WO2021130756A1

WO2021130756A1 - Combinations of non-protein amino acids and acetolactate synthase enzyme inhibitors

Info

Publication number: WO2021130756A1
Application number: PCT/IL2020/051330
Authority: WO
Inventors: Liron KLIPCAN; Ruth VAN-OSS PINHASI
Original assignee: The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Center)
Priority date: 2019-12-25
Filing date: 2020-12-24
Publication date: 2021-07-01

Abstract

The present invention provides compositions and methods for disrupting protein synthesis in plant cells.

Description

COMBINATIONS OF NON-PROTEIN AMINO ACIDS AND ACETOLACTATE

SYNTHASE ENZYME INHIBITORS

[0001] BACKGROUND OF THE DISCLOSURE

[0002] Non-Protein Amino Acids (NPAAs) are amino acids, which are not encoded by the genetic code of any organism. Despite the use of only 23 amino acids (21 in eukaryotes) by the translational machinery to assemble proteins (i.e. the proteinogenic amino acids), over 500 natural non-protein amino acids are known, and thousands of more combinations of coded and non-coded amino acids are possible.

[0003] In addition to the NPAAs that are naturally produced in plants, other NPAAs can be either designed synthetically or produced in vivo by the oxidation of amino acid side-chains. Certain structural analogues of the protein amino acids can escape detection by the cellular machinery of protein synthesis and therefore be mis-incorporated into the elongating polypeptide chain of proteins to generate non-native proteins. Several non-proteinogenic amino acids possess important biological roles, as they may be incorporated into proteins via biosynthetic pathways or introduced post-translationally into the proteome, and thus affect protein functions. Some may possess a defined physiological role (e.g., as neurotransmitters and toxins). Importantly, these natural or synthetic compounds can be utilized in the pharmaceutical industry and agriculture.

[0004] The acetolactate synthase (ALS) enzyme (also known as acetohydroxy acid synthase, or AHAS) is a protein found in plants, fungi, and micro-organisms. ALS catalyzes the first step in the synthesis of the branched-chain amino acids (valine, leucine, and isoleucine). Inhibitors of ALS are used as herbicides that slowly starve affected plants of these amino acids, which eventually leads to inhibition of DNA synthesis. The ALS inhibitor family includes sulfonylureas (SUs), imidazolinones (IMIs), triazolopyrimidines (TPs), pyrimidinyl oxybenzoates (POBs), and sulfonylamino carbonyl triazolinones (SCTs).

[0005] There is a need for highly potent herbicides as they can be used in very low concentrations in the field and are of the highest food safety for humans.

[0006] SUMMARY OF THE DISCLOSURE [0007] The present disclosure is based on the surprising finding that sub-effective amounts of non-protein amino acid (NPAA) and acetolactate synthase (ALS) enzyme inhibitors are effective together in disrupting regular protein synthesis in plant cells.

[0008] The present disclosure, in some embodiments thereof, relates to methods to inhibit the growth of weeds, and, more particularly, but not exclusively, to the use of branched amino acids (Leucine, Valine and Isoleucine) analogues, for killing weeds. The present invention further relates in some embodiments to using branch side amino acids analogues to increase the phytotoxic effect to plants by their synergistic effect with at least one ALS inhibitor.

[0009] The present disclosure provides, in one aspect, a method of preventing, slowing or disrupting protein synthesis in a cell of a weed plant, the method comprising the steps of: (i) increasing the level of at least one Non-Protein Amino Acid (NPAA) in the cell; and (ii) inhibiting an acetolactate synthase (ALS) enzyme in the cell.

[00010] In certain embodiments, the weed plant is an angiosperm.

[00011] In certain embodiments, the weed plant is selected from the group consisting of

Alopecurus aequalis (Shortawn Foxtail), Alopecurus japonicus (Japanese Foxtail), Alopecurus myosuroides (Blackgrass), Apera spica-venti (Silky Windgrass), Avena fatua (Wild Oat), Avena sterilis (Sterile Oat), Avena sterilis ssp. Ludoviciana (Sterile oat), Beckmannia syzigachne (American Sloughgrass), Brachypodium distachyon (Annual Falsebrome), Bromus diandrus (Ripgut Brome), Bromus diandrus ssp. rigidus (Rigid Brome), Bromus sterilis (Poverty Brome), Bromus tectorum (Downy Brome, Cheatgrass), Cynosurus echinatus (Hedgehog Dogtailgrass), Digitaria ciliaris (Southern Crabgrass), Digitaria insularis (Sourgrass), Digitaria ischaemum (Smooth Crabgrass), Digitaria sanguinalis (Large Crabgrass), Echinochloa colona (Junglerice), Echinochloa crus-galli var. crus-galli (Barnyardgrass), Echinochloa crus-galli var. formosensis (Taiwan barnyardgrass), Echinochloa oryzoides (Early Watergrass), Echinochloa phyllopogon (E. oryzicola, Late Watergrass), Ehrharta longiflora (Longflowered veldtgrass), Eleusine indica (Goosegrass), Eriochloa punctate (Pasto Amargo), Hordeum murinum ssp. Glaucum (Smooth Barley), Hordeum murinum ssp. Leporinum (Hare Barley), Ischaemum rugosum (Saramollagrass), Leptochloa chinensis (Chinese Sprangletop), Leptochloa panicoides (Amazon Sprangletop), Lolium perenne (Perennial Ryegrass), Lolium perenne ssp. Multiflorum (Italian Ryegrass), Lolium persicum (Persian Darnel), Lolium rigidum (Rigid Ryegrass), Phalaris brachystachys (Shortspike Canary grass), Phalaris minor (Little seed Canary grass), Phalaris paradoxa (Hood Canarygrass), Polypogon fugax (Asia Minor bluegrass), Rottboellia cochinchinensis (R. exaltata, Itchgrass), Sclerochloa kengiana (Keng Stiffgrass), Setaria faberi (Giant Foxtail), Setaria viridis (Green Foxtail), Setaria viridis var. major (var. robusta-alba, var. robustapurpurea, Giant Green Foxtail), Snowdenia polystachya (Ethiopian grass), Sorghum bicolor ssp. drummondii (Sorghum Sudanese, Sudangrass), Sorghum halepense (Johnsongrass), and Urochloa plantaginea (Brachiaria plantaginea, Alexandergrass).

[00012] As used herein, the term "weed plant" generally refers to undesirable vegetative matter. As used herein, the term "weed plant" further generally refers to an unwanted plant that is growing in a place or in a manner that is detrimental to a plant of interest. In certain embodiments, the weed plant prevents or decreases the viability of a domesticated plant. In certain embodiments, the domesticated plant is a crop plant. In certain embodiments, the domesticated plant is an ornamental plant.

[00013] As used herein, the term "domesticated plant" refers to an individual living plant or population of same, a species, subspecies, variety, cultivar or strain, that has been subjected to natural or artificial selection pressure and developed a commercially or aesthetically relevant trait.

[00014] In certain embodiments, the domesticated plant is selected from the group consisting of Acacia spp., Acer spp., Actinidia spp., Aesculus spp., Agathis australis, Albizia amara, Alsophila tricolor, Andropogon spp., Arachis spp, Areca catechu, Astelia fragrans, Astragalus cicer, Baikiaea plurijuga, Betula spp., Brassica spp., Bruguiera gymnorrhiza, Burkea africana, Butea frondosa, Cadaba farinosa, Calliandra spp, Camellia sinensis, Canna indica, Capsicum spp., Cassia spp., Centroema pubescens, Chacoomeles spp., Cinnamomum cassia, Coffea arabica, Colophospermum mopane, Coronillia varia, Cotoneaster serotina, Crataegus spp., Cucumis spp., Cupressus spp., Cyathea dealbata, Cydonia oblonga, Cryptomeria japonica, Cymbopogon spp., Cynthea dealbata, Cydonia oblonga, Dalbergia monetaria, Davallia divaricata, Desmodium spp., Dicksonia squarosa, Dibeteropogon amplectens, Dioclea spp, Dolichos spp., Dorycnium rectum, Echinochloa pyramidalis, Ehraffia spp., Eleusine coracana, Eragrestis spp., Erythrina spp., Eucalypfus spp., Euclea schimperi, Eulalia villosa, Pagopyrum spp., Feijoa sellowlana, Fragaria spp., Flemingia spp, Freycinetia banksli, Geranium thunbergii, GinAgo biloba, Glycine javanica, Gliricidia spp, Gossypium hirsutum, Grevillea spp., Guibourtia coleosperma, Hedysarum spp., Hemaffhia

B altissima, Heteropogon contoffus, Hordeum vulgare, Hyparrhenia rufa, Hypericum erectum, Hypefflielia dissolute, Indigo incamata, Iris spp., Leptarrhena pyrolifolia, Lespediza spp., Lettuca spp., Leucaena leucocephala, Loudetia simplex, Lotonus bainesli, Lotus spp., Macrotyloma axillare, Malus spp., Manihot esculenta, Medicago saliva, Metasequoia glyptostroboides, Musa sapientum, Nicotianum spp., Onobrychis spp., Ornithopus spp., Oryza spp., Peltophorum africanum, Pennisetum spp., Per sea gratissima, Petunia spp., Phaseolus spp., Phoenix canariensis, Phormium cookianum, Photinia spp., Picea glauca, Pinus spp., Pisum sativam, Podocarpus totara, Pogonarthria fleckii, Pogonaffhria squarrosa, Populus spp., Prosopis cineraria, Pseudotsuga menziesii, Pterolobium stellatum, Pyrus communis, Quercus spp., Rhaphiolepsis umbellata, Rhopalostylis sapida, Rhus natalensis, Ribes grossularia, Ribes spp., Robinia pseudoacacia, Rosa spp., Rubus spp., Salix spp., Schyzachyrium sanguineum, Sciadopitys vefficillata, Sequoia sempervirens, Sequoiadendron giganteum, Sorghum bicolor, Spinacia spp., Sporobolus fimbriatus, Stiburus alopecuroides, Stylosanthos humilis, Tadehagi spp, Taxodium distichum, Themeda triandra, Trifolium spp., Triticum spp., Tsuga heterophylla, Vaccinium spp., Vicia spp., Vitis vinifera, Watsonia pyramidata, Zantedeschia aethiopica , Zea mays, amaranth, artichoke, asparagus, broccoli, Brussels sprouts, cabbage, canola, carrot, cauliflower, celery, collard greens, flax, kale, lentil, oilseed rape, okra, onion, potato, rice, soybean, straw, sugar beet, sugar cane, sunflower, tomato, squash tea, maize, wheat, barley, rye, oat, peanut, pea, lentil and alfalfa, cotton, rapeseed, canola, pepper, sunflower, tobacco, eggplant, eucalyptus, a perennial grass and a forage crop.

[00015] In certain embodiments, the method comprises decreasing the production of a branched amino acid in the cell. In certain embodiments, the branched amino acid is selected from the group consisting of valine, isoleucine, and leucine. In certain embodiments, the branched amino acid is valine. In certain embodiments, the branched amino acid is isoleucine. In certain embodiments, the branched amino acid is leucine.

[00016] In certain embodiments, the method comprises contacting the plant cell with at least one NPAA. In certain embodiments, the method comprises contacting the plant cell with 1 to 500 mM NPAA. In certain embodiments, the method comprises contacting the plant cell with 50 pM to 100 pM NPAA. In certain embodiments, the method comprises contacting the plant cell with 50 pM or 100 pM NPAA. [00017] In certain embodiments, the NPAA is a branched amino acid analogue. In certain embodiments, the NPAA is selected from the group consisting of Norvaline, Norleucine, Fluoro- Valine, Thia-isoleucine, o-methyl-Threonine, 3 -Methyl-valine, L-allo- Threonine, L-cyclohexyl-Glycine, 3-Fluoro-Leucine, and 3-Cyclopentyl-DL-alanine. In certain embodiments, the NPAA is Norvaline, Norleucine, Fluoro-valine, Thia-isoleucine, or o-methyl-Threonine.

[00018] In certain embodiments, the NPAA is Norvaline. In certain embodiments, the NPAA is Norleucine. In certain embodiments, the NPAA is Fluoro-valine. In certain embodiments, the NPAA is Thia-isoleucine. In certain embodiments, the NPAA is o-methyl- Threonine.

[00019] In certain embodiments, the NPAA is at least as active as Norvaline, Norleucine, Fluoro-valine, Thia-isoleucine, or o-methyl-Threonine. A person of average skill in the art would understand that the activity of the NPAA is inhibitory effect on plant growth. In certain embodiments, the NPAA has an IC50 value of Norvaline, Norleucine, Fluoro-valine, Thia-isoleucine, or o-methyl-Threonine or lower. In certain embodiments, the NPAA has an IC50 value of Norvaline, Norleucine, Fluoro-valine, Thia-isoleucine, or o-methyl-Threonine, or lower, on plant root length. In certain embodiments, the NPAA has an IC50 value of Norvaline, Norleucine, Fluoro-valine, Thia-isoleucine, or o-methyl-Threonine, or lower, on plant fresh weight. A person of average skill in the art would be familiar with many alternative ways known in the art to determine IC50 values of NPAAs, e.g. for plant root length and/or plant fresh weight, e.g. as the methods described herein.

[00020] In certain embodiments, the method comprises contacting the plant cell with at least one ALS enzyme inhibitor. In certain embodiments, the method comprises contacting the plant cell with 1 to 100 mM ALS enzyme inhibitor. In certain embodiments, the method comprises contacting the plant cell with 0.5 pM to 10 pM ALS enzyme inhibitor. In certain embodiments, the method comprises contacting the plant cell with 2.5 pM to 10 pM ALS enzyme inhibitor.

[00021] In certain embodiments, the ALS enzyme inhibitor is a sulfonylurea. In certain embodiments, the ALS enzyme inhibitor is selected from the group consisting of Amidosulfuron, Azimsulfuron, Bensulfuron-Methyl, Bispyribac, Chlorimuron-Ethyl, Chlorsulfuron, Cinosulfuron, Cloransulam-Methyl, Cyclosulfamuron, Diclosulam, Ethametsulfuron-Methyl, Ethoxysulfuron, Flazasulfuron, Florasulam, Flucarbazone-Sodium, Flucetosulfuron, Flumetsulam, Flupyrsulfuron-Methyl, Foramsulfuron, Halosulfuron -Methyl, Imazamethabenz-Methyl, Imazamox, Imazapic, Imazapyr, Imazaquin, Imazethapyr, Imazosulfuron, Iodosulfuron, Mesosulfuron, Metazosulfuron, Metosulam, Metsulfuron- Methyl, NC-330, Nicosulfuron, Orthosulfamuron, Oxasulfuron, Penoxsulam, Primisulfuron- Methyl, Propoxycarbazone-Sodium, Prosulfuron, Pyrazosulfuron-Ethyl, Pyribenzoxim, Pyriftalid, Pyriminobac-Methyl, Pyrithiobac, Pyroxsulam, Rimsulfuron, Sulfometuron- Methyl, Sulfosulfuron, Thifensulfuron-Methyl, Triasulfuron, Tribenuron-Methyl, Trifloxysulfuron, Triflusulfuron-Methyl, and Tritosulfuron. In certain embodiments, the ALS enzyme inhibitor is Tribenuron-Methyl.

[00022] In certain embodiments, an ALS enzyme inhibitor, an agent which decreases the transcription of an ALS gene, or an agent which decreases the translation of an ALS mRNA, is at least as active as Tribenuron-Methyl. A person of average skill in the art would understand that the activity of the Tribenuron-Methyl is inhibitory effect on plant growth. In certain embodiments, the ALS enzyme inhibitor, agent which decreases the transcription of an ALS gene, or agent which decreases the translation of an ALS mRNA, has an IC50 value of Tribenuron-Methyl or lower. In certain embodiments, the ALS enzyme inhibitor, agent which decreases the transcription of an ALS gene, or agent which decreases the translation of an ALS mRNA, has an IC50 value of Tribenuron-Methyl, or lower, on plant root length. In certain embodiments, the ALS enzyme inhibitor, agent which decreases the transcription of an ALS gene, or agent which decreases the translation of an ALS mRNA, has an IC50 value of Tribenuron-Methyl, or lower, on plant fresh weight. A person of average skill in the art would be familiar with many alternative ways known in the art to determine IC50 values of ALS enzyme inhibitors, agents which decrease the transcription of an ALS gene, or agents which decrease the translation of an ALS mRNA, e.g. for plant root length and/or plant fresh weight, e.g. as the methods described herein.

[00023] In certain embodiments, the method comprises contacting the plant cell with at least one agent which decreases the transcription of an ALS gene.

[00024] In certain embodiments, the method comprises contacting the plant cell with at least one agent which decreases the translation of an ALS mRNA. [00025] In certain embodiments, the ALS enzyme inhibitor, ALS mRNA translation- inhibiting agent, or ALS gene transcription-inhibiting agent, is applied to the cell of the weed plant before growing a plant of interest in the vicinity of the weed plant. In certain embodiments, the ALS enzyme inhibitor is applied to the cell of the weed plant before growing a plant of interest in the vicinity of the weed plant.

[00026] In certain embodiments, the ALS enzyme inhibitor, ALS mRNA translation- inhibiting agent, or ALS gene transcription-inhibiting agent, is applied to the cell of the weed plant while growing a plant of interest in the vicinity of the weed plant. In certain embodiments, the ALS enzyme inhibitor is applied to the cell of the weed plant while growing a plant of interest in the vicinity of the weed plant.

[00027] The present invention provides, in another aspect, a composition, comprising: (i) at least one NPAA, and (ii) at least one ALS enzyme inhibitor, at least one ALS mRNA translation-inhibiting agent, or at least one ALS gene transcription-inhibiting agent.

[00028] The present invention provides, in another aspect, a kit, comprising: (i) a composition comprising at least one NPAA, and (ii) a composition comprising at least one ALS enzyme inhibitor, at least one ALS mRNA translation-inhibiting agent, or at least one ALS gene transcription-inhibiting agent.

[00029] In certain embodiments, the composition comprising at least one NPAA and the composition comprising at least one ALS enzyme inhibitor are different compositions.

[00030] In certain embodiments, the composition comprising at least one NPAA and the composition comprising at least one ALS enzyme inhibitor are the same composition.

[00031 ] BRIEF DESCRIPTION OF THE DRAWINGS

[00032] Figure l is a schematic representation of the potential effects of ALS inhibitors in combination with branched amino acids analogues.

[00033] Figure 2 are panel images depicting the effects of branched side amino acids analogs, ALS inhibitor (Tribenuron-methyl), and their combination on Arabidopsis thaliana (var. Columbia) seed-germination and seedlings establishment. [00034] Figure 3 are panel images depicting the effects of the branch side amino acid analog Norvaline, ALS inhibitor (Tribenuron-methyl), and their combination on the monocot plant Teff ( Eragrostis abyssinicd) seed-germination and seedlings establishment.

[00035] Figure 4 is a bar graph depicting the effects of ALS inhibitors in combination with branched amino acids analogues on plant root length (black) and plant fresh weight (gray).

[00036] Figure 5 are panel images depicting the effects of branched side amino acids analogs, ALS inhibitors, and their combinations.

[00037] Figure 6 is a box graph depicting the effects of ALS inhibitors in combination with branched amino acids analogues on plant root length (black) and plant fresh weight (gray).

[00038] Figure 7 is a box graph depicting the expected effects (black) and observed effects (gray) of ALS inhibitors in combination with branched amino acids analogues on plant root length.

[00039] Figure 8 is a box graph (left panel) and dot graph (right panel) depicting the effects of ALS inhibitors in combination with branched amino acids analogues on plant fresh weight.

[00040] Figure 9 is a bar graph depicting the effects of branched side amino acids analogs, ALS inhibitors, and their combinations, on proteome incorporation of the branched side amino acids analogs.

[00041] Figure 10 is a bar graph (left panel - Norvaline; right panel - Norleucine) depicting the effects of ALS inhibitors in combination with branched amino acids analogues on wild-type and mutant plant fresh weight.

[00042] DETAILED DESCRIPTION OF THE DISCLOSURE

[00043] The present disclosure relates, inter alia, to mixtures and combinations of non protein amino acid (NPAA) and acetolactate synthase (ALS) enzyme inhibitors.

[00044] The Norvaline analog is a naturally occurring non-protein amino acid. Experimental data indicates that Norvaline is produced by nonspecific side reactions in main biosynthesis pathways of branched amino acids biosynthesis due to broad substrate specificity of the enzymes involved. In bacterial cells, it can be incorporated into proteins instead of leucine, especially under conditions of oxygen limitation, which induce intracellular Norvaline accumulation. It was reported that Norvaline can be incorporated into heterologous proteins, expressed in E. coli , at positions normally occupied by leucine. Norvaline is an efficient substrate for leucine tRNA synthetase (LeuRS) and may represent a significant threat to quality control at leucine codons.

[00045] Valine, isoleucine, and leucine are synthesized in plants and microbes by a common pathway, which involves several enzymes. A compound from the substance class of the ALS inhibitors inhibits the enzyme acetolactate synthase (ALS), which is responsible for the biosynthesis of branched amino acids. Inspired by the idea that inhibitory effects of ALS could be reversed by adding branched amino acids to the media, we propose that the reduction of branched amino acids biosynthesis opens way to more efficient incorporation of analogues. As a result of misincorporation, ALS in combination with branched amino acids analogues will lead to a strong phytotoxic effect on plants. The invention provides a method for induction of mis-incorporation of branch side non-protein amino acids in place of leucine, valine and isoleucine in plant proteins (Figure 1). The method can be utilized in the production of analogues damaged proteins. The present disclosure concerns a synergistic herbicidal mixture comprising a herbicidally sub-effective amounts of ALS inhibitor and amino acid analogues.

[00046] There are five chemical families of ALS-inhibiting herbicides that have been commercialized including; imidazolinones, sulfonylureas, triazolopyrimidines, pyrimidinylthio(or oxy)-benzoates, and sulfonylamino-carbonyltriazolinones. About 30 commercial herbicides are from the family of sulfonylureas, but only 2-6 are from each of the rest chemical families.

[00047] It is demonstrated herein that the application of sub-effective dose of branched amino acids analogues in parallel with ALS inhibitors may have a profound inhibitory effect on A. thaliana root growth (Figure 2).

[00048] Compared to herbicides applied alone, the performance of herbicide mixtures can be either synergistic, or additive. Additivity is the combined action, which is equal to the total response predicted by considering the response of each herbicide applied alone. Synergism is the combined action of two herbicides where the observed response to their joint application is greater than the response predicted by Colby method. The Colby method makes use of the equation Ei= Xi_*Yi/100, where Ei represents the expected growth as a percent-of- control, Xi represents the growth as a percent-of-control with herbicide A, and Y i represents the growth as a percent-of-control with herbicide. The value for the herbicide mixture is then compared to the ‘expected’ value, and if it is less than the ‘expected’ value it implies an antagonistic effect, if it is equal to the ‘expected’ value it implies an additive effect, and if it is greater than the ‘expected’ value it implies a synergistic effect.

[00049] However, in many cases, the synergistic total increase in action is so high that Colby’s criterion can be dispensed. The application of different branched amino acids analogues coupled with ALS inhibitors clearly have a synergistic effect (Figure 2).

[00050] The data provided herein indicates that seed germination was strongly affected by the presence of Norvaline, 3F-Valine and Thia-isoleucine. These results show that the branch side amino acids analogues tested herein, modified at branch side position, have phytotoxic effects alone or in combination with ALS inhibitors, influencing seed germination and plants development (Figure 2). It should be noted that many more branched amino acids analogues can be easily synthesized chemically based on the present teachings. In addition to the compounds already tested, other ALS inhibitors and various branch side amino acids derivatives are useful for different types of weeds, achieving synergistic effect to e.g. overcome resistance to either one of these agents alone (Figure 3).

[00051] The following is a list of herbicides, which may act as ALS inhibitors: Amidosulfuron (CAS Number: 120923-37-7), Azimsulfuron (CAS Number: 120162-55-2), Bensulfuron-Methyl (CAS Number: 83055-99-6), Bispyribac (CAS Number: 125401-75-4), Chlorimuron-Ethyl (CAS Number: 90982-32-4), Chlorsulfuron (CAS Number: 64902-72-3), Cinosulfuron (CAS Number: 94593-91-6), Cloransulam-Methyl (CAS Number: 147150-35- 4), Cyclosulfamuron (CAS Number: 136849-15-5), Diclosulam (CAS Number: 145701-21-9), Ethametsulfuron-Methyl (CAS Number: 97780-06-8), Ethoxysulfuron (CAS Number: 126801-58-9), Flazasulfuron (CAS Number: 104040-78-0), Florasulam (CAS Number: 145701-23-1), Flucarbazone-Sodium (CAS Number: 181274-17-9), Flucetosulfuron (CAS Number: 412928-75-7), Flumetsulam (CAS Number: 98967-40-9), Flupyrsulfuron-Methyl (CAS Number: 144740-53-4), Foramsulfuron (CAS Number: 173159-57-4), Halosulfuron- Methyl (CAS Number: 100784-20-1), Imazamethabenz-Methyl (CAS Number: 81405-85-8), Imazamox (CAS Number: 114311-32-9), Imazapic (CAS Number: 104098-48-8), Imazapyr (CAS Number: 81334-34-1), Imazaquin (CAS Number: 81335-37-7), Imazethapyr (CAS Number: 81335-77-5), Imazosulfuron (CAS Number: 122548-33-8), Iodosulfuron (CAS Number: 185119-76-0), Mesosulfuron (CAS Number: 400852-66-6), Metazosulfuron (CAS Number: 868680-84-6), Metosulam (CAS Number: 139528-85-1), Metsulfuron-Methyl (CAS Number: 74223-64-6), NC-330 (CAS Number: 104770-29-8), Nicosulfuron (CAS Number: 111991-09-4), Orthosulfamuron (CAS Number: 213464-77-8), Oxasulfuron (CAS Number: 144651-06-9), Penoxsulam (CAS Number: 219714-96-2), Primisulfuron-Methyl (CAS Number: 86209-51-0), Propoxycarbazone-Sodium (CAS Number: 181274-15-7), Prosulfuron (CAS Number: 94125-34-5), Pyrazosulfuron-Ethyl (CAS Number: 93697-74-6), Pyribenzoxim (CAS Number: 168088-61-7), Pyriftalid (CAS Number: 135186-78-6), Pyriminobac-Methyl (CAS Number: 147411-69-6), Pyrithiobac (CAS Number: 123342-93-8), Pyroxsulam (CAS Number: 422556-08-9), Rimsulfuron (CAS Number: 122931-48-0), Sulfometuron-Methyl (CAS Number: 74222-97-2), Sulfosulfuron (CAS Number: 141776-32- 1), Thifensulfuron-Methyl (CAS Number: 79277-27-3), Triasulfuron (CAS Number: 82097- 50-5), Tribenuron-Methyl (CAS Number: 101200-48-0), Trifloxysulfuron (CAS number: 145099-21-4), Triflusulfuron-Methyl (CAS Number: 135990-29-3), and Tritosulfuron (CAS Number: 142469-14-5).

[00052] The term “plant” as used herein encompasses whole plants, ancestors and progeny of the plants and plant parts, including seeds, shoots, stems, roots (including tubers), and plant cells, tissues and organs. The plant may be in any form including suspension cultures, embryos, meristematic regions, callus tissue, leaves, gametophytes, sporophytes, pollen, and microspores.

[00053] Plants that are particularly useful in the methods of the invention include all plants which belong to the superfamily Viridiplantae, in particular monocotyledonous and dicotyledonous plants including a fodder or forage legume, ornamental plant, food crop, tree, or shrub selected from the list consisting of Acacia spp., Acer spp., Actinidia spp., Aesculus spp., Agathis australis, Albizia amara, Alsophila tricolor, Andropogon spp., Arachis spp., Areca catechu, Astelia fragrans, Astragalus cicer, Baikiaea plurijuga, Betula spp., Brassica spp., Bruguiera gymnorrhiza, Burkea africana, Butea frondosa, Cadaba farinosa, Calliandra spp., Camellia sinensis, Canna indica, Capsicum spp., Cassia spp., Centroema pubescens, Chacoomeles spp., Cinnamomum cassia, Coffea arabica, Colophospermum mopane, Coronillia varia, Cotoneaster serotina, Crataegus spp., Cucumis spp., Cupressus spp., Cyathea dealbata, Cydonia oblonga, Cryptomeria japonica, Cymbopogon spp., Cynthea dealbata, Cydonia oblonga, Dalbergia monetaria, Davallia divaricata, Desmodium spp., Dicksonia squarosa, Dibeteropogon amplectens, Dioclea spp., Dolichos spp., Dorycnium rectum, Echinochloa pyramidalis, Ehraffia spp., Eleusine coracana, Eragrestis spp., Erythrina spp., Eucalypfus spp., Euclea schimperi, Eulalia villosa, Pagopyrum spp., Feijoa sellowlana, Fragaria spp., Flemingia spp., Freycinetia banksli, Geranium thunbergii, GinAgo biloba, Glycine javanica, Gliricidia spp., Gossypium hirsutum, Grevillea spp., Guibourtia coleosperma, Hedysarum spp., Hemaffliia altissima, Pieter opogon contoffus, Hordeum vulgare, Hyparrhenia rufa, Hypericum erectum, Hypefflielia dissolute, Indigo incamata, Iris spp., Leptarrhena pyrolifolia, Lespediza spp., Lettuca spp., Leucaena leucocephala, Loudetia simplex, Lotonus bainesli, Lotus spp., Macrotyloma axillare, Malus spp., Manihot esculenta, Medicago saliva, Metasequoia glyptostroboides, Musa sapientum, Nicotianum spp., Onobrychis spp., Ornithopus spp., Oryza spp., Peltophorum africanum, Pennisetum spp., Per sea gratissima, Petunia spp., Phaseolus spp., Phoenix canariensis, Phormium cookianum, Photinia spp., Picea glauca, Pinus spp., Pisum sativam, Podocarpus totara, Pogonarthria fleckii, Pogonaffhria squarrosa, Populus spp., Prosopis cineraria, Pseudotsuga menziesii, Pterolobium stellatum, Pyrus communis, Quercus spp., Rhaphiolepsis umbellata, Rhopalostylis sapida, Rhus natalensis, Ribes grossularia, Ribes spp., Robinia pseudoacacia, Rosa spp., Rubusspp., Salixspp., Schyzachyrium sanguineum, Sciadopitys vefficillata, Sequoia sempervirens, Sequoiadendron giganteum, Sorghum bicolor, Spinacia spp., Sporobolus fimbriatus, Stiburus alopecuroides, Stylosanthos humilis, Tadehagi spp., Taxodium distichum, Themeda triandra, Trifolium spp., Triticum spp., Tsuga heterophylla, Vaccinium spp., Vicia spp., Vitis vinifera, Watsonia pyramidata, Zantedeschia aethiopica, Zea mays, amaranth, artichoke, asparagus, broccoli, Brussels sprouts, cabbage, canola, carrot, cauliflower, celery, collard greens, flax, kale, lentil, oilseed rape, okra, onion, potato, rice, soybean, straw, sugar beet, sugar cane, sunflower, tomato, squash tea, maize, wheat, barley, rye, oat, peanut, pea, lentil and alfalfa, cotton, rapeseed, canola, pepper, sunflower, tobacco, eggplant, eucalyptus, a perennial grass and a forage crop.

[00054] The present disclosure provides, in one aspect, a method of disrupting protein synthesis in a cell of a weed plant, comprising the steps of: (a) increasing the level of at least one Non-Protein Amino Acid (NPAA) in the cell; and (b) inhibiting an acetolactate synthase (ALS) enzyme in the cell.

[00055] In certain embodiments, the weed plant is an angiosperm. [00056] In certain embodiments, the weed plant is selected from the group consisting of Alopecurus aequalis (Shortawn Foxtail), Alopecurus japonicus (Japanese Foxtail), Alopecurus myosuroides (Blackgrass), Apera spica-venti (Silky Windgrass), Avena fatua (Wild Oat), Avena sterilis (Sterile Oat), Avena sterilis ssp. Ludoviciana (Sterile oat), Beckmannia syzigachne (American Sloughgrass), Brachypodium distachyon (Annual Falsebrome), Bromus diandrus (Ripgut Brome), Bromus diandrus ssp. rigidus (Rigid Brome), Bromus sterilis (Poverty Brome), Bromus tectorum (Downy Brome, Cheatgrass), Cynosurus echinatus (Hedgehog Dogtailgrass), Digitaria ciliaris (Southern Crabgrass), Digitaria insularis (Sourgrass), Digitaria ischaemum (Smooth Crabgrass), Digitaria sanguinalis (Large Crabgrass), Echinochloa colona (Junglerice), Echinochloa crus-galli var. crus-galli (Barnyardgrass), Echinochloa crus-galli var. formosensis (Taiwan barnyardgrass), Echinochloa oryzoides (Early Watergrass), Echinochloa phyllopogon (E. oryzicola, Late Watergrass), Ehrharta longiflora (Longflowered veldtgrass), Eleusine indica (Goosegrass), Eriochloa punctate (Pasto Amargo), Hordeum murinum ssp. Glaucum (Smooth Barley), Hordeum murinum ssp. Leporinum (Hare Barley), Ischaemum rugosum (Saramollagrass), Leptochloa chinensis (Chinese Sprangletop), Leptochloa panicoides (Amazon Sprangletop), Lolium perenne (Perennial Ryegrass), Lolium perenne ssp. Multiflorum (Italian Ryegrass), Lolium persicum (Persian Darnel), Lolium rigidum (Rigid Ryegrass), Phalaris brachystachys (Shortspike Canarygrass), Phalaris minor (Little seed Canary grass), Phalaris paradoxa (Hood Canarygrass), Polypogon fugax (Asia Minor bluegrass), Rottboellia cochinchinensis (R. exaltata, Itchgrass), Sclerochloa kengiana (Keng Stiffgrass), Setaria faberi (Giant Foxtail), Setaria viridis (Green Foxtail), Setaria viridis var. major (var. robusta-alba, var. robustapurpurea, Giant Green Foxtail), Snowdenia polystachya (Ethiopian grass), Sorghum bicolor ssp. drummondii (Sorghum Sudanese, Sudangrass), Sorghum halepense (Johnsongrass), and Urochloa plantaginea (Brachiaria plantaginea, Alexandergrass).

[00057] In certain embodiments, the weed plant is Alopecurus aequalis (Shortawn Foxtail). In certain embodiments, the weed plant is Alopecurus japonicus (Japanese Foxtail). In certain embodiments, the weed plant is Alopecurus myosuroides (Blackgrass). In certain embodiments, the weed plant is Apera spica-venti (Silky Windgrass). In certain embodiments, the weed plant is Avena fatua (Wild Oat). In certain embodiments, the weed plant is Avena sterilis (Sterile Oat). In certain embodiments, the weed plant is Avena sterilis ssp. Ludoviciana (Sterile oat). In certain embodiments, the weed plant is Beckmannia syzigachne (American Sloughgrass). In certain embodiments, the weed plant is Brachypodium distachyon (Annual

IB Falsebrome). In certain embodiments, the weed plant is Bromus diandrus (Ripgut Brome). In certain embodiments, the weed plant is Bromus diandrus ssp. rigidus (Rigid Brome). In certain embodiments, the weed plant is Bromus sterilis (Poverty Brome). In certain embodiments, the weed plant is Bromus tectorum (Downy Brome. In certain embodiments, the weed plant is Cheatgrass). In certain embodiments, the weed plant is Cynosurus echinatus (Hedgehog Dogtailgrass). In certain embodiments, the weed plant is Digitaria ciliaris (Southern Crabgrass). In certain embodiments, the weed plant is Digitaria insularis (Sourgrass). In certain embodiments, the weed plant is Digitaria ischaemum (Smooth Crabgrass). In certain embodiments, the weed plant is Digitaria sanguinalis (Large Crabgrass). In certain embodiments, the weed plant is Echinochloa colona (Junglerice). In certain embodiments, the weed plant is Echinochloa crus-galli var. crus-galli (Bamyardgrass). In certain embodiments, the weed plant is Echinochloa crus-galli var. formosensis (Taiwan bamyardgrass). In certain embodiments, the weed plant is Echinochloa oryzoides (Early Watergrass). In certain embodiments, the weed plant is Echinochloa phyllopogon (E. oryzicola. In certain embodiments, the weed plant is Late Watergrass). In certain embodiments, the weed plant is Ehrharta longiflora (Longflowered veldtgrass). In certain embodiments, the weed plant is Eleusine indica (Goosegrass). In certain embodiments, the weed plant is Eriochloa punctate (Pasto Amargo). In certain embodiments, the weed plant is Hordeum murinum ssp. Glaucum (Smooth Barley). In certain embodiments, the weed plant is Hordeum murinum ssp. Leporinum (Hare Barley). In certain embodiments, the weed plant is Ischaemum rugosum (Saramollagrass). In certain embodiments, the weed plant is Leptochloa chinensis (Chinese Sprangletop). In certain embodiments, the weed plant is Leptochloa panicoides (Amazon Sprangletop). In certain embodiments, the weed plant is Lolium perenne (Perennial Ryegrass). In certain embodiments, the weed plant is Lolium perenne ssp. Multiflorum (Italian Ryegrass). In certain embodiments, the weed plant is Lolium persicum (Persian Darnel). In certain embodiments, the weed plant is Lolium rigidum (Rigid Ryegrass). In certain embodiments, the weed plant is Phalaris brachystachys (Shortspike Canarygrass). In certain embodiments, the weed plant is Phalaris minor (Little seed Canary grass). In certain embodiments, the weed plant is Phalaris paradoxa (Hood Canarygrass). In certain embodiments, the weed plant is Polypogon fugax (Asia Minor bluegrass). In certain embodiments, the weed plant is Rottboellia cochinchinensis (R. exaltata. In certain embodiments, the weed plant is Itchgrass). In certain embodiments, the weed plant is Sclerochloa kengiana (Keng Stiffgrass). In certain embodiments, the weed plant is Setaria faberi (Giant Foxtail). In certain embodiments, the weed plant is Setaria viridis (Green Foxtail). In certain embodiments, the weed plant is Setaria viridis var. major (var. robusta-alba. In certain embodiments, the weed plant is var. robustapurpurea. In certain embodiments, the weed plant is Giant Green Foxtail). In certain embodiments, the weed plant is Snowdenia polystachya (Ethiopian grass). In certain embodiments, the weed plant is Sorghum bicolor ssp. drummondii (Sorghum Sudanese. In certain embodiments, the weed plant is Sudangrass). In certain embodiments, the weed plant is Sorghum halepense (Johnsongrass). In certain embodiments, the weed plant is Urochloa plantaginea (Brachiaria plantaginea, Alexandergrass).

[00058] In certain embodiments, the weed plant prevents or decreases the viability of a domesticated plant. In certain embodiments, the weed plant prevents the viability of a domesticated plant. In certain embodiments, the weed plant decreases the viability of a domesticated plant.

[00059] In certain embodiments, the domesticated plant possess tolerance to sulfonylurea herbicides. In certain embodiments, the domesticated plant possess a gene encoding for acetolactate synthase (ALS) that contains a point mutation for possessing tolerance to sulfonylurea herbicides. In certain embodiments, the mutation resulting in tolerance is Prol97 to Serl97. In certain embodiments, the mutation resulting in tolerance is substitution of an amino acid for Ala (position 117), Pro (position 192) or Try (position 586).

[00060] In certain embodiments, the domesticated plant possess tolerance to imidazolinones herbicides. In certain embodiments, the domesticated plant possess a gene encoding for acetolactate synthase (ALS) that contains a point mutation for possessing tolerance to imidazolinones herbicides. In certain embodiments, the mutation resulting in tolerance is Ser653 to Asn653.

[00061] In certain embodiments, the domesticated plant is a crop plant.

[00062] In certain embodiments, the domesticated plant is an ornamental plant.

[00063] In certain embodiments, the domesticated plant is selected from the group consisting of Acacia spp., Acer spp., Actinidia spp., Aesculus spp., Agathis australis, Albizia amara, Alsophila tricolor, Andropogon spp., Arachis spp., Areca catechu, Astelia fragrans, Astragalus cicer, Baikiaea plurijuga, Betula spp., Brassica spp., Bruguiera gymnorrhiza, Burkea africana, Butea frondosa, Cadaba farinosa, Calliandra spp., Camellia sinensis, Canna indica, Capsicum spp., Cassia spp., Centroema pubescens, Chacoomeles spp., Cinnamomum cassia, Coffea arabica, Colophospermum mopane, Coronillia varia, Cotoneaster serotina, Crataegus spp., Cucumis spp., Cupressus spp., Cyathea dealbata, Cydonia oblonga, Cryptomeria japonica, Cymbopogon spp., Cynthea dealbata, Cydonia oblonga, Dalbergia monetaria, Davallia divaricata, Desmodium spp., Dicksonia squarosa, Dibeteropogon amplectens, Dioclea spp., Dolichos spp., Dorycnium rectum, Echinochloa pyramidalis, Ehraffia spp., Eleusine coracana, Eragrestis spp., Erythrina spp., Eucalypfus spp., Euclea schimperi, Eulalia villosa, Pagopyrum spp., Feijoa sellowlana, Fragaria spp., Flemingia spp., Freycinetia banksli, Geranium thunbergii, GinAgo biloba, Glycine javanica, Gliricidia spp., Gossypium hirsutum, Grevillea spp., Guibourtia coleosperma, Hedysarum spp., Hemaffhia altissima, Heteropogon contoffus, Hordeum vulgare, Hyparrhenia rufa, Hypericum erectum, Hypeffhelia dissolute, Indigo incamata, Iris spp., Leptarrhena pyrolifolia, Lespediza spp., Lettuca spp., Leucaena leucocephala, Loudetia simplex, Lotonus bainesli, Lotus spp., Macrotyloma axillare, Malus spp., Manihot esculenta, Medicago saliva, Metasequoia glyptostroboides, Musa sapientum, Nicotianum spp., Onobrychis spp., Ornithopus spp., Oryza spp., Peltophorum africanum, Pennisetum spp., Perseagratissima, Petunia spp., Phaseolus spp., Phoenix canariensis, Phormium cookianum, Photinia spp., Picea glauca, Pinus spp., Pisum sativam, Podocarpus totara, Pogonarthria fleckii, Pogonaffhria squarrosa, Populus spp., Prosopis cineraria, Pseudotsuga menziesii, Pterolobium stellatum, Pyrus communis, Quercus spp., Rhaphiolepsis umbellata, Rhopalostylis sapida, Rhus natalensis, Ribes grossularia, Ribes spp., Robinia pseudoacacia, Rosa spp., Rubus spp., Salix spp., Schyzachyrium sanguineum, Sciadopitys vefficillata, Sequoia sempervirens, Sequoiadendron giganteum, Sorghum bicolor, Spinacia spp., Sporobolus fimbriatus, Stiburus alopecuroides, Stylosanthos humilis, Tadehagi spp., Taxodium distichum, Themeda triandra, Trifolium spp., Triticum spp., Tsuga heterophylla, Vaccinium spp., Vicia spp., Vitis vinifera, Watsonia pyramidata, Zantedeschia aethiopica, Zea mays, amaranth, artichoke, asparagus, broccoli, Brussels sprouts, cabbage, canola, carrot, cauliflower, celery, collard greens, flax, kale, lentil, oilseed rape, okra, onion, potato, rice, soybean, straw, sugar beet, sugar cane, sunflower, tomato, squash tea, maize, wheat, barley, rye, oat, peanut, pea, lentil and alfalfa, cotton, rapeseed, canola, pepper, sunflower, tobacco, eggplant, eucalyptus, a perennial grass and a forage crop.

[00064] In certain embodiments, the domesticated plant is Acacia spp.. In certain embodiments, the domestic plant is Acer spp.. In certain embodiments, the domestic plant is Actinidia spp.. In certain embodiments, the domestic plant is Aesculus spp.. In certain embodiments, the domestic plant is Agathis australis. In certain embodiments, the domestic plant is Albizia amara. In certain embodiments, the domestic plant is Alsophila tricolor. In certain embodiments, the domestic plant is Andropogon spp.. In certain embodiments, the domestic plant is Arachis spp. In certain embodiments, the domestic plant is Areca catechu. In certain embodiments, the domestic plant is Astelia fragrans. In certain embodiments, the domestic plant is Astragalus cicer. In certain embodiments, the domestic plant is Baikiaea plurijuga. In certain embodiments, the domestic plant is Betula spp.. In certain embodiments, the domestic plant is Brassica spp.. In certain embodiments, the domestic plant is Bruguiera gymnorrhiza. In certain embodiments, the domestic plant is Burkea africana. In certain embodiments, the domestic plant is Butea frondosa. In certain embodiments, the domestic plant is Cadaba farinosa. In certain embodiments, the domestic plant is Calliandra spp. In certain embodiments, the domestic plant is Camellia sinensis. In certain embodiments, the domestic plant is Canna indica. In certain embodiments, the domestic plant is Capsicum spp.. In certain embodiments, the domestic plant is Cassia spp.. In certain embodiments, the domestic plant is Centroema pubescens. In certain embodiments, the domestic plant is Chacoomeles spp.. In certain embodiments, the domestic plant is Cinnamomum cassia. In certain embodiments, the domestic plant is Coffea arabica. In certain embodiments, the domestic plant is Colophospermum mopane. In certain embodiments, the domestic plant is Coronillia varia. In certain embodiments, the domestic plant is Cotoneaster serotina. In certain embodiments, the domestic plant is Crataegus spp.. In certain embodiments, the domestic plant is Cucumis spp.. In certain embodiments, the domestic plant is Cupressus spp.. In certain embodiments, the domestic plant is Cyathea dealbata. In certain embodiments, the domestic plant is Cydonia oblonga. In certain embodiments, the domestic plant is Cryptomeria japonica. In certain embodiments, the domestic plant is Cymbopogon spp.. In certain embodiments, the domestic plant is Cynthea dealbata. In certain embodiments, the domestic plant is Cydonia oblonga. In certain embodiments, the domestic plant is Dalbergia monetaria. In certain embodiments, the domestic plant is Davallia divaricata. In certain embodiments, the domestic plant is Desmodium spp.. In certain embodiments, the domestic plant is Dicksonia squarosa. In certain embodiments, the domestic plant is Dibeteropogon amplectens. In certain embodiments, the domestic plant is Dioclea spp. In certain embodiments, the domestic plant is Dolichos spp.. In certain embodiments, the domestic plant is Dorycnium rectum. In certain embodiments, the domestic plant is Echinochloa pyramidalis. In certain embodiments, the domestic plant is Ehraffia spp.. In certain embodiments, the domestic plant is Eleusine coracana. In certain embodiments, the domestic plant is Eragrestis spp.. In certain embodiments, the domestic plant is Erythrina spp.. In certain embodiments, the domestic plant is Eucalypfus spp.. In certain embodiments, the domestic plant is Euclea schimperi. In certain embodiments, the domestic plant is Eulalia villosa. In certain embodiments, the domestic plant is Pagopyrum spp.. In certain embodiments, the domestic plant is Feijoa sellowlana. In certain embodiments, the domestic plant is Fragaria spp.. In certain embodiments, the domestic plant is Flemingia spp. In certain embodiments, the domestic plant is Freycinetia banksli. In certain embodiments, the domestic plant is Geranium thunbergii. In certain embodiments, the domestic plant is GinAgo biloba. In certain embodiments, the domestic plant is Glycine javanica. In certain embodiments, the domestic plant is Gliricidia spp. In certain embodiments, the domestic plant is Gossypium hirsutum. In certain embodiments, the domestic plant is Grevillea spp.. In certain embodiments, the domestic plant is Guibourtia coleosperma. In certain embodiments, the domestic plant is Hedysarum spp.. In certain embodiments, the domestic plant is Hemaffhia altissima. In certain embodiments, the domestic plant is Heteropogon contoffus. In certain embodiments, the domestic plant is Hordeum vulgare. In certain embodiments, the domestic plant is Hyparrhenia rufa. In certain embodiments, the domestic plant is Hypericum erectum. In certain embodiments, the domestic plant is Hypeffhelia dissolute. In certain embodiments, the domestic plant is Indigo incamata. In certain embodiments, the domestic plant is Iris spp.. In certain embodiments, the domestic plant is Leptarrhena pyrolifolia. In certain embodiments, the domestic plant is Lespediza spp.. In certain embodiments, the domestic plant is Lettuca spp.. In certain embodiments, the domestic plant is Leucaena leucocephala. In certain embodiments, the domestic plant is Loudetia simplex. In certain embodiments, the domestic plant is Lotonus bainesli. In certain embodiments, the domestic plant is Lotus spp.. In certain embodiments, the domestic plant is Macrotyloma axillare. In certain embodiments, the domestic plant is Malus spp.. In certain embodiments, the domestic plant is Manihot esculenta. In certain embodiments, the domestic plant is Medicago saliva. In certain embodiments, the domestic plant is Metasequoia glyptostroboides. In certain embodiments, the domestic plant is Musa sapientum. In certain embodiments, the domestic plant is Nicotianum spp.. In certain embodiments, the domestic plant is Onobrychis spp.. In certain embodiments, the domestic plant is Omithopus spp.. In certain embodiments, the domestic plant is Oryza spp.. In certain embodiments, the domestic plant is Peltophorum africanum. In certain embodiments, the domestic plant is Pennisetum spp.. In certain embodiments, the domestic plant is Persea gratissima. In certain embodiments, the domestic plant is Petunia spp.. In certain embodiments, the domestic plant is Phaseolus spp.. In certain embodiments, the domestic plant is Phoenix canariensis. In certain embodiments, the domestic plant is Phormium cookianum. In certain embodiments, the domestic plant is Photinia spp.. In certain embodiments, the domestic plant is Picea glauca. In certain embodiments, the domestic plant is Pinus spp.. In certain embodiments, the domestic plant is Pisum sativam. In certain embodiments, the domestic plant is Podocarpus totara. In certain embodiments, the domestic plant is Pogonarthria fleckii. In certain embodiments, the domestic plant is Pogonaffhria squarrosa. In certain embodiments, the domestic plant is Populus spp.. In certain embodiments, the domestic plant is Prosopis cineraria. In certain embodiments, the domestic plant is Pseudotsuga menziesii. In certain embodiments, the domestic plant is Pterolobium stellatum. In certain embodiments, the domestic plant is Pyrus communis. In certain embodiments, the domestic plant is Quercus spp.. In certain embodiments, the domestic plant is Rhaphiolepsis umbellata. In certain embodiments, the domestic plant is Rhopalostylis sapida. In certain embodiments, the domestic plant is Rhus natalensis. In certain embodiments, the domestic plant is Ribes grossularia. In certain embodiments, the domestic plant is Ribes spp.. In certain embodiments, the domestic plant is Robinia pseudoacacia. In certain embodiments, the domestic plant is Rosa spp.. In certain embodiments, the domestic plant is Rubus spp.. In certain embodiments, the domestic plant is Salix spp.. In certain embodiments, the domestic plant is Schyzachyrium sanguineum. In certain embodiments, the domestic plant is Sciadopitys vefficillata. In certain embodiments, the domestic plant is Sequoia sempervirens. In certain embodiments, the domestic plant is Sequoiadendron giganteum. In certain embodiments, the domestic plant is Sorghum bicolor. In certain embodiments, the domestic plant is Spinacia spp.. In certain embodiments, the domestic plant is Sporobolus fimbriatus. In certain embodiments, the domestic plant is Stiburus alopecuroides. In certain embodiments, the domestic plant is Stylosanthos humilis. In certain embodiments, the domestic plant is Tadehagi spp. In certain embodiments, the domestic plant is Taxodium distichum. In certain embodiments, the domestic plant is Themeda triandra. In certain embodiments, the domestic plant is Trifolium spp.. In certain embodiments, the domestic plant is Triticum spp.. In certain embodiments, the domestic plant is Tsuga heterophylla. In certain embodiments, the domestic plant is Vaccinium spp.. In certain embodiments, the domestic plant is Vicia spp.. In certain embodiments, the domestic plant is Vitis vinifera. In certain embodiments, the domestic plant is Watsonia pyramidata. In certain embodiments, the domestic plant is Zantedeschia aethiopica. In certain embodiments, the domestic plant is Zea mays. In certain embodiments, the domestic plant is amaranth. In certain embodiments, the domestic plant is artichoke. In certain embodiments, the domestic plant is asparagus. In certain embodiments, the domestic plant is broccoli. In certain embodiments, the domestic plant is Brussels sprouts. In certain embodiments, the domestic plant is cabbage. In certain embodiments, the domestic plant is canola. In certain embodiments, the domestic plant is carrot. In certain embodiments, the domestic plant is cauliflower. In certain embodiments, the domestic plant is celery. In certain embodiments, the domestic plant is collard greens. In certain embodiments, the domestic plant is flax. In certain embodiments, the domestic plant is kale. In certain embodiments, the domestic plant is lentil. In certain embodiments, the domestic plant is oilseed rape. In certain embodiments, the domestic plant is okra. In certain embodiments, the domestic plant is onion. In certain embodiments, the domestic plant is potato. In certain embodiments, the domestic plant is rice. In certain embodiments, the domestic plant is soybean. In certain embodiments, the domestic plant is straw. In certain embodiments, the domestic plant is sugar beet. In certain embodiments, the domestic plant is sugar cane. In certain embodiments, the domestic plant is sunflower. In certain embodiments, the domestic plant is tomato. In certain embodiments, the domestic plant is squash tea. In certain embodiments, the domestic plant is maize. In certain embodiments, the domestic plant is wheat. In certain embodiments, the domestic plant is barley. In certain embodiments, the domestic plant is rye. In certain embodiments, the domestic plant is oat. In certain embodiments, the domestic plant is peanut. In certain embodiments, the domestic plant is pea. In certain embodiments, the domestic plant is lentil and alfalfa. In certain embodiments, the domestic plant is cotton. In certain embodiments, the domestic plant is rapeseed. In certain embodiments, the domestic plant is canola. In certain embodiments, the domestic plant is pepper. In certain embodiments, the domestic plant is sunflower. In certain embodiments, the domestic plant is tobacco. In certain embodiments, the domestic plant is eggplant. In certain embodiments, the domestic plant is eucalyptus. In certain embodiments, the domestic plant is a perennial grass. In certain embodiments, the domestic plant is a forage crop.

[00065] In certain embodiments, the method comprises decreasing the production of branched amino acids in the cell.

[00066] In certain embodiments, the branched amino acids are valine, isoleucine, or leucine. In certain embodiments, the branched amino acid is valine. In certain embodiments, the branched amino acid is isoleucine. In certain embodiments, the branched amino acid is leucine.

[00067] In certain embodiments, the method comprises contacting the plant cell with at least one NPAA. [00068] In certain embodiments, the method comprises contacting the plant cell with 1 to 500 mM NPAA. In certain embodiments, the method comprises contacting the plant cell with 1 to 400 pM NPAA. In certain embodiments, the method comprises contacting the plant cell with 1 to 300 pM NPAA. In certain embodiments, the method comprises contacting the plant cell with 1 to 200 pM NPAA. In certain embodiments, the method comprises contacting the plant cell with 1 to 100 pM NPAA.

[00069] In certain embodiments, the method comprises contacting the plant cell with 5 pM to 50 pM NPAA. In certain embodiments, the method comprises contacting the plant cell with 10 pM to 50 pM NPAA. In certain embodiments, the method comprises contacting the plant cell with 15 pM to 50 pM NPAA. In certain embodiments, the method comprises contacting the plant cell with 20 pM to 50 pM NPAA. In certain embodiments, the method comprises contacting the plant cell with 25 pM to 50 pM NPAA. In certain embodiments, the method comprises contacting the plant cell with 30 pM to 50 pM NPAA. In certain embodiments, the method comprises contacting the plant cell with 35 pM to 50 pM NPAA. In certain embodiments, the method comprises contacting the plant cell with 40 pM to 50 pM NPAA. In certain embodiments, the method comprises contacting the plant cell with 45 pM to 50 pM NPAA.

[00070] In certain embodiments, the method comprises contacting the plant cell with 50 pM NPAA.

[00071] In certain embodiments, the NPAA is a branched amino acid analogue.

[00072] In certain embodiments, the NPAA is selected from the group consisting of Norvaline, Fluoro- Valine, Thia-isoleucine, Norleucine, 3-Methyl-Valine, L-allo-Threonine, L- cyclohexyl-Glycine, 3-Fluoro-Leucine, and 3-Cyclopentyl-DL-alanine.

[00073] In certain embodiments, the NPAA is Norvaline. In certain embodiments, the NPAA is Fluoro-Valine. In certain embodiments, the NPAA is Thia-isoleucine. In certain embodiments, the NPAA is Norleucine. In certain embodiments, the NPAA is 3-Methyl- Valine. In certain embodiments, the NPAA is L-allo-Threonine. In certain embodiments, the NPAA is L-cyclohexyl-Glycine. In certain embodiments, the NPAA is 3-Fluoro-Leucine. In certain embodiments, the NPAA is 3-Cyclopentyl-DL-alanine. [00074] In certain embodiments, the NPAA is Norvaline, Fluoro-Valine or Thia- isoleucine. In certain embodiments, the NPAA is Norvaline. In certain embodiments, the NPAA is Fluoro-Valine. In certain embodiments, the NPAA Thia-isoleucine.

[00075] In certain embodiments, the NPAA is Norleucine in a concentration of 100 mM to 400 mM. In certain embodiments, the NPAA is Norleucine in a concentration of 100 mM. In certain embodiments, the NPAA is Norleucine in a concentration of 200 mM. In certain embodiments, the NPAA is Norleucine in a concentration of 300 mM. In certain embodiments, the NPAA is Norleucine in a concentration of 400 mM.

[00076] In certain embodiments, the NPAA is Norvaline in a concentration of 100 mM to 400 mM. In certain embodiments, the NPAA is Norvaline in a concentration of 100 mM. In certain embodiments, the NPAA is Norvaline in a concentration of 200 mM. In certain embodiments, the NPAA is Norvaline in a concentration of 300 mM. In certain embodiments, the NPAA is Norvaline in a concentration of 400 mM.

[00077] In certain embodiments, the NPAA is Fluoro-Valine in a concentration of 50 mM to 200 mM. In certain embodiments, the NPAA is Fluoro-Valine in a concentration of 50 mM. In certain embodiments, the NPAA is Fluoro-Valine in a concentration of 100 mM. In certain embodiments, the NPAA is Fluoro-Valine in a concentration of 150 mM. In certain embodiments, the NPAA is Fluoro-Valine in a concentration of 200 mM.

[00078] In certain embodiments, the NPAA is Thia-isoleucine in a concentration of 2 mM to 25 mM. In certain embodiments, the NPAA is Thia-isoleucine in a concentration of 2 mM. In certain embodiments, the NPAA is Thia-isoleucine in a concentration of 5 mM. In certain embodiments, the NPAA is Thia-isoleucine in a concentration of 10 mM. In certain embodiments, the NPAA is Thia-isoleucine in a concentration of 25 mM.

[00079] In certain embodiments, the NPAA is o-Methylthreonine in a concentration of 2 mM to 25 mM. In certain embodiments, the NPAA is o-Methylthreonine in a concentration of 2 mM. In certain embodiments, the NPAA is o-Methylthreonine in a concentration of 5 mM. In certain embodiments, the NPAA is o-Methylthreonine in a concentration of 10 mM. In certain embodiments, the NPAA is o-Methylthreonine in a concentration of 25 mM.

[00080] In certain embodiments, the ALS inhibitor is in a concentration of 1 mM to 10 mM. In certain embodiments, the ALS inhibitor is in a concentration of 1 mM. In certain embodiments, the ALS inhibitor is in a concentration of 2 mM. In certain embodiments, the ALS inhibitor is in a concentration of 3 mM. In certain embodiments, the ALS inhibitor is in a concentration of 4 mM. In certain embodiments, the ALS inhibitor is in a concentration of 5 mM. In certain embodiments, the ALS inhibitor is in a concentration of 10 mM.

[00081] In certain embodiments, the ALS inhibitor is in a concentration of 5 mM, and the NPAA is in a concentration of 50 mM. In certain embodiments, the ALS inhibitor is in a concentration of 10 mM, and the NPAA is in a concentration of 50 mM.

[00082] In certain embodiments, the ALS inhibitor is in a concentration of 3 mM or 6 mM, and the NPAA is in a concentration of 100 mM. In certain embodiments, the ALS inhibitor is in a concentration of 3 mM, and the NPAA is in a concentration of 100 mM. In certain embodiments, the ALS inhibitor is in a concentration of 6 mM, and the NPAA is in a concentration of 100 mM.

[00083] In certain embodiments, the ALS inhibitor is in a concentration of 2.5 mM, and the NPAA is in a concentration of 100 mM.

[00084] In certain embodiments, the ALS inhibitor is in a concentration of 2.5 mM to 250 mM, and the NPAA is in a concentration of 50 mM to 5000 mM. In certain embodiments, the ALS inhibitor is in a concentration of 2.5 mM to 25 mM, and the NPAA is in a concentration of 50 mM to 500 mM.

[00085] In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 1 mM to 10 mM. In certain embodiments, the ALS inhibitor is Tribenuron- methyl in a concentration of 1 mM. In certain embodiments, the ALS inhibitor is Tribenuron- methyl in a concentration of 2 mM. In certain embodiments, the ALS inhibitor is Tribenuron- methyl in a concentration of 3 mM. In certain embodiments, the ALS inhibitor is Tribenuron- methyl in a concentration of 4 mM. In certain embodiments, the ALS inhibitor is Tribenuron- methyl in a concentration of 5 mM. In certain embodiments, the ALS inhibitor is Tribenuron- methyl in a concentration of 10 mM.

[00086] In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 5 mM, and the NPAA is Norvaline in a concentration of 50 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 5 mM, and the NPAA is Fluoro-valine in a concentration of 50 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 5 mM, and the NPAA is Thia-isoleucine in a concentration of 50 mM.

[00087] In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 10 mM, and the NPAA is Norvaline in a concentration of 50 mM.

[00088] In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 5 mM, and the NPAA is Norvaline, Norleucine, Fluoro-valine, Thia- isoleucine, or o-methyl-Threonine in a concentration of 50 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 5 mM, and the NPAA is Norvaline in a concentration of 50 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 5 mM, and the NPAA is Norleucine in a concentration of 50 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 5 mM, and the NPAA is Fluoro-valine in a concentration of 50 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 5 mM, and the NPAA is Thia-isoleucine in a concentration of 50 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 5 mM, and the NPAA is o-methyl-Threonine in a concentration of 50 mM.

[00089] In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 3 mM or 6 mM, and the NPAA is Norvaline or Norleucine in a concentration of 100 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 3 mM, and the NPAA is Norvaline in a concentration of 100 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 3 mM, and the NPAA is Norleucine in a concentration of 100 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 6 mM, and the NPAA is Norvaline in a concentration of 100 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 6 mM, and the NPAA is Norvaline or Norleucine in a concentration of 100 mM.

[00090] In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 2.5 mM, and the NPAA is Norvaline or Norleucine in a concentration of 100 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 2.5 mM, and the NPAA is Norvaline in a concentration of 100 mM. In certain embodiments, the ALS inhibitor is Tribenuron-methyl in a concentration of 2.5 mM, and the NPAA is Norleucine in a concentration of 100 mM. [00091] In certain embodiments, the method comprises contacting the plant cell with at least one ALS inhibitor.

[00092] In certain embodiments, the method comprises contacting the plant cell with at least one ALS inhibitor, and contacting the plant cell with at least one NPAA. In certain embodiments, the method comprises contacting the plant cell with at least one ALS inhibitor and least one NPAA in the same time. A person skilled in the art would understand the phrase “at the same time” includes simultaneous events, and plurality of events separated by an hour or less. In certain embodiments, the method comprises first contacting the plant cell with at least one ALS inhibitor, and then contacting the plant cell with at least one NPAA. In certain embodiments, the method comprises first contacting the plant cell with at least one NPAA, and then contacting the plant cell with at least one ALS inhibitor.

[00093] In certain embodiments, the method comprises contacting the plant cell with 1 to 5 mM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 1 to 10 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 1 to 20 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 1 to 30 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 1 to 40 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 1 to 50 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 1 to 60 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 1 to 70 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 1 to 80 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 1 to 90 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 1 to 100 pM ALS inhibitor.

[00094] In certain embodiments, the method comprises contacting the plant cell with 0.5 pM to 100 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 1 pM to 50 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 2.5 pM to 25 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 3 pM to 15 pM ALS inhibitor.

[00095] In certain embodiments, the method comprises contacting the plant cell with 5 pM to 10 pM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 5 mM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 6 mM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 7 mM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 8 mM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 9 mM ALS inhibitor. In certain embodiments, the method comprises contacting the plant cell with 10 mM ALS inhibitor.

[00096] In certain embodiments, the ALS inhibitor is selected from the group consisting of Amidosulfuron, Azimsulfuron, Bensulfuron-Methyl, Bispyribac, Chlorimuron-Ethyl, Chlorsulfuron, Cinosulfuron, Cloransulam-Methyl, Cyclosulfamuron, Diclosulam, Ethametsulfuron-Methyl, Ethoxysulfuron, Flazasulfuron, Florasulam, Flucarbazone-Sodium, Flucetosulfuron, Flumetsulam, Flupyrsulfuron-Methyl, Foramsulfuron, Halosulfuron -Methyl, Imazamethabenz-Methyl, Imazamox, Imazapic, Imazapyr, Imazaquin, Imazethapyr, Imazosulfuron, Iodosulfuron, Mesosulfuron, Metazosulfuron, Metosulam, Metsulfuron- Methyl, NC-330, Nicosulfuron, Orthosulfamuron, Oxasulfuron, Penoxsulam, Primisulfuron- Methyl, Propoxycarbazone-Sodium, Prosulfuron, Pyrazosulfuron-Ethyl, Pyribenzoxim, Pyriftalid, Pyriminobac-Methyl, Pyrithiobac, Pyroxsulam, Rimsulfuron, Sulfometuron- Methyl, Sulfosulfuron, Thifensulfuron-Methyl, Triasulfuron, Tribenuron-Methyl, Trifloxysulfuron, Triflusulfuron-Methyl, and Tritosulfuron.

[00097] In certain embodiments, the ALS inhibitor is Amidosulfuron. In certain embodiments, the ALS inhibitor is Azimsulfuron. In certain embodiments, the ALS inhibitor is Bensulfuron-Methyl. In certain embodiments, the ALS inhibitor is Bispyribac. In certain embodiments, the ALS inhibitor is Chlorimuron-Ethyl. In certain embodiments, the ALS inhibitor is Chlorsulfuron. In certain embodiments, the ALS inhibitor is Cinosulfuron. In certain embodiments, the ALS inhibitor is Cloransulam-Methyl. In certain embodiments, the ALS inhibitor is Cyclosulfamuron. In certain embodiments, the ALS inhibitor is Diclosulam. In certain embodiments, the ALS inhibitor is Ethametsulfuron-Methyl. In certain embodiments, the ALS inhibitor is Ethoxysulfuron. In certain embodiments, the ALS inhibitor is Flazasulfuron. In certain embodiments, the ALS inhibitor is Florasulam. In certain embodiments, the ALS inhibitor is Flucarbazone-Sodium. In certain embodiments, the ALS inhibitor is Flucetosulfuron. In certain embodiments, the ALS inhibitor is Flumetsulam. In certain embodiments, the ALS inhibitor is Flupyrsulfuron-Methyl. In certain embodiments, the ALS inhibitor is Foramsulfuron. In certain embodiments, the ALS inhibitor is Halosulfuron- Methyl. In certain embodiments, the ALS inhibitor is Imazamethabenz-Methyl. In certain embodiments, the ALS inhibitor is Imazamox. In certain embodiments, the ALS inhibitor is Imazapic. In certain embodiments, the ALS inhibitor is Imazapyr. In certain embodiments, the ALS inhibitor is Imazaquin. In certain embodiments, the ALS inhibitor is Imazethapyr. In certain embodiments, the ALS inhibitor is Imazosulfuron. In certain embodiments, the ALS inhibitor is Iodosulfuron. In certain embodiments, the ALS inhibitor is Mesosulfuron. In certain embodiments, the ALS inhibitor is Metazosulfuron. In certain embodiments, the ALS inhibitor is Metosulam. In certain embodiments, the ALS inhibitor is Metsulfuron-Methyl. In certain embodiments, the ALS inhibitor is NC-330. In certain embodiments, the ALS inhibitor is Nicosulfuron. In certain embodiments, the ALS inhibitor is Orthosulfamuron. In certain embodiments, the ALS inhibitor is Oxasulfuron. In certain embodiments, the ALS inhibitor is Penoxsulam. In certain embodiments, the ALS inhibitor is Primisulfuron-Methyl. In certain embodiments, the ALS inhibitor is Propoxycarbazone-Sodium. In certain embodiments, the ALS inhibitor is Prosulfuron. In certain embodiments, the ALS inhibitor is Pyrazosulfuron- Ethyl. In certain embodiments, the ALS inhibitor is Pyribenzoxim. In certain embodiments, the ALS inhibitor is Pyriftalid. In certain embodiments, the ALS inhibitor is Pyriminobac- Methyl. In certain embodiments, the ALS inhibitor is Pyrithiobac. In certain embodiments, the ALS inhibitor is Pyroxsulam. In certain embodiments, the ALS inhibitor is Rimsulfuron. In certain embodiments, the ALS inhibitor is Sulfometuron-Methyl. In certain embodiments, the ALS inhibitor is Sulfosulfuron. In certain embodiments, the ALS inhibitor is Thifensulfuron- Methyl. In certain embodiments, the ALS inhibitor is Triasulfuron. In certain embodiments, the ALS inhibitor is Tribenuron-Methyl. In certain embodiments, the ALS inhibitor is Trifloxysulfuron. In certain embodiments, the ALS inhibitor is Triflusulfuron-Methyl. In certain embodiments, the ALS inhibitor is Tritosulfuron.

[00098] In certain embodiments, the ALS inhibitor is a sulfonylurea.

[00099] In certain embodiments, the ALS inhibitor is Tribenuron-Methyl.

[000100] In certain embodiments, the ALS enzyme is inhibited by contacting the plant cell with at least one agent which decreases the translation of the mRNA of the ALS enzyme.

[000101] In certain embodiments, the ALS enzyme is inhibited by contacting the plant cell with at least one agent which decreases the transcription of the ALS gene. [000102] In certain embodiments, the ALS inhibitor, ALS translation-inhibiting agent or ALS transcription-inhibiting agent is applied to the cell of the weed plant before growing a plant of interest in the vicinity of the weed plant.

[000103] In certain embodiments, the ALS inhibitor, ALS translation-inhibiting agent or ALS transcription-inhibiting agent is applied to the cell of the weed plant while growing a plant of interest in the vicinity of the weed plant.

[000104] The term “plant of interest” as used herein generally refers to any plant the growth of which is desired and/or beneficial, including domesticated plants, crop plants and/or ornamental plants.

[000105] EXAMPLES

[000106] Example 1.

[000107] Figure 1 is a hypothetical representation of possible mode of action of analogues in combination with AHAS inhibitors.

[000108] Example 2.

[000109] Efficiency of the different combinations was analyzed on Arabidopsis (Figure 2). Seeds were sown on MS plates supplemented with concentrations of 5 mM Tribenuron- methyl (CAS Number 101200-48-0), 50 pM Norvaline (CAS Number 6600-40-4), 50 pM Fluoro-valine (CAS Number 43163-94-6), 50 pM Thia-isoleucine (CAS Number 443-80-1), and their combinations. Plants were grown in a controlled growth room at 25°C in 16/8 h light/dark cycle. After 10 days of growth, the root length of seedlings was measured.

[000110] Example 3.

[000111] Efficiency of different combinations was analyzed on Teff ( Eragrostis abyssinicd) (Figure 3). Seeds were sown on MS plates supplemented with concentrations of 10 pM Tribenuron-methyl, 50 pM Norvaline, and their combinations. Plants were grown in a controlled growth room at 25°C in 16/8 h light/dark cycle. After 10 days of growth, the root length of seedlings was measured.

[000112] Example 4. Phytotoxic effect of branch chain NPAA analogues and AHAS inhibitor. [000113] Norvaline, Norleucine (CAS Number 327-57-1), Fluoro-valine, Thia-isoleucine and o-Methylthreonine (CAS Number 4144-02-9) are structural analogues of branch-chain amino acids. Though the effect of some of these branched-chain NPAA analogues was reported in E. coli , there were no clear studies on higher organisms, especially in plants. The phytotoxic effects of these analogues on Arabidopsis were determined by growing the seeds on medium supplementation with individual analogues at varying concentrations (Figure 4). Norvaline and Norleucine have been considered as structural isomers of leucine and potential substrates for LeuRS.

[000114] It has been observed that Norvaline at lower concentrations (100 mM) has no phytotoxic effect on Arabidopsis seedlings and minimal impact even at higher concentrations (200, 300, and 400 pM). It may be due to the post-transfer editing capacity of LeuRS. The CPI editing domain of E. Coli LeuRS has rapidly hydrolyzed the Nva-tRNA^Leu, thereby avoiding or minimizing Norvaline participation in protein synthesis at a very low rate of frequency (10-²).

[000115] Another analogue, Fluoro-valine (isomer of valine), has shown no phytotoxic effect either on root growth or biomass on all the tested concentrations, however it helps in inducing the root plant growth.

[000116] In contrast, isoleucine analogues Thia-isoleucine and o-methyl-Threonine were confirmed as having profound phytotoxic effects on Arabidopsis seedlings' growth in terms of shortened roots and reduced biomass. It might be due to the accumulation of these analogues in the newly synthesized proteome and eventually leading to the production of analogues damaged proteins.

[000117] The AHAS enzyme (EC 2.2.1.6) catalyzes the first common step in the biosynthesis of the BCAAs (valine, leucine, and isoleucine). The enzyme is relatively prevalent in archaea, bacteria, fungi, algae, and plants, but is absent from animals. The AHAS inhibitor used in this study (Tribenuron-methyl) demonstrates a clear dose-dependent phytotoxic effect on Arabidopsis seedlings. Further, IC50 (Inhibitory Concentration to achieve 50% growth reduction) values for all the branch chain NPAA analogues and AHAS inhibitor were determined based on the hill reaction model for further studies (Table 1).

[000118] Table 1.

[000119] As evident in Figure 4 and Table 1, when applied alone, Norvaline, Norleucine, and Fluoro-valine have little effect on plant growth. In contrast, the AHAS inhibitor, Thia- isoleucine, and o-methyl-Threonine have strong phytotoxic activity.

[000120] Example 5. Synergistic effect of branch chain NPAA analogues with AHAS inhibitor.

[000121] Applying a sub-lethal dose (IC50) of branched amino acid analogues in parallel with an AHAS inhibitor may have a profound phytotoxic effect on Arabidopsis root growth and biomass accumulation, compared to individual treatments (Figure 5 and Figure 6).

[000122] In the combination treatments of Norvaline, Norleucine, and Fluoro-valine with the AHAS inhibitor, severe phytotoxic effect represented as reduced root lengths and biomass was observed. Growth arrest of the seedlings was observed soon after germination in these treatments.

[000123] Seedlings grown with either Thia-isoleucine or o-methyl-Threonine in combination with the AHAS inhibitor also exhibit growth retardation, but not as severe as Norvaline or Norleucine treatments. In all the cases, profound growth retardation in the combined treatments rather than the individual treatments was noted.

[000124] When the Colby method was applied to calculate synergism, it was demonstrated that only Norvaline and Norleucine have synergistic effects together with the AHAS inhibitor. In contrast, Thia-isoleucine and o-methyl-threonine had an antagonistic effect on plant growth when applied together with the AHAS inhibitor. The activity of Fluoro-valine was very additive and did not drastically improve the performance of the AHAS inhibitor (Figure 7). [000125] Example 6. Synergism between Norvaline, Norleucine, and AHAS inhibitor, using two different concentration of AHAS inhibitor.

[000126] Efficiency of the different combinations was analyzed on Arabidopsis (Figure 8). Seeds were sown on MS plates supplemented with concentrations of 3 mM and 6 mM Tribenuron-methyl and its combinations with 100 pM Norvaline or Norleucine. Plants were grown in a controlled growth room at 25°C in 16/8 h light/dark cycle. After 10 days of growth, the biomass of seedlings was measured. The molecule herbicide mixture interactions were identified using Colby’s method on the percent control data and biomass data. For Colby’s method, an expected value (E) is calculated using the Equation E = (X+Y)-(XY)/100, where E is the expected level of control (or biomass reduction) of a given species when two moieties are applied in a mixture, and variables X and Y represent the level of control of a given weed species provided by each molecule applied individually. If E was significantly smaller than the observed value for a given mixture, it was deemed synergistic.

[000127] Example 7. Norvaline is incorporated into plant proteome.

[000128] Previous work already demonstrates that Norvaline probably formed during plant treatment with AHAS inhibitors. Other results in yeast and bacteria show that Norvaline is a substrate of LeuRS. Thus, the hypothesis that plants treated with AHAS inhibitors should incorporate Norvaline and Norleucine in their proteome was tested.

[000129] Plant proteins were isolated, hydrolyzed, and Norvaline and Norleucine incorporation into proteome was quantified using LC-MS analysis. Surprisingly, the results clearly show that plants treated with AHAS inhibitor accumulate Norvaline in the proteome.

[000130] It was also demonstrated that plants treated with Norvaline or Norleucine alone also replace native Leucine with analogues in their proteome. However, the most significant accumulation of Norvaline and Norleucine was observed in plants treated with AHAS inhibitor and Norvaline or Norleucine in parallel (Figure 9).

[000131] Example 8. LeuRS as a target of Norvaline and Norleucine.

[000132] Arabidopsis plants which overexpressed native LeuRS were created. The data presented in Figure 10 shows that plants which overexpress normal or mutated LeuRS forms demonstrate normal growth. However, the mutant plants are more susceptible to the effect of the combination of 2.5 mM AHAS inhibitor (Tribenuron-methyl) and 100 mM Norvaline or Norleucine in comparison to non-mutant plants.

[000133] The foregoing description of the specific embodiments will so fully reveal the general nature of the invention that others can, by applying current knowledge, readily modify and/or adapt for various applications such specific embodiments without undue experimentation and without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments. It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. The means, materials, and steps for carrying out various disclosed functions may take a variety of alternative forms without departing from the invention.

Claims

WE CLAIM

1. A method of preventing, slowing or disrupting protein synthesis in a cell of a weed plant, the method comprising the steps of:

(a) increasing the level of at least one Non-Protein Amino Acid (NPAA) in the cell; and

(b) inhibiting an acetolactate synthase (ALS) enzyme in the cell.

2. The method of claim 1, wherein the weed plant is an angiosperm.

3. The method of claim 1 or claim 2, wherein the weed plant is selected from the group consisting of Alopecurus aequalis (Shortawn Foxtail), Alopecurus japonicus (Japanese Foxtail), Alopecurus myosuroides (Blackgrass), Apera spica-venti (Silky Windgrass), Avena fatua (Wild Oat), Avena sterilis (Sterile Oat), Avena sterilis ssp. Ludoviciana (Sterile oat), Beckmannia syzigachne (American Sloughgrass), Brachypodium distachyon (Annual Falsebrome), Bromus diandrus (Ripgut Brome), Bromus diandrus ssp. rigidus (Rigid Brome), Bromus sterilis (Poverty Brome), Bromus tectorum (Downy Brome, Cheatgrass), Cynosurus echinatus (Hedgehog Dogtailgrass), Digitaria ciliaris (Southern Crabgrass), Digitaria insularis (Sourgrass), Digitaria ischaemum (Smooth Crabgrass), Digitaria sanguinalis (Large Crabgrass), Echinochloa colona (Junglerice), Echinochloa crus-galli var. crus-galli (Barnyardgrass), Echinochloa crus-galli var. formosensis (Taiwan barnyardgrass), Echinochloa oryzoides (Early Watergrass), Echinochloa phyllopogon (E. oryzicola, Late Watergrass), Ehrharta longiflora (Longflowered veldtgrass), Eleusine indica (Goosegrass), Eriochloa punctate (Pasto Amargo), Hordeum murinum ssp. Glaucum (Smooth Barley), Hordeum murinum ssp. Leporinum (Hare Barley), Ischaemum rugosum (Saramollagrass), Leptochloa chinensis (Chinese Sprangletop), Leptochloa panicoides (Amazon Sprangletop), Lolium perenne (Perennial Ryegrass), Lolium perenne ssp. Multiflorum (Italian Ryegrass), Lolium persicum (Persian Darnel), Lolium rigidum (Rigid Ryegrass), Phalaris brachystachys (Shortspike Canary grass), Phalaris minor (Little seed Canary grass), Phalaris paradoxa (Hood Canarygrass), Polypogon fugax (Asia Minor bluegrass), Rottboellia cochinchinensis (R. exaltata, Itchgrass), Sclerochloa kengiana (Keng Stiffgrass), Setaria faberi (Giant Foxtail), Setaria viridis (Green Foxtail), Setaria viridis var. major (var. robusta-alba, var. robustapurpurea, Giant Green Foxtail), Snowdenia polystachya (Ethiopian grass), Sorghum bicolor ssp. drummondii (Sorghum Sudanese, Sudangrass), Sorghum halepense (Johnsongrass), and Urochloa plantaginea (Brachiaria plantaginea, Alexandergrass).

BB

4. The method of any one of claims 1 to 3, wherein the weed plant prevents or decreases the viability of a domesticated plant.

5. The method of claim 4, wherein the domesticated plant is a crop plant.

6. The method of claim 4, wherein the domesticated plant is an ornamental plant.

7. The method of claim 4, wherein the domesticated plant is selected from the group consisting of Acacia spp., Acer spp., Actinidia spp., Aesculus spp., Agathis australis, Albizia amara, Alsophila tricolor, Andropogon spp., Arachis spp, Areca catechu, Astelia fragrans, Astragalus cicer, Baikiaea plurijuga, Betula spp., Brassica spp., Bruguiera gymnorrhiza, Burkea africana, Butea frondosa, Cadaba farinosa, Calliandra spp, Camellia sinensis, Canna indica, Capsicum spp., Cassia spp., Centroema pubescens, Chacoomeles spp., Cinnamomum cassia, Coffea arabica, Colophospermum mopane, Coronillia varia, Cotoneaster serotina, Crataegus spp., Cucumis spp., Cupressus spp., Cyathea dealbata, Cydonia oblonga, Cryptomeria japonica, Cymbopogon spp., Cynthea dealbata, Cydonia oblonga, Dalbergia monetaria, Davallia divaricata, Desmodium spp., Dicksonia squarosa, Dibeteropogon amplectens, Dioclea spp, Dolichos spp., Dorycnium rectum, Echinochloa pyramidalis, Ehraffia spp., Eleusine coracana, Eragrestis spp., Erythrina spp., Eucalypfus spp., Euclea schimperi, Eulalia villosa, Pagopyrum spp., Feijoa sellowlana, Fragaria spp., Flemingia spp, Freycinetia banksli, Geranium thunbergii, GinAgo biloba, Glycine javanica, Gliricidia spp, Gossypium hirsutum, Grevillea spp., Guibourtia coleosperma, Hedysarum spp., Hemaffliia altissima, Heteropogon contoffus, Hordeum vulgare, Hyparrhenia rufa, Hypericum erectum, Hypefflielia dissolute, Indigo incamata, Iris spp., Leptarrhena pyrolifolia, Lespediza spp., Lettuca spp., Leucaena leucocephala, Loudetia simplex, Lotonus bainesli, Lotus spp., Macrotyloma axillare, Malus spp., Manihot esculenta, Medicago saliva, Metasequoia glyptostroboides, Musa sapientum, Nicotianum spp., Onobrychis spp., Ornithopus spp., Oryza spp., Peltophorum africanum, Pennisetum spp., Per sea gratissima, Petunia spp., Phaseolus spp., Phoenix canariensis, Phormium cookianum, Photinia spp., Picea glauca, Pinus spp., Pisum sativam, Podocarpus totara, Pogonarthria fleckii, Pogonaffhria squarrosa, Populus spp., Prosopis cineraria, Pseudotsuga menziesii, Pterolobium stellatum, Pyrus communis, Quercus spp., Rhaphiolepsis umbellata, Rhopalostylis sapida, Rhus natalensis, Ribes grossularia, Ribes spp., Robinia pseudoacacia, Rosa spp., Rubus spp., Salix spp., Schyzachyrium sanguineum, Sciadopitys vefficillata, Sequoia sempervirens, Sequoiadendron giganteum, Sorghum bicolor, Spinacia spp., Sporobolus fimbriatus, Stiburus alopecuroides, Stylosanthos humilis, Tadehagi spp, Taxodium distichum, Themeda triandra, Trifolium spp., Triticum spp., Tsuga heterophylla, Vaccinium spp., Vicia spp., Vitis vinifera, Watsonia pyramidata, Zantedeschia aethiopica, Zea mays, amaranth, artichoke, asparagus, broccoli, Brussels sprouts, cabbage, canola, carrot, cauliflower, celery, collard greens, flax, kale, lentil, oilseed rape, okra, onion, potato, rice, soybean, straw, sugar beet, sugar cane, sunflower, tomato, squash tea, maize, wheat, barley, rye, oat, peanut, pea, lentil and alfalfa, cotton, rapeseed, canola, pepper, sunflower, tobacco, eggplant, eucalyptus, a perennial grass and a forage crop.

8. The method of any one of claims 1 to 7, wherein the method comprises decreasing the production of a branched amino acid in the cell.

9. The method of claim 8, wherein the branched amino acid is selected from the group consisting of valine, isoleucine, and leucine.

10. The method of any one of claims 1 to 9, wherein the method comprises contacting the plant cell with at least one NPAA.

11. The method of claim 10, comprising contacting the plant cell with 1 to 500 mM NPAA.

12. The method of claim 11, comprising contacting the plant cell with 50 pM to 100 pM NPAA.

13. The method of claim 12, comprising contacting the plant cell with 50 pM or 100 pM NPAA.

14. The method of any one of claims 1 to 13, wherein the NPAA is a branched amino acid analogue.

15. The method of any one of claims 1 to 14, wherein the NPAA is selected from the group consisting of Norvaline, Norleucine, Fluoro- Valine, Thia-isoleucine, o-methyl-Threonine, 3- Methyl -valine, L-allo-Threonine, L-cyclohexyl-Glycine, 3-Fluoro-Leucine, and 3- Cyclopentyl-DL-alanine.

16. The method of claim 15, wherein the NPAA is Norvaline, Norleucine, Fluoro-valine, Thia-isoleucine, or o-methyl-Threonine.

17. The method of any one of claims 1 to 16, wherein the method comprises contacting the plant cell with at least one ALS enzyme inhibitor.

18. The method of claim 17, comprising contacting the plant cell with 1 to 100 mM ALS enzyme inhibitor.

19. The method of claim 18, comprising contacting the plant cell with 0.5 pM to 10 pM ALS enzyme inhibitor.

20. The method of claim 19, comprising contacting the plant cell with 2.5 pM to 10 pM ALS enzyme inhibitor.

21. The method of any one of claims 17 to 20, wherein the ALS enzyme inhibitor is a sulfonylurea.

22. The method of any one of claims 17 to 21, wherein the ALS enzyme inhibitor is selected from the group consisting of Amidosulfuron, Azimsulfuron, Bensulfuron-Methyl, Bispyribac, Chlorimuron-Ethyl, Chlorsulfuron, Cinosulfuron, Cloransulam-Methyl, Cyclosulfamuron, Diclosulam, Ethametsulfuron-Methyl, Ethoxysulfuron, Flazasulfuron, Florasulam, Flucarbazone-Sodium, Flucetosulfuron, Flumetsulam, Flupyrsulfuron-Methyl, Foramsulfuron, Halosulfuron-Methyl, Imazamethabenz-Methyl, Imazamox, Imazapic, Imazapyr, Imazaquin, Imazethapyr, Imazosulfuron, Iodosulfuron, Mesosulfuron, Metazosulfuron, Metosulam, Metsulfuron-Methyl, NC-330, Nicosulfuron, Orthosulfamuron, Oxasulfuron, Penoxsulam, Primisulfuron-Methyl, Propoxycarbazone-Sodium, Prosulfuron, Pyrazosulfuron-Ethyl, Pyribenzoxim, Pyriftalid, Pyriminobac-Methyl, Pyrithiobac, Pyroxsulam, Rimsulfuron, Sulfometuron-Methyl, Sulfosulfuron, Thifensulfuron-Methyl, Triasulfuron, Tribenuron- Methyl, Trifloxysulfuron, Triflusulfuron-Methyl, and Tritosulfuron.

23. The method of any one of claims 17 to 22, wherein the ALS enzyme inhibitor is Trib enur on-Methyl .

24. The method of any one of claims 1 to 16, wherein the method comprises contacting the plant cell with at least one agent which decreases the transcription of an ALS gene.

25. The method of any one of claims 1 to 16, wherein the method comprises contacting the plant cell with at least one agent which decreases the translation of an ALS mRNA.

26. The method of any one of claims 17 to 25, wherein the ALS enzyme inhibitor, ALS mRNA translation-inhibiting agent, or ALS gene transcription-inhibiting agent, is applied to the cell of the weed plant before growing a plant of interest in the vicinity of the weed plant.

27. The method of any one of claims 17 to 26, wherein the ALS enzyme inhibitor, ALS mRNA translation-inhibiting agent, or ALS gene transcription-inhibiting agent, is applied to the cell of the weed plant while growing a plant of interest in the vicinity of the weed plant.

28. A composition, comprising:

(i) at least one NPAA, and

(ii) at least one ALS enzyme inhibitor, at least one ALS mRNA translation- inhibiting agent, or at least one ALS gene transcription-inhibiting agent.

29. The composition of claim 28, wherein the NPAA is a branched amino acid analogue.

30. The composition of claim 28 or claim 29, wherein the NPAA is selected from the group consisting of Norvaline, Norleucine, Fluoro- Valine, Thia-isoleucine, o-methyl-Threonine, 3- Methyl-valine, L-allo-Threonine, L-cyclohexyl-Glycine, 3-Fluoro-Leucine, and 3- Cyclopentyl-DL-alanine.

31. The composition of claim 30, wherein the NPAA is Norvaline, Norleucine, Fluoro- valine, Thia-isoleucine, or o-methyl-Threonine.

32. The composition of any one of claims 28 to 31, comprising 1 to 500 mM NPAA.

33. The composition of claim 32, comprising 50 pM to 100 pM NPAA.

34. The composition of claim 33, comprising 50 pM or 100 pM NPAA.

35. The composition of any one of claims 28 to 34, wherein the ALS enzyme inhibitor is a sulfonylurea.

36. The composition of any one of claims 28 to 35, wherein the ALS enzyme inhibitor is selected from the group consisting of Amidosulfuron, Azimsulfuron, Bensulfuron-Methyl, Bispyribac, Chlorimuron-Ethyl, Chlorsulfuron, Cinosulfuron, Cloransulam-Methyl, Cyclosulfamuron, Diclosulam, Ethametsulfuron-Methyl, Ethoxysulfuron, Flazasulfuron, Florasulam, Flucarbazone-Sodium, Flucetosulfuron, Flumetsulam, Flupyrsulfuron-Methyl, Foramsulfuron, Halosulfuron -Methyl, Imazamethabenz-Methyl, Imazamox, Imazapic, Imazapyr, Imazaquin, Imazethapyr, Imazosulfuron, Iodosulfuron, Mesosulfuron, Metazosulfuron, Metosulam, Metsulfuron -Methyl, NC-330, Nicosulfuron, Orthosulfamuron, Oxasulfuron, Penoxsulam, Primisulfuron-Methyl, Propoxycarbazone-Sodium, Prosulfuron, Pyrazosulfuron-Ethyl, Pyribenzoxim, Pyriftalid, Pyriminobac-Methyl, Pyrithiobac, Pyroxsulam, Rimsulfuron, Sulfometuron-Methyl, Sulfosulfuron, Thifensulfuron-Methyl, Triasulfuron, Tribenur on-Methyl, Trifloxysulfuron, Triflusulfuron-Methyl, and Tritosulfuron.

37. The composition of any one of claims 28 to 36, wherein the ALS enzyme inhibitor is Trib enur on-Methyl .

38. The composition of any one of claims 28 to 37, comprising 1 to 100 mM ALS enzyme inhibitor.

39. The composition of claim 38, comprising 0.5 pM to 10 pM ALS enzyme inhibitor.

40. The composition of claim 39, comprising 2.5 pM to 10 pM ALS enzyme inhibitor.

41. A kit, comprising:

(i) a composition comprising at least one NPAA, and

(ii) a composition comprising at least one ALS enzyme inhibitor, at least one ALS mRNA translation-inhibiting agent, or at least one ALS gene transcription- inhibiting agent.

42. The kit of claim 41, wherein the NPAA is a branched amino acid analogue.

43. The kit of claim 41 or claim 42, wherein the NPAA is selected from the group consisting of Norvaline, Norleucine, Fluoro- Valine, Thia-isoleucine, o-methyl-Threonine, 3 -Methyl- valine, L-allo-Threonine, L-cyclohexyl-Glycine, 3-Fluoro-Leucine, and 3-Cyclopentyl-DL- alanine.

44. The kit of claim 43, wherein the NPAA is Norvaline, Norleucine, Fluoro-valine, Thia- isoleucine, or o-methyl-Threonine.

45. The kit of any one of claims 41 to 44, comprising 1 to 500 pM NPAA.

46. The kit of claim 45, comprising 50 pM to 100 pM NPAA.

47. The kit of claim 46, comprising 50 pM or 100 pM NPAA.

48. The kit of any one of claims 41 to 47, wherein the ALS enzyme inhibitor is a sulfonylurea.

49. The kit of any one of claims 41 to 48, wherein the ALS enzyme inhibitor is selected from the group consisting of Amidosulfuron, Azimsulfuron, Bensulfuron-Methyl, Bispyribac, Chlorimuron-Ethyl, Chlorsulfuron, Cinosulfuron, Cloransulam-Methyl, Cyclosulfamuron, Diclosulam, Ethametsulfuron-Methyl, Ethoxysulfuron, Flazasulfuron, Florasulam, Flucarbazone-Sodium, Flucetosulfuron, Flumetsulam, Flupyrsulfuron-Methyl, Foramsulfuron, Halosulfuron-Methyl, Imazamethabenz-Methyl, Imazamox, Imazapic, Imazapyr, Imazaquin, Imazethapyr, Imazosulfuron, Iodosulfuron, Mesosulfuron, Metazosulfuron, Metosulam, Metsulfuron-Methyl, NC-330, Nicosulfuron, Orthosulfamuron, Oxasulfuron, Penoxsulam, Primisulfuron-Methyl, Propoxycarbazone-Sodium, Prosulfuron, Pyrazosulfuron-Ethyl, Pyribenzoxim, Pyriftalid, Pyriminobac-Methyl, Pyrithiobac, Pyroxsulam, Rimsulfuron, Sulfometuron-Methyl, Sulfosulfuron, Thifensulfuron-Methyl, Triasulfuron, Tribenuron- Methyl, Trifloxysulfuron, Triflusulfuron-Methyl, and Tritosulfuron.

50. The kit of any one of claims 41 to 49, wherein the ALS enzyme inhibitor is Tribenuron- Methyl.

51. The kit of any one of claims 41 to 50, comprising 1 to 100 mM ALS enzyme inhibitor.

52. The kit of claim 51, comprising 0.5 pM to 10 pM ALS enzyme inhibitor.

53. The kit of claim 52, comprising 2.5 pM to 10 pM ALS enzyme inhibitor.