacco plant, tobacco seed, or plant part is of a tobacco variety selected from the group consisting of flue-cured, bright, burley, Virginia, Maryland, dark, Galpao, Oriental, and Turkish. 9. The modified plant, seed, or part thereof of any one of aspects 1-8, wherein said plant is male sterile or cytoplasmically male sterile. 10. The modified plant, seed or part thereof of any one of aspects 1-6, wherein said modified plant, seed or plant part is a cannabis plant, cannabis seed or cannabis plant part. 11. The modified plant, seed, or plant part of any one of embodiments 1-10, wherein the plant or plant part comprises at least one terpene in an amount that is increased by at least 5% compared to a control plant lacking the recombinant nucleic acid molecule. 12. The modified plant, seed, or plant part of any one of embodiments 1-11, wherein the heterologous promoter is selected from the group consisting of a constitutive promoter, an inducible promoter, and a tissue-preferred or tissue-specific promoter. 13. The modified plant, seed, or plant part of any one of embodiments 1-12, wherein at least one leaf of the modified plant comprises a greater average trichome density compared to a leaf of a control plant grown under equivalent conditions. 14. The modified plant, seed, or plant part of any one of embodiments 1-12, wherein at least one leaf of the modified plant comprises a greater average trichome density on the abaxial surface of the leaf compared to the abaxial surface of a leaf of a control plant grown under equivalent conditions. 15. The modified plant, seed, or plant part of any one of embodiments 1-12, wherein at least one leaf of the modified plant comprises a greater average trichome density on the adaxial surface of the leaf as compared to the adaxial surface of the leaf of a control plant grown under equivalent conditions. 16. A cured tobacco material derived from a modified tobacco plant or tobacco plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. 17. The cured tobacco material of embodiment 16, wherein the cured tobacco material is selected from the group consisting of flue-cured tobacco material, air-cured tobacco material, flue-cured tobacco material, and sun-cured tobacco material. 18. A tobacco product comprising the dried tobacco material of embodiment 16. 19. The tobacco product of embodiment 18 or 23, selected from the group consisting of kretek, bidi cigarette, cigarillo, non-ventilated recessed filter cigarette, ventilated recessed filter cigarette, cigar, pipe tobacco, cigar tobacco, cigarette tobacco, chewing tobacco, moist snuff, nasal snuff, dry snuff, snus, leaf tobacco, water tobacco, shredded tobacco, and cut tobacco. 20. The tobacco product of embodiment 19, which is a smokeless tobacco product. 21. The tobacco product of embodiment 20, wherein the smokeless tobacco product is selected from the group consisting of loose-leaf chewing tobacco, solid chewing tobacco, moist snuff, nasal snuff, dry snuff, and snus. 22. A reconstituted tobacco comprising the dried tobacco material of embodiment 16. 23. A tobacco product comprising material derived from a modified tobacco plant, tobacco seed or tobacco plant part, comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. 24. A cannabis product comprising material from a modified cannabis plant, cannabis seed or cannabis plant part, comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. 25. A recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33, wherein the nucleic acid sequence is operably linked to a heterologous promoter. 26. The recombinant nucleic acid molecule of embodiment 25, wherein the nucleic acid sequence encodes a protein that is at least 90% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. 27. The recombinant nucleic acid molecule of embodiment 25 or 26, wherein the nucleic acid sequence is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30, and 34-36. 28. The recombinant nucleic acid molecule of any one of aspects 25-27, wherein the nucleic acid sequence encodes a protein that is at least 95% identical or similar to a sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33. 29. The recombinant nucleic acid molecule of any one of aspects 25-28, wherein the nucleic acid sequence is selected from the group consisting of SEQ ID NOs:13-17, 23-30, and 34-36. 30. The recombinant nucleic acid molecule of any one of aspects 25-29, wherein the nucleic acid sequence encodes a protein selected from the group consisting of SEQ ID NOs:18-22 and 31-33. 31. A method for producing a plant, comprising the steps of: (a) obtaining at least one plant comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter; (b) crossing the at least one plant with at least one plant of a second variety to produce at least one progeny seed; and (c) selecting the at least one progeny seed produced in step (b), or a plant germinated therefrom, wherein the at least one progeny seed or a plant germinated therefrom contains the recombinant nucleic acid molecule. 32. The method of embodiment 31, further comprising the steps of: (f) harvesting plant material from a plant germinated from said at least one progeny seed; and (g) producing a product consisting of or derived from the harvested plant material. 33. The method of embodiment 31 or 32, wherein a plant grown from the at least one progeny seed further comprises at least one terpene in an amount that is increased by at least 5% relative to a control plant lacking the recombinant nucleic acid molecule. 34. The method of any one of aspects 31-33, wherein a plant grown from the at least one progeny seed further comprises at least one terpene in an amount that is increased by at least 10% relative to a control plant lacking the recombinant nucleic acid molecule. 35. The method of any one of aspects 31-34, wherein the at least one plant is from a genus selected from the group consisting of Nicotiana and Cannabis. 36. The method of any one of aspects 31-35, wherein the at least one plant is from a genus selected from the group consisting of tobacco and cannabis. 37. The method of any one of aspects 31-36, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence encoding a protein at least 80% identical or similar to a sequence selected from the group consisting of SEQ ID NOs:18-22 and 31-33. 38. The method of any one of aspects 31-37, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs:13-27 and 34-36. 39. The method of any one of aspects 31-38, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence encoding a protein that is at least 90% identical or similar to a sequence selected from the group consisting of SEQ ID NOs:18-22 and 31-33. 40. The method of any one of aspects 31-39, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs:13-27 and 34-36. 41. The method of any one of aspects 31-40, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence encoding a protein selected from the group consisting of SEQ ID NOs:18-22 and 31-33. 42. A method for producing a modified plant, comprising the steps of: (a) introducing into at least one plant cell a recombinant nucleic acid molecule comprising a nucleic acid sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell contains the recombinant nucleic acid molecule; and (c) regenerating a modified plant from at least one plant cell selected in step (b). 43. A method comprising preparing a tobacco product using cured tobacco material from a modified tobacco plant or a part derived therefrom, wherein the modified tobacco plant or a part derived therefrom comprises in at least one plant cell a recombinant nucleic acid molecule comprising a nucleic acid sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter. 44. A method comprising preparing a cannabis product using material from a modified cannabis plant or part derived therefrom, wherein the modified cannabis plant or part derived therefrom comprises in at least one plant cell a recombinant nucleic acid molecule comprising a nucleic acid sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, wherein the nucleic acid sequence is operably linked to a heterologous promoter. 45. A method comprising transforming a plant cell with at least one recombinant nucleic acid molecule for the plant cell, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-17, 23-30 and 34-36, and the nucleic acid sequence is operably linked to a heterologous promoter. 46. ​​A method for producing a plant having increased trichome density, comprising the steps of: (a) providing to at least one plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, the polynucleotide encoding an amino acid sequence at least 80% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-22 and 31-33; and (b) regenerating a modified plant from at least one plant cell of step (a), wherein the modified plant comprises increased trichome density in at least one leaf compared to a leaf from a control plant lacking the recombinant nucleic acid when grown under equivalent conditions.

[0188] Having now generally described the present disclosure, the same will be more readily understood by reference to the following examples, which are provided by way of illustration and are not intended to limit the disclosure, unless specifically stated. EXAMPLES

[0189] Example 1. Identification of candidate transcription factors for trichome density modification Transcription factors involved in regulating trichome formation and development were used as queries in a search of the National Center for Biotechnology Information (NCBI) database. A similar approach was utilized to identify lipid transport proteins in tobacco. The resulting sequences were then extracted from a second internal database. This methodology identified the transcription factor family members MYB (Figures 3A and 3B) and C2H2 (Figure 4). From these families, three transcription factor candidates are selected for the modification of trichome density in plants. This approach also identified 86 members of the lipid transfer protein (LTP) family in tobacco, which subsequently led to the identification of two variant isoforms of LTP1: isoform 2 (SEQ ID NO: 27) has a 5' extension compared to isoform 1 (SEQ ID NO: 26). See Figures 8A and 8B. This approach allowed the curation of a list of candidate genes that are trichome-specific and involved in metabolite transport (SEQ ID NO: 13-27 and 34-36). See Table 1.

[0190] Example 2. Vector construction The candidate genes identified in Example 1 (e.g., SEQ ID NOs: 13-27 and 34-36) are cloned using PCR Cloning System with GATEWAY Technology (ThermoFisher Scientific, Catalog Number 12535029). The cloned genes are then subcloned into a GATEWAY expression vector, in which the subcloned genes are fused to GREEN Fluorescence Protein (G3GFP) operably linked to the CaMV 35S promoter. See FIG. 2.

[0191] Example 3. Transformation and regeneration of modified tobacco plants Each of the vector constructs made in Example 2 is transformed into tobacco cells separately in separate experiments. Briefly, the vector is introduced into tobacco leaf disks via Agrobacterium transformation. See, for example, Mayo et al., Nat. Protoc., 1:1105-1111 (2006); and Horsch et al., Science, 227:1229-1231 (1985).

[0192] Transformed tobacco plants (varieties "TN90" and "Izmir Ego") and N. benthamiana are grown in Magenta™ GA-7 boxes and leaf disks are cut and placed in Petri dishes. Agrobacterium tumefaciens cells containing the transformation vector are harvested by centrifuging 20 mL of cell suspension in a 50 mL centrifuge tube at 3500 RPM for 10 minutes. The supernatant is removed and the Agrobacterium tumefaciens cell pellet is resuspended in 40 mL of liquid resuspension medium. Tobacco leaves, avoiding the midrib, are cut into eight 0.6 cm disks with a #15 razor blade and placed upside down in a Petri dish. A thin layer of Murashige & Skoog (MS) containing B5 vitamin liquid resuspension medium is added to the Petri dish and the leaf disks are poked evenly with a fine-tipped needle. Add approximately 25 mL of Agrobacterium tumefaciens suspension to the Petri dish and incubate the leaf discs in the suspension for 10 min.

[0193] Transfer the leaf discs to a co-cultivation Petri dish (1 / 2 MS medium) and place the discs upside down in contact with a filter paper laid on co-cultivation TOM medium (MS medium containing 30 g / L sucrose; 0.1 mg / L 1-naphthaleneacetic acid (NAA); and 1 mg / L 6-benzylaminopurine (BAP)). Seal the Petri dish with parafilm and incubate in the dark for 2 days.

[0194] After incubation, the leaf discs are transferred to regeneration / selection TOM K medium Petri dishes (TOM medium + 200 mg / L cefotaxime and 50 mg / L hygromycin). The calli arising from the leaf discs are subcultured to fresh TOM-Hyg medium once every 2 weeks at 24 °C in dim light (60-80 mE / ms) with a photoperiod of 18 hours on and 6 hours off until shoots (plantlets) can be cut. The plantlets arising from the calli are removed with forceps and subcultured to MS rooting medium (MS medium containing 3 g / L sucrose, 7 g / L dextrose with 200 mg / L cefotaxime and 50 mg / L hygromycin). Shoots on MS basal medium containing 50 mg / L hygromycin are incubated at 24°C under the same illumination (approximately 60-80 mE / ms) with a photoperiod of 18 hours on and 6 hours off to induce rooting.

[0195] Once the plantlets, including both shoots and roots, are large enough (e.g., greater than half the height of a Magenta™ GA-7 box), they are transferred to Jiffy peat pellets for acclimation in a growth chamber. Once rooted, the seedlings are transferred to the greenhouse for further growth, propagation and analysis.

[0196] Example 4. Confirmation of expression of candidate genes in modified tobacco plants During the vegetative growth period, RNA is extracted from young leaves of the modified tobacco plants produced in Example 3, and from control tobacco plants lacking the recombinant nucleic acid construct grown under equivalent conditions. The extracted RNA is used to generate cDNA. Gene expression of NtMYB and NtGIS is quantified in the T0 transgenic plants using quantitative real-time PCR (qRT-PCR) (see Figures 10 and 11). Expression of NtMYB and NtGIS in the modified plants is compared to control tobacco plants to confirm that the constructs are expressing the recombinant nucleic acids.

[0197] Gene expression of NtMYB and NbGIS is also quantified in T1 transgenic plants, see Figures 16 and 17, respectively.

[0198] Example 5. Confirmation of altered trichome density in modified tobacco plants During the vegetative growth period, samples of NtMYB86, NtGIS and NbGIS transgenic lines from various tobacco backgrounds are examined under a stereomicroscope (model / type) to identify changes in trichome density in various plant parts compared to control plants. Results demonstrate that transgenic plants containing NtMYB86, NtGIS or NbGIS exhibit an increased number of glandular trichomes per unit area (i.e., increased trichome density) in leaf and stem tissues (see Figures 5-7).

[0199] Example 6. Measurement of terpene levels in engineered tobacco plants During the vegetative growth period, young leaves are collected from the modified tobacco plants from Examples 3, 4 and 5, and from control tobacco plants lacking the recombinant nucleic acid construct grown under equivalent conditions, for use in qualitative metabolic profile analysis following the protocol outlined by Jiang et al. (Curr Protoc Plant Biol. 2016;1:345-358). Leaf samples are ground in liquid nitrogen, and the samples are then mixed with 60:40 hexane:ethyl acetate (v / v) supplemented with heptadecanol (internal standard) and incubated overnight with shaking.

[0200] The solvent extract is concentrated in a chilled SpeedVac™ (ThermoFisher Scientific) and loaded onto a silica column. The column is washed with hexane and drained into a collection tube. An aliquot of the sample is taken from the collection tube and used for gas chromatography-mass spectrometry (GC-MS) of metabolites.

[0201] To identify secondary metabolites secreted from leaf trichomes, 1 gram of leaf sample from transgenic lines is cut into small sections, immersed in hexane supplemented with heptadecanol (internal standard) and incubated overnight with shaking. Extracts are filtered and used for analysis of metabolic profiles by gas chromatography-mass spectrometry (GC-MS). See Figures 12-15.

Claims

1. A modified tobacco plant, tobacco seed or tobacco plant part, comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 18-20, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

2. 2. The modified tobacco plant, tobacco seed or tobacco plant part of claim 1, wherein the nucleic acid sequence is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-15 and 23-25.

3. 2. The modified tobacco plant, tobacco seed or tobacco plant part of claim 1, wherein the nucleic acid sequence encodes a protein that is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 18-20.

4. 2. The modified tobacco plant, tobacco seed or tobacco plant part of claim 1, wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 13-15 and 23-25.

5. 2. The modified tobacco plant, tobacco seed or tobacco plant part of claim 1, wherein the amino acid sequence is selected from the group consisting of SEQ ID NOs: 18-20.

6. 6. The modified tobacco plant, seed or tobacco plant part thereof of any one of claims 1 to 5, wherein the tobacco plant, seed or tobacco plant part is of a tobacco variety selected from the group consisting of flue-cured variety, bright variety, Burley variety, Virginia variety, Maryland variety, dark variety, Galpao variety, Oriental variety, and Turkish variety.

7. 7. The modified tobacco plant, tobacco seed or tobacco plant part thereof according to any one of claims 1 to 6, wherein the tobacco plant is male sterile or cytoplasmically male sterile.

8. 8. The modified tobacco plant, tobacco seed or tobacco plant part of any one of claims 1 to 7, wherein the tobacco plant or tobacco plant part comprises at least one terpene in an amount that is increased by at least 5% compared to a control tobacco plant or control tobacco plant part lacking the recombinant nucleic acid molecule.

9. 9. The modified tobacco plant, tobacco seed or tobacco plant part of any one of claims 1 to 8, wherein the heterologous promoter is selected from the group consisting of a constitutive promoter, an inducible promoter, and a tissue-preferred or tissue-specific promoter.

10. 10. The modified tobacco plant, tobacco seed or tobacco plant part of any one of claims 1 to 9, wherein at least one leaf of the modified tobacco plant comprises a greater average trichome density compared to a leaf of a control tobacco plant grown under equivalent conditions.

11. 10. The modified tobacco plant, tobacco seed or tobacco plant part of any one of claims 1 to 9, wherein at least one leaf of the modified tobacco plant comprises a greater average trichome density on the abaxial surface of the leaf compared to the abaxial surface of the leaf of a control tobacco plant grown under equivalent conditions.

12. 10. The modified tobacco plant, tobacco seed or tobacco plant part of any one of claims 1 to 9, wherein at least one leaf of the modified tobacco plant comprises a greater average trichome density on the adaxial surface of the leaf compared to the adaxial surface of the leaf of a control tobacco plant grown under equivalent conditions.

13. A cured tobacco material derived from a modified tobacco plant or tobacco plant part comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 18-20, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

14. 14. The cured tobacco material of claim 13, wherein the cured tobacco material is selected from the group consisting of flue-cured tobacco material, air-cured tobacco material, flue-cured tobacco material, and sun-cured tobacco material.

15. A tobacco product comprising the dried tobacco material of claim 13 or 14.

16. 16. The tobacco product of claim 15, wherein the tobacco product is selected from the group consisting of kreteks, bidi cigarettes, cigarillos, non-ventilated recess filter cigarettes, vented recess filter cigarettes, cigars, pipe tobacco, cigar tobacco, cigarette tobacco, chewing tobacco, moist snuff, nasal snuff, dry snuff, snus, leaf tobacco, water tobacco, shredded tobacco, and cut tobacco.

17. 16. The tobacco product of claim 15, which is a smokeless tobacco product.

18. 18. The tobacco product of claim 17, wherein the smokeless tobacco product is selected from the group consisting of loose-leaf chewing tobacco, solid chewing tobacco, moist snuff, nasal snuff, dry snuff, and snus.

19. 15. A reconstituted tobacco comprising the dry tobacco material of claim 13 or 14.

20. A tobacco product comprising material derived from a modified tobacco plant, tobacco seed, or tobacco plant part, comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 18-20, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

21. A recombinant nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 18-20, wherein the nucleic acid sequence is operably linked to a heterologous promoter.

22. 22. The recombinant nucleic acid molecule of claim 21, wherein the nucleic acid sequence is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-15 and 23-25.

23. 22. The recombinant nucleic acid molecule of claim 21, wherein the nucleic acid sequence encodes a protein that is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 18-20.

24. 22. The recombinant nucleic acid molecule of claim 21, wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 13-15 and 23-25.

25. 22. The recombinant nucleic acid molecule of claim 21, wherein the nucleic acid sequence encodes a protein selected from the group consisting of SEQ ID NOs: 18-20.

26. 1. A method for producing a tobacco plant, comprising the steps of: (a) obtaining at least one tobacco plant comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-15 and 23-25, wherein the nucleic acid sequence is operably linked to a heterologous promoter; (b) crossing the at least one tobacco plant with at least one tobacco plant of a second variety to produce at least one progeny tobacco seed; and (c) selecting the at least one progeny tobacco seed produced in step (b), or a tobacco plant germinated therefrom, wherein the at least one progeny tobacco seed or a tobacco plant germinated therefrom contains the recombinant nucleic acid molecule.

27. 27. The method of claim 26, further comprising the steps of: (d) harvesting plant material from a tobacco plant germinated from said at least one progeny tobacco seed; and (e) producing a product consisting of or derived from the plant material harvested in step (d).

28. 27. The method of claim 26, wherein a tobacco plant grown from said at least one progeny tobacco seed further comprises at least one terpene in an amount that is increased by at least 5% compared to a control tobacco plant lacking said recombinant nucleic acid molecule.

29. 27. The method of claim 26, wherein a tobacco plant grown from said at least one progeny tobacco seed further comprises at least one terpene in an amount that is increased by at least 10% compared to a control tobacco plant lacking said recombinant nucleic acid molecule.

30. The method of any one of claims 26 to 29, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence encoding a protein that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 18 to 20.

31. The method of any one of claims 26 to 29, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence that is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-15 and 23-25.

32. The method of any one of claims 26 to 29, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence encoding a protein that is at least 95% identical to a sequence selected from the group consisting of SEQ ID NOs: 18 to 20.

33. The method of any one of claims 26 to 29, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 13-15 and 23-25.

34. The method of any one of claims 26 to 29, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence encoding a protein selected from the group consisting of SEQ ID NOs: 18 to 20.

35. 1. A method for producing a modified tobacco plant, comprising the steps of: (a) introducing into at least one tobacco plant cell a recombinant nucleic acid molecule comprising a nucleic acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-15 and 23-25, wherein the nucleic acid sequence is operably linked to a heterologous promoter; (b) selecting at least one plant cell from step (a), wherein the at least one plant cell contains the recombinant nucleic acid molecule; and (c) regenerating a modified tobacco plant from at least one plant cell selected in step (b).

36. 1. A method comprising: preparing a tobacco product using cured tobacco material from a modified tobacco plant or a part derived therefrom, wherein the modified tobacco plant or a part derived therefrom comprises a recombinant nucleic acid molecule comprising a nucleic acid sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 13-15 and 23-25, and operably linked to a heterologous promoter.

37. A method comprising the step of transforming a tobacco plant cell with a recombinant nucleic acid molecule for at least one plant cell, wherein the recombinant nucleic acid molecule comprises a nucleic acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NO:13-15 and 23-25 ​​and is operably linked to a heterologous promoter.

38. 1. A method for producing a tobacco plant having increased trichome density, comprising the steps of: (a) providing to at least one tobacco plant cell a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide, the polynucleotide encoding an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-20; and (b) regenerating a modified tobacco plant from at least one tobacco plant cell of step (a), wherein the modified tobacco plant comprises increased trichome density in at least one leaf compared to a leaf from a control tobacco plant lacking the recombinant nucleic acid when grown under equivalent conditions.

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