WO2018094008A1

WO2018094008A1 - Cellulose synthase inhibitors effective as herbicides

Info

Publication number: WO2018094008A1
Application number: PCT/US2017/061959
Authority: WO
Inventors: Steven Gutteridge; Il-Ho Kang
Original assignee: Fmc Corporation
Priority date: 2016-11-21
Filing date: 2017-11-16
Publication date: 2018-05-24
Also published as: EP3541188A1

Abstract

This invention relates to identification and production of herbicidal compounds and compositions which inhibit cellulose synthase.

Description

TITLE

CELLULOSE SYNTHASE INHIBITORS EFFECTIVE AS HERBICIDES

FIELD OF THE INVENTION

BACKGROUND OF THE INVENTION

Cellulose is the primary structural component and major load bearing polymer of plant cell walls. In growing cells cellulose microfibrils are laid down transversely reinforcing the cell laterally allowing longitudinal expansion. In plants, cellulose synthesis requires a coordinated transport of substrates across membranes and orientation of the membrane- associated synthase complexes. Cellulose is synthesized by a rosette protein complex composed of cellulose synthases (CESAs). It is thought that the rosette complexes in the primary and secondary cell walls each contain at least three different non-redundant cellulose synthases. In Arabidopsis the three cellulose synthases that form a rosette complex and responsible for primary cell wall deposition are coded for by CESAl, CESA3, and CESA6 genes. Although inhibitors of cellulose synthase at CESA3 and CESA6 are known, an inhibitor of plant cellulose synthase specifically at CESAl is a previously undescribed site of action for herbicides of practical application.

SUMMARY OF THE INVENTION

One aspect of the present invention is a method for producing a herbicidal composition comprising the following steps:

(a) screening a candidate compound in a cellulose synthase inhibition assay; and

(b) if the candidate compound is active in the cellulose synthase inhibition assay, testing the compound for activity against a plant; and

(c) preparing a herbicidal composition comprising the compound identified in step (a) and verified in step (b).

Another aspect of the invention is a method for producing a herbicidal compound comprising testing a candidate compound in a cellulose synthase activity inhibition assay wherein the assay utilizes cellulose synthase from a weed to be controlled.

Another aspect of the invention is a method of controlling weeds comprising applying a herbicidally effective amount of a cellulose synthase inhibitor produced by the method described herein to a locus in need of such treatment.

A further aspect of the invention is a herbicidal composition comprising a cellulose synthase inhibitor produced by the method described herein as the active ingredient in combination with a carrier. Yet another aspect of the invention is a herbicidal composition comprising a cellulose synthase inhibitor produced by the method described herein in combination with another herbicide.

DETAILS OF THE INVENTION

As used herein, the terms "comprises," "comprising," "includes," "including," "has,"

"having," "contains", "containing," "characterized by" or any other variation thereof, are intended to cover a non-exclusive inclusion, subject to any limitation explicitly indicated. For example, a composition, mixture, process or method that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, mixture, process, or method.

The transitional phrase "consisting of excludes any element, step, or ingredient not specified. If in the claim, such would close the claim to the inclusion of materials other than those recited except for impurities ordinarily associated therewith. When the phrase "consisting of appears in a clause of the body of a claim, rather than immediately following the preamble, it limits only the element set forth in that clause; other elements are not excluded from the claim as a whole.

The transitional phrase "consisting essentially of is used to define a composition, method that includes materials, steps, features, components, or elements, in addition to those literally disclosed, provided that these additional materials, steps, features, components, or elements do not materially affect the basic and novel characteristic(s) of the claimed invention. The term "consisting essentially of occupies a middle ground between "comprising" and "consisting of.

Where applicants have defined an invention or a portion thereof with an open-ended term such as "comprising," it should be readily understood that (unless otherwise stated) the description should be interpreted to also describe such an invention using the terms "consisting essentially of or "consisting of."

Further, unless expressly stated to the contrary, "or" refers to an inclusive or and not to an exclusive or. For example, a condition A or B is satisfied by any one of the following: A is true (or present) and B is false (or not present), A is false (or not present) and B is true (or present), and both A and B are true (or present).

Also, the indefinite articles "a" and "an" preceding an element or component of the invention are intended to be nonrestrictive regarding the number of instances (i.e. occurrences) of the element or component. Therefore "a" or "an" should be read to include one or at least one, and the singular word form of the element or component also includes the plural unless the number is obviously meant to be singular. As referred to herein a cellulose synthase inhibition assay refers to assays which measure inhibition of enzymatic activity as well as binding assays and in-silico methods of computer aided molecular design.

"Indirect inhibitors" are relatively inactive compounds that inhibit cellulose synthase following metabolism to an active inhibitor in plants.

As used herein, the term a "herbicidally effective amount of cellulose synthase inhibitor" refers to an amount of cellulose synthase inhibitor sufficient to kill or inhibit the growth of the weed it is desired to control.

The term "weed(s)" relates to any unwanted vegetation and includes, for example, undesired carry-over or "rogue" or "volunteer" crop plants in a field of desired crop plants.

Isoxaben (N-[3-(l-ethyl-l-methylpropyl)-5-isoxazolyl]-2,6,dimethoxybenzamide CAS Registry No. 82558-50-7) is a preemergence, broad leaf herbicide used primarily on small grain, turf and ornamental cro s.

isoxaben

This compound is extremely active with an IC50 for oilseed rape {Bras ska napus) of

20 nM (Lefebvre et al., Weed Res. 1987, 27, 125-134). Isoxaben inhibits the incorporation of glucose into the cellulose-rich, acid-insoluble fraction of isolated walls and is an extremely powerful and specific inhibitor of cell wall biosynthesis. Cell wall-fractionation studies have revealed that the herbicidal action of isoxaben can be explained entirely by its effect on cellulose biosynthesis (Corio-Costet et al., Pest. Biochem. Phys. 1991, 40, 255-265). The probable mode of action is to directly inhibit cellulose synthesis, because resistant cell lines show an unaltered uptake or detoxification of the herbicide (Heim et al., Pest. Biochem. Phys. 1991, 39, 93-99) and only two genetic loci in Arabidopsis thaliana, termed ixrA (i.e. ixrl) and ixrB (i.e. ixr2), have been shown to confer resistance. Exhaustive studies have revealed that other cellular processes are unaffected by isoxaben (e.g. seed germination, mitosis, respiration, photosynthesis, and lipid and RNA synthesis. Treated cells fail to elongate with high fidelity and consequently grow isodiametrically (Lefebvre et al., Weed Res. 1987, 27, 125-134). Hence, this is the only commercial herbicidal compound which is an unequivocal inhibitor of plant cellulose synthase, specifically inhibiting isozymes CESA3 and CESA6. Although cellulose synthesis is an attractive commercial mode of herbicidal action given its specificity for plants, unfortunately no major commercial herbicide has been identified that combines the advantages of weed control and safety to crops that a successful and selective compound should command.

It is now discovered that Compound 1 (i.e. [3-(3,5-dichlorophenyl)-lH-pyrazol-4- yl]phenyl-methanone described as prepared in U.S. Pat. No. 5,939,559, CAS Registry No. 182141-92-0) is a potent inhibitor of plant cellulose synthase specifically at CESA1, a previously undescribed site of action for herbicides of practical application.

1 IC₅₀ (based on root length) = 0.01 ~ 0.1 μΜ

Validation of Target Site

Experimental Determination of CESA1 as the Target for Compound 1

Arabidopsis Landsberg erecta (Ler-0) seeds were sterilized in chlorine gas and plated on 0.5x Murashige and Skoog salts (MS) plant media (containing 0.05% 2-(N-Morpholino)- ethanesulfonic acid (MES), 0.5% sucrose, and 0.8% Phytagar in a 24-well plate. Each well contained 0.01, 0.1, 1, 5, 10 and 100 μΜ of Compound 1, respectively and the IC50 was determined by measuring fresh weights as well as root lengths compared to untreated control seedlings. The concentration of Compound 1 at which 50% of the fresh weights of untreated seedlings was determined to be the IC₅₀ concentration. For Compound 1 the IC₅₀ was determined to be 0.05 μΜ and each experiment was performed in triplicate.

Ethyl methanesulfonate (EMS)-mutagenized M₂ seeds of Arabidopsis Ler-0 (M2E- 04-07, Lehle Seeds. Round Rock, TX) were screened, in order to screen resistant mutant Arabidopsis plants to Compound 1, about 12,000 sterilized M₂ seeds were plated on MS plates containing 10 times of the IC₅₀ concentration of Compound 1 (0.5 μΜ). Resistant seedlings were selected after seven to 10 d on selective medium and transferred to soil. Soil-transferred plants were grown in growth chambers in 4-inch (10 cm) pots using Metromix 360 potting soil under cool-white fluorescent light using a 16 h day/8 h night photoperiod at 22 °C. Compound 1 resistance was confirmed in the next generation with the same concentration of herbicide used in the selection. In order to identify the mutated gene linked to the resistant trait to Compound 1, PCR-based mapping as well as SNP (Single Nucleotide Polymorphism) analysis was conducted.

Crosses

Crosses of the mutant to a diverged genome are required in order to reduce unlinked genetic background within selected resistant mutants. In order to create back-cross 1 (BC1) Fl seeds and out-cross 1 (OC1) Fl seeds, the selected Compound 1 resistant mutant males were crossed with wild-type Ler-0 females and wild-type Columbia (Col-0) females, respectively. In the same manner, BC2 and OC2 were created from BC1 with wild-type Ler-0 and OC1 with wild-type Col-0 plants, respectively. In every cross case, Compound 1-resistant Fl seeds were generated by pollinating emasculated flowers of wild-type plants with pollen from Compound 1-resistant mutant plants. The generated Fl (BC1 and OC1) and F2 (BC2 and OC2) seeds were plated on MS media containing same concentration of Compound 1 as was used in the original screen.

Seedlings showing significantly reduced growth in these concentrations of Compound 1 were used to provide samples for PCR mapping of the F2 genome. Using a combination of chromosomal markers the rough position of the allele responsible for resistance to the compound was mapped to a partial segment of chromosome 4 of the Arabidopsis genome. Cellulose synthase 1 (ATCESA1, AT4G324210) was one of 8 genes that harbored single nucleotide polymorphisms (SNPs) in the relevant segment of chromosome 4. The gene coding for Arabidopsis CESA1 is shown as SEQ ID NO: 1.

Cellulose Synthase Inhibition Assays

The method for production of potential herbicides does not require use of any particular cellulose synthase inhibition assay. Suitable assays are described hereinafter, but those skilled in the art can readily substitute functionally equivalent test methods. For example, although the in-vitro screening assay described hereinafter uses A. thaliana seedlings, seedlings of other plants may be substituted. For example a commercially significant weed may be used. The assays to be employed include but are not limited to the group selected from one or more of the following assays: in-vitro activity assays or binding assays.

In- Vitro Inhibition Assays

The herbicidal site of action at CESA1 and its discovery opens up the opportunity of identifying novel compounds that inhibit the same enzyme target either directly or following metabolism to an active inhibitor in plants ("indirect inhibitors").

Compounds that are active in the cellulose synthase inhibition assay are then tested using any desired herbicide whole plant activity test. In this context, "active in the cellulose synthase inhibition assay" means that a reduction in cellulose synthase activity is observed.

Our herbicidal method and herbicide composition require use of a "cellulose synthase inhibitor". As used in describing and claiming the herbicidal method and herbicide composition, the term "cellulose synthase inhibitor" encompasses any compound that: (a) produces measurable inhibition in a cellulose synthase inhibition assay using cellulose synthase from a plant; (b) is not a general enzyme inhibitor, and (c) a compound that was known to have herbicidal activity prior to the filing date of this application. It should be understood that no herbicide was known to have cellulose synthase inhibition at CESA1 as its site of action at the time the present invention was made. One embodiment of the invention is wherein step (b) above is "not an inhibitor of CESA3 or CESA6".

Preferred cellulose synthase inhibitors are those which produce at least a measurable reduction in cellulose synthase activity when tested at 10 μg/mL in the A. thaliana seedling assay described hereinafter.

Example 1

Cellulose Synthase Assay

Inhibition of cellulose synthase activity and thus cellulose deposition in plants often induces ectopic lignin formation. A positive stain for lignin, using phloroglucinol-HCl solution as described previously with slight modification (Cano-Delgado et ai, Development 2000, 727, 3395-3405) was thus indicative for disruption of cellulose deposition. Preparation of phloroglucinol-HCl staining solution (phloroglucinol in 20% ethanol and concentrated HC1 (12 N)) was carried out in the fume hood. Compound 1-treated Arabidopsis seedlings as well as untreated seedlings were incubated in phloroglucinol-HCl staining solution for one day and de-staining step was followed for 1 h with 70% ethanol. Images were taken from stained Arabidopsis roots on a bright-field stereomicroscope (Leica).

Example 2

High Throughput Screening

The assay method described in Example 1 was modified to accommodate multiple compounds in microtiter plates by methods well known in the art. The assay was conducted using Falcon® 96- well, flat bottom polystyrene plates having wells arrayed in 12 columns and 8 rows.

The inhibition of cellulose synthase by isoxaben was used to standardize the effects of other test compounds on cellulose synthase activity. Certain well(s) were used for determining background and total activity.

Preparation of the plates was automated using robotic workstations to dilute the stock compounds and add appropriate volumes of reaction solutions and compounds to the individual wells of the 96-well microtiter plate. Arabidopsis seedings (7-day) were dispensed into wells containing test compounds in MS media and incubated under a 16 h day/8 h night photoperiod at 22 °C. An aqueous solution of phloroglucinol was then added to each well for for 24 h. The stain was removed by washing the wells with 70% ethanol for 1 h and the seedlings inspected under a microscope for staining in the vasculature. Those compounds identified by this approach were then assessed for their relative efficacy by determination of their IC50 values in an Arabidopsis growth study also in plates with particular focus on root development. Those compounds rating above a certain level were further assessed for their efficacy on resistant Arabidopsis seed lines (Table 1). The ratio of the IC₅₀ on resistant lines and wild-type plants was used as a measure of the resistance factor. The table indicates that compounds structurally similar to Compound 1 were incapable of controlling Arabidopsis lines carrying a CESA1 mutation that results in a change from alanine (A) (see SEQ ID NO: 2) to valine (V) (see SEQ ID NO: 4) specifically at position 903.

Table 1: Cross-Resistance Study Using Arabidopsis Lines Carrying Mutations in Individual

Cellulose Synthase Genes.

ixr = isoxaben resistance; txr = thaxtomin resistance; irx = irregular xylem; rsw = radially swollen (effect on root growth); 46R4 = CESA1 resistance (line 4).

There was a strong correlation between activity of a compound in the cellulose synthase assay and herbicidal activity.

The present invention is directed to herbicidal use of compounds that inhibit cellulose synthase (i.e. at CESA1), as opposed to herbicidal use of compounds that inhibit enzymes generally. An example of a compound that inhibits enzymes generally is diethyl pyrocarbonate, which reacts preferentially with protein thiol and amino groups. To eliminate the possibility that a compound active in the CESA1 assay is a general enzyme inhibitor, the active compound may be tested in a second enzyme assay. A suitable assay for this purpose is, for example, the E. coli alkaline phosphatase assay described by Garen and Levinthal, Biochim. Biophys. ACTA 1960, 38, 470. If the compound is not inhibitory in the second enzyme assay, it may generally be safely concluded that the compound does not inhibit enzymes generally.

Biological efficacy of a herbicidal compound in whole organisms is influenced by many factors, including not only intrinsic activity of the compound, i.e. efficiency of its interaction with the target molecule, but also stability of the compound and ability of the compound to be translocated to the target site. The cellulose synthase inhibition assay using seedlings provides a measure of the ability of the compound to translocate and interfere with cellulose synthesis. It will be appreciated by those skilled in the art that once a potential herbicide is detected using the cellulose synthase assay, conventional techniques must be used to determine the usefulness of the compound in various environments.

A herbicidally effective amount of the compounds of this invention is determined by a number of factors. The exact concentration of compound required varies with the weed to be controlled, the type formulation employed, the method of application, formulation selected, the particular plant species, climate conditions and the like. Generally, a herbicidally effective amount of compounds of this invention is about 0.005 to 20 kg ha with a preferred range of about 0.01 to 1 kg/ha. One skilled in the art can easily determine the herbicidally effective amount necessary for the desired level of weed control. Weeds appropriate for the use of compositions that inhibit cellulose synthase (e.g., CESAl) include blackgrass (Alopecurus myosuroides), downy bromegrass (Bromus tectorum), green foxtail (Setaria viridis), Italian ryegrass (Lolium multiflorum), wild oat (Avenafatua), catchweed bedstraw (Galium aparine), bermudagrass (Cynodon dactylon), Surinam grass (Brachiaria decumbens), common cocklebur (Xanthium strumarium), large crabgrass (Digitaria sanguinalis), woolly cupgrass (Eriochloa villosa), giant foxtail (Setaria faberii), goosegrass (Eleusine indica), johnsongrass (Sorghum halepense), kochia (Kochia scoparia), lambsquarters (Chenopodium album), morningglory (Ipomoea coccinea), eastern black nightshade (Solarium ptycanthum), yellow nutsedge (Cyperus esculentus), pigweed (Amaranthus retroflexus), common ragweed (Ambrosia elatior), Russian thistle (Salsola kali), velvetleaf (Abutilon theophrasti), small- flower umbrella sedge (Cyperus difformis), ducksalad (Heteranthera limosa), barnyardgrass (Echinochloa crus-galli), wild poinsettia (Euphorbia heterophylla), palmer pigweed (Amaranthus palmeri), common waterhemp (Amaranthus rudis) (including ALS/Triazine -resistant and ALS/HPPD-resistant waterhemp), ladysthumb smartweed (Polygonum persicaria), Brazilian crabgrass (Digitaria horizontalis), fall panicum (Panicum dichotomiflorum), sandbur (southern sandbur, Cenchrus echinatus), arrowleaf sida (Sida rhombifolia), field bindweed (Convolvulus arvensis), hairy beggarticks (Bidens pilosa), annual bluegrass (Poa annua), canarygrass (Phalaris minor), common chickweed (Stellaria media), field poppy (Papaver rhoeas), field violet (Viola arvensis), henbit deadnettle (Lamium amplexicaule), scentless chamomile (Matricaria inodora), bird's-eye speedwell (Veronica persica), wild buckwheat (Polygonum convolvulus), wild mustard (Sinapis arvensis), wild oat (Avena fatua), wild radish (Raphanus raphanistrum), windgrass (Apera spica-venti) and Virginia dayflower (Commelina virginica).

As those skilled in the art will recognize the present invention contemplates the use of other plant cellulose synthase enzymes than that found in A. thaliana. Indeed the skilled artisan is capable of using known plant cellulose synthase sequences to create a consensus sequence useful in the plant cellulose synthase assay employed.

SEQUENCE LISTING TABLE

Those skilled in the art will recognize that the proteins and encoding DNA sequences depicted above are all capable of being used in the present invention. However the present invention is not limited to use of sequences such as those exemplified above but rather any plant cellulose synthase protein or encoding nucleic acids are intended. The invention includes the use of any cellulose synthase having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% homology; or having 100% homology to the amino acid identitified as SEQ ID NO: 2 (without taking into account mismatches at the N-terminus). Every integer value in between is intended.

Calculations of "homology" or "sequence identity" between two sequences (the terms are used interchangeably herein) are performed as follows. The sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and nonhomologous sequences can be disregarded for comparison purposes). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein amino acid or nucleic acid "identity" is equivalent to amino acid or nucleic acid "homology"). The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.

The comparison of sequences and determination of percent sequence identity between two sequences may be accomplished using a mathematical algorithm. In a preferred embodiment, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch (/. Mol. Biol. 1970, 48, 444-53) algorithm, which has been incorporated into the GAP program in the GCG software package (available at www.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. In yet another preferred embodiment, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (available at www.gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. A particularly preferred set of parameters (and the one that should be used if the practitioner is uncertain about what parameters should be applied to determine if a molecule is within a sequence identity or homology limitation of the invention) is a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5. The percent identity between two amino acid or nucleotide sequences can also be determined using the algorithm of Meyers and Miller (CABIOS 1989, 4, 11-17), which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.

The above sequences can be conventionally synthesized by the modified phosphotriester method using fully protected deoxyribonucleotide building blocks. Such synthetic methods are well known in the art and can be carried out in substantial accordance with the procedure of Itakura et al., Science 1977, 198, 1056 and Crea et al, Proc. Nat. Acad. Sci. U.S.A. 1978, 75, 5765. In addition, an especially preferred method is disclosed in Hsiung et al., Nucleic Acid Research 1983, 11, 3227 and Narang et al., Methods in Enzymology 1980, 68, 90. In addition to the manual procedures referenced above, the DNA sequence can be synthesized using automated DNA synthesizers, such as the ABS (Applied Biosystems, 850 Lincoln Centre Drive, Foster City, Calif. 94404) 380B DNA Synthesizer.

It is more convenient however to prepare a desired DNA sequence by the polymerase chain reaction. See, U.S. Pat. Nos. 4,800,159 and 4,683,202 and European Patent Publication No. 0258017 from any individual plant species of interest by procedures well known in the art. The amino acid sequences depicted above can be encoded by a multitude of different DNA sequences because most of the amino acid residues are encoded by more than one DNA triplet. Because these alternate DNA sequences would encode the same amino acid residue sequences of the present invention, the present invention further comprises these alternate sequences.

The following Example is provided to further illustrate and exemplify, but not limit the scope of the present invention.

Example 3

Cellulose synthase isozymic identity

There are known mutations in the genes coding for cellulose synthases. Isoxaben resistant Arabidopsis lines have been described before and harbor mutations in CESA3 and CESA6. The CESAl resistant line that emerged after selection with Compound 1 along with the CESA3 and CESA6 lines thus provide tools through cross-resistance studies to determine which isozyme is the target of any active compounds identified in the seedling assay.

Preparation of the Compositions

A compound of this invention will generally be used as a herbicidal active ingredient in a composition, i.e. formulation, with at least one additional component selected from the group consisting of surfactants, solid diluents and liquid diluents, which serves as a carrier. The formulation or composition ingredients are selected to be consistent with the physical properties of the active ingredient, mode of application and environmental factors such as soil type, moisture and temperature.

Useful formulations include both liquid and solid compositions. Liquid compositions include solutions (including emulsifiable concentrates), suspensions, emulsions (including microemulsions, oil-in-water emulsions, flowable concentrates and/or suspoemulsions) and the like, which optionally can be thickened into gels. The general types of aqueous liquid compositions are soluble concentrate, suspension concentrate, capsule suspension, concentrated emulsion, microemulsion, oil-in-water emulsion, flowable concentrate and suspo-emulsion. The general types of nonaqueous liquid compositions are emulsifiable concentrate, microemulsifiable concentrate, dispersible concentrate and oil dispersion.

The general types of solid compositions are dusts, powders, granules, pellets, prills, pastilles, tablets, filled films (including seed coatings) and the like, which can be water-dispersible ("wettable") or water-soluble. Films and coatings formed from film- forming solutions or flowable suspensions are particularly useful for seed treatment. Active ingredient can be (micro)encapsulated and further formed into a suspension or solid formulation; alternatively the entire formulation of active ingredient can be encapsulated (or "overcoated"). Encapsulation can control or delay release of the active ingredient. An emulsifiable granule combines the advantages of both an emulsifiable concentrate formulation and a dry granular formulation. High-strength compositions are primarily used as intermediates for further formulation.

Sprayable formulations are typically extended in a suitable medium before spraying. Such liquid and solid formulations are formulated to be readily diluted in the spray medium, usually water, but occasionally another suitable medium like an aromatic or paraffinic hydrocarbon or vegetable oil. Spray volumes can range from about from about one to several thousand liters per hectare, but more typically are in the range from about ten to several hundred liters per hectare. Sprayable formulations can be tank mixed with water or another suitable medium for foliar treatment by aerial or ground application, or for application to the growing medium of the plant. Liquid and dry formulations can be metered directly into drip irrigation systems or metered into the furrow during planting. The formulations will typically contain effective amounts of active ingredient, diluent and surfactant within the following approximate ranges which add up to 100 percent by weight.

Weight Percent

Active

Ingredient Diluent Surfactant

Water-Dispersible and Water- 0.001-90 0-99.999 0-15

soluble Granules, Tablets and

Powders

Oil Dispersions, Suspensions, 1-50 40-99 0-50

Emulsions, Solutions (including

Emulsifiable Concentrates)

Dusts 1-25 70-99 0-5

Granules and Pellets 0.001-99 5-99.999 0-15

High Strength Compositions 90-99 0-10 0-2

Solid diluents include, for example, clays such as bentonite, montmorillonite, attapulgite and kaolin, gypsum, cellulose, titanium dioxide, zinc oxide, starch, dextrin, sugars (e.g., lactose, sucrose), silica, talc, mica, diatomaceous earth, urea, calcium carbonate, sodium carbonate and bicarbonate, and sodium sulfate. Typical solid diluents are described in Watkins et al., Handbook of Insecticide Dust Diluents and Carriers, 2nd Ed., Dorland Books, Caldwell, New Jersey.

Liquid diluents include, for example, water, N,N-dimethylalkanamides (e.g.,

N,N-dimethylformamide), limonene, dimethyl sulfoxide, N-alkylpyrrolidones (e.g., N-methylpyrrolidinone), alkyl phosphates (e.g., triethyl phosphate), ethylene glycol, Methylene glycol, propylene glycol, dipropylene glycol, polypropylene glycol, propylene carbonate, butylene carbonate, paraffins (e.g., white mineral oils, normal paraffins, isoparaffins), alkylbenzenes, alkylnaphthalenes, glycerine, glycerol triacetate, sorbitol, aromatic hydrocarbons, dearomatized aliphatics, alkylbenzenes, alkylnaphthalenes, ketones such as cyclohexanone, 2-heptanone, isophorone and 4-hydroxy-4-methyl-2-pentanone, acetates such as isoamyl acetate, hexyl acetate, heptyl acetate, octyl acetate, nonyl acetate, tridecyl acetate and isobornyl acetate, other esters such as alkylated lactate esters, dibasic esters, alkyl and aryl benzoates and γ-butyrolactone, and alcohols, which can be linear, branched, saturated or unsaturated, such as methanol, ethanol, n-propanol, isopropyl alcohol, n-butanol, isobutyl alcohol, rc-hexanol, 2-ethylhexanol, n-octanol, decanol, isodecyl alcohol, isooctadecanol, cetyl alcohol, lauryl alcohol, tridecyl alcohol, oleyl alcohol, cyclohexanol, tetrahydrofurfuryl alcohol, diacetone alcohol, cresol and benzyl alcohol. Liquid diluents also include glycerol esters of saturated and unsaturated fatty acids (typically C6-C22 such as plant seed and fruit oils (e.g., oils of olive, castor, linseed, sesame, corn (maize), peanut, sunflower, grapeseed, safflower, cottonseed, soybean, rapeseed, coconut and palm kernel), animal- sourced fats (e.g., beef tallow, pork tallow, lard, cod liver oil, fish oil), and mixtures thereof. Liquid diluents also include alkylated fatty acids (e.g., methylated, ethylated, butylated) wherein the fatty acids may be obtained by hydrolysis of glycerol esters from plant and animal sources, and can be purified by distillation. Typical liquid diluents are described in Marsden, Solvents Guide, 2nd Ed., Interscience, New York, 1950.

The solid and liquid compositions of the present invention often include one or more surfactants. When added to a liquid, surfactants (also known as "surface-active agents") generally modify, most often reduce, the surface tension of the liquid. Depending on the nature of the hydrophilic and lipophilic groups in a surfactant molecule, surfactants can be useful as wetting agents, dispersants, emulsifiers or defoaming agents.

Surfactants can be classified as nonionic, anionic or cationic. Nonionic surfactants useful for the present compositions include, but are not limited to: alcohol alkoxylates such as alcohol alkoxylates based on natural and synthetic alcohols (which may be branched or linear) and prepared from the alcohols and ethylene oxide, propylene oxide, butylene oxide or mixtures thereof; amine ethoxylates, alkanolamides and ethoxylated alkanolamides; alkoxylated triglycerides such as ethoxylated soybean, castor and rapeseed oils; alkylphenol alkoxylates such as octylphenol ethoxylates, nonylphenol ethoxylates, dinonyl phenol ethoxylates and dodecyl phenol ethoxylates (prepared from the phenols and ethylene oxide, propylene oxide, butylene oxide or mixtures thereof); block polymers prepared from ethylene oxide or propylene oxide and reverse block polymers where the terminal blocks are prepared from propylene oxide; ethoxylated fatty acids; ethoxylated fatty esters and oils; ethoxylated methyl esters; ethoxylated tristyrylphenol (including those prepared from ethylene oxide, propylene oxide, butylene oxide or mixtures thereof); fatty acid esters, glycerol esters, lanolin- based derivatives, polyethoxylate esters such as polyethoxylated sorbitan fatty acid esters, polyethoxylated sorbitol fatty acid esters and polyethoxylated glycerol fatty acid esters; other sorbitan derivatives such as sorbitan esters; polymeric surfactants such as random copolymers, block copolymers, alkyd peg (polyethylene glycol) resins, graft or comb polymers and star polymers; polyethylene glycols (pegs); polyethylene glycol fatty acid esters; silicone-based surfactants; and sugar-derivatives such as sucrose esters, alkyl polyglycosides and alkyl polysaccharides.

Useful anionic surfactants include, but are not limited to: alkylaryl sulfonic acids and their salts; carboxylated alcohol or alkylphenol ethoxylates; diphenyl sulfonate derivatives; lignin and lignin derivatives such as lignosulfonates; maleic or succinic acids or their anhydrides; olefin sulfonates; phosphate esters such as phosphate esters of alcohol alkoxylates, phosphate esters of alkylphenol alkoxylates and phosphate esters of styryl phenol ethoxylates; protein-based surfactants; sarcosine derivatives; styryl phenol ether sulfate; sulfates and sulfonates of oils and fatty acids; sulfates and sulfonates of ethoxylated alkylphenols; sulfates of alcohols; sulfates of ethoxylated alcohols; sulfonates of amines and amides such as N,N- alkyltaurates; sulfonates of benzene, cumene, toluene, xylene, and dodecyl and tridecylbenzenes; sulfonates of condensed naphthalenes; sulfonates of naphthalene and alkyl naphthalene; sulfonates of fractionated petroleum; sulfosuccinamates; and sulfosuccinates and their derivatives such as dialkyl sulfosuccinate salts.

Useful cationic surfactants include, but are not limited to: amides and ethoxylated amides; amines such as N-alkyl propanediamines, tripropylenetriamines and dipropylenetetramines, and ethoxylated amines, ethoxylated diamines and propoxylated amines (prepared from the amines and ethylene oxide, propylene oxide, butylene oxide or mixtures thereof); amine salts such as amine acetates and diamine salts; quaternary ammonium salts such as quaternary salts, ethoxylated quaternary salts and diquaternary salts; and amine oxides such as alkyldimethylamine oxides and bis-(2-hydroxyethyl)-alkylamine oxides.

Also useful for the present compositions are mixtures of nonionic and anionic surfactants or mixtures of nonionic and cationic surfactants. Nonionic, anionic and cationic surfactants and their recommended uses are disclosed in a variety of published references including McCutcheon 's Emulsifiers and Detergents, annual American and International Editions published by McCutcheon' s Division, The Manufacturing Confectioner Publishing Co. ; Sisely and Wood, Encyclopedia of Surface Active Agents, Chemical Publ. Co., Inc., New York, 1964; and A. S. Davidson and B. Milwidsky, Synthetic Detergents, Seventh Edition, John Wiley and Sons, New York, 1987.

Compositions of this invention may also contain formulation auxiliaries and additives, known to those skilled in the art as formulation aids (some of which may be considered to also function as solid diluents, liquid diluents or surfactants). Such formulation auxiliaries and additives may control: pH (buffers), foaming during processing (antifoams such polyorganosiloxanes), sedimentation of active ingredients (suspending agents), viscosity (thixotropic thickeners), in-container microbial growth (antimicrobials), product freezing (antifreezes), color (dyes/pigment dispersions), wash-off (film formers or stickers), evaporation (evaporation retardants), and other formulation attributes. Film formers include, for example, polyvinyl acetates, polyvinyl acetate copolymers, polyvinylpyrrolidone-vinyl acetate copolymer, polyvinyl alcohols, polyvinyl alcohol copolymers and waxes. Examples of formulation auxiliaries and additives include those listed in McCutcheon's Volume 2: Functional Materials, annual International and North American editions published by McCutcheon's Division, The Manufacturing Confectioner Publishing Co.; and PCT Publication WO 03/024222. The cellulose synthase inhibitor or indirect inhibitor and any other active ingredients are typically incorporated into the present compositions by dissolving the active ingredient in a solvent or by grinding in a liquid or dry diluent. Solutions, including emulsifiable concentrates, can be prepared by simply mixing the ingredients. If the solvent of a liquid composition intended for use as an emulsifiable concentrate is water- immiscible, an emulsifier is typically added to emulsify the active-containing solvent upon dilution with water. Active ingredient slurries, with particle diameters of up to 2,000 μηι can be wet milled using media mills to obtain particles with average diameters below 3 μιη. Aqueous slurries can be made into finished suspension concentrates (see, for example, U.S. 3,060,084) or further processed by spray drying to form water-dispersible granules. Dry formulations usually require dry milling processes, which produce average particle diameters in the 2 to 10 μηι range. Dusts and powders can be prepared by blending and usually grinding (such as with a hammer mill or fluid-energy mill). Granules and pellets can be prepared by spraying the active material upon preformed granular carriers or by agglomeration techniques. See Browning, "Agglomeration", Chemical Engineering, December 4, 1967, pp 147-48, Perry's Chemical Engineer's Handbook, 4th Ed., McGraw-Hill, New York, 1963, pages 8-57 and following, and WO 91/13546. Pellets can be prepared as described in U.S. 4, 172,714. Water-dispersible and water-soluble granules can be prepared as taught in U.S. 4,144,050, U.S. 3,920,442 and DE 3,246,493. Tablets can be prepared as taught in U.S. 5, 180,587, U.S. 5,232,701 and U.S. 5,208,030. Films can be prepared as taught in GB 2,095,558 and U.S. 3,299,566.

Compositions of this invention can also be mixed with RNA to enhance effectiveness or to confer safening properties. Accordingly, a compositions containing Compound 1 can be mixed with polynucleotides including but not limited to DNA, RNA, and/or chemically modified nucleotides influencing the amount of a particular target through down regulation, interference, suppression or silencing of the genetically derived transcript that render a herbicidal effect. Alternatively, a composition containing a Compound 1 can be mixed with polynucleotides including but not limited to DNA, RNA, and/or chemically modified nucleotides influencing the amount of a particular target through down regulation, interference, suppression or silencing of the genetically derived transcript that render a safening effect.

For further information regarding the art of formulation, see T. S. Woods, "The Formulator's Toolbox - Product Forms for Modern Agriculture" in Pesticide Chemistry and Bioscience, The Food-Environment Challenge, T. Brooks and T. R. Roberts, Eds., Proceedings of the 9th International Congress on Pesticide Chemistry, The Royal Society of Chemistry, Cambridge, 1999, pp. 120-133. See also U.S. 3,235,361, Col. 6, line 16 through Col. 7, line 19 and Examples HMl; U.S. 3,309, 192, Col. 5, line 43 through Col. 7, line 62 and Examples 8, 12, 15, 39, 41, 52, 53, 58, 132, 138-140, 162-164, 166, 167 and 169-182; U.S. 2,891,855, Col. 3, line 66 through Col. 5, line 17 and Examples 1-4; Klingman, Weed Control as a Science, John Wiley and Sons, Inc., New York, 1961, pp 81-96; Hance et al., Weed Control Handbook, 8th Ed., Blackwell Scientific Publications, Oxford, 1989; and Developments in formulation technology, PJB Publications, Richmond, UK, 2000.

Preparation of the Mixtures

A mixture of one or more of the following herbicides with a compound or composition of this invention may be particularly useful for weed control: acetochlor, acifluorfen and its sodium salt, aclonifen, acrolein (2-propenal), alachlor, alloxydim, ametryn, amicarbazone, amidosulfuron, aminocyclopyrachlor and its esters (e.g., methyl, ethyl) and salts (e.g., sodium, potassium), aminopyralid, amitrole, ammonium sulfamate, anilofos, asulam, atrazine, azimsulfuron, beflubutamid, benazolin, benazolin-ethyl, bencarbazone, benfluralin, benfuresate, bensulfuron-methyl, bensulide, bentazone, benzobicyclon, benzofenap, bicyclopyrone, bifenox, bilanafos, bispyribac and its sodium salt, bromacil, bromobutide, bromofenoxim, bromoxynil, bromoxynil octanoate, butachlor, butafenacil, butamifos, butralin, butroxydim, butylate, cafenstrole, carbetamide, carfentrazone-ethyl, catechin, chlomethoxyfen, chloramben, chlorbromuron, chlorflurenol-methyl, chloridazon, chlorimuron-ethyl, chlorotoluron, chlorpropham, chlorsulfuron, chlorthal-dimethyl, chlorthiamid, cinidon-ethyl, cinmethylin, cinosulfuron, clacyfos, clefoxydim, clethodim, cyclopyrimorate, clodinafop-propargyl, clomazone, clomeprop, clopyralid, clopyralid-olamine, cloransulam-methyl, cumyluron, cyanazine, cycloate, cyclopyrimorate, cyclosulfamuron, cycloxydim, cyhalofop-butyl, 2,4-D and its butotyl, butyl, isoctyl and isopropyl esters and its dimethylammonium, diolamine and trolamine salts, daimuron, dalapon, dalapon-sodium, dazomet, 2,4-DB and its dimethylammonium, potassium and sodium salts, desmedipham, desmetryn, dicamba and its diglycolammonium, dimethylammonium, potassium and sodium salts, dichlobenil, dichlorprop, diclofop-methyl, diclosulam, difenzoquat metilsulfate, diflufenican, diflufenzopyr, dimefuron, dimepiperate, dimethachlor, dimethametryn, dimethenamid, dimethenamid-P, dimethipin, dimethylarsinic acid and its sodium salt, dinitramine, dinoterb, diphenamid, diquat dibromide, dithiopyr, diuron, DNOC, endothal, EPTC, esprocarb, ethalfluralin, ethametsulfuron-methyl, ethiozin, ethofumesate, ethoxyfen, ethoxysulfuron, etobenzanid, fenoxaprop-ethyl, fenoxaprop-P-ethyl, fenoxasulfone, fenquinotrione, fentrazamide, fenuron, fenuron-TCA, flamprop-methyl, flamprop-M-isopropyl, flamprop-M-methyl, flazasulfuron, florasulam, fluazifop-butyl, fluazifop-P-butyl, fluazolate, flucarbazone, flucetosulfuron, fluchloralin, flufenacet, flufenpyr, flufenpyr-ethyl, flumetsulam, flumiclorac-pentyl, flumioxazin, fluometuron, fluoroglycofen-ethyl, flupoxam, flupyrsulfuron-methyl and its sodium salt, flurenol, flurenol- butyl, fluridone, flurochloridone, fluroxypyr, flurtamone, fluthiacet-methyl, fomesafen, foramsulfuron, fosamine-ammonium, glufosinate, glufosinate-ammonium, glufosinate-P, glyphosate and its salts such as ammonium, isopropylammonium, potassium, sodium (including sesquisodium) and trimesium (alternatively named sulfosate), halauxifen, halauxifen-methyl, halosulfuron-methyl, haloxyfop-etotyl, haloxyfop-methyl, hexazinone, imazamethabenz-methyl, imazamox, imazapic, imazapyr, imazaquin, imazaquin- ammonium, imazethapyr, imazethapyr-ammonium, imazosulfuron, indanofan, indaziflam, iofensulfuron, iodosulfuron-methyl, ioxynil, ioxynil octanoate, ioxynil- sodium, ipfencarbazone, isoproturon, isouron, isoxaben, isoxaflutole, isoxachlortole, lactofen, lenacil, linuron, maleic hydrazide, MCPA and its salts (e.g., MCPA-dimethylammonium, MCPA-potassium and MCPA-sodium, esters (e.g., MCPA-2-ethylhexyl, MCPA-butotyl) and thioesters (e.g., MCPA-thioethyl), MCPB and its salts (e.g., MCPB -sodium) and esters (e.g., MCPB-ethyl), mecoprop, mecoprop-P, mefenacet, mefluidide, mesosulfuron-methyl, mesotrione, metam-sodium, metamifop, metamitron, metazachlor, metazosulfuron, methabenzthiazuron, methylarsonic acid and its calcium, monoammonium, monosodium and disodium salts, methyldymron, metobenzuron, metobromuron, metolachlor, S-metolachlor, metosulam, metoxuron, metribuzin, metsulfuron-methyl, molinate, monolinuron, naproanilide, napropamide, napropamide-M, naptalam, neburon, nicosulfuron, norflurazon, orbencarb, orthosulfamuron, oryzalin, oxadiargyl, oxadiazon, oxasulfuron, oxaziclomefone, oxyfluorfen, paraquat dichloride, pebulate, pelargonic acid, pendimethalin, penoxsulam, pentanochlor, pentoxazone, perfluidone, pethoxamid, pethoxyamid, phenmedipham, picloram, picloram-potassium, picolinafen, pinoxaden, piperophos, pretilachlor, primisulfuron-methyl, prodiamine, profoxydim, prometon, prometryn, propachlor, propanil, propaquizafop, propazine, propham, propisochlor, propoxycarbazone, propyrisulfuron, propyzamide, prosulfocarb, prosulfuron, pyraclonil, pyraflufen-ethyl, pyrasulfotole, pyrazogyl, pyrazolynate, pyrazoxyfen, pyrazosulfuron-ethyl, pyribenzoxim, pyributicarb, pyridate, pyriftalid, pyriminobac-methyl, pyrimisulfan, pyrithiobac, pyrithiobac-sodium, pyroxasulfone, pyroxsulam, quinclorac, quinmerac, quinoclamine, quizalofop-ethyl, quizalofop-P-ethyl, quizalofop-P-tefuryl, rimsulfuron, saflufenacil, sethoxydim, siduron, simazine, simetryn, sulcotrione, sulfentrazone, sulfometuron-methyl, sulfosulfuron, 2,3,6-TBA, TCA, TCA-sodium, tebutam, tebuthiuron, tefuryltrione, tembotrione, tepraloxydim, terbacil, terbumeton, terbuthylazine, terbutryn, thenylchlor, thiazopyr, thiencarbazone, thifensulfuron-methyl, thiobencarb, tiafenacil, tiocarbazil, topramezone, tralkoxydim, tri-allate, triafamone, triasulfuron, triaziflam, tribenuron-methyl, triclopyr, triclopyr-butotyl, triclopyr-triethylammonium, tridiphane, trietazine, trifloxysulfuron, trifluralin, triflusulfuron-methyl, tritosulfuron, vernolate, 3-(2- chloro-3,6-difluorophenyl)-4-hydroxy-l-methyl-l,5-naphthyridin-2(lH)-one, 5-chloro-3-[(2- hydroxy-6-oxo-l-cyclohexen-l-yl)carbonyl]-l-(4-methoxyphenyl)-2(lH)-quinoxalinone, 2- chloro-N-(l-methyl-lH-tetrazol-5-yl)-6-(trifluoromethyl)-3-pyridinecarboxamide, 7-(3,5- dichloro-4-pyridinyl)-5-(2,2-difluoroethyl)-8-hydroxypyrido[2,3-¾]pyrazin-6(5H)-one), 4- (2,6-diethyl-4-methylphenyl)-5-hydroxy-2,6-dimethyl-3(2H)-pyridazinone), 5-[[(2,6- difluorophenyl)methoxy]methyl]-4,5-dihydro-5-methyl-3-(3-methyl-2-thienyl)isoxazole (previously methioxolin), 3-[7-fluoro-3,4-dihydro-3-oxo-4-(2-propyn-l-yl)-2H-l,4- benzoxazin-6-yl]dihydro ,5-dim 4-(4- fluorophenyl)-6 (2-hydroxy-6-oxo -cyclohexen-l-yl)carbonyl]-2-metliyl-l,2,4-triazine- 3,5(2H,4H)-dione, methyl 4-amino-3-chloro-6-(4-chloro-2-fluoro-3-methoxyphenyl)-5- fluoro-2-pyridinecarboxylate, 2-methyl-3-(methylsulfonyl)-N-(l-methyl-lH-tetrazol-5-yl)-4- (trifluoromethyl)benzamide, and 2-methyl-N-(4-methyl-l,2,5-oxadiazol-3-yl)-3- (methylsulfinyl)-4-(trifluoromethyl)benzamide. Other herbicides also include bioherbicides such as Alternaria destruens Simmons, Colletotrichum gloeosporiodes (Penz.) Penz. & Sacc, Drechsiera monoceras (MTB-951), Myrothecium verrucaria (Albertini & Schweinitz) Ditmar: Fries, Phytophthora palmivora (Butl.) Butl. and Puccinia thlaspeos Schub.

Compounds of this invention can also be used in combination with plant growth regulators such as aviglycine, N-(phenylmethyl)-lH-purin-6-amine, epocholeone, gibberellic acid, gibberellin A₄ and A₇, harpin protein, mepiquat chloride, prohexadione calcium, prohydrojasmon, sodium nitrophenolate and trinexapac-methyl, and plant growth modifying organisms such as Bacillus cereus strain BP01.

General references for agricultural protectants (i.e. herbicides, herbicide safeners, insecticides, fungicides, nematocides, acaricides and biological agents) include The Pesticide Manual, 13th Edition, C. D. S. Tomlin, Ed., British Crop Protection Council, Farnham, Surrey, U.K., 2003 and The BioPesticide Manual, 2nd Edition, L. G. Copping, Ed., British Crop Protection Council, Farnham, Surrey, U.K., 2001.

For embodiments where one or more of these various mixing partners are used, the weight ratio of these various mixing partners (in total) to the compound or composition of the invention is typically between about 1:3000 and about 3000: 1. Of note are weight ratios between about 1:300 and about 300: 1 (for example ratios between about 1 :30 and about 30: 1). One skilled in the art can easily determine through simple experimentation the biologically effective amounts of active ingredients necessary for the desired spectrum of biological activity. It will be evident that including these additional components may expand the spectrum of weeds controlled beyond the spectrum controlled by the compound or composition of the invention alone.

In certain instances, combinations of a compound of this invention with other biologically active (particularly herbicidal) compounds or agents (i.e. active ingredients) can result in a greater-than-additive (i.e. synergistic) effect on weeds and/or a less-than-additive effect (i.e. safening) on crops or other desirable plants. Reducing the quantity of active ingredients released in the environment while ensuring effective pest control is always desirable. Ability to use greater amounts of active ingredients to provide more effective weed control without excessive crop injury is also desirable. When synergism of herbicidal active ingredients occurs on weeds at application rates giving agronomically satisfactory levels of weed control, such combinations can be advantageous for reducing crop production cost and decreasing environmental load. When safening of herbicidal active ingredients occurs on crops, such combinations can be advantageous for increasing crop protection by reducing weed competition.

Of note is a combination of a compound of the invention with at least one other herbicidal active ingredient. Of particular note is such a combination where the other herbicidal active ingredient has different site of action from the compound of the invention. In certain instances, a combination with at least one other herbicidal active ingredient having a similar spectrum of control but a different site of action will be particularly advantageous for resistance management. Thus, a composition of the present invention can further comprise (in a herbicidally effective amount) at least one additional herbicidal active ingredient having a similar spectrum of control but a different site of action.

Compounds of this invention can also be used in combination with herbicide safeners such as allidochlor, benoxacor, cloquintocet-mexyl, cumyluron, cyometrinil, cyprosulfonamide, daimuron, dichlormid, dicyclonon, dietholate, dimepiperate, fenchlorazole-ethyl, fenclorim, flurazole, fluxofenim, furilazole, isoxadifen-ethyl, mefenpyr- diethyl, mephenate, methoxyphenone naphthalic anhydride (1,8-naphthalic anhydride), oxabetrinil, N-(aminocarbonyl)-2-methylbenzenesulfonamide, N-(aminocarbonyl)- 2-fluorobenzenesulfonamide, l-bromo-4-[(chloromethyl)sulfonyl]benzene (BCS), 4- (dichloroacetyl)-l-oxa-4-azospiro[4.5]decane (ΜΟΝ 4660), 2-(dichloromethyl)-2-methyl- 1,3-dioxolane (MG 191), ethyl l,6-dihydro-l-(2-methoxyphenyl)-6-oxo-2-phenyl-5- pyrimidinecarboxylate, 2-hydroxy-N,N-dimethyl-6-(trifluoromethyl)pyridine-3-carboxamide, and 3-oxo-l-cyclohexen-l-yl l-(3,4-dimethylphenyl)-l,6-dihydro-6-oxo-2-phenyl-5- pyrimidinecarboxylate to increase safety to certain crops. Antidotally effective amounts of the herbicide safeners can be applied at the same time as the compounds of this invention, or applied as seed treatments. Therefore an aspect of the present invention relates to a herbicidal mixture comprising a compound of this invention and an antidotally effective amount of a herbicide safener. Seed treatment is particularly useful for selective weed control, because it physically restricts antidoting to the crop plants. Therefore a particularly useful embodiment of the present invention is a method for selectively controlling the growth of undesired vegetation in a crop comprising contacting the locus of the crop with a herbicidally effective amount of a compound of this invention wherein seed from which the crop is grown is treated with an antidotally effective amount of safener. Antidotally effective amounts of safeners can be easily determined by one skilled in the art through simple experimentation.

Of note is a composition comprising a compound of the invention (in a herbicidally effective amount), at least one additional active ingredient selected from the group consisting of other herbicides and herbicide safeners (in an effective amount), and at least one component selected from the group consisting of surfactants, solid diluents and liquid diluents. The ratios of active ingredients and administration rates to be employed are easily determined by the skilled artisan.

Claims

CLAIMS What is claimed is:

1. A method for producing a herbicidal composition comprising:

(a) screening a candidate compound in a cellulose synthase inhibition assay; and (b) if said candidate compound is active in the cellulose synthase inhibition assay, testing said compound for activity against a plant; and

2. The method of claim 1 wherein the screening step (a) is an in- vitro activity assay, or a binding assay.

3. The method of claim 1 wherein the screening step (a) is an in- vitro cellulose synthase activity inhibition assay.

4. The method of claim 3 wherein the screening step makes use of a cellulose synthase having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% homology; or having 100% homology to the amino acid sequence of SEQ ID NO: 2.

5. The method of claim 1 wherein the screening step (a) is a cellulose synthase binding assay.

6. The method of claim 1 wherein the herbicidal composition contains a carrier.

7. A method for producing a herbicidal compound comprising testing a candidate compound in a cellulose synthase activity inhibition assay wherein the assay utilizes cellulose synthase from a weed to be controlled.

8. A method of controlling weeds comprising applying a herbicidally effective amount of a cellulose synthase inhibitor produced by the method described herein to a locus in need of such treatment.

9. A herbicidal composition comprising a cellulose synthase inhibitor produced by the method of any of Claims 1 through 6 as the active ingredient in combination with a carrier.

10. A herbicidal composition comprising a cellulose synthase inhibitor produced by the method of any of Claims 1 through 6 in combination with another herbicide.