HERBICIDAL COMBINATIONS OF MICROBIAL FERMENTATION PRODUCTS AND CHEMICAL AGENTS

Abstract: A method is provided for controlling the growth of weeds by applying to the weeds a herbicide, or other chemical agent, and a culture medium in which microorganisms such as bacteria or fungi have grown. The herbicidal action of the chemical herbicide when combined with the conditioned medium is greater than the herbicidal action of the herbicide alone. This makes it possible to control a broad range of weed species using lower levels of herbicide than are needed when the herbicide is used alone. The medium may be processed to concentrate it or to partially purify it, but it is not necessary to identify or to purify the specific agents in the medium that act to enhance the actions of the chemical agent.

Description: Description HERBICIDAL COMBINATIONS OF MICROBIAL FERMENTATION PRODUCTS AND CHEMICAL AGENTS Technical Field This application is a continuation-in-part application of copending U.S. Patent Application Serial No. 07/614,118, filed November 16, 1990, the entire disclosure of which is relied upon and incorporated by reference herein. Background Art The present invention relates to a method for controlling the growth of weeds. Weeds treated with a chemical herbicide are additionally treated with constituents from culture media in which microorganisms have grown. This results in a greater degree of control of the weeds than is obtained by the same level of chemical herbicide alone. Widespread use of chemical herbicides has created concern about possible adverse ecological effects caused by the large quantities of chemical agents that are being introduced into the environment. There are additional fears that these chemicals might pose risks to human and animal health. Special concern has been directed to health effects on agricultural workers, who may be exposed to high levels of chemical herbicides. It has been necessary to withdraw from the market or severely restrict the use of some chemical herbicides after adverse environmental or health effects became evident. Disposal of containers for chemical herbicides is heavily regulated, and proper disposal of these containers can be inconvenient and expensive for farmers. Disposal sites for these containers are limited and illegal container disposal is a growing concern. Thus, there exists a need in the art for compositions and methods for controlling a broad range of weed species SUBSTITUTESHEET while decreasing the quantity of herbicidal chemicals currently used for this purpose. One approach to the development of less environmentally harmful methods of weed control is to attempt to identify and isolate herbicidal substances produced by microorganisms. Many microorganisms produce substances that are toxic to weeds. An assumption underlying this approach is that these naturally-occurring agents are less likely to have deleterious environmental effects than the synthetic chemicals presently used as herbicides. Effort has been directed at identifying and isolating such compounds made by microorganisms for possible use as herbicides. The strategy in these studies has been to identify and isolate specific substances produced by microorganisms that are toxic to plants. To discover naturally occurring herbicides, culture media in which microorganisms have grown have been tested for their effects on weeds. If the medium produced signs of injury in the weeds, then efforts were directed towards identifying and isolating the chemical constituents in the medium that have herbicidal actions. The isolation and identification of herbicidal compounds made by microorganisms faces numerous obstacles. The process of identifying, purifying, and isolating such compounds is lengthy and expensive. Media in which microorganisms have grown are highly complex mixtures and the active compounds with herbicidal activity may be present in extremely low concentrations. If the active compounds are labile, or if herbicidal actions are due to synergistic interactions of multiple components, then the activity may be lost during purification. Even where an active compound is identified, the production of commercially useful quantities of such compounds by industrial fermentation and purification or by alternative methods involving chemical synthesis may require considerable additional effort. SUBSTITUTE SHEET Perhaps because of the difficulties and expense of isolating microbially-derived herbicidal compounds, only a few natural products made by microorganisms have been developed to the stage where they could be tested as potential commercial herbicides. One example is bialaphos, L-2-amino-4-[ (hydroxy) (methyl)phosphinoyl]-butyl-L-alanyl-L- alanine. A synthetic version of a derivative of bialaphos, glufosinate (IGNITE®), is also used as a herbicide. Efforts to develop herbicides from microbial phytotoxins are described in Duke, S.O., "Microbially produced phytotoxins as herbicides — a perspective," in Putnam, A.R. and Tang, C.-S. (Eds.) The Science Of Allelopathy. New York, John Wiley & Sons, 1986, p.287. Toxins produced by both bacteria and fungi have been tested as potential herbicides. A second approach to using the products of microorganisms for weed control is to use the living microorganisms themselves as bioherbicides. Most of the living organisms that have been developed as bioherbicides are fungi (mycoherbicides) . Fungal plant pathogens are generally highly selective, infecting only a single species or a narrow range of plants. Therefore, mycoherbicides have been used mainly as selective herbicides directed at specific species of weeds. For example, U.S. Patent No. 3,999,973 to Templeton teaches the use of Colletotrichu malvarum as a mycoherbicide for controlling prickly sida (Sida spinosa L.). Fungi have not been developed as broad spectrum mycoherbicides for the control of multiple weed species. For a discussion of the use of living fungi as herbicides, see R. Charudattan, "The Use of Natural and Genetically Altered Strains of Pathogens for Weed Control" in M.A. Hoy and D.C. Herzog (eds. ), Biological Control in Agricultural IPM Systems. N.Y. : Academic Press, 1985, pp. 347-372 and G.E. Templeton, "Biological Control of Weeds," A er. J. Alternative Agr. 3: 69-72 (1989). TITUTE SHEET Bacterial plant pathogens have not been extensively developed as bioherbicides. Bacteria that may have some application for weed control are described in Cherrington, C. A. and Eiliott, L. F., Incidence of inhibitory pseudomonads in the Pacific Northwest, Plant and Soil 101; 159-165 (1987); Elliott, L. F. and Kennedy, A. C, Effect of deleterious rhizobacteria on the growth of downy brome (Bromus tectorum L. ) - Agronomy Abstracts p. 181 (1987); and Kremer, R. J., Identity and properties of bacteria inhabiting seeds of selected broadleaf weed seeds, Microbial Ecology 14; 29-37 (1987). While the use of living microorganisms as bioherbicides has potential advantages compared to the use of chemical herbicides, it entails numerous problems. Microorganisms must be kept viable during storage and application. Successful infection of the targeted weeds may be adversely affected by environmental conditions. While some fungi have been applied for the selective control of specific weeds, living microorganisms have not previously been useful as broad spectrum herbicides. United States Patent Application No. 07/601,653, incorporated herein by reference, describes a method for the control of weeds by using a chemical herbicide in combination with a bacterial plant pathogen. The corresponding PCT application, PCT/US90/05031, which was filed on September 10, 1990, and was published on March 21, 1991 (International Publication Number WO 91/013161), is also incorporated herein by reference. The actions of the chemical herbicide facilitate opportunistic infections of the weeds by the bacteria. This makes it possible to control a broad range of weed species using lower levels of herbicide than are needed when the herbicide is used alone. In addition, the range of weed species controlled may be greater than the spectrum controlled by the herbicide alone. SUBSTITUTE SHEET The present invention employs a different strategy. It does not entail the laborious isolation of specific toxins produced by microorganisms. It also avoids some of the problems inherent in the use of living microorganisms in the field as bioherbicides. Rather, media in which microorganisms have grown are used to potentiate the actions of chemical herbicides, making it possible to employ lower concentrations of the chemical herbicides. With this approach, it is unnecessary to engage in the lengthy and expensive process of isolating the active herbicidal factors in the conditioned media. Rather, the conditioned media can be used as is, or after processing involving simple procedures such as concentration and partial purification, to provide an economically useful method of effectively controlling weeds while reducing the amounts of conventional herbicides applied to the environment. Disclosure of the Invention It has been discovered that culture medium in which microorganisms have grown ("conditioned medium") can be combined with chemical herbicides to control weeds, wherein the combination of chemical herbicide and conditioned medium produces a greater degree of injury than is produced by the herbicide or conditioned medium alone. The conditioned medium need not be purified and phytotoxic materials in the conditioned medium need not be identified in order to use this approach for effective weed control. With this approach to the control of weeds, it is possible to reduce the amount of chemical herbicides that are introduced into the environment. The present invention overcomes the problems and disadvantages of the prior art by providing a method to control the growth of a broad spectrum of weeds while reducing the amount of chemical herbicides used to achieve this goal. By combining chemical agents, such as herbicides, with fermentation products in media in which microorganisms have grown, greater weed control is achieved than is obtained by the same amount of chemical alone. Thus it is possible to control the growth of weeds while reducing the amount of chemical introduced into the environment, thereby reducing the worker exposure, environmental burden and container disposal problems associated with the use of chemical herbicides. In addition, a general method for broad spectrum control of weed growth is provided that is safer than current methods, inexpensive and effective. The present invention comprises a method of controlling weeds by applying to the weeds a chemical herbicide combined with medium conditioned by the growth of microorganisms, wherein the action of the herbicide is enhanced by the conditioned medium and wherein conditioned medium is effective to enhance the activity of the chemical herbicide. The chemical agent and conditioned medium are applied in amounts wherein killing of the weeds or the inhibition of weed growth is greater than would be caused by the same amounts of the chemical herbicide or conditioned medium applied alone. Another embodiment of the invention comprises a method of controlling the growth of weeds by applying a chemical herbicide together with conditioned medium which has been further processed by techniques such as concentration, filtration or fractionation, wherein inhibition of weed growth or killing of the weeds is greater than would be obtained by the same amounts of herbicide or processed conditioned medium applied alone. Another embodiment of the invention comprises a method for controlling weeds to prevent or inhibit weed growth which comprises applying to soil containing the seeds of weeds a herbicide and a conditioned medium, wherein the action of the herbicide on the seeds is enhanced by the conditioned medium, and wherein the conditioned medium is effective to enhance the activity of the herbicide, so that the killing of the seeds or the inhibition of the growth of weeds developing SUBSTITUTE SHEET fro the seeds is greater than would be caused by the same amounts of the herbicide or conditioned medium applied alone. Another embodiment of the invention comprises a herbicidal composition comprising a chemical herbicide in combination with a conditioned medium or processed conditioned medium present in an amount sufficient to enhance the activity of the chemical herbicide. Additional objects and advantages of the invention will be set forth in part in the description which follows, and in part will be apparent from the description or may be learned by practice of the invention. The objects and advantages of the invention will be realized and attained by means of the methods and combinations particularly pointed out in the appended claims. The following general description is offered by way of explanation and illustration, and not by way of limitation. Brief Description of the Drawings Figure 1 shows the effects of conditioned medium on herbicidal actions of sulfosate applied at 0.06 lbs/acre (0.067 kg/ha). Figure 2 shows the effects of conditioned medium on herbicidal actions of sulfosate applied at 0.125 lbs/acre (0.140 kg/ha) . Figure 3 shows the effects of conditioned medium on herbicidal actions of sulfosate applied at 0.25 lbs/acre (0.28 kg/ha) . Best Mode for Carrying Out the Invention For purposes of this application, a weed is "any plant that is objectionable or interferes with the activities or welfare of man" at the location where it is growing. A herbicide (here also called a chemical herbicide) is a "chemical used to control, suppress, or kill plants, or to severely interrupt their normal growth processes. " Herbicide Handbook of the Weed Society of America, Fifth Edition (1983), xxi-xxiv. Culture medium or medium is used here to refer to compositions prepared to support the growth and maintenance of microorganisms. "Conditioned medium" refers to any such medium in which microorganisms have grown. Medium in which microorganisms have grown is sometimes referred to as "spent medium" or "spent broth." The term "conditioned medium" is used here because the intended meaning is analogous to the use of this term in other contexts such as the tissue culture of eukaryotic cells, where emphasis is placed on the fact that the biological properties of the medium have been changed as a result of the growth of cells in the medium. The term "processed conditioned medium" will be used here to refer to materials derived from conditioned medium by processing the conditioned medium with treatments that alter its composition, where the processed product is a relatively complex mixture of constituents resulting from the interaction of microorganisms with culture medium or derivatives of such constituents. Such treatments may include, but are not limited to, concentration, drying, lyophilization, filtration, ultrafiltration, dialysis, removal of sediment by centrifugation, pasteurization, freezing, boiling, adjustment of pH, and the addition of chemicals such as preservatives, stabilizers, and agents to kill the microorganisms. This definition also includes partial purification by means including, but not limited to concentration, extraction, fractionation, and precipitation. The term "processed conditioned medium" is intended to exclude highly purified compounds isolated from conditioned culture media. After processing, the resulting processed conditioned medium remains a type of conditioned medium, and the general term "conditioned medium" encompasses processed conditioned media. To practice the invention, a chemical herbicide and conditioned medium (which may be a processed conditioned medium), are both applied topically to weeds or to soil SUBSTITUTE SHEET containing seeds of weeds. The chemical agent and conditioned medium are applied by well-known methods of application such as spraying or as a soil drench. Application may be preemergent, postemergent or as a nonselective contact herbicide. In the embodiments described below, the application is postemergent. For preemergent application, the herbicide and the conditioned medium are applied to the seeds of weeds before the seeds have germinated, or at early stages of germination before the developing plant has emerged from the soil, typically by application to soil in which the seeds are located. The invention can be used to control weeds in an agricultural setting and can also be used for nonagricultural applications. For example, the invention can be used to control weeds in turf and as a contact bioherbicide for the management of roadside vegetation. The chemical herbicide and the conditioned medium are generally applied simultaneously, usually in a single mixture. However, the chemical and the conditioned medium can also be applied from separate tanks, and may be applied sequentially. Compositions for practice of the invention can be formulated in numerous ways, including flowables, dry flowables, water dispersible granules or emulsified concentrates. While the embodiments described below each entail the use of a single herbicide and a single conditioned medium, mixtures of media in which different species or pathovars of microorganisms have been cultured can be used to practice of this invention. The invention can also be practiced by applying a mixture of different chemical agents in conjunction with the conditioned medium. Furthermore, the invention can be practiced in conjunction with a bioherbicide, i.e., a medium conditioned by one microorganism can be used together with a different microorganism which acts as a bioherbicide. SUBSTITUTE SHEET Various chemical herbicides can be used to practice this invention. The embodiments described below employed sulfosate, glyphosate, glufosinate, paraquat, fluazifop, sethoxydim, imazapyr, chlorimuron, dicamba, bentazon, fomesafen, and imazethapyr as chemical agents. Some additional chemical herbicides that can be used to practice this invention are listed in Table 2. Further examples of additional herbicides that can be used to practice this invention are described in Herbicide Handbook of the Weed Society of America, Sixth Ed. (1989), Farm Chemicals Handbook, Willoughby, Ohio; Meister Publishing Co., (1990), and in H. J. Lorenzini and L.S. Jeffrey, Weeds of the United States and Their Control. N.Y. ; Van Nostrand 1987, Table 1.9. While known chemical herbicides can be used to practice this invention, it should be appreciated that chemicals which do not fit the conventional definition of herbicide may also be used. For example, plant hormone analogues that are not currently useful alone as herbicides may nevertheless effectively control the growth of weeds when applied in combination with conditioned medium or processed conditioned medium. Examples of such chemical agents include phytohormones such as indole acetic acid derivatives and also include enzyme cofactors, allelopathic compounds, and analogues of amino acids, nucleic acid bases and vitamins. Microorganisms that can be used to condition their culture medium include bacteria and fungi. The invention here described can be practiced using naturally occurring microorganisms that are readily obtained from public depositories or isolated from the soil, plants, air, water, insects, etc. , using techniques well known in the art. It should be appreciated that microorganisms can also be modified to produce more potent conditioned medium. Well known techniques for the isolation or selection of mutants, either naturally occurring or induced, can be used to obtain bacteria that produce more effective conditioned media. This SUBSTITUTESHEET can also be done by modifying the microorganisms using well known techniques of genetic engineering and recombinant DNA. For example, the methods of genetic engineering can be used to insert the genes for the production of known phytotoxins into microorganisms that condition media by the production of other, possibly unknown, toxins. Various species of bacteria can be used to practice this invention. Pseudomonas syringae pv. tabaci has been found to be useful for this purpose. Other species that are known to produce phytotoxins may be used. Some examples of phytopathogenic bacteria that may used to practice this invention are listed in Table 3A. Other phytopathogenic bacteria may be selected from several prokaryotic genera including, but not limited to, Pseudomonas. Xanthomonas. Agrobacteriu , Erwinia, Corvnebacterium, Streptomyces, and Nocardia. See N.W. Schaad, How phytopathogenic bacteria are classified, in M.S. Mount and G.H. Lacy (eds.), Phytopathogenic Prokarvotes, Vol. 1, N.Y., Academic Press, 1982, pp. 19-29. It should be appreciated that bacteria that do not usually cause injury to plants may produce conditioned medium which can potentiate the actions of chemical herbicides. See, for example. Table 3B. Some fungi that can be used to practice this invention are listed in Table 4. Both phytopathogenic fungi (Table 4A) and fungi that are not usually phytopathogenic (Table 4B) may be used. A wide variety of liquid media can be used. The compositions of media appropriate for growing various species of microorganisms are well known in the art, as are methods for growing microorganisms and liquid media on a commercial scale. See, e.g., E.L. Demain and N.A. Solomon (Eds.), Manual of Industrial Microbiology and Biotechnology, Washington, D.C., American Society for Microbiology, 1986. In general, any medium that supports growth of the microorganisms will be conditioned by the microorganisms. SUBSTITUTESHEET Although liquid media are especially convenient for the practice of this invention, in principle pulverized solid media or extracts of solid media may be used. A particular microorganism can be used to condition a variety of different medium compositions, all of which will . be effective in potentiating the actions of herbicides. Whether a particular medium is preferable to others can easily be determined by testing under greenhouse or field conditions. Optimization of other parameters such as amount of herbicide, temperature, time of incubation, degree of aeration and agitation, etc., can also be determined by a person of ordinary skill by means of simple comparative testing. In some instances the composition of the medium may be modified to further enhance the potency of the conditioned medium by applying information known in the art concerning specific microorganisms. For example, in the embodiment in Example 1, below, the medium was supplemented with zinc, which has been reported to enhance the production of toxin by Pseudomonas syringae pv. tabaci. Durbin, R.D., and Uchytil, T.F., The Role of Zinc in Regulating Tabtoxin Production, Experientia 41; 136-37 (1985). However, subsequent experiments have shown that inclusion of zinc in the medium is not essential for the production of conditioned medium that enhances the action of herbicides. Thus, it will be understood that this invention can be practiced with or without a toxin-enhancing amount of zinc in the medium. In specific instances, the growth conditions also may be modified to further enhance the potency of the conditioned medium. For example, U.S. Patent No. 4,273,571 to Berg discloses the production of a herbicidal compound under aerobic conditions. In general, optimal conditions for production of conditioned medium will vary with different organisms and media. These can be determined through routine comparative testing under greenhouse or field conditions. SUBSTITUTE SHEET After medium has been conditioned by the growth of microorganisms, the living microorganisms must be either killed or must be removed from the medium. The microorganisms can be removed by well known methods such as sedimentation and microfiltration or ultrafiltration. Alternatively, the organisms may be killed by well known methods such as heating, irradiation, or the use of bacteriocidal or fungicidal chemicals. Although certain chemical herbicides are not toxic to specific microorganisms, other herbicides can be used to kill microorganisms in the medium. For example. United States Patent Application No. 07/601,653 discloses that the herbicide paraquat effectively kills Pseudomonas syringae pv. tabaci within one hour. Thus, with this particular combination of herbicide and microorganism, mixing the herbicide with the medium in preparation for application provides a convenient means for eliminating the live bacteria. After the bacteria have been killed, they may be removed from the conditioned medium, by sedimentation, filtration or other means. However, the invention can also be practiced without removing the dead bacteria from the conditioned medium. In addition to the removal of the microorganisms, a conditioned medium may be processed in other ways for convenience or to enhance its efficacy. Such treatments may include, but are not limited to, concentration, drying, lyophilization, filtration, ultrafiltration, dialysis, removal of sediment by centrifugation, pasteurization, freezing, boiling, adjustment of pH, and the addition of chemicals such as preservatives, stabilizers and agents to kill the microorganisms. Such treatments also include partial purification by means including, but not limited to concentration, extraction, fractionation, and precipitation. The components in the conditioned medium may enhance the action of the chemical herbicide through various mechanisms. SUBSTITUTE SHEET The microorganisms may produce known or unknown phytotoxins. In addition, a combination of toxins in the medium may „act synergistically. Furthermore, the effective components in the medium may not necessarily be toxins. Compounds in the medium, not otherwise phytotoxic, may have synergistic interactions with the herbicide, and may also have synergistic interactions with other components in the medium. For example. United States Patent 4,762,547 to Iwasaki et al. describes the enhancement of herbicidal action by the combination with esterases. They suggest that the esterases ' destroy the waxy outer layers of plants, facilitating penetration of the herbicide into the plant. United States Patent No. 4,436,547 to Sampson states that a variety of growth-enhancing additives including carbohydrates, fatty acids and vitamins, can enhance the action of herbicides. Mixtures of more than one conditioned medium, each medium being conditioned by a different microorganism, may be advantageous. Such mixtures may be especially effective where the different conditioned media facilitate the actions of herbicides through different mechanisms of action involving different fermentation products. Where such mixtures are used, the use of processed conditioned media that has been concentrated may be advantageous. The formulation of the invention may also contain adjuvants such as surfactants and suspension agents. Examples of adjuvants that can be used include spreaders such as ORTHO-77®, ATTAGEL-40® and X-77®, and organosilicones such as SILWETT L-77®, SILWETT L-7607®, X2-5309 (Dow Corning), and X2-5395 (Dow Corning) . Crop oils that can be used as adjuvants include Sure Crop Agri, Crop Surf, and CIDE-KICK®. Other adjuvants that might be used include SOY-DEX®, AGRI- DEX®, TRITON AG-98®, STEROX®, or invert emulsions. In practicing this broad approach to weed control, it should be understood that each particular combination of chemical agent and conditioned medium will not necessarily be SUBSTITUTE SHEET effective in controlling particular species of weeds. This is consistent with the behavior of broad range chemical herbicides in general, which are rarely, if ever, effective in controlling all types of weeds. Whether a particular combination of herbicide and conditioned medium or processed conditioned medium will be useful for a particular application can readily be determined by those skilled in the art without undue experimentation by testing in the greenhouse or in the field. Examples 2-4 illustrate the type of experiment that can be performed to determine the effectiveness of a given combination of conditioned medium and herbicide, as well as to optimize parameters such as the application rate for the herbicide. Certain principles may guide the optimization of parameters. First, the effect of the conditioned media is most evident when the herbicide is applied at a level where the herbicide alone produces weak or marginal effects on the weeds. Where the herbicide at a particular dosage already produces a high degree of weed control, additional benefits from the addition of conditioned media may not be apparent. Control is more apparent when the weed shows partial response to the level of chemical herbicide employed. Preferred levels for the application of three herbicides (sulfosate, glyphosate, and glufosinate) are illustrated in Examples 2-5 below. Effects of varying the concentration of one herbicide, sulfosate, are shown in Examples 2-3. Second, it will be apparent to those skilled in the art that while the rate of application of the chemical can advantageously be less than the usual field application rate (e.g., one-eighth of the recommended minimum field application rate for sulfosate in Example 2, below), some level of the herbicide will be too low to have a useful effect. However, further reductions may be possible for particular applications. SUBSTITUTE SHEET Practice of this invention may make it possible to achieve further reductions in the amount of chemical herbicides employed by giving users the option to replace the traditional prophylactic preemergent herbicide application with more efficient postemergent application. A postemergent chemical herbicide used in conjunction with conditioned medium could be applied selectively only as required, providing an alternative to heavy preventative preemergent application of herbicides. This approach has not been generally feasible in the past due to the cost and limited spectrum of most postemergent herbicides. The constituents of conditioned medium are expected to be naturally occurring substances. Thus, it can be anticipated that they will be biodegradable and should not produce long-term deleterious effects. Practice of this invention would have the additional advantage of reducing the costs associated with the disposal of chemical herbicide containers. This reduction would be accomplished by a reduction in the total amount of chemical herbicide used. The following examples are offered by way of illustration of the preferred embodiments and not by way of limitation, with the true scope of the invention being indicated by the claims which follow. Example 1 describes the preparation of a conditioned medium. EXAMPLE 1 A pathovar of Pseudomonas syringae pv. tabaci, a bacterium, was used to produce conditioned medium. The strain of Pseudomonas syringae pv. tabaci used was isolated from a tobacco plant in Maryland by Dr. Arvydas Grybauskas of the Department of Botany, University of Maryland, College Park, Maryland. The strain has been deposited with the American Type Culture Collection (12301 Parklawn Drive, Rockville, Maryland 20852) (ATCC #55090). Tobacco plants SUBSTITUTE SHEET (Nicotiana tabacum L.) were used periodically to confirm the virulence of this strain of Pseudomonas syringae pv. tabaci. The bacteria were grown in liquid culture in modified King's B medium (Table 1) supplemented with zinc. The cells were incubated in 2 liter Erlenmeyer flasks at 28° C. in a rotary shaker (New Brunswick G10 gyroshaker) at 180-190 r.p.m. for about 48-64 hours until the cells reached a q concentration of approximately 10 CFU/ml. After incubation, medium was centrifuged to remove the cells using a Sorval RC5C centrifuge at 8000 r.p.m. for 8 minutes at 10°C with a GSA rotor. Any cells that remained suspended in the supernatant were removed by microfiltratioή through a 0.2μm filter. Example 2 illustrates the effects of combining conditioned medium with a chemical herbicide, sulfosate. EXAMPLE 2 The effects on plants of combining low levels of a herbicide, sulfosate (TOUCHDOWN®), in combination with medium conditioned by Pseudomonas syringae pv. tabaci as described in Example 1, was tested under greenhouse conditions. Sulfosate (trimethylsulfonium carboxymethylaminomethyl- phosphonate; see U.S. Patent No. 4,315,765) was obtained from ICI Americas, Inc., Agricultural Products Division, Wilmington, Delaware. Sulfosate was applied at 0.06 lbs/acre (0.067 kg/ha), which is one-eighth of the recommended minimum field application rate. Application rates expressed in lbs/ acre (kilograms/hectare) refer to pounds (kilograms) of active ingredient per acre (hectare). The effects were evaluated on nine different weed species. Broadleaf weeds tested were morningglory (a mixture of Ipomea purpurea and I_ j _ hederacea . , velvetleaf (Abutilon theophrasti ) , cocklebur (Xanthium strumariuπO, redroot pigweed (Amaranthus retroflexus ) and jimsonweed (Datura stramonium ) . The grasses that were tested were barnyardgrass fEchinochloa crusgalli), yellow foxtail (Setaria glauca ) , johnsongrass (Sorghum halepense) and fall panicum (Panicum dichotomiflorum) . These were selected as being weeds of economic importance in row crops and as representing a spectrum of plant families. Effects were also tested on two major crops, corn and soybean. Both of these plants may become weeds when they appear in fields that are being cultivated for other crops. Seedlings were grown in 4-inch (10.2 cm) pots in a greenhouse under ordinary greenhouse conditions. Four treatments were compared: water alone, sulfosate alone, conditioned medium alone, or sulfosate plus conditioned medium. Sulfosate was applied at 0.06 lb/A (0.067 kg/ha), which is one-eighth of the minimum rate recommended for application in the field. To standardize the treatments, all solutions were sprayed on the weeds using a Model SB8 Spray Booth manufactured by DeVries Manufacturing (Hollandale, Minnesota) that was purchased from R & D Sprayers, Inc. (Opelousas, Louisiana). This device permits highly reproducible spraying of the test solutions. The sprayer was calibrated to deliver 25 gallons per acre (234 liters/ hectare) with a nozzle pressure of 23-25 pounds per square inch (1.6-1.7 bars). The speed of the traveling spray nozzle was 3.0 miles per hour (4.83 km/hr) . All solutions (including the water control treatment) contained 0.25 % (v/v) of ORTHO X-77® as an adjuvant. X-77® is a spreading agent sold by Chevron Chemical Company. According to the manufacturer, active agents in X-77® are alkylarylpolyoxyethylene glycols, free fatty acids and isopropanol. The experimental design was a randomized complete block design with at least 2 replications per treatment and 2 plants per replication. Injury was rated weekly on a scale of 0-100 with 0 = no injury and 100 = plant death, as described in Research Methods in Weed Science, 3rd Edition, Southern Weed Science SUBSTITUTE SHEET Society, 1986, p. 37. This type of rating evaluation is widely used to evaluate the effects of herbicides on weeds. The results are presented in Table 5A and are shown graphically in Figure 1. In Figure 1, MG = morningglory; VL = velvetleaf; CBUR = cocklebur; JW = jimsonweed; BG = barnyardgrass; YF = yellow foxtail; FP = fall panicum; JG = johnsongrass; SOY = soybean; and CORN = corn. Solid bars show the effect of sulfosate alone and cross-hatched bars show the effect of sulfosate plus conditioned medium. Bars shaded with diagonal lines show the effect of conditioned medium alone. Where such a bar is absent the conditioned medium alone produced no measurable plant injury. Percent injury scores in Figure 1 for sulfosate treatment alone are the averages of 7 experiments and the percent injury scores for the combination of sulfosate plus conditioned medium are the average of 4 experiments involving several different days after treatment. Water alone produced essentially no plant injury (data not shown). The conditioned medium alone caused no or only minor injury (data not shown in Table 5). It can be seen that the addition of conditioned medium substantially increased percent injury scores, when compared to the herbicide alone, for all of the grasses and also for morningglory, pigweed, soybean and corn. Some enhancement in the degree of injury was also seen with cocklebur and jimsonweed. There was little effect on velvetleaf. Thus, in comparison to sulfosate alone, the addition of conditioned medium enhanced the degree of injury to almost all of the weeds tested. Example 3 shows the effect of combining conditioned medium with higher levels of sulfosate (0.125 and 0.25 lbs/ acre) (0.14 and 0.28 kg/ha). EXAMPLE 3 The effects of conditioned medium was determined for sulfosate applied at 0.125 and 0.25 lbs/acre (0.14 and 0.28 SUBSTITUTE SHEET kg/ha) . Conditioned medium was produced as described in Example 1. Experiments were performed and plant injury was evaluated as described in Example 2. Table 5B and Figure 2 show the effect of conditioned medium on sulfosate applied at 0.125 lbs/acre (0.14 kg/ha), which is one-fourth of the minimum recommended field application rate. Symbols in Figure 2 are the same as in Figure 1. Conditioned medium enhanced the injury scores for all of the plants tested. The degree of enhancement was greatest for those plants where the sulfosate alone produced a percent injury score of 30% or less. In Figure 2, values for sulfosate alone are the average of 5 experiments and values for sulfosate plus conditioned medium are the average of 4 experiments. The effect of conditioned medium on sulfosate applied at 0.25 lbs/acre (0.28 kg/ha) (one-half of the minimum recommended field application rate) is shown in Table 5C and Figure 3. Symbols in Figure 3 are the same as in Figure 1. For some plants (most notably barnyardgrass, yellow foxtail and fall panicum) conditioned medium increased the injury scores. For other plants the sulfosate alone produced a high degree of injury and in those cases the addition of conditioned medium produced only modest or no enhancement. In Figure 3, values for sulfosate alone are the average of 2 experiments and values for sulfosate plus conditioned medium are from one experiment. These results show that the addition of conditioned medium to a chemical herbicide can produce enhanced injury to weeds over a range of herbicide concentrations. When compared to the effect of the herbicide alone, the enhancement in the degree of injury is greatest when the level of herbicide is chosen to produce a measurable but low degree of injury when applied alone. Example 4 shows the effect of combining conditioned medium with another herbicide, glyphosate. SUBSTITUTE SHEET EXAMPLE 4 The effects on plants of combining low levels of _ glyphosate (ROUNDUP®) with conditioned medium. Glyphosate was tested at two application rates, 0.06 lbs/acre (0.067 kg/ ha) and 0.125 lbs/acre (0.14 kg/ha). Recommended rates for application of glyphosate in the field range from 0.5 to 4.0 lbs/acre (0.56 to 4.48 kg/ha). Conditioned medium from Pseudomonas syringae pv. tabaci was prepared as described in Example.1. Experiments were performed and plant injury was evaluated as described in Example 2. The results are shown in Table 6. At both application rates, conditioned medium enhanced the percent injury scores for all of the plants tested. The effects were especially pronounced at the lower application rate (0.06 lbs/acre [0.067 kg/ha]). Example 5 shows the effect of combining conditioned medium with another herbicide, glufosinate. EXAMPLE 5 The effect of conditioned medium was tested on the actions of glufosinate (IGNITE®) applied at 0.06 and 0.125 lbs/acre (0.067 and 0.14 kg/ha). The minimum recommended field application rate for glufosinate is 0.5 lbs/acre (0.56 kg/ha). Conditioned medium was prepared as described in Example 1 and the methods were as described in Example 2. The selected levels of glufosinate produced very high degrees of injury in the broadleaf weeds ranging from 90 to 100%, and additional effects of conditioned medium could not be determined for those plants (Table 7) . Conditioned medium enhanced the effectiveness of the glufosinate on grasses. Example 6 shows the effect of combining conditioned medium with some other herbicides. EXAMPLE 6 Conditioned medium from Pseudomonas syringae pv. tabaci was prepared as described in Example 1. Experiments were performed and plant injury was evaluated as described in Example 2. In preliminary experiments, conditioned medium enhanced the effect of paraquat (applied over a range of 0.015-0.125 lbs/acre [0.017-0.14 kg/ha]) on grasses, but produced no consistent enhancement of injury to broadleaf weeds or the crop plants (soybean and corn) . Under the conditions tested, medium conditioned by Pseudomonas syringae pv. tabaci produced no enhancement of the effects of dicamba (0.06 and 0.125 lbs/acre [0.067 and 0.14 kg/ha]). The following Examples 7-28 show the effects of media conditioned by other species of microorganisms and the use of additional herbicides. King's B Medium was conditioned by various microorganisms as described in Example 1. In these experiments the medium was supplemented by zinc, although subsequent experiments suggest that zinc is probably not necessary for producing conditioned media that enhance the action of herbicides. To test the effects of herbicide alone, the herbicides were dissolved in water. To test the effects of the combination of herbicide and conditioned medium, the herbicide was dissolved in undiluted conditioned medium. Different herbicides were used in different tests. The herbicides tested fall into three broad classifications: Nonselective, broad spectrum herbicides included glufosinate, sulfosate, and imazapyr (ARSENAL®, American Cyanamid). Herbicides selective for grasses included fluazifop (FUSILADE 2000®, ICI Americas, Inc.), and sethoxydim (POAST®, BASF Corp.). Herbicides selective for broadleaf weeds included chlorimuron (CLASSIC®, E.I. DuPont De Nemours and Co.), dicamba (BANVEL®, Sandoz Crop Protection Corp.), bentazon (BASAGRAN®, BASF Corp.), fomesafen (REFLEX®, ICI Americas, Inc. ) , and imazethapyr (PURSUIT®, American Cyanamid) . Recommended field application rates for these herbicides SUBSTITUTE SHEET include: fluazifop - 0.063 lb/A (0.07 kg/ha); sethoxydim - 0.1 lb/A (0.12 kg/ha); imazapyr - 0.5 lb/A (0.55 kg/ha); imazethapyr - 0.03 lb/A (0.3 g/ha); bentazon - 0.75 lb/A (0.84 kg/ha); dicamba - 0.1 lb/A (0.12 kg/ha); chlorimuron - 0.5 oz/A (8 g/ha); and fomesafen - 0.125 lb/A (0.14 kg/ha). . Unless otherwise indicated in the tables, X-77® (0.25%) was added as an adjuvant. In a few experiments the adjuvant was Crop Oil Concentrate ("C.O.C."; Crop Surf Spray Oil, Universal Cooperatives, Inc., 7801 Metro Parkway, Minneapolis, MN 55440). Experiments were performed as generally described in Examples 1 and 2. The effect of herbicide alone on a panel of weeds was compared to the effect of herbicide plus conditioned medium. Broadleaf weeds that were tested included morningglory, velvetleaf, cocklebur, pigweed, jimsonweed, mustard (Brassica kaber). lambsquarters (Chenopodium album), and tomato (Lvcopersicon esculentum) . Although tomato is not a weed, it was viewed as representative of the reactions of broadleaf plants, and is sometimes used for screening herbicides. Grasses used included barnyardgrass, yellow foxtail, fall panicum, johnsongrass, and large crabgrass (Digitaria sanguinalis) . All of these weeds were not tested in every experiment. The specific weeds used in each test are indicated in the following tables. In addition, certain conditioned media were tested on various other weeds, as described below. Percent plant injury was rated between 13 and 18 days after treatment. Tables 8-27, were constructed by subtracting the percent injury score for herbicide alone from the percent injury score when herbicide was combined with conditioned medium. Thus, positive scores indicate enhancement of the action of the herbicide by the conditioned medium. In Tables 8-27, the following symbols have been used to represent the results: + = 1-10 percent injury score increase ++ = 11-20 percent injury score increase +++ = 21-30 percent injury score increase ++++ = 31-40 percent injury score increase +++++ - 41 or more percent injury score increase Thus, if the herbicide alone produced 30% injury, and the combination of herbicide plus conditioned medium produced 65% injury, then the 35% increase in percent injury would be represented as "++++" . Similarly, decreases in percent injury after the addition of conditioned medium are represented by minus signs (- = 1-10% injury score decrease; — = 11-20% injury score decrease, etc.). "0" indicates that there was no difference between the percent injury scores obtained with herbicide alone when compared to herbicide plus conditioned medium. An asterisk (*) indicates that the percent injury produced by the herbicide alone exceeded 90%, so that the addition of the conditioned medium could not be expected to produce large further increases in the injury score. "NA" indicates that the selective herbicide used is not active on the weed tested. "NT" indicates that the combination was not tested. In Tables 7-27, a "Y" in the column labeled "Other Weeds" indicates that the conditioned medium enhanced the effect of herbicides on other weeds that are not listed in the Tables. These are discussed in the text. All of the bacteria used to condition medium in Examples 7-28 were first found to have the ability to enhance the action of herbicides when applied as living bacteria in combination with the herbicide according to the method of U.S. Patent Application No. 07/601,653 and PCT Patent Application PCT/US90/05301. While prior testing of the activity of the living bacteria in combination with the herbicide according to the method of U.S. Patent Application No. 07/601,653 and PCT Patent Application PCT/US90/05301 is not necessary for practice of the instant invention, this may SUBSTITUTE SHEET be a useful strategy for the selection of bacteria that can be used to practice the instant invention. Examples 7-10 show the effect of medium conditioned by other pathovars of Pseudomonas syringae. Pathovars tested were Pseudomonas syringae pv. coronafaciens (ATCC #19607), Pseudomonas syringae pv. phaseolicola (ATCC #19304), Pseudomonas syringae pv. pisi (ATCC #11043), and Pseudomonas syringae pv. tagetis (ATCC #43128). EXAMPLE 7 Table 8 shows the effect of medium conditioned by Pseudomonas syringae pv. coronafaciens on the actions of several herbicides, some of which were tested at more than one concentration. The conditioned medium was effective in enhancing the action of glufosinate and sulfosate on both broadleaf weeds and grasses. The conditioned medium enhanced the action of the grass-selective herbicides (fluazifop and sethoxydim) on grasses, and there was modest enhancement of the action of the broadleaf herbicides on a few broadleaf weeds. In further tests, medium conditioned by Pseudomonas syringae pv. coronafaciens enhanced the action of herbicides as follows. Where more than one test was performed, the results are separated by commas: 1. Fluazifop (0.006 lb/A) (0.0067 kg/ha): large crabgrass (+++); broadleaf signal grass (++++); giant foxtail (++++); green foxtail (+++++); barnyard grass (++++); yellow foxtail (++++); johnsongrass (+++++); and red millet (+) . 2. Sethoxydim (0.015 lb/A) (0.017 kg/ha): large crabgrass (++, +++); broadleaf signal grass (+, ++); giant foxtail (++, ++); green foxtail (+, +); quack grass (++, 0); barnyard grass (+++, +); yellow foxtail (++, ++); johnsongrass (-, +++); tall fescue (+); and red millet (0) . EXAMPLE 8 Table 9 shows the effect of medium conditioned by t Pseudomonas syringae pv. phaseolicola (ATCC #19304) on the actions of herbicides. EXAMPLE 9 Table 10 shows the effect of medium conditioned by Pseudomonas syringae pv. pisi (ATCC #11043). EXAMPLE 10 Table 11 shows the effect of medium conditioned by Pseudomonas syringae pv. tagetis (ATCC #43128). Examples 11-20 show the effects of media conditioned by various pseudomonads on the action of herbicides. EXAMPLE 11 Table 12 shows the effect of medium conditioned by Pseudomonas viridiflava (ATCC #13223). EXAMPLE 12 Table 13 shows the effect of medium conditioned by Pseudomonas aureofaciens (ATCC #43051). In this experiment, the concentration of X-77® was 0.5%. EXAMPLE 13 Table 14 shows the effect of medium conditioned by Pseudomonas auricularis (ATCC #27319). Medium conditioned by Pseudomonas auricularis was also tested for its effect on other weeds. The results were as follows: 1. Sethoxydim (0.15 lb/A; (0.168 kg/ha) 1% C.O.C.): large crabgrass (++++); broadleaf signal grass (+); giant foxtail (+); green foxtail (+); red millet (+); and tall fescue (+). 2. Fluazifop (0.009 lb/A) (0.010 kg/ha): broadleaf signal grass (+++++); giant foxtail (++); green foxtail (+); quackgrass (+); red millet (+); and tall fescue (-) . 3. Sulfosate (0.13 lb/A) (0.146 kg/ha): quackgrass (+); purple nutsedge (+); Canada thistle (0); SUBSTTUTE SHEET bindweed (+); bermuda grass (+++++); and rhizome johnsongrass (+++). 4. Glufosinate (0.09 lb/A) (0.101 kg/ha): quackgrass (++); purple nutsedge (++); Canada thistle (+++++); bindweed (+++++); bermuda grass (++++); and rhizome johnsongrass (++++). 5. Imazapyr (0.004 lb/A) (0.0045 kg/ha): quackgrass (++.); purple nutsedge (0); Canada thistle (0); bindweed (+); bermuda grass (+); and rhizome johnsongrass (-). EXAMPLE 14 Table 15 shows the effect of medium conditioned by Pseudomonas avenae (ATCC #19860) on the action of herbicides. EXAMPLE 15 Table 16 shows the effect of medium conditioned by Pseudomonas carvophylli (ATCC #25418) on the action of herbicides. The effects of this conditioned medium were also tested on some additional weeds. The results were as follows: 1. Glufosinate (0.09 lb/A) (0.101 kg/ha): quackgrass (0); purple nutsedge (-); Canada thistle (0); bindweed (+++++); bermuda grass (+++); and rhizome johnsongrass (++++). 2. Sulfosate (0.13 lb/A) (0.146 kg/ha): quackgrass (+++); purple nutsedge (+); bindweed (++); bermuda grass (+++++); and rhizome johnsongrass (++). 3. Imazapyr (0.004 lb/A) (0.0045 kg/ha): quackgrass (+); purple nutsedge (0); Canada thistle (0); bindweed (++); bermuda grass (-); and rhizome johnsongrass (++). 4. Fluazifop (0.006 lb/A) (0.0067 kg/ha): broadleaf signal grass (+++); giant foxtail (+++); green foxtail (+++); quackgrass (0); and red millet (++). 5. Sethoxydim (0.15 lb/A, (0.168 kg/ha) 1% C.O.C., 2 tests): broadleaf signal grass (++,+); giant foxtail (+++,++); green foxtail (+,++); quackgrass (0,+); and red millet (+,0). EXAMPLE 16 Table 17 shows the effect of Pseudomonas cichorii (ATCC #19857). EXAMPLE 17 Table 18 shows the effect of medium conditioned by Pseudomonas fluorescens (ATCC #13525). EXAMPLE 18 Table 19 shows the effect of medium conditioned by Pseudomonas marginalis (ATCC #10844) on the action of herbicides. EXAMPLE 19 Table 20 shows the effect of medium conditioned by Pseudomonas phenazinium (ATCC #33666) on the action of herbicides. This conditioned medium was also tested on other weeds and the results were as follows: 1. Glufosinate (0.09 lb/A) (0.101 kg/ha): quackgrass (0); purple nutsedge (-); bindweed (+++++); bermuda grass (+++); and rhizome johnsongrass (++++). 2. Sulfosate (0.13 lb/A) (0.146 kg/ha): quackgrass (+); purple nutsedge (—); bindweed (+); bermuda grass (+++); and rhizome johnsongrass (0). 3. Imazapyr (0.004 lb/A) (0.0045 kg/ha) : quackgrass (++); purple nutsedge (0); bindweed (+++); bermuda grass (—); and rhizome johnsongrass (++). 4. Fluazifop (0.009 lb/A) (0.010 kg/ha): broadleaf signal grass (++++); giant foxtail (+); green foxtail (+); quackgrass (-); and red millet (+). 5. Sethoxydim (0.15 lb/A) (0.168 kg/ha): broadleaf signal grass (++); giant foxtail (+); green foxtail (+); quackgrass (+); and red millet (-). EXAMPLE 20 Table 21 shows the effect of medium conditioned by Pseudomonas pyrocinia (ATCC #15958). SUBSTITUTE SHEET Examples 21-26 show the effect of medium conditioned by other types of bacteria on the action of herbicides. EXAMPLE 21 Table 22 shows the effect of medium conditioned by Azospirillum brasilense on the action of herbicides. Two different isolates were compared, ATCC #29710 and ATCC #29145. Overall, the results obtained with the two isolates were similar. The effects of these strains were also tested on other weeds. Table 22A shows that the conditioned media enhanced the effects of fluazifop and sethoxydim on a variety of grasses. Two separate experiments with sethoxydim are shown. Table 22B shows the effect of the conditioned media on the actions of glufosinate, sulfosate and imazapyr on some additional weeds. EXAMPLE 22 Table 23 shows the effect of medium conditioned by Bacillus subtilis (ATCC #14593) on the action of herbicides. EXAMPLE 23 Table 24 shows the effect of medium conditioned by Corvnebacterium herculis (ATCC #13868) on the action of herbicides. The effect of medium conditioned by this bacterium was also tested on other weeds. The results were as follows: 1. Glufosinate (0.10 lb/A) (0.112 kg/ha): barnyard grass (+++); quackgrass (+++++); leafy spurge (+++++); yellow nutsedge (0); Canada thistle (+++); jimsonweed (0*); bindweed (+++); morningglory (0*); yellow foxtail (++++); bermuda grass (+++++); and seedling johnsongrass (+++++). 2. Sulfosate (0.20 lb/A) (0.224 kg/ha): barnyard grass (++++); quackgrass (++++); leafy spurge (+++++); yellow nutsedge (+); Canada thistle (+++); jimsonweed SUBSTITUTESHEET (-*); bindweed (++); morningglory (++); yellow foxtail (++); bermuda grass (+++); and seedling johnsongrass (++++). 3. Imazapyr (0.05 lb/A) (0.056 kg/ha): barnyard grass (++); quackgrass (+); leafy spurge (0); yellow nutsedge (0); Canada thistle (++); jimsonweed (0*); bindweed (0*); morningglory (-); yellow foxtail (+); bermuda grass (+); and seedling johnsongrass (+++). EXAMPLE 24 Table 25 shows the effect of medium conditioned by Flavinomonas orvzihabitans (ATCC #43272) on the action of herbicides. EXAMPLE 25 Table 26 shows the effect of medium conditioned by Xanthomonas campestris pv. phaseoli (ATCC #11766) on the action of herbicides. EXAMPLE 26 Table 27 shows the effect of medium conditioned by Xanthomonas campestris pv. translucens (ATCC #10731). Other weeds were also tested. EXAMPLE 27 Table 28 summarizes the effects of media conditioned by the organisms of Examples 7-26 on the action of herbicides. With most combinations that were tested, the conditioned media plus herbicide caused more injury than herbicide alone on more than half of the weed species examined (indicated by "Y" in Table 28). The action of glufosinate and sethoxydim was enhanced by all of the conditioned media tested for more than half of the species of weeds. The action of sulfosate was enhanced by all but one of the conditioned media and the action of fluazifop was enhanced by all but two of the conditioned media for more than half of the weeds tested. It should be noted that even for combinations marked "N" in Table 28, the conditioned media enhanced the action of the herbicide on some weeds, as can be seen by reference to the SUBSTITUTE SHEET corresponding Examples. These examples thus demonstrate that the herbicide and conditioned medium together produced, greater killing of weeds and greater inhibition of weed growth than is caused by the same amount of herbicide alone. It will be apparent to those skilled in the art that various modifications and variations can be made in the practice of the present invention without departing from the scope or spirit of the invention. Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. Thus, it is intended that the present invention cover modifications and variations of this invention, provided they fall within the scope of the appended claims or their equivalents. Table 1 Modified King's B Medium with Zinc (per liter) Proteose peptone No. 3 (Difco*) 20 g Glycerol 15 ml K 2 HP0 4 .3H 2 0 1.5 g MgS0 4 .7H 2 0 1.5 g ZnS0..7H 2 0 (optional) .0075 σ pH 7.2 King, E.O., Ward, M.K. , and Raney, D.E. Two Simple Media for the Demonstration of Pyocyanin and Fluorescin. J. Lab. Clin. Med. 44: 301-307 (1954). * Difco Laboratories, Inc., Detroit, Michigan 48232. SUBSTITUTE SHEET Table 2 Examples of Herbicides that Can Be Used in Combination with Conditioned Medium Alachlor Atrazine Bentazon Bialaphos Butachlor Butylate Chlorimuron ethyl Chlorsulfuron Cinmethylin Cyanazine Cycloate Dicamba 2,4-D (2,4-dichlorophenoxyacetic acid) EPTC Ethephon Fenoxaprop Fluazifop-butyl Fomesafen Glufosinate Glyphosate Haloxyfop Hoelon Imazapyr Imazaquin Imazethapyr Linuron Mefluidide Metolachlor Metribuzin Metsulfuron Molinate Norflurazon Oryzalin Oxyfluorfen Paraquat Pendimethalin Picloram Propachlor Propanil Pyridate Sethoxydim Simazine S,S,S-tributyl phosphorothioate Sulfometuron Sulfosate Trifluralin Table 3 Examples of Bacteria That Can Be Used to Condition Medium PHYTOPATHOGENIC BACTERIA Clavibacter michiganense C. michiganense subsp. michiganense C. michiganense subsp. nebraskense Curtobacterium flaccumfaciens Erwinia amylovora E. carotovora E. crvsanthemi E. herbicola E. stewartii E. tracheophila Pseudomonas andropogonis P. avenae P. carvophilli P. cattleyae P. cichorii P. corrugata P. florida P. gladioli P. glumae P. marginalis P. pastinacae P. plantarii P. pseudoalcaligenes P. rubri1ineans P. setariae P. solanacearum Pseudomonas syringae P. syringae py. aceris P. syringae py. anthirrhini P. syringae py. apii P. syringae py. apata P. syringae py. atrofaciens P. syringae py. atropurpurea P. syringae py. cannabina P. syringae py. ciccaronei P. syringae py. coronafaciens P. syringae py. delphinii P. syringae py. dvsoxyli P. syringae py. eribotrvae P. syringae py. garcae P. syringae py. glvcinea P. syringae py. helianthi P. syringae py. iaponica P. syringae py. lachrvmans P. syringae py. lapsa P. syringae py. maculicola P. syringae py. mellea P. syringae py. mori P. syringae py. morsprunorum P. syringae py. papulans P. syringae py. persicae P. syringae py. passiflorae P. syringae py. phaseolicola P. syringae py. pisi P. syringae py. primulae P. syringae py. ribicola P. syringae py. savastanoi P. syringae py. sesami P. syringae py. striafaciens P. syringae py. syringae P. syringae py. tabaci P. syringae py. tagetis P. syringae py. theae P. syringae py. tomato P. syringae py. ulmi P. syringae py. viburni P. syringae py. zizaniae P. viridiflava P. woodsii Streptomvces ipo ea Xanthomonas albilineans x. campestris x. campestris py. arrhenatheri x. campestris py. cerealis x. campestris py. graminis x. campestris py. phlei x. campestris py. poae x. campestris py. phaseoli x. campestris py. translucens Xylella fastidiosa NON-PHYTOPATHOGENIC BACTERIA Bacillus cereus Bradyrhizobium iaponicum Pseudomonas atlantica P. cannae P. chlororaphis P. fluorescens P. lindbergii P. putida Streptomvces atratus S. flavogriseus S. fradiae S. galilaeus S. hvdroscopicus S. hydroscopicus subs . limoneus S. tovocaensis S. tubercidus S. viridochromogenes Table 4 Examples of Fungi That Can Be Used to Condition Medium PHYTOPATHOGENIC FUNGI Alternaria alternata A. tenuis A. mali A. porri A. solani A. zinniae Ceratocystis adiposum C. coerulescens C. fagacearum C. fimbriata C. major C. moniliformis C. paradoxa C. ulmi Cercosporara asparagi C. beticola C. kikuchii C. nicotianae C. zeae-mavdis Cladosporium phlei Fomes annosus Fusariu iavanicum F. martii F. oxysporum F. solani Fusicoccum amygdali Nectria galligena Pestalotia cryptomeriarcola Phoma betae P. exigua P. lingam Polyporus tumulosus P. hispidus Pyrenophora tritici-repentis Pyricularia orvzae Rhizoctonia leguminicola R. solani Sclerotinia sclerotiorum Septoria nodorum Stemphyllium botryosum S. radicinum Stereum subpileatum NON-PHYTOPATHOGENIC FUNGI SUBSTITUTESHEET Table 5 Effect of Conditioned Medium on Herbicidal Actions of Sulfosate A. Sulfosate 0.06 lbs/acre (0.067 kg/ha) Sulfosate Alone Average 28.0 20.5 41.0 49.5 46.5 Sulfosate with Conditioned Medium Average 41.0 37.0 55.0 91.0 59.0 1 DAT - Days After Treatment 2 1 « Per Cent Injury Score SUBSTITUTE SHEET Table 5 (Continued) Effect of Conditioned Medium on Herbicidal Actions of Sulfosate Barnyard Yellow Fall Johnson Grass Foxtail Panicum Grass Soybean Corn EXP. * DAT 1 Z 2 Z Z Z Z Z_ A. Sulfosate 0.06 lbs/acre (0.067 kg/ha) Sulfosate Alone Average 3.0 27.0 9.5 1.0 7.5 17.0 Sulfosate with Conditioned Medium Average 20.5 61.0 26.5 24.0 21.0 25.0 1 DAT ■ Days After Treatment 2 Z = Per Cent Injury Score SUBSTITUTE SHEET Average 59.0 45.5 60.0 100.0 68.0 Average 57.5 52.5 93.5 100.0 85.0 Sulfosate with Conditioned Medium 15B 15 70.0 55.0 82.5 100.0 90.0 1 DAT « Days After Treatment 2 Z ~ Per Cent Injury Score Average 67.5 74.5 58.0 63.0 39.5 73.5 Average 37.5 67.5 39.0 30.0 31.5 77.5 Sulfosate with Conditioned Medium 15B 15 95.0 100.0 77.5 37.5 47.5 99.0 1 DAT - Days After Treatment 2 Z - Per Cent Injury Score SUBSTIT Average 10.0 6.5 37.5 52.0 41.5 Glyphosate with Conditioned Medium Average 44.0 33.0 60.0 91.5 75.0 Gl hosate with Conditioned Medium Average 49.0 45.0 69.0 94.5 79.0 1 DAT - Days After Treatment 2 Z ~ Per Cent Injury Score T Average 4.0 17.5 15.0 0.0 5.0 19.0 Glyphosate with Conditioned Medium Average 7.5 47.5 36.5 22.5 29.0 31.5 Glyphosate with Conditioned Medium 13A 12 72.5 80.0 75.0 82.5 45.0 65.0 14A 9 50.0 75.0 45.0 52.5 22.5 40.0 Average 61.5 77.5 60.0 67.5 34.0 52.5 1 DAT = Days After Treatment 2 Z = Per Cent Injury Score SUBSTITUTESHEET DAT - Days After Treatment Z - Per Cent Injury Score 3 Applied at 0.125 lbs/acre (0.14 kg/ha) Applied at 0.06 lbs/acre (0.067 kg/ha) Table 7 (Continued) Effect of Conditioned Medium on Herbicidal Actions of Glufosinate (IGNITE®) Barnyard Yellow Fall Johnson Grass Foxtail Panicum Grass Soybean Corn EXP. * DAT 1 Z Z Z Z 17A/.125 3 11 67.5 70.0 50.0 62.5 85.0 70.0 20A/.125 3 14 92.0 87.5 85.0 72.5 100.0 77.5 Average 20A/.06 4 14 17A/.125 3 11 20A/.125 3 14 Average 100.0 94.0 80.5 85.0 92.5 98.5 20A/.06 4 14 95.0 80.0 87.5 62.5 70.0 91.5 1 DAT • Days After Treatment 2 Z » Per Cent Injury Score 3 Applied at 0.125 lbs/acre (0.14 kg/ha) 4 Applied at 0.06 lbs/acre (0.067 kg/ha) SUBSTITUTE SHEET Table 8 Effect of Medium Conditioned by Pseudomonas syringae" pv. coronafaciens on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glor Leaf Cocklebur Pi weed Weed t u ters Tomato Table 8 (continued) Effect of Medium Conditioned by Pseudomonas syringae pv. coronafaciens on Action of Herbicides GRASSES Barnyard Yellow Johnson Fall Large Other Grass Foxtail Grass Panicum Crabgrass Weeds Glufosinate 1 +++++ +++ ++ +++++ NT NT 2 Glufosinate -* + + NT Y • 3 Glufosinate ++++ +++++ +* NT NT Sulfosate ++++ ++++ +++++ NT ++++ NT Sulfosate +++ +++++ ++++ +++ NT NT c Sulfosate " ++++ +++ +++++ +* NT Y Fluazifop +++ + NT Y Fluazifop ++ ++ ++ +++++ NT NT Fluazifop 8 0 +++ NT + NT Sethoxydim ++++ + NT 0 Y 10 Bentazon ' NT 0(NA) 0(NA) NT NT NT 11 Bentazon NT 0(NA) 0(NA) NT NT NT Chlorimuron 12 0(NA) 0(NA) 0(NA) NT + NT Fomesafen NT 0(NA) 0(NA) NT NT NT .1 0.04 lb/A (0. 045 kg/ha) glufosinate 2 0.09 lb/A (0.101 kg/ha) glufosinate 3 0.10 lb/A (0.112 kg/ha) glufosinate 4 0.06 lb/A (0.067 kg/ha) sulfosate 5 0.125 lb/A (0.14 kg/ha) sulfosate 6 0.006 lb/A (0.0067 kg/ha) fluazifop 7 0.004 lb/A (0.0045 kg/ha) fluazifop 8 0.01 lb/A (0.011 kg/ha) fluazifop 9 0.012 lb/A (0.013 kg/ha) sethoxydim 10 0.03 lb/A (0.034 kg/ha) bentazon, IZ C.O.C. 11 0.06 lb/A (0.067 kg/ha) bentazon; IZ C.O.C. 12 1.0 g/A (2.5 g/ha) chlorimuron 13 0.0007 lb/A (0.00078 kg/ha) fomesafen SUBSTITUTESHEET Table 9 Effect of Medium Conditioned by Pseudomonas syringae pv. phasέolicola on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glory Leaf Cocklebur Pi weed Weed rd te Tomato Glufosinate 1 ++++ +++ 2. Sulfosate +++ Fluazifop 0(NA) 0(NA) Sethoxydim 4 0(NA) 0(NA) Chlorimuron" NT NT +* NT NT GRASSES Chlorimuron 0(NA) 0(NA) 0(NA) NT 0(NA) NT 0.09 lb/A (0.101 kg/ha) glufosinate 0.17 lb/A (0.19 kg/ha) sulfosate 0.01 lb/A (0.011 kg/ha) fluazifop 0.012 lb/A (0.013 kg/ha) sethoxydim, IZ C.O.C. 1.0 g/A (2.5 g/ha) chlorimuron Table 10 Effect of Medium Conditioned by Pseudomonas syringae pv. pisi on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glory Leaf Cocklebur Pi weed Weed Mustard uarters Tomato Glufosinate 1 ++++ +++ 2 Sulfosate + ++ Fluazifop 0(NA) 0(NA) Sethoxydim 4 0(NA) 0(NA) Chlorimuron " NT NT +* NT NT GRASSES Barnyard Yellow Johnson Fall Large Other Grass Foxtail Grass Panicum Crabgrass Weeds Glufosinate 1 +* +++ +++++ NT NT Sulfosate +++++ +++ +++++ NT +++ NT Fluazifop +++ ++++ NT ++ NT 4 Sethoxydim +++++ ++++ + NT ++ NT Chlorimuron 0(NA) 0(NA) 0(NA) NT 0(NA) NT 0.09 lb/A (0.101 kg/ha) glufosinate 0.17 lb/A (0.19 kg/ha) sulfosate 0.01 lb/A (0.011 kg/ha) ' fluazifop *4 0.12 lb/A (0.13 kg/ha) sethoxydim, IZ C.O.C, *table 9 has 0.012 lb/A 1.0 g/A (2.5 g/ha) chlorimuron SUBSTITUTE SHEET Table 11 Effect of Medium Conditioned by Pseudomonas syringae pv. tagetis on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glory Leaf Cocklebur Pi weed Weed Mustard uarters Tomato Glufosinate 1 +++++ ++++ +++ 0(NA) GRASSES Fluazifop" NT NT NT 0.04 lb/A (0.04 kg/ha) glufosinate 0.10 lb/A (0.112 kg/ha) glufosinate 0.06 lb/A (0.067 kg/ha) sulfosate 0.125 lb/A (0.14 kg/ha) sulfosate 0.006 lb/A (0.0067 kg/ha) fluazifop, IZ C.O.C. Table 12 E ect of Medium Conditioned by Pseudomonas viridiflava on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glory Leaf Cocklebur Pi weed Weed Mustard uarters Tomato Glufosinate 1 ++++ +++ 2 Sulfosate + Fluazifop 0(NA) 0(NA) Sethoxydim 4 0(NA) 0(NA) Chlorimuron" ++ NT NT +* NT NT +++ GRASSES Chlorimuron 0(NA) 0(NA) NT NT 0(NA) NT 0.09 lb/A (0.101 kg/ha) glufosinate 0.17 lb/A (0.19 kg/ha) sulfosate 0.01 lb/A (0.011 kg/ha) fluazifop 0.12 lb/A (0.13 kg/ha) sethoxydim, IZ C.O.C. 1.0 g/A (2.5 g/ha) chlorimuron ET Table 13 Effect of Medium Conditioned by Pseudomonas aureofacieits on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glor Leaf Cocklebur Pi weed Weed Mustard Quarters Tomato ._* NT NT 0* NT Fluazifop 0(NA) 0(NA) NT NT 0(NA) 0(NA) NT +++ Chlorimuron " Dicamba GRASSES Fluazifop +++++ +++ +++ NT +++ NT Chlorimuron 0(NA) 0(NA) 0(NA) NT 0(NA) NT 6 Dicamba 0(NA) 0(NA) 0(NA) NT 0(NA) NT 1 0.09 lb/A (0.101 kg/ha) glufosinate, 0.5Z X-77 2 0.13 lb/A (0.146 kg/ha) sulfosate, 0.5Z X-77 3 0.004 lb/A (0.0045 kg/ha) imazapyr, 0.5Z X-77 4 0.009 lb/A (0.010 kg/ha) fluazifop, 0.5Z X-77 5 0.01 oz/A (0.7 g/ha) chlorimuron, 0.5Z X-77 6 0.015 lb/A (0.017 kg/ha) dicamba, 0.5Z X-77 Table 14 Ef ect of Medium Conditioned by Pseudomonas auriculari's on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glor Leaf Cocklebur Pi weed Weed Mustard uarters Tomato GRASSES 0.009 lb/A (0.010 kg/ha) fluazifop 0.015 lb/A (0.017 kg/ha) sethoxydim 0.03 lb/A (0.034 kg/ha) bentazon 0.06 lb/A (0.067 kg/ha) bentazon 0.0007 lb/A (0.00078 kg/ha) fomesafen SUBSTITUTE SHEET Table 15 Effect of Medium Conditioned by Pseudomonas avenae on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glor Leaf Cocklebur Pigweed Weed Mustard uarters Tomato GRASSES Fluazifop +++++ ++++ NT NT 0.09 lb/A (0.101 kg/ha) glufosinate 0.13 lb/A (0.146 kg/ha) sulfosate 0.004 lb/A (0.0045 kg/ha) imazapyr 0.009 lb/A (0.010 kg/ha) fluazifop SUBSTITUTE SHEET Table 16 Effect of Ifedium Conditioned by Pseudomonas caryophylli on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glory Leaf Cocklebur Pi weed Weed Mustard uarters Tomato SUBSTITUTE SHEET Table 16 (continued) Effect of Medium Conditioned by Pseudomonas caryophylli on Action of Herbicides GRASSES Barnyard Yellow Johnson Fall Large Other Grass Foxtail Grass Panicum Crabgrass Weeds Glufosinate 1 NT +++++ +++++ +++++ NT NT 2 Glufosinate NT +++ _* NT Y Sulfosate 3 NT +++++ +++++ +++++ NT NT 4 Sulfosate NT +++++ +++++ NT Y Fluazifop * ++ ■;+++++ Y Sethoxydim ++;+ ++;+ +++,0 Y Bentazon NT 0(NA) 0(NA) NT Bentazon NT 0(NA) 0(NA) NT c Fomesafen ' NT 0(NA) 0(NA) NT 1 0.04 lb/A (0.04 kg/ha) glufosinate 2 0.10 lb/A (0.112 kg/ha) glufosinate 3 0.06 lb/A (0.067 kg/ha) sulfosate 0.125 lb/A (0.14 kg/ha) sulfosate 0.006 lb/A (0.0067 kg/ha) fluazifop 0.015 lb/A (0.017 kg/ha) sethoxydim, IZ C.O.C. 0.03 lb/A (0.034 kg/ha) bentazon 0.06 lb/A (0.067 kg/ha) bentazon 0.0007 lb/A (0.00078 kg/ha) fomesafen SUBST Table 17 Effect of Medium Conditioned by Pseudomonas cichorii' on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glor Leaf Cocklebur Pi weed Weed Mustard uarters Tomato GRASSES Fluazifop +++++ +++ NT NT Chlorimuro 0(NA) 0(NA) 0(NA) NT 0(NA) NT Dicamba 6 0(NA) 0(NA) 0(NA) NT 0(NA) NT 1 0.09 lb/A (0.101 kg/ha) glufosinate 2 0.13 lb/A (0.146 kg/ha) sulfosate 3 0.004 lb/A (0.0045 kg/ha) imazapyr 4 0.009 lb/A (0.010 kg/ha) fluazifop 5 0.01 oz/A (0.7 g/ha) chlorimuron 6 0.015 lb/A (0.017 kg/ha) dicamba SUBSTITUTE SHEET Table 18 Effect of Medium Conditioned by Pseudomonas fluorescens on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glor Leaf Cocklebur Pi wee Weed tard ters Tomato SUBSTITUTE SHEET Table 18 (continued) Effect of Medium Conditioned by Pseudomonas fluorescens on Action of Herbicides GRASSES Barnyard Yellow Johnson Fall Large Other Grass Foxtail Grass Panicum Crabgrass Weeds Glufosinate 1 ++ ++++ NT +++++ NT NT Glufosinate 2 +++++ +++++ NT +++++ NT NT Glufosinate 3 +++++ +++++ NT +* NT NT Glufosinate 4 +++++ ++++ NT 0 NT NT Sulfosate - ++ NT +++ NT NT Sulfosate +++++ +++++ NT ++++ NT NT Sulfosate — + NT ++ NT NT 8 Sulfosate ++ + NT NT NT Fluazifop ' NT + NT NT 10 Fluazifop ++++ NT + NT NT 1 0.04 lb/A (0.04 kg/ha) glufosinate 2 0.06 lb/A (0.067 kg/ha) glufosinate 3 0.10 lb/A (0.112 kg/ha) glufosinate 4 0.10 lb/A (0.112 kg/ha) glufosinate 5 0.06 lb/A (0.067 kg/ha) sulfosate 6 0.06 lb/A (0.067 kg/ha) sulfosate 7 0.125 lb/A (0.14 kg/ha) sulfosate 8 0.125 lb/A (0.14 kg/ha) sulfosate 9 0.006 lb/A (0.0067 kg/ha) fluazifop 10 0.006 lb/A (0.0067 kg/ha) fluazifop Table 19 Effect of Medium Conditioned by Pseudomonas τnm-prττιnlift on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glor Leaf Cocklebur Pi weed Weed Mustard uarters Tomato Table 19 (continued) Effect of Medium Conditioned by Pseudomonas marginalis on Action of Herbicides GRASSES 1 0.04 lb/A (0.04 kg/ha) glufosinate 2 0.10 lb/A (0.112 kg/ha) glufosinate 3 0.06 lb/A (0.067 kg/ha) sulfosate 4 0.125 lb/A (0.14 kg/ha) sulfosate 5 0.006 lb/A (0.0067 kg/ha) fluazifop 6 0.009 lb/A (0.010 kg/ha) fluazifop 7 0.15 lb/A (0.168 kg/ha) sethoxydim, IZ C.O.C. 8 0.03 lb/A (0.034 kg/ha) bentazon 9 0.06 lb/A (0.067 kg/ha) bentazon 10 0.0007 lb/A (0.00078 kg/ha) fomesafen SHEET Table 20 Effect of Medium Conditioned by Pseudomonas phenazinium on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glor Leaf Cocklebur P wee W a e s Tomato GRASSES Fluazifop NT NT +++++ NT +++++ NT 0.09 lb/A (0.101 kg/ha) glufosinate 0.13 lb/A (0.146 kg/ha) sulfosate 0.004 lb/A (0.0045 kg/ha) imazapyr 0.009 lb/A (0.010 kg/ha) fluazifop Table 21 Effect of Medium Conditioned by Pseudomonas pyrocinia" on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glor Leaf Cocklebur Pi weed Weed Musta d ua ters Tomato Fluazifop 0(NA) 0(NA) NT NT 0(NA) 0(NA) NT NT Chlorimuron " NT NT ++ NT NT Dicamba NT NT NT NT GRASSES Fluazifop +++++ +++ ++++ NT NT Chlorimuron 0(NA) 0(NA) 0(NA) NT 0(NA) NT 6 Dicamba 0(NA) 0(NA) 0(NA) NT 0(NA) NT 1 0.09 lb/A (0.101 kg/ha) glufosinate 2 0.13 lb/A (0.146 kg/ha) sulfosate 3 0.004 lb/A (0.0045 kg/ha) imazapyr 4 0.009 lb/A (0.010 kg/ha) fluazifop 5 0.01 oz/A (0.7 g/ha) chlorimuron 6 0.015 lb/A (0.0171 kg/ha) dicamba TE SHEET Table 22 Effect of Media Conditioned by Two Strains Azospirillum brasilense (ATCC 29710 and 29145) on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glory Leaf Cocklebur Pigweed Weed Mustard uarters Tomato NT NT +++++ +++ NT NT ++++ NT NT + NT NT 0 + NT NT 0 NT NT _* Fluazifop 0(NA) 0(NA) NT NT 0(NA) 0(NA) Fluazifop 8 0(NA) 0(NA) NT NT 0(NA) 0(NA) Bentazon " NT NT + Bentazo 10 NT NT 0 Table 22 (continued) Effect of Media Conditioned by Two Strains Azospirillum brasilense (ATCC 29710 and 29145) on Action of Herbicides GRASSES Johnson Fall Large Other Grass Panicum Crabgrass Weeds 0 NT +++ Y 0 NT + Y + NT +++++ Y + NT +-_ ■ +++ Y NT Y NT Y Fluazifop + +++ ++ NT +++++ Y Fluazifop ++ +++ + NT + Y Bentazon * 0(NA) 0(NA) 0(NA) NT 0(NA) NT 10 Bentazon 0(NA) 0(NA) 0(NA) NT 0(NA) NT 1 0.06 lb/A (0.067 kg/ha) glufosinate, ATCC 29710 2 0.06 lb/A (0.067 kg/ha) glufosinate, ATCC 29145 3 0.10 lb/A (0.112 kg/ha) sulfosate, ATCC 29710 4 0.10 lb/A (0.112 kg/ha) sulfosate, ATCC 29145 5 0.003 lb/A (0.0034 kg/ha) imazapyr, ATCC 29710 6 0.003 lb/A (0.0034 kg/ha) imazapyr, ATCC 29145 7 0.006 lb/A (0.0067 kg/ha) fluazifop, ATCC 29710 8 0.006 lb/A (0.0067 kg/ha) fluazifop, ATCC 29145 9 0.05 lb/A (0.056 kg/ha) bentazon, ATCC 29710 10 0.05 lb/A (0.056 kg/ha) bentazon, ATCC 29145 SUBSTITUTE SHEET Broadleaf Signal Grass Fluazifop 1 + Fluazifop ++ Sethoxydim " + 4 Sethoxydim + Sethoxydim ++ Sethoxydim + 1 0.006 lb/A (0.0067 kg/ha) fluazifop, ATCC 29710 2 0.006 lb/A (0.0067 kg/ha) fluazifop, ATCC 29145 3 0.015 lb/A (0.017 kg/ha) sethoxydim, ATCC 29710, IZ C.O.C. 0.015 lb/A (0.017 kg/ha) sethoxydim, ATCC 29145, IZ C.O.C. 0.015 lb/A (0.017 kg/ha) sethoxydim, ATCC 29710, IZ C.O.C. 0.015 lb/A (0.017 kg/ha) sethoxydim, ATCC 29145, IZ C.O.C. SUBSTITUTE SHEET Table 22B Effect of Media Conditioned by Two Strains Azospirillum brasilense (ATCC 29710 and 29145) on Action of Herbicides BROAD LEAF WEEDS Jimson Weed Bindweed ._* +++ 0* +++ _* + _* + o* 0* _* 0* GRASSES Barnyard Yellow Quack Bermuda Johnsongrass Grass Foxtail Grass Grass (seedling) Glufosinate 1 ++++ +++ ++++ ++++ ++++ Glufosinate 2 ++++ +++ +++++ +++++ ++++ Sulfosate 3 ++++ ++ ++++ ++ +++ Sulfosate +++ + ++++ + +++ Imazapyr + + + + +++ Imazapyr 0 + + + + 1 0.10 lb/A (0.112 kg/ha) glufosinate, ATCC 29710 2 0.10 lb/A (0.112 kg/ha) glufosinate, ATCC 29145 3 0.20 lb/A (0.224 kg/ha) sulfosate, ATCC 29710 4 0.20 lb/A (0.224 kg/ha) sulfosate, ATCC 29145 5 0.005 lb/A (0.0056 kg/ha) imazapyr, ATCC 29710 6 0.005 lb/A (0.0056 kg/ha) imazapyr, ATCC 29145 SUBSTITUTESHEET Table 23 E fect of Medium Conditioned by Bacillus subtilis on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glory Leaf Cocklebur Pi weed Weed Mustard uarters Tomato Glufosinate 1 ++++ +++ 2 Sulfosate ++ Fluazifop 0(NA) 0(NA) Sethoxydim 4 0(NA) 0(NA) Chlorimuron " NT NT +* NT NT GRASSES Chlorimuron 0(NA) 0(NA) 0(NA) NT 0(NA) NT 1 0.09 lb/A (0.101 kg/ha) glufosinate 2 0.17 lb/A (0.19 kg/ha) sulfosate 3 0.01 lb/A (0.011 kg/ha) fluazifop 4 0.12 lb/A (0.13 kg/ha) sethoxydim, IZ C.O.C. 5 1.0 g/A (2.5 g/ha) chlorimuron SUBSTITUTE SHEET Table 24 Effect of Medium Conditioned by Corynebacterium herculls on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glor Leaf Cocklebur Pi weed W ters Tomato Fluazifop 0(NA) NT NT 0(NA) 0(NA) NT NT Bentazon " ++ NT NT NT NT GRASSES Fluazifop ++++ +++++ NT +++++ NT Bentazon 0(NA) 0(NA) 0(NA) NT 0(NA) NT 1 0.06 lb/A (0.067 kg/ha) glufosinate 2 0.10 lb/A (0.112 kg/ha) sulfosate 3 0.003 lb/A (0.0034 kg/ha) imazapyr 4 0.006 lb/A (0.0067 kg/ha) fluazifop 5 0.05 lb/A (0.056 kg/ha) bentazon SUBSTITUTE SHEET Table 25 Effect of Medium Conditioned by Flavinomonas oryzihabitans on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glory Leaf Cocklebur Pi weed Weed Mustard uarters Tomato Glufosinate 1 ++++ +++ 2 Sulfosate 0 ++ Fluazifop 0(NA) 0(NA) Sethoxydim 4 0(NA) 0(NA) Chlorimuron " NT NT +* NT NT GRASSES Chiorimuron 0(NA) 0(NA) 0(NA) NT +(NA) NT 1 0.09 lb/A (0.101 kg/ha) glufosinate 2 0.17 lb/A (0.19 kg/ha) sulfosate 3 0.01 lb/A (0.011 kg/ha.) fluazifop 4 0.012 lb/A (0.013 kg/ha) sethoxydim 5 1.0 g/A (2.5 g/ha) chlorimuron B TITUTE SHEET Table 26 Effect of Medium Conditioned by Xanthomonas campestris pv. phaseoli on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glory Leaf Cocklebur Pi weed Weed Mustard uarters Tomato Fluazifop 0(NA) NT NT NT 0(NA) 0(NA) NT NT Bentazon NT NT NT 0* ++ NT NT Imazethapyr NT ++ NT 0 SUBSTITUTE SHEET Table 26 (continued) Effect of Medium Conditioned by Xanthomonas campestris pv. phaseoli on Action of Herbicides GRASSES Fluazifop +++++ NT NT NT Bentazon Imazethapyr 1 0.04 lb/A (0.04 kg/ha) glufosinate 2 0.13 lb/A (0.146 kg/ha) glufosinate 3 0.08 lb/A (0.09 kg/ha) sulfosate 0.004 lb/A (0.0045 kg/ha) imazapyr 0.005 lb/A (0.0056 kg/ha) imazapyr 0.004 lb/A (0.0045 kg/ha) fluazifop 0.05 lb/A (0.056 kg/ha) bentazon 0.30 lb/A (0.336 kg/ha) imazethapyr SUBSTITUTE SHEET Table 27 Effect of Medium Conditioned by Xanthomonas campestris pv. translucens on Action of Herbicides BROAD LEAF WEEDS Morning Velvet Jimson Lambs Glor Leaf Cocklebur Pi weed Weed Mustard arters Tomato Fluazifop 0(NA) NT NT NT 0(NA) 0(NA) NT NT Bentazon NT NT NT 0* NT NT Imazethapyr NT + NT SUBSTITUTESHEET Table 27 (continued) Effect of Medium Conditioned by Xanthomonas i—mp s r s pv. translucens on Action of Herbicides GRASSES Fluazifop +++++ ++++ ++++ NT NT NT Bentazon Imazethapyr 1 0.04 lb/A (0.04 kg/ha) glufosinate 0.13 lb/A (0.146 kg/ha) glufosinate 3 0.08 lb/A (0.09 kg/ha) sulfosate 0.005 lb/A (0.0056 kg/ha) imazapyr 0.004 lb/A (0.0045 kg/ha) imazapyr 0.004 lb/A (0.0045 kg/ha) fluazifop 0.05 lb/A (0.056 kg/ha) bentazon 0.30 lb/A (0.336 kg/ha) imazethapyr SUBSTITUTE SHEET Table 2A SUHMΛRT COMBINATION TABLE (il.UFOSIKATB FI.1IAZ I POP. 1 SBTHOITOIN IMAZAPTR CIIIXJRIHUHON DICAMBA BENTAZON POMESAPEN IMAXBTHAPTH Y Y Y Y CO Y c Y Y N co I H Y Y α H I C Y N -I m Y co Y I m Y Y Y Y Y N YES "V - Conditioned media » herbicide caused more irι|ιιιy > IIΛII herbicide alυnβ wllh half or more υf the apeciuβ in the same study. NO "N' - Tha Conditioned meilia » herbliride tiuatmeiit di nut merl the above condit ion. Blank * The corobi nnt ion was nut I out oil T.ib l <_• 20 I C<>»t I IIHV«I > SUMMARY COMB INATION TABLE A 3 A C I H -4 Pl -J I (A X m YES "Y" " Conditioned media + herbicide caused more injury than herbicide alona with half or more of the species in the same study. NO "M" » The Conditioned media « herbicide treatment did not meet the above condlt ion. Blank - The ι-ombination was not tested

Claims: WHAT IS CLAIMED IS; 1. A method for controlling weeds which comprises applying to said weeds an amount effective to inhibit weed growth or to kill weeds of a chemical agent, and a medium which has been conditioned by the growth of microorganisms therein, said chemical agent, in the presence of said conditioned medium, being effective to inhibit the growth or to kill at least one weed species, and said conditioned medium being effective to enhance the activity of said chemical agent, said chemical agent and said conditioned medium being applied in amounts wherein killing of said weeds or the inhibition of the growth of said weeds is greater than would be caused by the same amounts of said chemical agent or said conditioned medium applied alone. 2. The method of claim 1 wherein said chemical agent is a herbicide. 3. The method of claim 1 wherein said medium conditioned by the growth of microorganisms is further processed to make a processed conditioned medium. 4. The method of claim 1 wherein said chemical agent and said conditioned medium are applied to said weeds simultaneously. 5. The method of claim 1, wherein said microorganism is a bacterium. 6. The method of claim 1, wherein such microorganism is a fungus. 7. The method of claim 5 wherein said bacterium is a species from the genus Pseudomonas. 8. The method of claim 7 wherein said bacterium is selected from the group consisting of Pseudomonas aureofaciens, Pseudomonas auricularis, Pseudomonas avenae, Pseudomonas caryophylli, Pseudomonas cichorii, Pseudomonas SUBSTITUTE SHEET fluorescens, Pseudomonas marσinalis, Pseudomonas phenazinium, and Pseudomonas viridiflava. 9. The method of claim 7 wherein said bacterium is a pathovar of the species Pseudomonas syringae. 10. The method of claim 9 wherein said bacterium is Pseudomonas syrinσae pv. tabaci. 11. The method of claim 9 wherein said bacterium is selected from the group consisting of Pseudomonas syrinσae pv. coronafaciens, Pseudomonas syringae pv. phaseolicola, Pseudomonas syringae pv. pisi, and Pseudomonas syringae pv. tagetis. 12. The method of claim 5 wherein said bacterium is selected from the group consisting of Azospirillum brasilense. Bacillus subtilis, Corynebacterium herculis, Flavinomonas oryzihabitans. Xanthomonas campestris pv. phaseoli, and Xanthomonas campestris pv. translucens. 13. The method of claim 2 wherein said herbicide is sulfosate. 14. The method of claim 2 wherein said herbicide is glufosinate. 15. The method of claim 2 wherein said herbicide is glyphosate. 16. The method of claim 2 wherein said herbicide is selected from the group consisting of fluazifop, sethoxydim, imazapyr, chlorimuron, dicamba, bentazon, fomesafen, and imazethapyr. 17. The method of claim 1 wherein said microorganism is genetically modified. 18. The method of claim 17 wherein said genetically modified microorganism contains recombinant DNA. 19. A herbicidal composition comprising a chemical herbicide in combination with a medium, wherein said medium has been conditioned by the growth of microorganisms therein which render the medium effective to enhance the activity of said chemical herbicide. EET said medium being present in said composition in amount sufficient to enhance the herbicidal activity of said chemical herbicide against at least one weed species. 20. A composition as claimed in claim 19 wherein said composition contains said herbicide. 21. A composition as claimed in claim 19 wherein said conditioned medium is produced by growth of a bacterium that is a species from the genus Pseudomonas. 22. A composition as claimed in claim 21 wherein said bacterium is selected from the group consisting of Pseudomonas aureofaciens, Pseudomonas auricularis, Pseudomonas avenae, Pseudomonas caryophylli, Pseudomonas cichorii, Pseudomonas fluorescens, Pseudomonas marginalis, Pseudomonas phenazinium, and Pseudomonas viridiflava. 23. A composition as claimed in claim 21 wherein said bacterium is a pathovar of the species Pseudomonas syringae. 24. A composition as claimed in claim 23 wherein said bacterium is Pseudomonas syringae pv. tabaci. 25. A composition as claimed in claim 23 wherein said bacterium is selected from the group consisting of Pseudomonas syringae pv. coronafaciens, Pseudomonas syringae pv. phaseolicola, Pseudomonas syringae pv. pisi, and Pseudomonas syringae pv. tagetis. 26. A composition as claimed in claim 19 wherein said herbicide is sulfosate. 27. A composition as claimed in claim 19 wherein said herbicide is glyphosate. 28. A composition as claimed in claim 19 wherein said herbicide is glufosinate. 29. A composition as claimed in claim 19 wherein said- herbicide is selected from the group consisting of fluazifop, sethoxydim, imazapyr, chlorimuron, dicamba, bentazon, fomesafen, and imazethapyr. SUBSTITUTE SHEET 30. A composition as claimed in claim 19 wherein said medium conditioned by the growth of microorganisms is further processed to make a processed conditioned medium. 31. A method for controlling weeds which comprises applying to the seeds of said weeds an amount effective to prevent or inhibit weed growth of a chemical agent,, and a medium which has been conditioned by the growth of microorganisms therein, said chemical agent, in the presence of said conditioned medium, being effective to kill or inhibit the growth of weeds from said seeds of at least one weeds species, and said conditioned medium being effective to enhance the activity of said chemical agent, such chemical agent and conditioned medium being applied in amounts wherein killing of said seeds or the inhibition of the growth of said weeds from said seeds is greater than would be caused by the same amounts of said chemical agent or said conditioned medium applied alone. SUBSTITUTESHEET

Inventors: CARLSON PETER S, HERBST KATHLEEN, KOSTKA STANLEY J, CARLSON, PETER, S., HERBST, KATHLEEN, KOSTKA, STANLEY, J.
Applicants: CROP GENETICS INT, CROP GENETICS INTERNATIONAL CORPORATION
Application year: 1991
Main classification: A01N63
IPC codes: A01N35, A01N57, A01N63