Protection of rice against group 15 herbicides

Abstract

The present invention provides methods for reducing rice injury and achieving weed control by treating rice with both a safener and a microencapsulated Group 15 herbicide. Herein, we demonstrate that tolerance of fenclorim-protected rice to the Group 15 herbicide acetochlor is substantially improved when applied as a microencapsulated formulation, particularly when applied pre-emergence or delayed pre-emergence. Rice plants produced by these methods and seeds produced by the rice plants are also provided.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This patent application claims the benefit of priority to U.S. Provisional Patent Application No. 62 / 906,902, filed September 27, 2019, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] Rice is an ancient agricultural crop and remains one of the major food crops in the world. There are two cultivars of rice: Asian rice (Oryza sativa L.) and African rice (O. glaberrima Steud.). The Sativa species constitutes virtually all of the cultivated rice in the world and is the species cultivated in the United States. Weedy rice (Oryza sativa L. var. sylvatica), also known as red rice (Oryza sativa L.), is one of the most problematic weeds in rice production. Weedy rice plants generally have a faster growth rate and more tillers than cultivated rice, but produce much less grain. However, weedy rice is closely related to cultivated rice, making it difficult to selectively control without harming the surrounding rice. Even low levels of contamination can cause serious losses in the yield and quality of cultivated rice, as extra milling is required to remove contaminated weedy rice. Thus, weedy rice increases production costs and reduces farm profits. Unfortunately, weedy rice is a major culprit in the CO2 emissions from cultivated rice. 2 Weedy rice responds more aggressively to elevated levels of arsenic than cultivated rice. Thus, the competitive ability of weedy rice is likely to increase as climate change continues, becoming an even more serious problem than it is today (Agron. J. (2010) 102:118-123).

[0003] Prior to the introduction of imidazolinone-resistant rice (Clearfield® BASF Corporation, Research Triangle Park, NC) in 2002, weedy rice was primarily controlled using direct seeding in flooded fields and in rotation with soybeans, corn, and sorghum (Weed Technology crop rotationy (2008) 22:200-208). Clearfield® technology was rapidly adopted in the mid-south United States because it allowed growers to selectively control weedy rice and troublesome grasses such as barnyard grass (Echinochloa crus-galli (L.) Beauv.) using acetolactate synthase (ALS)-inhibiting herbicides. In 2014, approximately 49% of rice acreage in Arkansas was planted with Clearfield® rice (AAES Research Series (2014) 626:11-22), although in recent years that percentage has declined somewhat, in part due to resistance of weedy rice and barnyard grass to these herbicides. In the mid-2000s, widespread use of ALS inhibitors such as imazethapyr and imazamox, in part due to lack of adherence to proper use guidelines, rapidly led to resistance in several weed populations. To date, resistance to ALS site of action (SOA) has been identified in 11 species in Arkansas, including weedy rice, barnyard grass, Chinese yarrow, Japanese yarrow, and Palmer amaranth (The International Survey of Herbicide Resistant Weeds (2018)). However, natural hybridization and resulting outcrossing between weedy rice and cultivated rice is largely responsible for the increase in ALS-resistant weedy rice populations (Crop Protection (2007) 26:349-356).

[0004] It has been shown that repeated use of the same herbicide SOA can quickly lead to herbicide resistance. When the same SOA is repeatedly targeted, the frequency of resistance alleles increases in the population in response to selection pressure, thereby reducing the effectiveness of the herbicide and limiting control options (Weed Science (1996) pp: 176-193). However, the evolution of resistance in problematic weeds such as barnyard grass and weedy rice can be delayed by rotating or mixing different herbicide SOAs (Weed Science (2012) 60:31-62). In recent years, no herbicides containing new SOAs have been commercialized, limiting growers' options for effective control. Therefore, there is a need to explore alternative herbicides that can be used to delay resistance and control resistant weeds in rice. Very long chain fatty acid (VLCFA) inhibitor herbicides (WSSA Group 15) are used in row crops for the control of annual grasses and small-seeded broadleaf species (Weed Technology (2000) 14:161-166; American Journal of Plant Sciences (2014) 5:2040). However, VLCFA inhibitor herbicides are not labeled for rice production in the United States because they are primarily harmful to rice. However, if rice tolerance could be established, VLCFA inhibitor herbicides would be alternative herbicide SOAs. Certain application methods, such as fall application before planting and slow-release microencapsulated herbicide substrates, have been shown to reduce injury induced by these herbicides. However, currently available methods do not reduce injury to commercially acceptable levels. Thus, there is a need in the art for new methods to selectively remove weedy rice and other weeds from cultivated rice without significantly damaging the crop. Summary of the Invention

[0005] The present invention provides methods of cultivating rice, the methods including treating the rice with a safener and applying to the soil a microencapsulated Group 15 herbicide. In a preferred embodiment, the safener is fenclorim and the Group 15 herbicide is acetochlor. The present invention further provides rice plants produced by the methods disclosed herein, as well as seeds produced by said rice plants. [Brief description of the drawings]

[0006] [Figure 1] 1 is a set of photographs from a greenhouse trial comparing weedy rice with cultivated rice simultaneously treated with 1,050 grams active ingredient / hectare of microencapsulated (ME) acetochlor (Warrant®, Bayer CropScience, St. Louis, Mo.), where the seeds were planted alone or with cultivated rice seeds treated with a safener provided herein, when acetochlor was added to the soil three weeks prior to planting. Reduced injury was also observed when the herbicide was applied prior to planting and when the seeds were pretreated with the safener. [Diagram 2] FIG. 2 is a bar graph showing the results of the greenhouse trial of FIG. 1 as a percentage of injury to the plants. [Diagram 3] 3 is a bar graph showing percent rice injury 21 days after treatment (DAT) from preemergence applications of the safener fenclolim in combination with the herbicide ME acetochlor. Acetochlor was applied at rates of 313, 632, 1,262, and 2,524 grams active ingredient / hectare, formulated as an emulsifiable concentrate (EC; dark grey bars) or microencapsulated (ME; light grey bars). [Figure 4] 4 is a bar graph showing percent rice injury 21 days after treatment (DAT) from a combination of the safener fenclorim and the herbicide acetochlor applied late preemergence, 4 days after planting. Acetochlor formulated as an emulsifiable concentrate (EC; dark grey bars) or microencapsulated (ME; light grey bars) was applied at rates of 313, 632, and 1,262 grams active ingredient per hectare. [Diagram 5] Figure 5 is a set of photographs of acetochlor treated rice (left), untreated rice (center), and rice treated with fenclorim seed treatment followed by acetochlor (right) 4 weeks after application. ME acetochlor was a delayed pre-emergence application. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0007] In the present invention, it has been discovered that treating rice with both a safener and a microencapsulated Group 15 herbicide reduces rice injury to commercially acceptable levels while still providing effective weed rice control. In some embodiments, the rice is treated with a combination of a microencapsulated formulation of the safener fenclolim and the Group 15 herbicide acetochlor. Importantly, the inventors show that application of either fenclolim or microencapsulated acetochlor alone provides only partial protection to the rice. Thus, a combination of these two treatments is necessary to achieve sufficient tolerance to commercial use rates of the Group 15 herbicide. The present invention provides a method of growing rice comprising treating the rice with a combination of a safener and a microencapsulated Group 15 herbicide.

[0008] Herbicides As used herein, the term "herbicide" is used to refer to a substance used to destroy or inhibit undesirable vegetation. When a herbicide is generally referred to herein by name, unless otherwise specified, the herbicide includes all commercially available forms known in the art, such as salts, esters, free acids and free bases, and stereoisomers thereof. Herbicides are commonly used as emulsifiable concentrates (ECs). However, in the present invention, the primary herbicides are also utilized in microencapsulated (ME) formulations in which the herbicide molecules are protected from degradation processes by a porous polymer shell. See, for example, U.S. Pat. No. 9,877,478, the entire contents of which are incorporated herein by reference. Upon exposure to soil moisture (e.g., by activating rainfall), the polymer shell dissolves, allowing for a sustained release of the herbicide. This delayed release of the herbicide allows time for the rice plant to absorb soil water during germination and grow uninhibited for a period of time after application. Furthermore, the gradual release of the herbicide over time allows the herbicide to provide longer residual control of target weeds compared to EC formulations. Here and elsewhere, we have demonstrated that rice is more tolerant to ME herbicide formulations than EC, likely due to the potential for immediate absorption of the herbicide from EC formulations after rainfall (Weed Technology (2018) 33:239-245).

[0009] In microencapsulated formulations, the release rate of the core material can be controlled by the selection of several parameters, including shell wall composition, mass ratio of herbicide to shell wall material, core material components, average microcapsule particle size, processing conditions such as mixing shear and time, and combinations thereof. In some formulations, diluents, such as solvents, may be added to change the solubility characteristics of the core material to increase or decrease the release rate of the active ingredient from the microcapsules. The diluent may be selected from essentially any known in the art, so long as it is compatible with the core material and shell. In some formulations, the core material may include a blend of a first population and a second population of particulate microencapsulated herbicides to provide a multimodal (e.g., bimodal) release rate. Additional components may be added to the core material to improve its properties. Exemplary components include, but are not limited to, thickeners, stabilizers, anti-filling agents, drift control agents, biocides or preservatives, antifreeze agents, and defoamers.

[0010] Group 15 herbicides are very long chain fatty acid (VLCFA) inhibitors. Thus, the terms "Group 15 herbicides", "VLCA inhibitors", and "VLCFA-inhibiting herbicides" are used interchangeably throughout this application. These soil-applied herbicides are primarily absorbed by shoots and roots, where they inhibit cell development and cell division. Only five weed species worldwide are resistant to VLCFA-inhibiting herbicides, suggesting a lower risk of resistance to this class of herbicides compared to other rice herbicides. VLCFA-inhibiting herbicides include chloroacetanilide chemistry (e.g., acetochlor, alachlor, metolachlor, dimethenamid, petoxamide, pretilachlor, butachlor) and isoxazoline chemistry (e.g., pyroxasulfone). Although the VLCFA inhibitors pretilachlor and butachlor are commonly used in Asian rice production, they provide only marginal weed control. Thus, in a preferred embodiment, the herbicide used in the present invention provides weed control at commercially acceptable levels. Suitable herbicides include, but are not limited to, acetolachlor, metolachlor, and dimethenamid. In a particularly preferred embodiment, an acetochlor herbicide is utilized. Acetochlor is a widely used VLCFA inhibitor that belongs to the chloroacetamide family. Currently, in the United States, it is labeled for use in corn, cotton, soybean, and sorghum. Acetochlor is generally applied pre-emergence for the control of annual grasses and small-seeded broadleaf species. Acetochlor is commercially available in a microencapsulated (ME) formulation (Warrant®, Monsanto Co., St. Louis, MO).

[0011] In the methods of the invention, the Group 15 herbicide may be applied either before or after the rice is planted. In some embodiments, the herbicide is applied to the field 1-250 days prior to planting. In other embodiments, the herbicide is applied after planting, and application may be either pre-emergence or post-emergence. Compared to pre-emergence plants, emergent plants are generally relatively unaffected by seedling growth inhibiting herbicides. Therefore, the prior art practice has been to apply herbicides after crop emergence and before weed emergence. Post-emergence treatments include early post-emergence (EPOST) applications, as well as applications at the spiking stage, 1-2 leaf stage, or 3-4 leaf stage. In some embodiments, the herbicide application is pre-emergence. "Pre-emergence" refers to any time between planting of the crop plant and emergence (i.e., before dehiscence or spiking) of the crop plant, but not including the time of emergence. Pre-emergence treatment includes both treatment of the crop area before sowing (i.e., pre-planting incorporation) and treatment of the sown crop area where the plants have not yet emerged. In particular, the activity of herbicides such as acetochlor is highly dependent on rainfall, and earlier application times may increase the likelihood of damage to rice. Therefore, in another embodiment, Group 15 herbicides are applied as delayed pre-emergence. "Late pre-emergence" generally refers to a time at least 4 days after planting and up to 14 days after planting, when the seeds have absorbed water and germinated, but before the shoots have emerged. The effective amount of microcapsules applied to agricultural fields depends on the attributes of the herbicide encapsulated, the release rate of the microcapsules, the crop being treated, and environmental conditions, particularly the type and moisture of the soil. In general, the application rate of the herbicide, such as acetochlor, is about 0.1, 0.2, 0.5, 1, 2, 3, 4, or 5 kilograms / hectare of herbicide, or ranges thereof, such as 0.1 to 5 kilograms / hectare, 0.2 to 4 kilograms / hectare, 0.25 to 2 kilograms / hectare, or 0.5 to 1 kilogram / hectare. Preferably, application rates of about 0.25 to about 2 kilograms / hectare are used. In some embodiments, the herbicide is applied at a rate of at least 250 grams / hectare of active ingredient. In other embodiments, the herbicide is applied at a rate of at least 1,260 grams / hectare of active ingredient.

[0012] The method may be applied in combination with additional herbicides. Applying several herbicides with different modes of action may be useful, for example, to treat fields with herbicide-resistant weeds, such as barnyard grass. Exemplary adjuvant herbicides include, but are not limited to, ACCase inhibitors (e.g., aryloxyphenoxypropionics), enolpyruvylshikimate-3-phosphate synthase (EPSPS) inhibitors (e.g., glyphosate), glutamine synthetase inhibitors (e.g., glufosinate), synthetic auxins (e.g., aromatic acid, phenoxy and pyridine herbicides), photosystem II (PSII) inhibitors (e.g., urea and triazines), ALS or AHAS inhibitors (e.g., sulfonylureas, triazolopyrimidynins and imidazolinones), photosystem I (PSI) inhibitors (e.g., paraquat), protoporphyrinogen oxidase (PPO) inhibitors (e.g., diphenoxypropionic acid, ... Herbicides that may be used include, for example, aryl ethers, phenylpyrazoles, aryl triazones, and oxadiazoles), mitotic inhibitors (e.g., anilides, amides, certain organophosphorus and carbanilic acid herbicides), cellulose inhibitors (e.g., nitrile and oxazole herbicides), oxidative phosphorylation uncouplers, dihydropteroate synthase inhibitors, fatty acid and lipid biosynthesis inhibitors (e.g., thiocarbamates and certain organophosphorus herbicides), auxin transport inhibitors (e.g., amide and urea herbicides), and carotenoid biosynthesis inhibitors (e.g., isoxazolidinones, benzoylcyclohexanediones, and benzoylpyrazole herbicides), salts and esters thereof, and mixtures thereof. In some embodiments of the present invention, one or more of these adjuvant herbicides are not encapsulated.

[0013] Safeners As used herein, the term "safener" is used to refer to a compound that antagonizes the harmful effects of herbicides on cultivated plants. Safeners were previously called "antidotes" and the terms can be used interchangeably. Preferably, these compounds protect (meaning control) cultivated plants without significantly affecting the action of the herbicide on the weed. The method of the present invention may utilize any safener that protects rice plants from injury from Group 15 herbicides. In a preferred embodiment, the safener is fenclorim (4,6-dichloro-2-phenylpyrimidine). Safeners are most effective when applied before or at the same time as the herbicides whose damage they prevent. Depending on their properties, safeners can be used to pre-treat the seeds of cultivated plants (dressed seeds or shoots), can be incorporated into the soil (e.g., in furrows) before or after sowing the seeds, or can be applied before or after the emergence of the plants, alone or together with the herbicide (e.g., as a tank mix). Thus, the treatment of the plants or seeds with safeners can be carried out independently of the application of the herbicide, or the treatments can be carried out simultaneously. In a preferred embodiment, the safeners are applied to the seeds before planting (i.e., by coating the seeds with the safeners).

[0014] The rate at which safeners are applied in relation to herbicides depends largely on the mode of application. When safeners are applied as field treatments, either alone or as tank mixtures with herbicides, the ratio of safener to herbicide is usually 1:100 to 10:1, but more commonly 1:5 to 8:1. However, when safeners are applied as seed dressings, much smaller amounts are required per hectare of crop area than when applied later. For seed dressings, usually 0.1 to 10 g of safener per kg of seed is required, with the preferred amount being 0.1 to 3 g per kg of seed. The safeners can be used in unmodified form or as compositions with conventional auxiliaries and carriers.The safeners can be formulated in any known manner, for example, as emulsifiable concentrates, directly sprayable or dilutable solutions, dilute emulsifiable concentrates, wettable powders, soluble powders, dusts, granules, and encapsulation in, for example, polymeric substances.The compositions can also contain further ingredients, such as stabilizers, defoamers, viscosity regulators, binders, adhesives, and fertilizers or other active compounds, to achieve special effects. Safener formulations are prepared in known manner, for example by mixing and / or grinding the active ingredient with extenders, such as solvents, solid carriers and, where appropriate, surface-active compounds (surfactants). Suitable safener solvents include, but are not limited to, aromatic hydrocarbons, preferably fractions containing 8 to 12 carbon atoms, such as xylene mixtures or substituted naphthalenes, phthalates such as dibutyl phthalate or dioctyl phthalate, aliphatic hydrocarbons such as cyclohexane, or paraffins, alcohols and glycols and their ethers and esters, for example ethanol, ethylene glycol, ethylene glycol monomethyl or monoethyl ether; ketones such as cyclohexanone, highly polar solvents such as N-methyl-2-pyrrolidone, dimethyl sulfoxide or dimethylformamide; and epoxidized vegetable oils such as epoxidized coconut oil or soybean oil; or water. Suitable solid carriers, for example used for dusts and dispersible powders, are usually natural mineral fillers, such as calcite, talc, kaolin, montmorillonite or attapulgite. Also, highly dispersed silicic acid or highly dispersed absorbent polymers can be added to improve the physical properties. Suitable granulated adsorptive carriers are of the porous type, such as pumice, crumbled brick, sepiolite or bentonite, and suitable non-adsorbent carriers are materials such as calcite or sand. In addition, a large number of pre-granulated materials of inorganic or organic nature can be used, such as dolomite or crushed plant residues in particular. Depending on the nature of the safener to be formulated, suitable surface-active compounds are non-ionic, cationic and / or anionic surfactants with good emulsifying, dispersing and wetting properties. Surfactants commonly used in such formulations are described, for example, in the following publications: "McCutcheon's Detergents and Emulsifiers Annual", MC Publishing Corp., Ringwood, NJ, 1979; Sisely and Wood, "Encyclopedia of Surface Active Agents", Chemical Publishing Co. Inc., New York, 1964.

[0015] At the molecular level, herbicides and their respective safeners are usually quite similar. Thus, safeners may act as "biomodulators" that affect the amount of herbicide that reaches its target site in an active form, or as "antagonists" of the herbicide at a similar site of action. Although some safeners reduce the amount of herbicide that reaches its site of action by slowing its rate of uptake and / or translocation, most safeners currently developed function by increasing the rate of metabolic detoxification. Safeners belonging to several chemical classes (i.e., phenylpyrimidines, dichloroacetamides, oxime ethers, and thiazoles) are believed to protect plants against injury from chloroacetanilide herbicides (a class of Group 15 herbicides) by enhancing the binding of these herbicides to the reduced form of the thiol glutathione. After the herbicide is bound to glutathione in the cytoplasm, it is sequestered in the vacuole for degradation and, therefore, detoxification. Glutathione S-transferase enzymes (GSTs) catalyze the conjugation of glutathione to a variety of substrates, and therefore these safeners may function by either increasing the levels of reduced glutathione or by inducing the activity of GSTs.

[0016] grant The method of the present invention can be used to grow any cultivated rice variety. Cultivated rice is generally from the Oryza genus, most commonly from the Sativa and Glaberima species. Rice utilized in the present invention may include both non-traited and traited rice varieties. As used herein, the term "traited rice" is used to refer to rice having a herbicide resistance trait. Commonly used trait rice varieties include imidazolinone-resistant (Clearfield®, BASF Corporation, Research Triangle Park, NC) rice and quizalofop-resistant (Provisia®, BASF Corporation, Research Triangle Park, NC) rice. Although these traited varieties are very popular, widespread use of certain inhibitors used with these varieties and lack of adherence to proper use guidelines has led to rapid resistance development in some weed populations. Thus, the methods of the present invention utilizing herbicides with different sites of action are particularly useful for cultivating trait-imparted varieties that suffer from resistant weeds.Specific cultivars that may be used with the methods provided herein include, but are not limited to, rice cultivars Diamond, Titan, FP7521, FP7321, PVL01, and PVL02.

[0017] Rice treated with the method of the invention can be planted using several techniques. In the United States, rice production is broadly classified as either dry or flooded direct seeding. In dry direct seeding, rice is sown in a prepared seedbed with a row seeder or by scattering the seeds and working them in with a disk or harrow. Moisture for seed germination is then provided by irrigation or rainfall. In a preferred embodiment, the safener is applied to the rice seeds as a dressing or concentrated formulation, and the seeds are planted by dry direct seeding. In contrast, in flooded direct seeding, rice seeds are soaked for 12-36 hours to induce germination, and the seeds are sprayed by airplane into flooded fields. Shoots germinate by shallow flooding, or the fields may be drained for a short period of time to enhance shoot establishment. Thus, in embodiments where flooded direct seeding is used, the safener may be applied to the soaking solution used to induce germination. Furthermore, rice treated with the method of the present invention can be grown in unfavorable environmental conditions. Specifically, the inventors have demonstrated that the method can be used in cool and humid conditions (see Example 5). Thus, in some embodiments, rice is grown under cool and humid environmental conditions. Temperatures in the range of 10-15°C can cause chilling damage to the rice plant depending on the reproductive stage of the plant. Thus, a climate with an average night temperature of 10°C to 16°C and / or an average day temperature of 20°C to 27°C can be considered "cool" for growing rice. As used herein, "humid" conditions for growing rice include those with soil moisture of 70% to 90% on average. The experiment was conducted in a growth chamber at 13.8°C (night) and 23.8°C (day) with a 10-h night and 14-h day photoperiod. Diamond rice was planted in pots filled with the same mass of soil to maintain 80% soil moisture throughout the experiment. Treatments were 0 and 1,050 g ai / ha. -1 ME acetochlor at 0 and 2.5 g ai / kg -1 The seed consisted of fenclorim seed treatment applications. Data were analyzed using R and subjected to ANOVA. Means were separated using Fisher's LSD (α=0.05).

[0018] The methods of the invention can be useful for controlling a variety of weeds, ie, plants that are considered pests or competitors of commercially important crop plants. Examples of weeds that may be controlled according to the method of the present invention include, but are not limited to, Echinochloa crus-galli and other weed species in the genus Echinochloa, crabgrass in the genus Digitaria, Amaranthus palmeri and other weed species in the genus Amaranthus, Portulaca oleracea and other weed species in the genus Portulaca, Chenopodium album and other Chenopodium spp., Setaria lutescens and other Setaria spp., Solanum nigrum and other Solanum spp., Lolium multiflorum and other Lolium spp., Brachiaria platyphylla and other Braehiaria spp., Conyza canadensis and other Conyza spp., and Eleusine indica. In certain preferred embodiments, the weed species is Oryza sativa L. var. sylvatica (weedy rice) or Oryza sativa L. (red rice).

[0019] As further used herein, "weed control" refers to any observable measure of control of plant growth, which may include one or more of: (1) death; (2) inhibition of growth, reproduction or proliferation; and (3) removal, destruction, or otherwise reduction of plant development and activity. Weed control can be measured by any of a variety of methods known in the art. For example, weed control can be determined as a percentage compared to untreated plants according to standard procedures in which plant mortality and growth reduction are visually assessed by a person skilled in the art. Control can be defined, for example, in terms of average plant mass reduction or the percentage of plants that fail to germinate after pre-emergence herbicide application. "Commercially acceptable weed control rates" vary depending on the weed species, the degree of infestation, environmental conditions, and the crop plant involved. Commercially effective weed control can be defined as destruction (or inhibition) of at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, or even at least 85%, or even at least 90%. Generally, destruction of 80% or more of the weeds is preferred from a commercial point of view, but commercially acceptable weed control can only occur at much lower destruction or inhibition levels, especially for some highly harmful herbicide-resistant plants. Advantageously, the herbicidal microcapsules used according to the present invention achieve commercially acceptable weed control within an application period of 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 weeks after application of the herbicidal microcapsules. Crop injury can be measured by any means known in the art, such as those described above for weed control determination. The "commercially acceptable crop injury rate" of the present invention similarly varies with crop plant species. In general, commercially acceptable crop injury rates are defined as less than about 20%, 18%, 16%, 15%, 13%, 12%, 11%, 10% or even less than about 5%. The method of the present invention limits crop injury to a commercially acceptable rate as measured from about 24 hours (about 1 DAT) to 3 weeks (about 21 DAT) after application.

[0020] The disclosure is not limited to the specific details of the configurations, arrangement of components, or method steps shown herein. The compositions and methods disclosed herein can be made, implemented, used, performed, and / or formed in a variety of ways that will be apparent to one of skill in the art in light of the following disclosure. The phrases and terms used herein are merely for descriptive purposes and should not be considered as limiting the scope of the claims. Ordinal indicators such as first, second, and third, when used in the description and claims to refer to various structures or method steps, are not meant to be construed as indicating any particular structure or step, or any particular order or arrangement for such structures or steps. All methods described herein can be performed in any suitable order unless otherwise noted herein or clearly contradicted by context. Any and all examples provided herein, or the use of exemplary language (e.g., "etc.") are intended merely to facilitate disclosure and do not imply any limitation on the scope of the disclosure unless specifically stated otherwise. No language in this specification should be construed as indicating that a structure shown in a drawing is essential to the practice of the disclosed subject matter. Use of the terms "including," "comprising," or "having" and variations thereof herein is meant to encompass the elements listed thereafter and equivalents thereof, as well as additional elements. Embodiments recited as including, comprising, or having particular elements are also contemplated as "consisting essentially of" and "consisting of" those particular elements.

[0021] The recitation of ranges of values ​​herein is intended to serve only as a shorthand method of individually referring to each individual value falling within the range, unless otherwise stated herein, and each individual value is incorporated herein as if it were individually recited herein. For example, if a concentration range is stated as 1% to 50%, it is intended that values ​​such as 2% to 40%, 10% to 30%, or 1% to 3%, etc. are expressly recited herein. These are only examples of what is specifically intended, and all possible combinations of values ​​therebetween, including the lowest and highest recited values, should be considered to be expressly stated in this disclosure. The use of the word "about" to describe a particular recited amount or range of amounts is meant to indicate that the amount includes values ​​that are very close to the recited amount, such as values ​​that may or would be accounted for due to manufacturing tolerances, equipment and human error in making measurements, and the like. All percentages referring to amounts are by weight unless otherwise stated.

[0022] No admission is made that any reference, including any non-patent or patent literature cited herein constitutes prior art. In particular, unless otherwise noted, it will be understood that the reference of any document herein is not an admission that any of these documents form part of the common general knowledge in the art in the United States or any other country. The discussion of any reference states what their authors assert, and applicants reserve the right to challenge the accuracy and pertinence of the documents cited herein. All references cited herein are incorporated by reference in their entirety unless expressly noted otherwise. In the event of any discrepancy between definitions and / or explanations found in the cited references, the present disclosure shall control. The following examples are meant to be illustrative only and are not meant to limit the scope of the invention or the claims appended hereto. The present invention also provides rice plants cultivated by the methods disclosed herein, as well as seeds produced by these rice plants. EXAMPLES

[0023] In previous work, the inventors determined that rice resistance to the Group 15 herbicide acetochlor was improved by providing this herbicide in a microencapsulated (ME), controlled release formulation (Weed Technology (2018) 33:239-245). In the following examples, rice was treated with a combination of ME acetochlor and a safener. Also, Ferenfrom was applied as a seed treatment and ME acetochlor was applied either pre-emergence or delayed post-emergence. Yet another aspect of the present invention may be as follows. [1] A method for cultivating rice, comprising treating rice seeds with a safener and applying to the soil a herbicide, wherein the herbicide is a Group 15 herbicide, and the herbicide is microencapsulated. [2] The method according to [1], wherein the herbicide is selected from the group consisting of acetochlor, metolachlor, dimethenamid, and alachlor. [3] The method according to any one of [1] to [2] above, wherein the herbicide is applied before or after rice is planted. [4] The method according to [3] above, wherein the herbicide application is pre-emergence or delayed pre-emergence. [5] The method according to any one of [1] to [4], wherein the herbicide is applied at a rate of at least 250 grams of active ingredient per hectare. [6] The method according to [5], wherein the herbicide is applied at a rate of at least 1,260 grams of active ingredient per hectare. [7] The method according to any one of [1] to [6], wherein the safener is fenclorim. [8] The method according to any one of [1] to [7], wherein the safener is applied to rice seeds before planting. [9] The method according to [8], wherein the rice seeds are coated with the safener before planting.

[10] The method according to any one of [1] to [9], wherein the rice is a sativa species (Oryza sativa L.).

[11] The method according to any one of [1] to

[10] above, wherein the rice is sown using a seed drill.

[12] The method according to any one of [1] to

[11] above, wherein weeds are controlled by the herbicide.

[13] The method according to

[12] , wherein the herbicide controls barnyard grass and / or weedy rice.

[14] The method according to

[12] above, wherein the weed control rate is at least 50%.

[15] The method according to any one of [1] to

[14] , which reduces crop damage and / or reduces upright drop caused by the herbicide compared to a crop treated with the herbicide alone.

[16] The method according to

[15] above, wherein the crop damage rate is 20% or less and / or the upright drop rate is 20% or less.

[17] The method according to any one of [1] to

[16] above, wherein a commercially acceptable weed control rate and a commercially acceptable crop damage rate are simultaneously achieved.

[18] The method according to [1], wherein the herbicide is acetochlor, the safener is fenclorim, and the safener is applied to rice seeds before planting.

[19] The method according to any one of [1] to

[18] , wherein the rice is grown under cool and humid environmental conditions.

[20] A rice plant produced by the method according to any one of [1] to

[19] above.

[21] Seeds produced by the rice plant according to

[20] above.

[0024] Example 1: Greenhouse Study To determine whether the timing of herbicide application or any other method would allow the use of Group 15 herbicides in rice, we conducted several greenhouse trials. Figure 1 is a set of photographs from a greenhouse trial showing that the Group 15 herbicide acetochlor can effectively reduce the growth of weedy rice when applied at planting. However, when microencapsulated (ME) acetochlor was applied to the rice crop at planting, it caused a significant amount of injury to the rice (over 80% injury), as shown in Figure 2. In contrast, when acetochlor was applied 3 weeks before planting, the injury to the rice crop was reduced to about 50%, which is still too much injury for commercial use. Finally, cultivated rice seeds were treated with a safener and ME acetochlor herbicide was applied to the soil 3 weeks before planting. In this trial, the injury to the rice was only about 10% (see Figure 2), which is acceptable for commercial use.

[0025] Example 2: Field study Materials and Methods: Field trials were conducted to evaluate the tolerance of fenclorim-protected rice to different rates, timing, and formulations of acetochlor. All trials were conducted in silty loam soil. Prior to the trials, rice cultivar Diamond seeds were soaked in water at 0, 0.25, or 2.5 grams per kilogram of seed (g kg seed) using standard procedures. -1 ) of fenclorim. The trial was set up using a split plot design with the acetochlor rates and formulations as main plots and the fenclorim-treated seeds as subplots. Treatments were replicated with four times. Each plot was 2.2m wide by 1.5m long and surrounded by a 1.5m border. In each plot, rice seeds were hand planted at 22 seeds per 0.3m in furrows spaced 15cm apart using a 10-row cone drill planter. Seeds were dosed at 0, 0.25 and 2.5g kg seed. -1 The seeds treated with fenclorim were planted in furrows 2, 3, 5, 6, 8 and 9, respectively.

[0026] result: Treatment 1: Preemergence application of acetochlor Acetochlor, formulated as an emulsifiable concentrate (EC, Harness®) or microencapsulated (ME, Warrant®), was administered at doses of 313, 632, 1,262 and 2,524 grams of active ingredient per hectare (g ai ha -1 The application was done pre-emergence at a rate of acetochlor equivalent to 1000 ml of CO2 per hectare using a 110015AIXR nozzle calibrated to deliver 140 liters per hectare. 2 A pressurized backpack sprayer was used to spray at 4.8 kilometers per hour (km hr -1 The plots were then irrigated using an overhead side irrigation system to incorporate the herbicide into the soil solution. The injury to the rice plants was recorded 21 days after treatment, and the results are shown in Figure 3. Treatment 2: Delayed preemergence application of acetochlor Acetochlor, formulated as an emulsifiable concentrate (EC, Harness®) or microencapsulated (ME, Warrant®), was administered at 313, 632, and 1,262 g ai ha -1 The equivalent acetochlor rate was applied as delayed pre-emergence. Application was at 140 L ha -1 CO fitted with an 110015AIXR nozzle calibrated to deliver 2 4.8km hr using a pressurized backpack sprayer -1 After planting the rice seeds, the plots were irrigated from above using a side irrigation system to promote germination. Herbicide treatments were applied 4 days later, and the herbicide was taken up into the soil solution by natural rainfall 2 days later. At this time, the coleoptiles of the rice plants began to protrude from the soil (spiking stage). At 21 days after treatment, injury to the rice plants was recorded and the results are shown in Figure 4.

[0027] Conclusion: Rice treated with a combination of microencapsulated acetochlor and the safener fenclorim showed increased herbicide tolerance, allowing the use of the potent Group 15 herbicide acetochlor to control grass weeds, including weedy rice. The combination treatment was effective when acetochlor was applied both preemergence and delayed preemergence, with the most dramatic reduction in rice injury observed with delayed application.

[0028] Example 3: Tolerance of rice to pre-emergence (PRE) and delayed pre-emergence (DPRE) applications of microencapsulated (ME) acetochlor A trial was conducted in spring 2020 to determine whether a fenclorim rice seed treatment would increase the tolerance of rice crops to microencapsulated (ME) acetochlor under typical planting conditions. The rice cultivar ‘Diamond’ was grown at 72 seeds per metre row (m -1 Rice plots were treated using standard miniplot research methods. Treatments included two application times [preemergence (PRE) and delayed preemergence (DPRE)] and three ME acetochlor rates (0, 1,260, and 1,890 g ai ha). -1 ), and fenclorim seed treatments (0 and 2.5 g ai kg -1 Data were analyzed using R, and analysis of variance (ANOVA) was performed using Fisher's LSD (α = 0.05) for separation.

[0029] [Table 1] [Table 2]

[0030] Conclusion: Fenclorim seed treatments demonstrated a mitigating effect in the form of reduced stand loss from ME acetochlor. When averaged over application times and acetochlor rates, loss of stand from acetochlor was reduced by more than 50% (Table 1). Although not significant, loss of stand was greater in plots without fenclorim seed treatment when comparing treatments with PRE and DPRE application times and acetochlor rates (Table 2). Loss of stand was less than 16% for treatments containing fenclorim seed treatment with the PRE application time and less than 13% for treatments with the DPRE application time.

[0031] Example 4: Tolerance of several rice cultivars to ME-acetochlor when using fenclorim seed treatment Trials were conducted in spring 2020 to determine cultivar response to fenchlorim rice seed treatment and ME acetochlor. Two inbred cultivars ('Diamond' and 'Titanium') and two hybrid cultivars ('FP7521' and 'FP7321') were grown for 72 and 36 seed m, respectively. -1 The rice plots were treated using standard small plot research methods. Treatments were 0 and 1,260 g ai ha -1 ME acetochlor at 0, 2.5, and 5.0 g ai kg -1 The seeds consisted of a fenclorim seed treatment. Cultivars were analyzed separately using R, with ANOVA performed to separate means using Fisher's LSD (α=0.05).

[0032] [Table 3] [Table 4]

[0033] Conclusion: Results from the cultivar trials show the protective effect of fenclorim seed treatment. The protective effect can be observed from ground cover data 6 weeks after treatment (Table 3). As the fenclorim seed treatment rate increased, upright loss and injury generally decreased (Table 4). Fenclorim seed treatment produced a positive response in canopy closure for rice cultivar Titan without acetochlor treatment. Furthermore, in the acetochlor treated plots, fenclorim seed treatment improved canopy closure, indicating protection from the seed treatment for all cultivars.

[0034] Example 5: Tolerance of rice to ME acetochlor under cool and wet soil conditions A trial was conducted in fall 2019 to determine rice tolerance to ME acetochlor when treated with a fenchlorim rice seed treatment under cool and wet environmental conditions. The trial was conducted in a growth chamber at 13.8°C (night) and 23.8°C (day) using a 10-h night and 14-h day photoperiod. Diamond rice was planted in pots filled with the same mass of soil to maintain 80% soil moisture throughout the trial. Treatments were 0 and 1,050 g ai ha. -1 of ME acetochlor and 0 and 2.5 g ai kg -1 The seeds consisted of fenclorim seed treatments. Data were analyzed using R and ANOVA was performed. Means were separated using Fisher's LSD (α = 0.05).

[0035] [Table 5] Under cool, moist conditions, fenclorim seed treatments and ME acetochlor provided sufficient safety (Table 5). Seeds treated with both fenclorim seed treatments and ME acetochlor were not significantly different from untreated rice when assessed at 3 weeks after application (WAA) and 4WAA. Figure 5 shows the protective effects seen in the growth chamber.

[0036] Example 6: Weed control and rice tolerance with ME acetochlor and fenclorim seed treatments A trial was conducted in spring 2020 to determine the control of barnyardgrass and weedy rice in cultivated rice for ME acetochlor when applied in a fenchlorim rice seed treatment. The trial was conducted under standard small plot research methods. Diamond rice was grown in 72 seed m -1 Treatments were applied at four application times (preemergence, delayed preemergence, spiking, and 1-leaf) and three application rates (630, 1,260, and 1,890 g ai ha -1 ) and 0 and 2.5 g ai kg -1 The seed consisted of fenclorim seed treatments at 100-2000 s. Data were analyzed using R and ANOVA was performed. Means were separated using Fisher's LSD (α=0.05).

[0037] [Table 6] [Table 7] ME acetochlor (1,260 and 1,890 g ai ha -1 For the PRE and DPRE applications of ME-FENCLORIM, the herbicide provided greater than 80% control of knotweed and greater than 50% control of weedy rice (Table 6). In the absence of fenclorim seed treatment, it is not possible to eliminate weedy rice from cultivated rice. Furthermore, fenclorim seed treatment also reduced the level of injury observed from ME-Acetochlor applications, and the seed treatment did not adversely affect weed control (Table 7).

Claims

1. 1. A method of cultivating rice comprising treating rice seeds with a safener and applying to the soil a herbicide, said herbicide being a Group 15 herbicide, said herbicide being microencapsulated, said herbicide being selected from the group consisting of acetochlor, metolachlor, dimethenamid, and alachlor, and said safener being fenclorim.

2. 10. The method of claim 1, wherein the herbicide is applied before or after the rice is planted.

3. 3. The method of claim 2, wherein the herbicide application is pre-emergence or delayed pre-emergence.

4. 4. The method according to any one of claims 1 to 3, wherein the herbicide is applied at a rate of at least 250 grams of active ingredient per hectare.

5. 5. The method of claim 4, wherein the herbicide is applied at a rate of at least 1,260 grams active ingredient per hectare.

6. The method according to any one of claims 1 to 5, wherein the safener is applied to rice seeds before planting.

7. The method of claim 6 , wherein the rice seeds are coated with the safener before planting.

8. The method according to any one of claims 1 to 7, wherein the rice is Oryza sativa L.

9. The method according to any one of claims 1 to 8, wherein the rice is sown with a seed drill.

10. 10. The method according to any one of claims 1 to 9, wherein weeds are controlled by said herbicide.

11. 11. The method of claim 10, wherein the herbicide controls barnyard grass and / or weedy rice.

12. 11. The method of claim 10, wherein the weed control rate is at least 50%.

13. 12. The method of any one of claims 1 to 11, which results in reduced crop injury and / or reduced upright drop caused by the herbicide compared to a crop treated with the herbicide alone.

14. 14. The method of claim 13, wherein the crop injury rate is 20% or less and / or the upright drop rate is 20% or less.

15. 15. The method of any one of claims 1 to 14, wherein a commercially acceptable rate of weed control and a commercially acceptable rate of crop damage are simultaneously achieved.

16. 2. The method of claim 1, wherein the herbicide is acetochlor, the safener is fenclorim, and the safener is applied to rice seeds prior to planting.

17. 17. The method according to any one of claims 1 to 16, wherein the rice is grown in cool and humid environmental conditions.

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