Patent Document

PRIORITY CLAIM 
     This application is a continuation of Ser. No. 13/553,677, filed on Jul. 19, 2012. application Ser. No. 13/553,677 is a continuation-in-part of application Ser. No. 12/650,315 filed on Dec. 30, 2009. application Ser. No. 12/650,315 claims the benefits of priorities from U.S. provisional application Ser. No. 61/142,179, filed on Dec. 31, 2008, and 61/261,504 filed on Nov. 16, 2009 under 35 U.S.C. 119 (e), and Taiwan patent application number 098144895 filed on Dec. 25, 2009 under 35 U.S.C. 119(a)-(d). The contents of all of which are incorporated herein by reference in their entirety. 
    
    
     REFERENCE TO GOVERNMENT GRANT 
     This invention was supported in part by funds obtained from the U.S. Government (USDA SBIR Grant Number: 2011-33610-30455). The U.S. Government may have certain rights in the invention. 
    
    
     FIELD OF THE INVENTION 
     This invention relates to compositions and methods for controlling the germination and growth of broadleaf, sedge and grass weeds using compounds comprising thaxtomin, a cyclic dipeptide produced by  Streptomyces  sp., as an active ingredient. 
     BACKGROUND OF THE INVENTION 
     Natural products are substances produced by microbes, plants, and other organisms. Microbial natural products offer an abundant source of chemical diversity, and there is a long history of utilizing natural products for pharmaceutical purposes. However, secondary metabolites produced by microbes can also be successfully used for weed and pest control in agricultural applications. 
     Thaxtomins (4-nitroindol-3-yl-containing 2,5-dioxopiperazines) are a family of dipeptide phytotoxins produced by plant-pathogenic  Streptomyces  sp. ( S. scabies, S. acidiscabies ) that cause scab diseases in potato ( Solanum tuberosum ) (King, Lawrence et al. 1992). Toxin production occurs in diseased tissue and can also be elicited in vitro in an optimal growth medium containing oat bran (Loria, Bukhalid et al. 1995; Beauséjour, Goyer et al. 1999). King and her coworkers (King, Lawrence et al. 2001) demonstrated that all plant pathogenic species in the  Streptomyces  family produce one or more thaxtomins with herbicidal activity. Hiltunen et al. (Hiltunen, Laakso et al. 2006) purified four thaxtomin analogs (thaxtomin A, thaxtomin A ortho isomer, thaxtomin B and thaxtomin D) from cultures of  S. scabies  and  S. turbidiscabies  and showed that all four compounds induced similar symptoms of reduced shoot and root growth, root swelling, (at 10-200 ppb) and necrosis (at 200-1000 ppb) on micropropagated in vitro cultures of potato. In addition, thaxtomins applied in combinations, showed additive effects, but no synergism (Hiltunen, Laakso et al. 2006). According to Duke et al. (Duke, Baerson et al. 2003), both thaxtomin A ( FIG. 1 ) and thaxtomin D have marked activity as pre and post emergent, non-systemic herbicides, and concentrations of less than 1 uM of thaxtomin A causes cell swelling, necrosis and growth inhibition in mono and dicotyledonous seedlings (Healy, Wach et al. 2000). Thaxtomin has been evaluated as an herbicide by Dow Agro Sciences, Inc., and while active, it lacked systemic action (King, Lawrence et al. 2001). The presence of the nitro group in the indole ring required for an L,L-configuration of the diketopiperazine appears to be the minimal requirement for phytotoxicity. The position of the nitro group in the indole ring is very site specific, and the phenyl portion of the phenylalanine plays a necessary role in structural requirements of phytotoxicity (King, Lawrence et al. 1989; King, Lawrence et al. 1992; King, Lawrence et al. 2003). The herbicidal mode of action is based on disruption of cell wall synthesis (Fry and Loria 2002), with inhibition of cellulose biosynthesis being the main target (King et al., 2001; Duval et al., 2005; Johnson et al. 2007). Recently, Kang et al. (Kang, Semones et al. 2008) have described the use of thaxtomin and thaxtomin compositions as algaecides to control algae in water environments. 
     SUMMARY OF THE INVENTION 
     The present invention discloses the use of thaxtomin as a pre or post-emergence herbicide against most common weeds in the cereal, pasture grass, Timothy grasses and turf grass, residential gardens, vineyards, orchards and park growth systems. A “growth system” may be any ecosystem for growing cereal, pasture grass, Timothy grass and turf grass. For example, a “cereal growth system” may be a cereal growth culture or may be a field containing planted cereal crops or cereal seeds. Similarly, a “turf grass growth system” may be a turf grass growth culture or may be a field, lawn or golf course containing planted turf grass or turf grass seeds. It can serve as a safer alternative to synthetic herbicides now on the market. A primary object of the invention is to provide novel herbicidal compositions against both broadleaf, sedge and grassy weeds, which include but are not limited to  Chenopodium  sp. (e.g.,  Chenopodium album ),  Abutilon  sp. (e.g.,  Abutilon theophrasti ),  Helianthus  sp. (e.g.,  Helianthus annuus ),  Ambrosia  sp. (e.g.,  Ambrosia artemesifolia ),  Amaranthus  sp. (e.g.,  Amaranthus retroflexus ),  Convolvulu  sp. (e.g.,  Convolvulus arvensis ),  Brassica  sp. (e.g.,  Brassica kaber ),  Taraxacum  sp. (e.g.,  Taraxacum officinale ),  Solanum  sp. (e.g.,  Solanum nigrum ),  Malva  sp. (e.g.,  Malva neglect ),  Setaria  sp. (e.g.,  Setaria lutescens ),  Bromus  sp. (e.g.,  Bromus tectorum ),  Poa  sp. (e.g.,  Poa annua, Poa pratensis ),  Lollium  sp. (e.g.,  Lolium perenne  L. var. Pace),  Festuca  sp. (e.g.,  Festuca arundinaceae )  Schreb . Sp. (e.g.,  Schreb . var. Aztec II, Anthem II, LS 1100),  Echinochloa  sp. (e.g.,  Echinochloa crus - galli ), and particularly, Lambsquarter— Chenopodium album , Redroot Pigweed— Amaranthus retroflexus , Wild Mustard— Brassica kaber , Dandelion— Taraxacum officinale , and Black Nightshade— Solanum nigrum , that contains thaxtomin as an active ingredient. Another object is to provide a safe, non-toxic herbicidal composition that does not harm cereal crops, pasture grass, Timothy grass or turf grass and a method that will not harm the environment. 
     The above and other objects are accomplished by the present invention which is directed to herbicidal compositions containing at least one herbicidal agent, e.g., thaxtomin with optionally certain carriers to control the growth and germination of weeds in the cereal growth system and/or turf grass growth system and/or Timothy grass growth system and/or pasture grass growth system. In particular, the invention is further directed to an herbicidal composition for use in modulating the germination and growth of monocotyledonous and/or dicotyledenous and/or sedge weeds in a cereal growth system. In a particular embodiment, the cereal growth system is a non-rice cereal growth system comprising at least one herbicide in which said herbicide is thaxtomin. The compositions of the present invention may further comprise a carrier and/or diluent. In a particular embodiment, the composition is an aqueous composition. In another particular embodiment, the thaxtomin in the composition is dissolved in a diluent comprising an organic solvent such as ethanol, isopropanol, or an aliphatic ketone such as acetone, methyl ethyl ketone, methyl isobutyl ketone. 
     In a related aspect, the invention is directed to the use of at least one herbicidal agent, e.g., thaxtomin, in the formulation of an herbicide for modulating monocotyledonous and/or dicotyledenous and/or sedge weeds in a cereal growth system, e.g., a non-rice cereal growth system. Similarly, the invention is directed to the use of at least one herbicidal agent in formulation of an herbicide for modulating monocotyledonous and/or dicotyledenous and/or sedge weeds in a turf grass growth system and/or Timothy grass growth system and/or pasture grass growth system, wherein at least one herbicidal agent is thaxtomin. 
     The compositions of the present invention may comprise in addition to thaxtomin, at least one or more herbicides. Thus the invention may comprise a thaxtomin and a chemical herbicide and/or bioherbicide. Compositions comprising thaxtomin and at least a second herbicide may be used in cereal growth systems (e.g., wheat, triticale, barley, oats, rye, corn,  sorghum , sugarcane, rice or millet) and/or turf grass growth systems and/or Timothy grass growth systems and/or pasture grass growth systems and/or residential gardens, vineyards, orchards and park systems and/or aquatic systems. In a particular embodiment these compositions may be used to modulate growth of broadleaf, sedge and grassy weeds, which include but are not limited to  Chenopodium  sp. (e.g.,  Chenopodium album ),  Abutilon  sp. (e.g.,  Abutilon theophrasti ),  Helianthus  sp. (e.g.,  Helianthus annuus ),  Ambrosia  sp. (e.g.,  Ambrosia artemesifolia ),  Amaranthus  sp. (e.g.,  Amaranthus retroflexus ),  Convolvulu  sp. (e.g.,  Convolvulus arvensis ),  Brassica  sp. (e.g.,  Brassica kaber ),  Taraxacum  sp. (e.g.,  Taraxacum officinale ),  Solanum  sp. (e.g.,  Solanum nigrum ),  Malva  sp. (e.g.,  Malva neglect ),  Setaria  sp. (e.g.,  Setaria lutescens ),  Bromus  sp. (e.g.,  Bromus tectorum ),  Poa  sp. (e.g.,  Poa annua, Poa pratensis ),  Lollium  sp. (e.g.,  Lollium perenne  L. var. Pace,  Lollium arundinaceum  ( Schreb .) var. Atec II or Anthem II),  Festuca  sp. (e.g.,  Festuca arundinaceae )  Schreb . Sp.,  Echinochloa  sp. (e.g.,  Echinochloa crus - galli ), The compositions may also be used to modulate growth of aquatic weeds which include but are not limited to  Ammania  sp.,  Alisma plantago - aquatica, Cyperus  sp.,  Leptochloa  sp. 
     Given that the invention is directed to the use of thaxtomin as a pre- or post-emergence herbicide, the invention is directed to a method for selectively modulating germination and growth of monocotyledonous, dicotyledonous and sedge weeds in a cereal crop growth system. In a particular embodiment, the cereal growth system is a non-rice cereal crop growth system comprising applying to said weeds or soil in said cereal crop growing system at least one herbicidal agent, wherein said herbicidal agent is thaxtomin, in an amount of effective to modulate germination and growth of said weeds but not modulate growth of cereal crop in said cereal crop growth system. The cereal crop may include but is not limited to corn, wheat, triticale, barley, rye, oats,  sorghum , sugarcane, and millet. The invention is further directed to a method for modulating germination and growth of monocotyledonous, dicotyledonous and sedge weeds in a turf, pasture and/or Timothy grass growth system comprising applying to said weeds or soil in said turf grass growing system at least one herbicidal agent, wherein said herbicidal agent is thaxtomin, in an amount of effective to modulate growth of said weeds but not modulate germination and growth of turf grass in said turf grass growth system, pasture grass in said pasture grass growth system and/or Timothy grass in said Timothy grass growth system. The turf grass may be selected from the group consisting of  Festuca  sp.,  Poa  sp.,  Bromus  sp.,  Lolium  sp.,  Agrostis  sp.,  Zoysia  sp.,  Cynodon  sp. 
     Further, the invention is directed to a method for modulating germination and growth of weeds selected from the group consisting of  Chenopodium  sp. (e.g.,  Chenopodium album ),  Abutilon  sp. (e.g.,  Abutilon theophrasti ),  Helianthus  sp. (e.g.,  Helianthus annuus ),  Ambrosia  sp. (e.g.,  Ambrosia artemesifolia ),  Amaranthus  sp. (e.g.,  Amaranthus retroflexus ),  Convolvulu  sp. (e.g.,  Convolvulus arvensis ),  Brassica  sp. (e.g.,  Brassica kaber ),  Taraxacum  sp. (e.g.,  Taraxacum officinale ),  Solanum  sp. (e.g.,  Solanum nigrum ),  Malva  sp. (e.g.,  Malva neglect ),  Setaria  sp. (e.g.,  Setaria lutescens ),  Bromus  sp. (e.g.,  Bromus tectorum ),  Poa  sp. (e.g.,  Poa annua, Poa pratensis ),  Lollium  sp. (e.g.,  Lollium perenne  L. var. Pace,  Lollium arundinaceum  ( Schreb .) var. Atec II or Anthem II),  Festuca  sp. (e.g.,  Festuca arundinaceae )  Schreb . Sp.,  Echinochloa  sp. (e.g.,  Echinochloa crus - galli ), comprising applying to said weeds or soil an amount of thaxtomin or salt thereof and optionally a second herbicidal agent effective to modulate said germination and growth of said weeds. 
     As noted above, the method of the present invention may also involve the use of at least a second herbicidal agent. The two herbicidal agents may be applied together in one formulation or separately in two formulations. Control of weeds can be achieved by using thaxtomin A in a tank mix or rotation with other herbicidally active compounds known to have good activity against grass weeds but no or low phytotoxicity against cereal crops and/or turf grass and/or, pasture grass and/or Timothy grasses. In particular, the invention relates to a method for modulating growth of monocotyledonous, dicotyledonous and sedge weeds comprising applying to said weeds an amount of thaxtomin and amount of at least a second herbicidal agent to modulate growth of said weeds. The two herbicidal agents may be applied together in one formulation or separately in two formulations. The thaxtomin and second herbicidal agent may be applied in a cereal growth system (e.g., wheat, triticale, barley, oats, rye, corn,  sorghum , sugarcane, rice or millet) and/or turf grass growth system and/or pasture grass growth system and/or Timothy grass growth system. 
    
    
     
       BRIEF DESCRIPTION OF THE FIGURES 
         FIG. 1  shows the structure of Thaxtomin A. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges is also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either both of those included limits are also included in the invention. 
     Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are now described. 
     It must be noted that as used herein and in the appended claims, the singular forms “a,” “and” and “the” include plural references unless the context clearly dictates otherwise. Thaxtomin utilized in this invention may be derived in fermentation of the following actinomycetes cultures:  S. scabies —ATCC 49173,  S. acidiscabies —ATCC 49003 and BL37-EQ-010—or it can be purchased from commercial sources. 
     The thaxtomin utilized in the invention include but are not limited to agents described as cyclic dipeptides having the basic structure cyclo-(L-4-nitrotryptophyl-L-phenylalanyl). In embodiments, suitable diketopiperazne moieties may be N-methylated, and include congeners carrying phenylalanyl alpha andring-carbon hydroxyl groups. The chemical in a particular embodiment comprises: 
                                
wherein R 1  is methyl or H, R 2  is hydroxy or H, R 3  is methyl or H, R 4  is hydroxy or H, R 5  is hydroxy or H, R 6  is hydroxy or H, and combinations thereof.
 
     Non limiting examples of suitable thaxtomin is for use in accordance with the present invention include but are not limited to thaxtomin A, thaxtomin A ortho isomer, thaxtomin B, thaxtomin C, hydroxythaxtomin C, thaxtomin A p-isomer, hydroxythaxtomin A and des-N-methylthaxtomin C and derivatives of any of these (See  FIG. 1 ). 
     The compositions of the present invention may be sprayed on the plant or applied to soil. Particular embodiments are described in the Examples, infra. These compositions may be in the form of dust, coarse dust, micro granules, granules, wettable powder, emulsifiable concentrate, liquid preparation, suspension concentrate, water degradable granules or oil suspension. 
     The compositions of the invention do comprise a carrier and/or diluent. The term, ‘carrier’ as used herein means an inert, organic or inorganic material, with which the active ingredient is mixed or formulated to facilitate its application to plant or other object to be treated, or its storage, transport and/or handling. Examples of diluents or carriers for the pre- and post-emergence herbicides include, but are not limited to, water, milk, ethanol, mineral oil, glycerol. 
     The compositions of the present invention may comprise at least two herbicidal agents. One herbicidal agent is thaxtomin set forth above. It may be present in one embodiment thaxtomin is present in an amount ranging from about 0.01 to about 5.0 mg/mL. The other herbicidal agent may be a bioherbicide and/or a chemical herbicide. The bioherbicide may be derived from a plant or may be a microbial bioherbicide. In particular, the bioherbicide derived from a plant may be derived from pepper (e.g., sarmentine) or may be a plant essential oil (e.g., lemongrass oil). The microbial bioherbicide may be derived from bacteria (e.g.  Streptomyces  sp.) or fungus. The bioherbicide may be selected from the group consisting of clove, cinnamon, lemongrass, citrus oils, orange peel oil, bialaphos, cornexistin, AAL-toxin, leptospermone, sarmentine, sarmentine analog momilactone B, sorgoleone, ascaulatoxin, manuka oil, Phoma macrostoma, m-tyrosine, chelated iron and ascaulatoxin aglycone. In a particular embodiment, the composition may comprise thaxtomin, lemongrass oil and optionally a surfactant and/or vegetable oil. In another embodiment, the composition may comprise thaxtomin, sarmentine and optionally a nonionic surfactant and/or vegetable oil. In another particular embodiment, the composition may comprise thaxtomin, bialaphos (also known as bialafos) and optionally a nonionic surfactant and/or vegetable oil. The bioherbicide such as lemongrass oil, bialaphos (bialfos) or sarmentine may be present in an amount ranging from about 0.1 mg/mL to about 100 mg/mL and more preferably between about 0.5 mg/mL to about 50 mg/mL 
     The chemical herbicide may be selected from the group consisting diflufenzopyr and salts thereof, dicamba and salts thereof, topramezone, tembotrione, S-metolachlor, atrazine, mesotrione, primisulfuron-methyl, 2,4-dichlorophenoxyacetic acid, nicosulfuron, thifensulfuron-methyl, asulam, metribuzin, diclofop-methyl, fluazifop, fenoxaprop-p-ethyl, asulam, oxyfluorfen, rimsulfuron, mecoprop, and quinclorac, thiobencarb, clomazone, cyhalofop, propanil, bensulfuron-methyl, penoxsulam, triclopyr and triclopyr-ester, trifloxysulfuron-sodium, imazethapyr, halosulfuron-methyl, pendimethalin, bispyribac-sodium, carfentrazone ethyl, sodium bentazon/sodium acifluorfen, glufosinate, glyphosate and orthosulfamuron, as well as a member of the dinitroaniline family, which includes but is not limited to pendimenthalin, oryzalin, trifuralin, etc., as well as members of the pyridine, phenylurea, chloroacetamide and triazine families, among others. 
     The chemical herbicide such as pendimethalin or clomazone, atrazine, oryzalin, trifluralin and metolachlor may be present in a pre-emergent weed control application in an amount ranging from about 0.1 mg/mL to about 50 mg/mL and a chemical herbicide such as cyhalofop, S-metolachlor, bispyribac-sodium, glyophosate, glufosinate, mesotione, penoxsulam, carfentrazone, quinclorac, triclopyr-ester, trioxysulfuron-sodium, thiobencarb, propanil, 2,4-D, dicamba in a post-emergent application from about 0.1 mg/mL to about 50 mg/mL The composition may further comprise an adjuvant which may be vegetable oil comprising ethyl oleate, polyethylene dialkyl ester and ethoxylated nonylphenol. The composition may additionally comprise a surfactant to be used for the purpose of emulsification, dispersion, wetting, spreading, integration, disintegration control, stabilization of active ingredients, improvement of fluidity or rust inhibition. The choice of dispersing and emulsifying agents, such as non-ionic, anionic, amphoteric and cationic dispersing and emulsifying agents, and the amount employed is determined by the nature of the composition and the ability of the agent to facilitate the dispersion of the herbicidal compositions of the present invention. 
     For post-emergent formulations, the formulation components used may contain smectite clays, attapulgite clays and similar swelling clays, thickeners such as xanthan gums, gum Arabic and other polysaccharide thickeners as well as dispersion stabilizers such as nonionic surfactants (for example polyoxyethylene (20) monolaurate or polysorbate 60 POE (20) sorbitan monostearate, ethylene glycol monostearate). The concentration of the clays may vary between about 0-2.5% w/w of the total formulation, the polysaccharide thickeners may range between about 0-0.5% w/w of the total formulation and the surfactants may range from about 0-5% w/w of the total formulation. 
     EXAMPLES 
     The composition and method of the present invention will be further illustrated in the following, non-limiting Examples. The examples are illustrative of various embodiments only and do not limit the claimed invention regarding the materials, conditions, weight ratios, process parameters and the like recited herein. 
     Example 1 
     In a pot study test in greenhouse conditions, 6-inch corn plants ( Zea mays  var. Sunglow) were sprayed with increasing concentrations of thaxtomin A mixed in a carrier 4% ethanol, 0.02% polysorbate 60 POE (20) sorbitan monostearate solution. The spraying solutions contained 0.125, 0.25, 0.5 and 1.0 mg thaxtomin A/mL, and the plants are sprayed until total coverage. Each treatment was done in three replicates, and a control solution consists of water with 4% ethanol and 0.02% polysorbate 60 POE (20) sorbitan monostearate as a surfactant. Prior to and after treatments, plants are grown in a greenhouse under artificial lights (12-h light/dark cycle) at 25° C. 
     Plants are evaluated in one-week intervals starting at 7 days after treatment. The final evaluation is done three weeks after treatment, at which time point, no phytotoxicity is observed in any of the test plants even at the highest thaxtomin A concentration. 
     Example 2 
     A pot study is conducted to test the phytotoxicity of thaxtomin A on corn ( Zea mays  var. Early Sunglow) and wheat ( Triticum aestivum  var. PR1404). To confirm the activity on broadleaf weeds, pigweed ( Amaranthus  sp.) is planted in the same pot with either three corn or five wheat seeds, and sprayed simultaneously with the cereal test plants. The less than 3-inch tall plants grown under growth lights (12-h light/12-h dark) at 28° C. are sprayed with thaxtomin A solutions derived from a liquid culture of  S. acidiscabies  containing 0.5, and 1.0 mg thaxtomin A per mL of solvent (4% ethanol and 0.2% non-ionic surfactant). A solution of 4% ethanol+0.2% non-ionic surfactant without thaxtomin A is used as a control treatment. Each treatment is conducted in three replicates. Treated plants are kept at 28° C. under growth lights and observed at three time points—7, 14 and 21 days after treatment—for visual symptoms of phytotoxicity on corn and wheat and % control of pigweed. 
     At each time point, no symptoms of phytotoxicity are observed in the cereal plants treated with thaxtomin A. The highest concentration of thaxtomin A (1.0 mg/mL) results in a complete control of pigweed grown in the same pots with corn and wheat. 
     Example 3 
     To test the phytotoxicity of thaxtomin A on  sorghum  plants, five seeds of  sorghum  ( Sorghum bicolor ) are planted in each 4″×41″ plastic pot filled with soil. Plants were grown under optimal conditions in a greenhouse before and after treatment with solutions containing 0.5 and 1.0 mg thaxtomin A/mL. At the time of the treatment, the plants are about 3 inches tall. Each treatment is applied in three replicates, and a control treatment included plants treated with just the carrier (4% EtOH, 0.02% polysorbate 60 POE (20) sorbitan monostearate). Evaluations for phytotoxicity are performed at 7-day intervals starting one week after treatment. The last evaluation is performed three weeks after the treatment at which point, no phytotoxicity is observed in the treated plants in any treatment concentration. 
     Example 4 
     A strain of  S. acidiscabies  (ATCC-49003) is grown in oat bran broth for 5 days (25° C., 200 rpm). The whole cell broth with thaxtomin A is extracted using XAD resin. The dried crude extract was resuspended in 4% ethanol and 0.02% non-ionic surfactant at a concentration of 10 mg/mL, and the solutions with two different concentrations of thaxtomin A (0.5 and 1.0 mg/mL) are tested the following broadleaf weed species:
         Lambsquarter— Chenopodium album      Velvetleaf— Abutilon theophrasti      Sunflower— Helianthus annuus      Ragweed, Common— Ambrosia artemesifolia      Pigweed, Redroot— Amaranthus retroflexus      Bindweed, Common— Convolvulus arvensis      Mustard, Wild— Brassica kaber      Dandelion— Taraxacum officinale      Nightshade, Black— Solanum nigrum      Mallow, Common— Malva neglecta  
 
and on the following grass weed species:
   Foxtail— Setaria lutescens      Brome, Downy— Bromus tectorum      Bluegrass, Annual— Poa annua      Bluegrass, Kentucky— Poa pratensis      Ryegrass, Perennial—( Lolium perenne  L. var. Pace)   Fescue, Tall—( Festuca arundinaceae Schreb . var. Aztec II, Anthem II, LS1100)   Barnyard Grass— Echinochloa crus - galli          

     All plant species are tested in 4″×4″ plastic pots in three replicates. The untreated control plants are sprayed with the carrier solution (4% Ethanol, 0.02% glycosperse) and the positive control plants with Roundup at a rate corresponding to 1 fl. oz/acre. Treated plants are kept in a greenhouse under 12 h light/12 h dark conditions. Data for broadleaf species from weekly evaluations are presented in Table 1. 
     
       
         
               
             
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Weed control efficacy of a  S. acidiscabies  extract containing thaxtomin A on different weed species. Rating 
               
               
                 scale: 0 - no control, 1 - 10% control, 2 - 25% control, 3 - 50% control, 4 - 75% control, 5 - 100% control. 
               
             
          
           
               
                   
                   
                 THAXTOMIN 
                 THAXTOMIN 
               
               
                 Weed 
                 UTC 
                 SOLUTION 0.5 mg/mL 
                 SOLUTION 1.0 mg/mL 
               
             
          
           
               
                 species 
                 7 DAYS 
                 14 DAYS 
                 21 DAYS 
                 7 DAYS 
                 14 DAYS 
                 21 DAYS 
                 7 DAYS 
                 14 DAYS 
                 21 DAYS 
               
               
                   
               
               
                 Dandelion 
                 0.0 
                 0.0 
                 0.0 
                 2.0 
                 2.3 
                 4.0 
                 2.0 
                 2.0 
                 3.7 
               
               
                 Nightshade 
                 0.0 
                 0.0 
                 0.0 
                 2.7 
                 2.2 
                 2.3 
                 2.7 
                 2.0 
                 2.3 
               
               
                 Lambsquarter 
                 0.0 
                 0.0 
                 0.0 
                 2.0 
                 2.0 
                 2.0 
                 2.0 
                 2.0 
                 2.0 
               
               
                 Ragweed 
                 0.0 
                 0.0 
                 0.0 
                 1.0 
                 0.5 
                 0.0 
                 1.0 
                 0.5 
                 0.0 
               
               
                 Velvetleaf 
                 0.0 
                 0.0 
                 0.0 
                 1.7 
                 1.0 
                 1.0 
                 2.0 
                 1.0 
                 0.3 
               
               
                 Bindweed 
                 0.0 
                 0.0 
                 0.0 
                 1.0 
                 1.0 
                 0.0 
                 1.2 
                 1.0 
                 0.0 
               
               
                 Mustard 
                 0.0 
                 0.0 
                 0.0 
                 3.3 
                 4.0 
                 4.5 
                 3.5 
                 2.8 
                 3.5 
               
               
                 Sunflower 
                 0.0 
                 0.0 
                 0.0 
                 1.0 
                 2.0 
                 0.5 
                 1.0 
                 1.7 
                 0.5 
               
               
                 Mallow 
                 0.0 
                 0.0 
                 0.0 
                 1.0 
                 1.0 
                 1.0 
                 1.2 
                 1.0 
                 1.0 
               
               
                 Pigweed 
                 0.0 
                 0.0 
                 0.0 
                 3.5 
                 4.0 
                 4.0 
                 4.2 
                 3.0 
                 3.7 
               
               
                   
               
             
          
         
       
     
     The extract from a bacterial culture of  S. acidiscabies  with a thaxtomin A concentration of 0.5 mg/mL or higher showed good efficacy (&gt;50%) against at least three of the most common broadleaf weed species (dandelion, mustard and pigweed) in both cereal and turf growing systems. Control of some weeds such as Black nightshade and Common lambsquarter was not complete but thaxtomin A even at the lower concentration (0.5 mg/mL) results in severe stunting of these weeds. In this same study, no adverse effects are observed in grass species treated with either 0.5 or 1.0 mg/mL thaxtomin A. In all tested grass species, no phytotoxic effects were visible at even the higher thaxtomin A concentration. 
     Example 5 
     The combined effect of thaxtomin A and two commercial herbicides (Bipyribac-sodium formulated as Regiment and Lemongrass oil formulated as GreenMatch EX) on small-flower umbrella sedge and watergrass is tested in a field study using small (1-sq foot) plots. All single product treatments and tank mix combinations were sprayed at 57 gal per acre. Evaluation of % control was done 14 days after treatment and the results are presented in Table 2 below. Means in each column marked with the same letter in Table 2 are not statistically different from each other at p&lt;0.05 
     According to the results, lemongrass oil at 1.25% weight does not improve the efficacy of thaxtomin A (at 0.25 mg/mL) on sedge but it significantly increases the efficacy on grass weeds such as watergrass (field test) and sprangletop (greenhouse test). 
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 Effect of thaxtomin A alone and in combination with  
               
               
                 bispyribac-sodium and lemongrass oil on two rice weeds,  
               
               
                 small-flower umbrella sedge and watergrass. 
               
             
          
           
               
                   
                   
                 Watergrass  
               
               
                 Treatment 
                 Sedge control (%) 
                 control (%) 
               
               
                   
               
               
                 Thaxtomin 0.25 mg/mL 
                  95a 
                  5d 
               
               
                 Thaxtomin 0.5 mg/mL 
                 100a 
                  5d 
               
               
                 Bispyribac-sodium (12 g/acre) 
                 87.5a  
                 32.5a   
               
               
                 (12 g/acre) 
                   
                   
               
               
                 Bispyribac-sodium ½ (6 g/acre) 
                 47.5c  
                 15c 
               
               
                 Bispyribac-sodium ½ + 
                 67.5b  
                  25ab 
               
               
                 Thaxtomin 0.5 mg/mL 
                   
                   
               
               
                 Bispyribac-sodium ½ + 
                 55bc 
                 7.5c  
               
               
                 Thaxtomin 0.25 mg/mL 
                   
                   
               
               
                 Lemongrass oil 5% 
                  15d 
                 10c 
               
               
                 Lemongrass oil 2.5% 
                 12.5d  
                 10c 
               
               
                 Lemongrass oil 1.25% 
                  20d 
                  5d 
               
               
                 Lemongrass oil 1.25% + 
                 100a 
                 10c 
               
               
                 Thax 0.25 mg/mL 
                   
                   
               
               
                 Lemongrass oil 1.25% + 
                 100a 
                 20b 
               
               
                 Thaxtomin 0.5 mg/mL 
               
               
                   
               
             
          
         
       
     
     According to the results, lemongrass oil at 1.25% does not improve the efficacy of thaxtomin A (at 0.25 mg/mL) on sedge but it significantly increases the efficacy on grass weeds such as watergrass (field test) and sprangletop. Thaxtomin A (at 0.5 mg/mL) improves the efficacy of an ALS inhibitor, bipyribac sodium; used at half label rate on both sedge and grasses. 
     Example 6 
     The efficacy of thaxtomin A derived from a liquid culture of  S. acidiscabies  is tested in a field study on rice using 4.9 sq-ft plots surrounded by a metal ring. Treatments with either thaxtomin A or thaxtomin A in combination with lemongrass oil (formulated as GreenMatch EX) or cyhalofop (formulated as Clincher CA) were done using a hand-held sprayer with a water volume corresponding to 57 gallons per acre. Rice (variety M209) was grown until maturity and harvested by hand for yield and weed count assessment. Results of yield (kg/ha), and numbers of redstem, small-flower umbrella sedge, and sprangletop in each plot are presented in Table 3 below. 
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 Effect of thaxtomin A alone and in combination with lemongrass  
               
               
                 oil and cyhalofop on rice yield and weed control. 
               
             
          
           
               
                 Treatment 
                 Yield (kg/ha)  
                 # of redstem 
                 # of sedge 
                 # of sprangletop 
               
               
                   
               
             
          
           
               
                 1 
                 7516b 
                 10.3  
                 0.8a 
                 86.0a 
               
               
                 2 
                 7876b 
                  0.5b 
                 1.0a 
                 76.0a 
               
               
                 3 
                 9054ab 
                  0.3b 
                 0.5a 
                 69.3a 
               
               
                 4 
                 11296a 
                 12.8a 
                 0.5a 
                  4.0b 
               
               
                   
               
               
                 1. UTC; 
               
               
                 2. Thaxtomin A (180 g/acre); 
               
               
                 3. Lemongrass oil 1.25% + thaxtomin A (90 g/acre); 
               
               
                 4. Cyhalofop (half label rate; 52 g/acre) + thaxtomin A (90 g/acre) + veg oil 2.5% Means in each column marked with the same letter are not statistically different from each other at p &lt; 0.05. 
               
             
          
         
       
     
     Results indicate that thaxtomin at 180 g/acre significantly reduced the number of sedges but had no effect on sprangletop or yield. When used at half rate (thaxtomin A 90 g/acre), a combination with lemongrass oil had better effect on sedges than a combination with cyhalofop (used at half label rate 52 g/acre). Good grass weed (sprangletop) control is achieved when thaxtomin (90 g/acre) is combined with cyhalofop at half the label rate—this combination also improves the yield significantly. 
     Example 7 
     Cyhalofop (2-[4-(4-cyano-2-fluorophenoxy)phenoxy]propanoic acid, butyl ester) is also mixed together with adjuvant containing ethyl oleate, polyethylene dialky ester and ethoxylated nonylphenol (2.5% v/v) and increasing concentrations of thaxtomin A (purified from the ATCC strain 49003) at concentrations 0.1, 0.2 and 0.4 mg/ml. The concentrations of the 2-[4-(4-cyano-fluorophenoxy)phenoxy]propanoic acid, butyl ester before dilution are 29.6% (2.38 lb/gal) and 21.7% (2 lb/gal), respectively. The effect of these mixtures on the growth of common water plantain, red stem, smallflower sedge and sprangletop is determined in the greenhouse. Similarly, rice plants of variety M104 are grown and tested for phytotoxic effects, and all plants are evaluated 7, 14, and 21 days after treatment. Results of from the study with cyhalofop formulated as Clincher CA at the 21-day evaluation point are presented in Table 4 below. 
                                                                 TABLE 4                   Effect of thaxtomin A alone and with        cyhalofop on rice yield and weed control            Cyhalofop                        (6.5 oz/acre) +                       Thaxtomin A    Redstem   Waterplantain   Sedge   Sprangletop       (mg/mL)   % control   % control   % control   % control                    UTC   0   0   0   0       0—no thx A   75   8   0   90       0.1   100   85   87   100       0.2   97   87   88   100       0.4   100   85   100   100                    
As a conclusion, Clincher CA (29.6% cyhalofop by weight) applied at half label rate (6.5 oz/acre) has good efficacy against grass weeds—not so good on broadleaves and poor on sedges. A combination of Clincher CA (cyhalofop) and thaxtomin A provides good control of all rice weeds tested in this study. Efficacy of thaxtomin A against grass weeds is substantially improved if combined with Clincher. Combination of thaxtomin A with Clincher CA did not cause phytotoxicity on rice at any tested concentration.
 
     Example 8 
     Penoxsulam(2-(2,2-difluoroethoxy)-N-(5,8-dimethoxyl[1,2,4]triazolo[1,5c]pyrimidin-2-yl)-6-trifluoromethyl)benzenesulfonamide) is mixed together with adjuvant containing ethyl oleate, polyethylene dialky ester and ethoxylated nonylphenol (2.5% v/v) and increasing concentrations of thaxtomin A (purified from the ATCC strain 49003) at concentrations 0.1, 0.2 and 0.4 mg/ml. The concentrations of the 2-[4-(4-cyano-fluorophenoxy)phenoxy]propanoic acid, butyl ester or 2-(2,2-difluoroethoxy)-N-(5,8-dimethoxy[1,2,4]triazolo[1,5c]pyrimidin-2-yl)-6-trifluoromethyl)benzenesulfonamide before dilution are 29.6% (2.38 lb/gal) and 21.7% (2 lb/gal), respectively. The effect of these mixtures on the growth of common water plantain, red stem, smallflower sedge and sprangletop is determined in the greenhouse. Similarly, rice plants of variety M104 are grown and tested for phytotoxic effects, and all plants are evaluated 7, 14, and 21 days after treatment. 
     Example 9 
     A strain of  S. acidiscabies  was grown in oat bran broth for 5 days (25° C., 200 rpm). The whole cell broth was extracted using XAD resin, and the dried crude extract was resuspended in 4% ethanol and 0.2% non-ionic surfactant at a concentration of 10 mg/mL. The diluted extracts containing 0.2 and 0.4 mg thaxtomin A per mL were tested on three weed species (redstem;  Ammania  spp., smallflower umbrella sedge;  Cyperus difformis  and sprangletop:  Leptochloa uninervia ). Other treatments included sarmentine at 2.5 and 5.0 mg/mL, and a combination treatment containing 0.2 mg thaxtomin A and 2.5 mg sarmentine per mL. Each treatment was applied in three replicates. Treated plants were kept in a greenhouse under 12 h light/12 h dark conditions. Results from an evaluation performed 25 days after treatment are presented in Table 5. 
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 Efficacy of herbicidal treatments using thaxtomin A  
               
               
                 (0.2 and 0.4 mg/mL) or sarmentine (2.5 or 5.0 mg/mL)  
               
               
                 alone or in combination (0.2 + 2.5 mg/mL) to control  
               
               
                 broadleaf, sedge and grass weeds in a greenhouse study. 
               
             
          
           
               
                   
                   
                 Control  
                 Control  
                 Control of 
               
               
                   
                   
                 of redstem 
                 of sedge 
                 sprangletop  
               
               
                   
                 Treatment 
                 (%) 
                 (%) 
                 (%) 
               
               
                   
                   
               
             
          
           
               
                   
                 UTC 
                   0a 
                   0a 
                   0a 
               
               
                   
                 Thaxtomin A 0.2 mg/mL 
                 5.0b 
                 48.3b 
                 8.3ab 
               
               
                   
                 Thaxtomin A 0.4 mg/mL 
                 11.7c* 
                 91.7d 
                 10.0b 
               
               
                   
                 Thaxtomin 0.2 + 
                 11.7c* 
                 61.7c 
                 73.3c 
               
               
                   
                 Sarmentine 2.5 mg/mL 
                   
                   
                   
               
               
                   
                 Sarmentine 2.5 mg/mL 
                   0a 
                  8.3a 
                 80.0c 
               
               
                   
                 Sarmentine 5.0 mg/mL 
                 2.5ab  
                  6.7a 
                 92.3d 
               
               
                   
                   
               
               
                   
                 *stunted. In a column, Means followed by the same letter are not statistically different from each other at p &lt; 0.05. 
               
             
          
         
       
     
     Thaxtomin A at the highest concentration of 0.4 mg/mL provides excellent control of sedge but poor control of the grass weed (sprangletop). When combined with sarmentine, the efficacy against grass weeds improves significantly. Also, efficacy against sedge is improved with the combination treatment compared with the single application of thaxtomin A alone at the corresponding concentration. In this study, the control of the broadleaf weed (redstem) is poor with all treatments. 
     Example 10 
     Bialaphos is produced by bacteria  Streptomyces  spp. and its synthesized active ingredient glufosinate is marketed as Rely® 200 (Bayer CropScience, Research Triangle Park, N.C.). Bialaphos has a non-selective property and inhibits the activity of glutamine synthetase, an enzyme involved in the synthesis of the amino acid glutamine. 
     The MBI-005 and bialaphos were prepared at various concentrations either as single ingredients or in combination with bialaphos. The treatments were applied at approximately ⅔ ml per replicate with a hand-help spray nozzle to barnyard grass or sedge. There were 3 replicates per treatment which were randomized after spray and were kept in a greenhouse at 25° C. for evaluation of phytotoxicity (% control). 
     When MBI-005 was mixed with bialaphos, the efficacy was increased several times more than when they were used alone (Table 6, 7, and 8). At higher rates of the mixtures, 100% control was achieved (Table 8). Synergy was observed when bialaphos at 0.178 mg/mL was mixed with MBI-005 at 0.25 mg/mL, and about 42% efficacy was achieved when the rate of bialaphos was increased close to 1.0 mg/mL from 10% control with bialaphos alone (Table 7). 
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                 Effects of bialaphos, MBI-005 (thaxtomin A), and the combinations  
               
               
                 of bialaphos with MBI-005 in controlling barnyard grass. 
               
             
          
           
               
                   
                 Bialaphos 
                 MBI-005 
                 % Control 
                   
               
               
                 Treatment 
                 (mg/mL) 
                 (mg/L) 
                 (14 days) 
                 E/Ee #   
               
               
                   
               
             
          
           
               
                 Untreated Control 
                   
                   
                 0.0 a* 
                   
               
               
                 (deionized water) 
                   
                   
                   
                   
               
               
                 Bialaphos 
                 0.089 
                   
                 0.0 a  
                   
               
               
                 Bialaphos 
                 0.178 
                   
                 0.0 a  
                   
               
               
                 Bialaphos 
                 0.356 
                   
                 0.0 a  
                   
               
               
                 Bialaphos 
                 0.534 
                   
                 0.0 a  
                   
               
               
                 Bialaphos 
                 0.712 
                   
                 0.0 a  
                   
               
               
                 Bialaphos 
                 0.890 
                   
                 1.3 ab 
                   
               
               
                 Bialaphos 
                 1.068 
                   
                  5.0 abc 
                   
               
               
                 MBI-005 (thaxtomin A) 
                   
                 0.25 
                 1.3 ab 
                   
               
               
                 Bialaphos + MBI-005 
                 0.089 
                 0.25 
                 1.3 ab 
                 1.0 
               
               
                 Bialaphos + MBI-005 
                 0.178 
                 0.25 
                  3.8 abc 
                 3.0 
               
               
                 Bialaphos + MBI-005 
                 0.356 
                 0.25 
                 11.9 c  
                 9.5 
               
               
                 Bialaphos + MBI-005 
                 0.534 
                 0.25 
                 29.4 d  
                 23.5 
               
               
                 Bialaphos + MBI-005 
                 0.712 
                 0.25 
                 34.4 d  
                 27.5 
               
               
                 Bialaphos + MBI-005 
                 0.890 
                 0.25 
                 59.4 e  
                 23.9 
               
               
                 Bialaphos + MBI-005 
                 1.068 
                 0.25 
                 62.5 e  
                 10.1 
               
               
                   
               
               
                 *Treatment means in each column marked with the same letter are not statistically different at LSD at p = 0.05 level. 
               
               
                   # Synergy is calculated from Colby&#39;s formula (Colby, 1967. Weeds 15: 20-22): Ee = X + Y − (XY/100) (Where E is the observed efficacy of product A + B, Ee is expected efficacy of A + B, and X and Y are the efficacy of product A or B when used alone. If E/Ee &lt; 1 the combination is antagonistic; if E/Ee = 1 the combination is additive; if E/Ee &gt; 1 the combination is synergistic). 
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 7 
               
             
             
               
                   
               
               
                 Effects of bialaphos, MBI-005 (thaxtomin A), and the combinations  
               
               
                 of bialaphos with MBI-005 in controlling barnyard grass. 
               
             
          
           
               
                   
                 Bialaphos 
                 MBI-005/011 
                 % Control 
                   
               
               
                 Treatment 
                 (mg/mL) 
                 (mg/mL) 
                 (14 days) 
                 E/Ee 
               
               
                   
               
             
          
           
               
                 Untreated Control 
                   
                   
                 0.0 a* 
                   
               
               
                 (deionized water) 
                   
                   
                   
                   
               
               
                 Bialaphos 
                 0.18 
                   
                 6.7 bc 
                   
               
               
                 Bialaphos 
                 0.53 
                   
                 3.3 ab 
                   
               
               
                 Bialaphos 
                 1.07 
                   
                 10.0 c  
                   
               
               
                 MBI-005 (thaxtomin A) 
                   
                 0.25 
                 6.7 bc 
                   
               
               
                 Bialaphos + MBI-005 
                 0.18 
                 0.25 
                  5.0 abc 
                 0.4 
               
               
                 Bialaphos + MBI-005 
                 0.53 
                 0.25 
                 25.0 d  
                 2.6 
               
               
                 Bialaphos + MBI-005 
                 1.07 
                 0.25 
                 41.7 e  
                 2.6 
               
               
                   
               
               
                 *Treatment means in each column marked with the same letter are not statistically different with LSD test at p = 0.05 level 
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 8 
               
             
             
               
                   
               
               
                 Effects of bialaphos, MBI-005 (thaxtomin A), and the combinations of  
               
               
                 bialaphos with MBI-005 or MBI-011 in controlling barnyard grass. 
               
             
          
           
               
                   
                   
                   
                 %  
                 %  
                   
               
               
                   
                   
                   
                 Control 
                 Control 
                 E/Ee 
               
               
                   
                 Bialaphos 
                 MBI-005 
                 (7  
                 (14  
                 (7  
               
               
                 Treatment 
                 (mg/mL) 
                 (mg/mL) 
                 days) 
                 days) 
                 days) 
               
               
                   
               
             
          
           
               
                 Untreated Control 
                   
                   
                   
                  0.0 a 
                   
               
               
                 (deionized water) 
                   
                   
                  0.0 a* 
                   
                   
               
               
                 Bialaphos 
                 1.1 
                   
                  5.0 ab 
                  3.7 a 
                   
               
               
                 Bialaphos 
                 1.4 
                   
                 10.8 b  
                 23.3 b 
                   
               
               
                 Bialaphos 
                 1.8 
                   
                 62.5 c  
                 66.7 c 
                   
               
               
                 MBI-005  
                   
                 0.38 
                  6.7 ab 
                 21.7 b 
                   
               
               
                 (thaxtomin A) 
                   
                   
                   
                   
                   
               
               
                 Bialaphos +  
                 1.1 
                 0.38 
                 87.5 d  
                 100.0 d  
                 7.7 
               
               
                 MBI-005 
                   
                   
                   
                   
                   
               
               
                 Bialaphos +  
                 1.4 
                 0.38 
                 87.5 d  
                 100.0 d  
                 5.2 
               
               
                 MBI-005 
                   
                   
                   
                   
                   
               
               
                 Bialaphos +  
                 1.8 
                 0.38 
                 87.5 d  
                 100.0 d  
                 1.3 
               
               
                 MBI-005 
               
               
                   
               
               
                 *Treatment means in each column marked with the same letter are not statistically different with LSD at p = 0.05 level. 
               
             
          
         
       
     
     Example 11 
     The test species barnyard grass, ragweed, sedge, and broad-leaf mustard were used for the valuation of synergy between MBI-005 and the rice herbicides clomazone, penoxsulam, cyhalofop, fenoxaprop-p-ethyl, bispyribac-sodium, thiobencarb, and propanil. 
     The common turf weeds dandelion and plantain were used in testing for synergy between MBI-005 and 2,4- or dicamba, two common turf herbicides. 
     Three other herbicides commonly used for field crops, glyphosate, glufosinate, synthetic version of bialaphos, and mesotrione were also tested for synergy with MBI-005 on crabgrass and ragweed. 
     There were 3 replicates per treatment which were sprayed with approximately ⅔ ml per replicate. The treatments were completely randomized and kept in a greenhouse at 25° C. The efficacy was rated at 7 and 14 days post treatment. The results are shown in Table 9 to 11. For barnyard grass control, MBI-005 had synergistic effects when combined with clomazone, (penoxsulam, bispyribac-sodium, thiobencarb, and propanil (Table 9). MBI-005 had additive effects when combined with cyhalofop, and fenoxaprop-p-ethyl (Table 10). 
     MBI-005 showed great synergy with glyphosate for controlling ragweed and also showed synergy with both turf herbicides. The synergistic effect of MBI-005 with glufosinate (synthetic bialaphos) (Table 11) on crabgrass was likely less since the rate of MBI-005 was too low. 
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 9 
               
             
             
               
                   
               
               
                 Summary of synergistic or additive effects between MBI-005 (thaxtomin 
               
               
                 A) and commercial products for rice weed control. The data are means 
               
               
                 of percentage control of three replicates 14 days post treatment. 
               
             
          
           
               
                   
                   
                 Product 
                 MBI-005 
                 % 
                 % 
                 % Control 
                   
               
               
                   
                 Test 
                 Rate 
                 Rate 
                 Control 
                 Control 
                 Product + 
               
               
                 Active Ingredient 
                 Species 
                 (mg/mL) 
                 (mg/mL) 
                 Product 
                 MBI-005 
                 MBI-005 
                 E/Ee 
               
               
                   
               
             
          
           
               
                 clomazone 
                 Barnyard 
                 0.513 
                 0.25 
                 37.5 
                 28.3 
                 91.7 
                 1.7 
               
               
                   
                 grass 
               
               
                   
                 Mustard 
                 0.513 
                 0.25 
                 25.0 
                 58.3 
                 70.8 
                 1.0 
               
               
                   
                 Sedge 
                 0.501 
                 0.02 
                 0.0 
                 37.5 
                 50.0 
                 1.3 
               
               
                 penoxsulam 
                 Barnyard 
                 0.051 
                 0.125 
                 25.0 
                 17.5 
                 75.0 
                 2.0 
               
               
                   
                 grass 
               
               
                   
                 Sedge 
                 0.047 
                 0.01 
                 66.7 
                 20.0 
                 87.5 
                 1.2 
               
               
                 cyhalofop 
                 Mustard 
                 0.051 
                 0.125 
                 11.7 
                 13.3 
                 15.0 
                 0.6 
               
               
                   
                 Sedge 
                 1.176 
                 0.01 
                 3.3 
                 20.0 
                 8.3 
                 0.4 
               
               
                 fenoxaprop-p-ethyl 
                 Barnyard 
                 0.006 
                 0.25 
                 91.7 
                 66.7 
                 87.5 
                 0.9 
               
               
                   
                 grass 
               
               
                   
                 Mustard 
                 0.006 
                 0.125 
                 0.0 
                 45.8 
                 33.3 
                 0.7 
               
               
                   
                 Sedge 
                 0.116 
                 0.02 
                 0.0 
                 70.8 
                 66.7 
                 0.9 
               
               
                 bispyribac-sodium 
                 Barnyard 
                 0.032 
                 0.125 
                 0.0 
                 3.3 
                 62.5 
                 18.8 
               
               
                   
                 grass 
               
               
                   
                 Mustard 
                 0.0216 
                 0.25 
                 53.3 
                 37.5 
                 95.8 
                 1.4 
               
               
                   
                 Ragweed 
                 0.0216 
                 0.125 
                 5.0 
                 23.3 
                 50.0 
                 1.8 
               
               
                 thiobencarb 
                 Barnyard 
                 1.743 
                 0.25 
                 41.7 
                 41.7 
                 79.2 
                 1.2 
               
               
                   
                 grass 
               
               
                   
                 Mustard 
                 1.743 
                 0.125 
                 10.0 
                 54.2 
                 54.2 
                 0.9 
               
               
                   
                 Sedge 
                 3.15 
                 0.02 
                 58.3 
                 37.5 
                 70.8 
                 1.0 
               
               
                 propanil 
                 Barnyard 
                 0.365 
                 0.125 
                 32.5 
                 10.0 
                 79.2 
                 2.0 
               
               
                   
                 grass 
               
               
                   
                 Mustard 
                 0.036 
                 0.25 
                 1.67 
                 58.3 
                 50.0 
                 0.9 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 10 
               
             
             
               
                   
               
               
                 Summary of synergistic or additive effects between MBI-005 (thaxtomin 
               
               
                 A) and commercial products for turf weed control. The data are means 
               
               
                 of percentage control of three replicates 14 days post treatment. 
               
             
          
           
               
                   
                   
                   
                 MBI-005 
                   
                   
                 % Control 
                   
               
               
                 Active 
                 Weed 
                 Product Rate 
                 Rate 
                 % Control 
                 % Control 
                 Product + 
               
               
                 Ingredient 
                 Species 
                 (mg/mL) 
                 (mg/mL) 
                 Product 
                 MBI-005 
                 MBI-005 
                 E/Ee 
               
               
                   
               
             
          
           
               
                 2,4-D 
                 Dandelion 
                 0.176 
                 0.1 
                 25.0 
                 20.0 
                 91.7 
                 2.3 
               
               
                   
                 Plantain 
                 2.340 
                 0.24 
                 50.0 
                 25.0 
                 83.3 
                 1.3 
               
               
                 dicamba 
                 Dandelion 
                 0.121 
                 0.1 
                 45.8 
                 20.0 
                 75.0 
                 1.3 
               
               
                   
                 Plantain 
                 6.025 
                 0.12 
                 50.0 
                 10.0 
                 79.2 
                 1.4 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 11 
               
             
             
               
                   
               
               
                 Summary of synergistic or additive effects between MBI-005 (thaxtomin 
               
               
                 A) and commercial products with broad spectrum. The data are means 
               
               
                 of percentage control of three replicates 14 days post treatment. 
               
             
          
           
               
                   
                   
                   
                 MBI-005 
                   
                   
                 % Control 
                   
               
               
                 Active 
                 Weed 
                 Product Rate 
                 Rate 
                 % Control 
                 % Control 
                 Product + 
               
               
                 Ingredient 
                 Species 
                 (mg/mL) 
                 (mg/mL) 
                 Product 
                 MBI-005 
                 MBI-005 
                 E/Ee 
               
               
                   
               
             
          
           
               
                 glyphosate 
                 Crabgrass 
                 0.754 
                 0.125 
                 45.8 
                 28.3 
                 83.3 
                 1.4 
               
               
                   
                 Ragweed 
                 2.198 
                 0.125 
                 15.0 
                 6.7 
                 29.2 
                 1.4 
               
               
                 glufosinate 
                 Crabgrass 
                 0.151 
                 0.125 
                 83.3 
                 20.0 
                 75.0 
                 0.9 
               
               
                 (bialaphos) 
                 Ragweed 
                 0.194 
                 0.125 
                 22.5 
                 37.5 
                 75.0 
                 1.5 
               
               
                 mesotrione 
                 Crabgrass 
                 0.24 
                 0.125 
                 62.5 
                 24.2 
                 75.0 
                 1.1 
               
               
                   
                 Ragweed 
                 0.96 
                 0.125 
                 25.0 
                 15.0 
                 45.8 
                 1.3 
               
               
                   
               
             
          
         
       
     
     Although this invention has been described with reference to specific embodiments, the details thereof are not to be construed as limiting, as it is obvious that one can use various equivalents, changes and modifications and still be within the scope of the present invention. 
     Various references are cited throughout this specification, each of which is incorporated herein by reference in its entirety. 
     CITED REFERENCES 
     Beauséjour, J., C. Goyer, et al. (1999). “Production of thaxtomin A by  Streptomyces scabies  strains in plant extract containing media.”  Can J Microbiol  45: 764-768. 
     Duke, S. O., S. R. Baerson, et al. (2003). “United States Department of Agriculture-Agricultural Research Service research on natural products for pest management.”  Pest Manag Sci  59: 708-717. 
     Duke, S. O., F. E. Dayan, et al. (2000). “Natural products as sources of herbicides: current status and future trends.”  Weed Research  40: 99-111. 
     Fry, B. A. and R. Loxia (2002). “Thaxtomin A: Evidence for a plant cell wall target.”  Physiological and Molecular Plant Pathology  60: 1-8. 
     Gerwick, B. C., P. R. Graupner, et al. (2005). Methylidene mevalonates and their use as herbicides. U. p. 7393812: 16. 
     Healy, F. G., M. J. Wach, et al. (2000). “The txtAB genes of the plant pathogen  Streptomyces  acidiscabies encode a peptidesynthetase required for phytotoxin thaxtomin A production and pathogenicity.”  Molecular Microbiology  38: 794-804. 
     Hiltunen, L. H., I. Laakso, et al. (2006). “Influence of thaxtomins in different combinations and concentrations on growth of micropropagated potato shoot cultures.”  J Agric Food Chem  54: 3372-3379. 
     Hoagland, R. E. (2001). “Microbial allelochemicals and pathogens as bioherbicidal agents.”  Weed Technology  15: 835-857. 
     Kang, Y., S. Semones, et al. (2008). Methods of controlling algae with thaxtomin and thaxtomin compositions. USA, Novozymes Biologicals, Inc. 
     King, R. R., C. H. Lawrence, et al. (1992). “Chemistry of phytotoxins associated with  Streptomyces scabies , the causal organism of potato common scab.”  J. Agric. Food Chem  40: 834-837. 
     King, R. R., C. H. Lawrence, et al. (1989). “Isolation and characterization of phytotoxin associated with  Streptomyces scabies.” Journal of the Chemical Society, Chemical Communications  13: 849-850. 
     King, R. R., C. H. Lawrence, et al. (2003). “More chemistry of the thaxtomin phytotoxins.”  Phytochemistry  64: 1091-1096. 
     King, R. R., C. H. Lawrence, et al. (2001). “Herbicidal properties of the thaxtomin group of phytotoxins.”  J Agric Food Chem  49: 2298-2301. 
     Loxia, R., R. A. Bukhalid, et al. (1995). “Differential production of thaxtomins by pathogenic  Streptomyces  species in vitro”  Phytopathology  85: 537-541.

Technology Category: 1