Nematode suppression

Abstract

The present disclosure relates to methods for controlling nematodes. Also provided are methods for increasing the yield of nematode-susceptible plants.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority pursuant to 35 U.S.C. §1.19(e) of U.S. Provisional Patent Application No. 63 / 174,191, filed April 13, 2021, the entirety of which is incorporated herein by reference.

[0002] The present disclosure relates to methods for controlling nematodes. Also provided are methods for increasing the yield of nematode-susceptible plants.

[0003] Incorporation by Reference of Material Submitted Electronically This application contains a sequence listing in computer readable form, which is incorporated by reference in its entirety as a separate part of this disclosure and is identified as follows: Filename: 202662_Seqlisting.txt; Size: 15,423 bytes; Creation date: March 21, 2022. [Background technology]

[0004] Nematodes are active, adaptable, elongated organisms that live on moist surfaces or in liquid environments, including water films in soil and moist tissues within other organisms. Many species of nematodes have evolved to be highly successful parasites of plants and animals, responsible for significant economic losses in agriculture and livestock, and morbidity and mortality in humans (Whitehead (1998) Plant Nematode Control. CAB International, New York).

[0005] Parasitic nematodes are estimated to cost the horticulture and agricultural industry over $78 billion annually worldwide, based on an estimated average annual loss of 12% across all major crops. For example, nematodes are estimated to be responsible for approximately $3.2 billion in soybean losses worldwide each year (Barker et al. (1994) Plant and Soil Nematodes: Societal Impact and Focus for the Future. The Committee on National Needs and Priorities in Nematology. Cooperative State Research Service, US Department of Agriculture and Society of Nematologists). Plant parasitic nematodes are major pests of all food crops worldwide, including corn, barley, sorghum, oats, rye, rice, potato, cassava, sweet potato, wheat, soybean, rapeseed, and sunflower (Nicol et al. (2011)), in addition to being important pests of fruits and vegetables, fiber crops (e.g., cotton), ornamentals, and turfgrass (Current Nematode Threats to World Agriculture. In: J. Jones et al. (eds.) Genomics and Molecular Genetics of Plant-Nematode Interactions, Springer Science+Business Media BV2011).

[0006] Nematodes are known to affect crop and plant yield, growth, and health. Physiological changes in the roots of host plants caused by larval and / or adult nematodes can cause the formation of galls, which leads to the destruction of the vascular system of the plant's roots. Root elongation can be completely halted, possibly resulting in insufficient supply of water and nutrients caused by the shrinkage of the root system, causing leaf yellowing and / or damping off, and stunted growth, both of which can result in low yield or death. In addition, nematodes can cause physiological effects that cause the plant's roots to become more susceptible to bacterial and / or fungal attack, including bacteria and / or fungi that the plant would normally resist. Such attack can cause widespread secondary decline and decay.

[0007] The root lesion nematode Pratylenchus brachyurus has become an increasingly important soybean pathogen. It has a wide host range and is widely distributed in tropical and subtropical regions, especially in Brazil, Africa, and the southern United States. Pratylenchus brachyurus is of concern among soybean, corn, and cotton growers in the Cerrado region of Brazil, where it is considered the main nematode pathogen of soybean. In soybean, the nematode can reduce yields by 30-50%, with greater losses observed in sandy soils.

[0008] Several methods spanning cultural, biological, and chemical control can be deployed for nematode management. Host-plant resistance (a form of cultural control) has consistently been the most effective and cost-effective management method. However, host-plant resistance is not available for many nematode species, especially mobile parasites such as Pratylenchus brachyurus. Host-plant resistance may also be limited in its adoption if the genetics are complex (i.e., involve multiple genes), undesirable agronomic traits are also imparted, or pest biotypes exist that can overcome the resistance. Other cultural control tactics include plant quarantine, crop rotation, and cultivation. Plant quarantine limits the import and movement of plant parts that may harbor or promote the growth of invasive pest species. Plant quarantine can keep nematodes out of a country or region, but once a nematode species has become widely distributed within a country or region, such as P. brachyurus in Brazil, such quarantine is not appropriate.

[0009] Crop rotation is the practice of alternating the cultivation of multiple crops over an entire season in a single location. Crop rotation is an effective management strategy for nematode pests with restricted host ranges. The host range of a nematode is defined as the plants that can support the survival and reproduction of the nematode species. Many mobile pests, such as P. brachyurus, have particularly wide host ranges, making rotation to non-hosts impractical. For P. brachyurus in Brazil, all major commodity crops rotated with soybean are suitable hosts. Only a limited number of cover crops, such as Crotalaria spectabilis and Crotalaria ochroleuca, are non-hosts. However, these cover crops do not provide marketable grain or forage yields, so they represent a cost of plants with no economic return. Leaving fields fallow for a season is another strategy similar to planting a non-host cover crop. While leaving fields fallow eliminates the cost of planting cover crops, it presents an inherent risk of potentially undesirable ecological outcomes such as soil erosion. Also, most nematodes can undergo a dormant state that allows for long-term survival in fallow fields until a suitable host is planted. In the case of P. brachyurus, survival can be maintained for more than 90 days in the absence of host plants (Ribeiro et al., Heliyon 6:e05075, 2020). Finally, tillage is a potential nematode control tactic, but is not available to farmers who practice no-tillage agriculture for soil health reasons. Tillage is the preparation of the soil for planting through physical disturbance. Most soybean fields in the Cerrado region of Brazil are tilled under no-tillage practices (i.e., no mechanical tillage occurs during the growing season or between planting seasons). Tillage can also have adverse effects on various nematode species. Soybean cyst nematode (Heterodera glycines) and P. brachyurus occur simultaneously in many fields in the Cerrado region.Tillage may increase the incidence (Workneh et al., Phytopathology 89:844-850, 1999), spread (Gavassoni et al., Phytopathology 91:534-545, 2001) and damage caused by the soybean cyst nematode (H. glycines), while two to three rounds of deep tillage are essential to reduce Pratylenchus spp. populations in some production systems (Khan et al., (2021) Emerging Important Nematode Problems in Field Crops and Their Management. In: Singh KP, Jahagirdar S., Sarma BK (eds) Emerging Trends in Plant Pathology. Springer, Singapore).

[0010] Biological control for nematodes is not easily managed. Fungal and bacterial seed treatments with nematode-suppressing properties have been recently developed. Typically, these seed treatments have several limitations, including additional cost, moderate efficacy, variable performance across environments, and a short window of protection, often limited to the early part of the growing season (Dias-Arieira et al., Journal of Phytopathology 166:722-728, 2018).

[0011] Chemical means of controlling plant parasitic nematodes remain essential for many crops that lack sufficient host plant resistance. However, the activity of chemical agents is often not selective and may adversely affect non-target organisms, including the temporary collapse of beneficial microbial populations. In recent years, the registration of many chemical nematicides has been withdrawn, revoked, or their use restricted, limiting the availability of in-furrow nematicides (Fosu-Nyarko and Jones,In Advances in Botanical Research v73 doi:10.1016 / bs.abr.2014.12.012,2015). Summary of the Invention [Problem to be solved by the invention]

[0012] Thus, there is a need for additional means to control nematode populations that damage and / or cause harm to agriculturally important plants. [Means for solving the problem]

[0013] In one aspect, described herein is a method of suppressing nematode populations at a locus, comprising growing a nematode resistant plant at the locus, wherein growing the nematode resistant plant suppresses nematode populations at the locus or maintains suppression of nematode populations at the locus during and / or beyond the growing of the nematode resistant plant. In some embodiments, the period extends to one or more growing seasons after the growing season in which the nematode resistant plant was grown. In some embodiments, the method further comprises growing a secondary plant at the locus at a time after growing the nematode resistant plant. In some embodiments, the method further comprises growing a secondary plant at the locus at a time prior to growing the nematode resistant plant. In some embodiments, the method further comprises growing a nematode resistant plant at the locus simultaneously with the secondary plant.

[0014] In some embodiments, the secondary plant is a nematode susceptible plant. In some embodiments, the nematode susceptible plant is a perennial or annual plant. In some embodiments, the nematode susceptible plant is a soybean plant, a corn plant, a cotton plant, a canola plant, a sugarcane plant, a potato plant, a wheat plant, a vegetable plant, a rice plant, an alfalfa plant, a barley plant, a sorghum plant, an oat plant, a rye plant, a cassava plant, a sweet potato plant, a sunflower plant, a fruit plant, an orchard plant (e.g., a fruit or nut tree), an ornamental plant, or a grapevine. In some embodiments, the nematode population is suppressed or maintained below the detection limit. In a further embodiment, the secondary plant is of the Brachiaria genus, and the suppression of the nematode population is achieved when the number of nematodes is 60, about 60, or less than about 60 per gram of root. In some embodiments, the secondary plant is corn and suppression of the nematode population is achieved when the number of nematodes is 300, about 300, or less than about 300 nematodes per gram of root. In further embodiments, the secondary plant is cotton and suppression of the nematode population is achieved when the number of nematodes is 60, about 60, or less than about 60 nematodes per gram of root. In some embodiments, the secondary plant is sorghum and suppression of the nematode population is achieved when the number of nematodes is 250, about 250, or less than about 250 nematodes per gram of root. In further embodiments, suppression of the nematode population at the location where the secondary plant is grown is achieved when there is a 5% reduction, about a 5% reduction, or at least about a 5% reduction in the number of nematodes per gram of root compared to the number of nematodes per gram of root at a location comparable to the location where the secondary plant is grown.

[0015] In another aspect, described herein is a method of nematode management for a locus comprising growing nematode resistant plants at the locus in a first growing season, wherein the nematode resistant plants suppress a nematode population at the locus or maintain suppression of a nematode population at the locus; growing nematode susceptible plants at the locus in the same or a subsequent growing season; and achieving improved health and / or yield of the nematode susceptible plants compared to the health and / or yield expected if the nematode population was not suppressed.

[0016] In some embodiments, the nematode resistant plant expresses a Cry protein, such as, but not limited to, a nematicidal Cry protein. In some embodiments, the nematode resistant plant is a soybean plant, a corn plant, a cotton plant, a canola plant, a sugarcane plant, a sugar beet plant, a potato plant, a wheat plant, a rice plant, an alfalfa plant, a barley plant, a sorghum plant, an oat plant, a rye plant, a cassava plant, a sweet potato plant, a sunflower plant, a vegetable plant, a fruit plant, an ornamental plant, an orchard plant (e.g., a fruit or nut-bearing tree), or a grape vine.

[0017] In some embodiments, the nematode is a nematode species selected from a Pratylenchus spp. nematode population, such as Pratylenchus brachyurus, Meloidogyne spp., Heterodera spp., such as Heterodera glycines, Globodera spp., Rotylenchulus reniformis, Helicotylenchus spp., such as Helicotylenchus dihystera, Scutellonema brachyurus, Tubixaba tuxaua, or Aphelencoides besseyi. In some embodiments, the Pratylenchus species is Pratylenchus brachyurus.

[0018] In some embodiments, improved health of nematode-susceptible plants includes one or more of the following: improved root development (e.g., improved root or root hair growth); improved yield; faster emergence; improved plant stress management, including increased stress tolerance and / or improved recovery from stress; increased mechanical strength; improved drought tolerance; reduced fungal, bacterial, and / or viral disease infection; or any combination thereof.

[0019] Also provided is a method of protecting nematode susceptible plants from nematode damage or damage, comprising growing a nematode resistant plant at a locus for at least one growing season prior to planting the nematode susceptible plant; and growing a nematode susceptible plant at a locus for at least one growing season after growing the nematode resistant plant. In some embodiments, protecting nematode susceptible plants from nematode damage comprises increasing the yield of harvested nematode susceptible plant material or revenue from the sale of harvested nematode susceptible plant material.

[0020] Also provided is a method of nematode management for a locus, the method comprising growing a nematode resistant plant at the locus; and growing a nematode susceptible plant at the locus at a time after growing the nematode resistant plant. In some embodiments, the nematode susceptible plant is a perennial. In some embodiments, the nematode susceptible plant is an annual.

[0021] In another aspect, the present disclosure provides a method of nematode management for a locus, comprising growing a nematode-resistant plant in the locus simultaneously with a nematode-susceptible plant. In some embodiments, the nematode-susceptible plant is a perennial plant. In some embodiments, the nematode-susceptible plant is an annual plant.

[0022] In another aspect, the disclosure provides a method of nematode management for a locus, the method comprising: planting a nematode-susceptible plant at the locus, the plant being an annual or perennial; and growing a nematode-resistant plant at the locus at a time after planting the nematode-susceptible plant. In some embodiments, the nematode-susceptible plant is a vegetable plant, a fruit plant, an orchard plant, an ornamental plant, or a vine.

[0023] In another aspect, the disclosure provides a locus having suppressed nematode population density, where the suppression of nematode population density is achieved by the methods described herein. Also provided are nematode-resistant plants grown in the locus, plant material harvested from the plants, and seeds produced by the plants, where the locus (i) does not require tillage; (ii) does not require a cover crop; or (iii) does not need to lie fallow each year or for one growing season per crop rotation cycle. In some embodiments, the locus provides one or more of the following benefits: a. the locus does not need to lie fallow for one growing season per crop rotation cycle; b. the locus does not need to be tilled for one growing season per crop rotation cycle; and c. the locus does not need to be planted with a cover crop. In some embodiments, the suppression of nematode population density is about 250, 200, 150, 100, 50, 20, or 10, or less than about 250, 200, 150, 100, 50, 20, or 10, per gram (g) of root.

[0024] The methods and systems described herein provide the benefits and value of a crop rotation system by (i) allowing for increased use of a locus that does not need to lie fallow, be cultivated, or have a cover crop grown on it for at least one growing season in a crop rotation cycle in an attempt to control nematodes, and (ii) allowing for more successful use of the locus by suppressing nematode populations at the locus, which results in increased successful growing and yield of inherently valuable nematode-susceptible plants. Also provided are nematode-susceptible plants grown at the locus, plant material harvested from the nematode-susceptible plants (e.g., in the case of cotton plants, cotton lint and cotton fiber), and seeds produced by the nematode-susceptible plants.

[0025] In another aspect, the disclosure provides a system for increasing use of a locus, the system comprising: growing a nematode resistant plant at the locus in a first growing season; and growing a nematode susceptible plant at the locus in a subsequent growing season. In some embodiments, the nematode resistant plant and the first nematode susceptible plant are each unique value crop plants. In some embodiments, over the subsequent growing season, the locus is not left fallow during the growing season. In some embodiments, no cover plant or cover crop is grown during the subsequent growing season. In some embodiments, the nematode resistant plant, the first nematode susceptible plant, and the second nematode susceptible plant are grown for three consecutive growing seasons. In some embodiments, the nematode resistant plant, the first nematode susceptible plant, and the second nematode susceptible plant are each unique value crop plants.

[0026] In yet another aspect, the disclosure provides a method of improving a crop rotation system, comprising: growing a nematode resistant plant at a locus during a first growing season; and growing a nematode susceptible plant at the locus during a subsequent growing season, wherein growing the nematode resistant plant at the locus during the first growing season results in suppression of nematode populations at the locus that enables or enhances the growing of the nematode susceptible plant in the subsequent growing season. In some embodiments, the improvement of the crop rotation system includes: Use of the location for at least one additional growing season each year; b. Reduction in cultivation at the location; c. Reduction in the treatment of nematode-susceptible crop seeds with nematicides; d. Reducing the area of ​​a site treated with a nematicide prior to or during the growing season of nematode-susceptible plants; e. A reduction in the rate of nematicides applied to nematode-susceptible plants and / or loci prior to or during the nematode-susceptible plant growing season; f. Reducing the number of nematicide applications made to nematode-susceptible plants and / or loci during the growing season; g. Increased location availability; h. Increase in value of location; i. Improving sustainable agricultural practices; and / or j. Increasing yields of nematode-susceptible crops The composition may further include one or more of the following:

[0027] In another aspect, the disclosure provides a method of nematode management for a locus comprising: growing a nematode resistant plant at the locus during a growing season; and growing a perennial plant at the locus prior to, during, and / or after the growing season, where growing the nematode resistant plant at the locus results in suppression of nematode populations at the locus that enables or enhances the growing of the perennial plant.

[0028] In some aspects, the disclosure provides a method of marketing a crop rotation system comprising: promoting the use of nematode resistant plants during a first growing season; and promoting the use of nematode susceptible plants at the locus during the same or a subsequent growing season, where growing nematode resistant plants at the locus during the first growing season results in suppression of nematode populations at the locus that enables or enhances growing nematode susceptible plants in the subsequent growing season. In some embodiments, the subsequent growing season is the closest to the first growing season.

[0029] In a further aspect, the disclosure provides marketing materials relating to a crop rotation system in which a nematode resistant plant is grown during a first growing season in conjunction with or followed by growing a nematode susceptible plant at the locus during a subsequent growing season, where growing the nematode resistant plant at the locus during the first growing season results in suppression of nematode populations at the locus that enables or enhances the growing of the nematode susceptible plant. In some embodiments, the marketing materials relate to promoting the crop rotation system.

[0030] In some aspects, a method of nematode management for a locus is provided, the method comprising: growing a nematode resistant plant at the locus prior to, simultaneously with, or after growing a secondary crop at the locus. In some embodiments, the nematode resistant plant expresses a nematocidal Cryl4Ab protein having at least 95, 96, 97, 98, or at least 99% sequence identity to SEQ ID NO: 1. In some embodiments, the nematode resistant plant comprises elite event EE-GM5. In further embodiments, the nematode resistant plant comprises elite event EE-GM4. In any of the aspects or embodiments of the present disclosure, the nematode resistant plant expresses Bacillus thuringiensis toxin Cry14Ab-1. In any of the aspects or embodiments of the present disclosure, the nematode resistant plant comprises elite event EE-GM5. In any of the aspects or embodiments of the present disclosure, the nematode resistant plant comprises elite event EE-GM4. [Brief description of the drawings]

[0031] [Figure 1] Figure 11 shows the effect of GMB151 soybean on Pratylenchus brachyurus population density. Population densities of Pratylenchus brachyurus were estimated for GMB151 homozygous and nullzygous soybean lines at three predicted high pressure sites and the Sinop2 site where P. brachyurus population densities were higher than predicted. The GMB151 transgenic soybean trait significantly reduced P. brachyurus populations. 95% confidence intervals are represented by error bars for the estimated differences between homozygous and nullzygous lines. [Diagram 2]Figure 14 shows soybean yield differences between GMB151 homozygous and nullzygous lines at each study site. Study sites are arranged from lowest to highest Pratylenchus brachyurus population density. Error bars for estimated yield differences represent 95% confidence intervals. The GMB151 trait had no effect on yield at the three locations with the lowest P. brachyurus population densities. At the three sites with the highest P. brachyurus population densities, GMB151 significantly improved soybean yield by an average of 7.0 bu / acre or 21%. [Diagram 3] Figure 3 shows a diagram of the field trial planting described in Example 3. Soybeans were planted in the safra (summer or first crop) soybean season. Five replicates were planted in the treatments in a randomized complete block design. Safrinha (winter or second crop) cotton and corn trials were planted after soybean harvest. Safrinha crop trials were planted on top of the soybean trial ground with the safrinha crop rows running perpendicular to the harvested safra soybean rows. The safrinha crop trial design also constituted a randomized complete block design, although there were only three replicates per crop. [Figure 4] Figure 14 shows the amount of Pratylenchus brachyurus control achieved by GMB151 soybean in the 2019 / 20 Brazilian growing season at the Rio Verde, Goias field site. Asterisks indicate significant differences at P<0.05. [Diagram 5] 13 is a graph showing safra soybean and safrinha corn yield and gross revenue for the 2019 / 20 growing season at the Rio Verde, Goias field site. [Figure 6]1 is a graph showing Pratylenchus brachyurus control (as amount of Pratylenchus brachyurus per gram of root in safrinha crop) achieved by GMB151 soybean in the 2019 / 20 Brazilian growing season across all field trial sites. [Figure 7] FIG. 1 is a graph showing safrinha maize and cotton yields for the 2019 / 20 growing season across all field trial sites. [Figure 8] 1 shows an exemplary crop rotation system. [Figure 9] Figure 13 is a graph showing the effect of GMB151 soybean on Pratylenchus brachyurus population density. Population density of Pratylenchus brachyurus was estimated for GMB151 homozygous and nullzygous soybean lines at 49 test sites. The GMB151 transgenic soybean trait significantly reduced P. brachyurus populations. Standard errors of the means for homozygous and nullzygous lines are represented by error bars. [Figure 10] 1 is a graph showing soybean yield differences between GMB151 homozygous and nullzygous lines across 48 test sites taken for yield. Error bars represent standard error of the treatment mean. The GMB151 trait significantly improved soybean yield by an average of 4.2 bu / acre or 9%. [Figure 11] Figure 13 is a graph showing alternative nematode management tools and their effect on Pratylenchus brachyurus population density in a safra soybean crop. Nematicide seed treatments and in-furrow nematicides were applied to both GMB151 and null soybeans. Significantly greater control was achieved with the GMB151 soybeans than with the seed treatments or in-furrow nematicides. [Figure 12]Figure 14 shows the effectiveness of the cultural nematode control tactic of tillage compared to the GMB151 soybean event and its effect on Pratylenchus brachyurus population density. The GMB151 soybean event achieved significantly greater control of Pratylenchus brachyurus than conventional tillage in safra soybean crops. [Figure 13] 1 is a graph showing Pratylenchus brachyurus control (as amount of Pratylenchus brachyurus per gram of root in a safrinha crop) provided by GMB151 soybean in a safrinha crop field trial. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0032] Plant parasitic nematodes can attack many species of plants (defined herein as "crops" or "agronomic crops") grown in farming or crop rotation systems and can build up large population densities over time. Cultivated plants that are viewed by farmers as economically beneficial ("cash crops") or are otherwise valuable or useful to farmers (such as livestock feed) (all "intrinsic value" crops), and non-beneficial plants or crops grown alone to protect fields from erosion or provide other soil health benefits ("cover crops") are nearly all susceptible to nematode damage to a greater or lesser extent. Depending on how a particular plant is used, it can be either an intrinsic value crop or a non-beneficial crop. One such plant is the genus Brachiaria, also known as signal grass. Signal grass can be grown as a pasture crop (hence an intrinsic value crop) or as a non-beneficial cover crop if not used for livestock grazing. Notably, commercial crops such as, but not limited to, soybeans, cotton, corn, wheat, sugarcane, potatoes, sugar beets, rice, alfalfa, barley, sorghum, oats, rye, cassava, sweet potato, sunflower, vegetables, fruit plants, fruit trees, nut trees, ornamentals, grapevines, and canola are susceptible to nematodes, and damage or injury caused by nematodes can significantly reduce yields and cause reduced income for farmers. Farmers must identify management options that are effective in controlling damage and injury in each crop in which nematode pests proliferate.

[0033] In addition, farmers may need to adjust their crop rotations or crop succession to avoid growing multiple susceptible crops in consecutive growing seasons. This problem can be difficult when farmers are presented with broad-host-range nematodes that can thrive on many agronomic crops. In the worst-case scenario, farmers may be forced to forgo planting economically beneficial cash crops in favor of planting unprofitable commercial crops, or even worse, leaving the fields fallow, due to the very wide host range of some nematodes, including most cover crops. Leaving the fields fallow provides limited relief to farmers, since most nematodes can survive for long periods (>9 months) without a host plant. The nematode pest can simply exist in a dormant state until the farmer plants a susceptible crop.

[0034] Farmers are also struggling to manage the agronomic challenges presented by nematodes. Farmers consider that such challenges must be met in the larger context of each farmer's need to adhere to other demands that are intended to protect the overall environment and address ecological issues such as soil erosion. Currently, there are cases where nematode-management practices and other agronomic practices and requirements are incompatible, leaving farmers with difficult choices. For example, Pratylenchus species can be managed through tillage, which is performed multiple times each year. However, tillage can promote soil erosion, water loss, organic carbon loss, and generally can be less beneficial to crop yields. In addition, even in situations where farmers may be prepared to plant unprofitable cover crops for nematode management, cover crops that limit nematode proliferation may not be the same as cover crops that achieve greater soil health benefits, such as increased organic carbon (Amorim et al., Journal of Agricultural Science 11:333-340, 2019). Nematode management methods that involve approaches that suppress or prevent the growth of nematode populations in a location while limiting interference with agronomic crop growth are most beneficial to farmers.

[0035] The present disclosure is based on the discovery that the benefits of nematicidal activity of nematode-resistant plants grown at a locus (e.g., a field or plot) extend beyond the period during which the nematode-resistant plants are grown. Growing a nematode-resistant plant at the locus suppresses nematode populations at the locus or maintains suppression of nematode populations at the locus, and the suppression of nematode populations or maintained suppression of nematode populations continues for a period beyond the growing of the nematode-resistant plant. Thus, in some embodiments, the present disclosure provides a method of nematode management for a locus, the method comprising: growing a nematode-resistant plant in a field before, simultaneously with, or after growing a secondary plant at the locus.

[0036] In some embodiments, the benefit of the nematicidal activity of the nematode-resistant plant grown in a location (e.g., a field or plot) in a first growing season (such nematicidal activity protects the nematode-resistant plant from nematode damage and injury) also extends nematode protection to any other plants (e.g., commercial crops with or without the nematode-resistant plant) by simply growing the other plants in the same growing season or in a subsequent growing season in the same location. The nematode-resistant plant can be grown periodically (e.g., in successive growing seasons, every other growing season, every third growing season, every fourth growing season, every fifth growing season, etc.) to reduce or control the overall population density of nematodes in a particular location. In some embodiments, the nematode-resistant plant is grown every growing season. In some embodiments, the nematode-resistant plant is grown in the growing season immediately preceding the growing season of the secondary plant (e.g., the nematode-susceptible plant). Thus, a nematode resistant plant not only protects itself from nematode damage, but also protects any plants, including nematode susceptible plants, that are grown simultaneously or at a later date in the same location.

[0037] The term "locus" as used herein refers to a location suitable for growing a plant. Exemplary loci include, but are not limited to, a pot or other container, a greenhouse or other contained location, a field, a hill, any plot of land, an orchard, a vineyard, or other environment suitable for growing a plant.

[0038] The term "nematode-resistant plant" as used herein refers to a plant that expresses a nucleic acid that results in the reduction of nematode movement, feeding, development, reproduction, or other functions if the nematode is in contact with the plant. In some embodiments, the nematode-resistant plant is engineered or derived from a plant that has been engineered via molecular biology techniques to express such a nucleic acid. As a non-limiting example of nematode reduction, the nematode may be killed by ingestion of a part of the nematode-resistant plant. In some embodiments, the nematode-resistant plant includes, but is not limited to, a soybean plant, a corn plant, a cotton plant, a canola plant, a sugarcane plant, a sugar beet plant, a potato plant, a wheat plant, a rice plant, alfalfa plant, a barley plant, a sorghum plant, an oat plant, a rye plant, a cassava plant, a sweet potato plant, a sunflower plant, a vegetable plant, a fruit plant, an ornamental plant, an orchard plant (e.g., a fruit or nut-bearing tree), or a grapevine.

[0039] The term "nematode-susceptible plant" as used herein refers to a plant that does not express a nucleic acid that results in the reduction of nematode movement, feeding, development, reproduction, or other functions if nematodes are in contact with the plant. Those skilled in the art will understand that any plant can be a nematode-resistant plant or a nematode-susceptible plant depending on whether the plant expresses a nucleic acid that results in the reduction of nematode movement, feeding, development, reproduction, or other functions if nematodes are in contact with the plant. In some embodiments, the nematode-resistant plant has been engineered or is derived from a plant that has been engineered via molecular biology techniques to express such a nucleic acid. In some embodiments, nematode-susceptible plants include, but are not limited to, soybean plants, corn plants, cotton plants, canola plants, sugarcane plants, potato plants, wheat plants, vegetable plants, rice plants, alfalfa plants, barley plants, sorghum plants, oat plants, rye plants, cassava plants, sweet potato plants, sunflower plants, fruit plants, orchard plants (e.g., fruit or nut-bearing trees), ornamental plants, or grape vines.

[0040] The present disclosure is also based on the discovery that growing a nematode-resistant plant at a locus results in a permanent suppression of nematode population density at the locus. Nematode suppression can be measured by a number of methods, including but not limited to, determining nematode population density at the locus, in plant roots growing at the locus or in an area of ​​other significance.

[0041] method The present disclosure relates to methods of suppressing nematode populations at a locus, the methods comprising: growing a nematode resistant plant at the locus, where growing the nematode resistant plant suppresses nematode populations at the locus for a period of time during and beyond the growing of the nematode resistant plant, in some embodiments, the period of time extending one or more growing seasons after the growing season in which the nematode resistant plant was grown.

[0042] In some embodiments, the method further comprises growing a secondary plant (which may be any plant) at the location at a time after growing the nematode resistant plant. In some embodiments, the method further comprises growing a secondary plant at the location at a time before growing the nematode resistant plant. In some embodiments, the method further comprises growing a nematode resistant plant at the location simultaneously with the secondary plant. In some embodiments, the secondary plant is a nematode susceptible plant. In some embodiments, the nematode resistant plant is grown in a furrow adjacent to the furrow in which the secondary plant is grown. In some embodiments, the nematode resistant plant is grown in the same furrow in which the secondary plant is grown. In some embodiments, the nematode resistant plant is grown both in a furrow adjacent to the furrow in which the secondary plant is grown and in the same furrow in which the secondary plant is grown. The present disclosure contemplates any configuration known in the art for growing the nematode-resistant plant and the secondary plant (e.g., grown in the same furrow or adjacent furrows; grown in narrow furrows vs. wide furrows; grown on the edge of the field vs. in the middle of the field; grown without intermixing with different furrows). In some embodiments, the secondary plant is a nematode-susceptible plant.

[0043] The disclosure also relates to a method of suppressing nematode populations at a locus, the method comprising: growing a nematode resistant plant at the locus, wherein growing the nematode resistant plant suppresses or maintains suppression of an already suppressed nematode population at the locus during and beyond the growing of the nematode resistant plant. In some embodiments, the period extends one or more growing seasons after the growing season in which the nematode resistant plant was grown. For example, the nematode resistant plant may be grown at a locus where a nematode resistant plant was previously grown and / or some other mechanism of suppressing nematode populations was previously utilized.

[0044] The present disclosure also relates to a method of suppressing nematode populations, the method comprising: growing a nematode-resistant plant at a location in a first growing season, where growing the nematode-resistant plant results in suppression of the nematode population at the location, and growing a secondary plant (which may be any plant) at the same location in the same or subsequent growing season (e.g., a second, third, fourth, or fifth growing season), where the secondary plant exhibits less nematode damage than would be observed if the nematode-resistant plant had not been grown at the location in the first growing season. In some embodiments, the subsequent growing season is the most recent growing season. In some embodiments, the nematode population exhibits little resurgence during the subsequent growing season. The first growing season may be any growing season throughout the year. The nematode-resistant plant may be grown for one or more growing seasons each year, including during consecutive growing seasons. In some embodiments, the nematode population at the location is already suppressed, and growing the nematode-resistant plant at the location maintains the suppression of the nematode population. The cultivation of nematode-resistant plants does not have to be for a specific amount of time or at a specific stage of development, as long as the nematode population at the location is suppressed or the suppression of the nematode population at the location is maintained. The suppression of the nematode population can be maintained by growing nematode-resistant plants at a location that previously hosted a nematode-resistant plant that suppressed the nematode population, and the maintenance of the suppression of the nematode population can be indicated by the presence of fewer nematodes at the location than at the same or comparable location. The suppression of the nematode population or the maintenance of the suppression of the nematode population can be indicated by the presence of fewer nematodes at the location than at the same or comparable location if no nematode management measures, including the cultivation of nematode-resistant plants, were taken. Alternatively or additionally, the suppression of the nematode population or the maintenance of the suppression of the nematode population can mean that the cultivation of nematode-resistant plants at the location achieves and / or maintains a level of nematodes at the location below the detection limit.

[0045] In some embodiments, suppression of nematode populations or maintenance of suppression of nematode populations is indicated by the presence of fewer nematodes in the location than in the same or comparable location if no nematode management measures, including the cultivation of nematode-resistant plants, were taken. In some embodiments, the comparable location is a location where nematode-resistant plants have not previously been grown. In some embodiments, the comparable location is a location where nematode-resistant plants have been grown prior to but not during the current crop rotation cycle.

[0046] In some embodiments, suppression of nematode population or maintaining suppression of nematode population when the secondary plant is of the genus Brachiaria is achieved when the number of nematodes is less than 60, about 60, or about 60 per gram of root. In further embodiments, suppression of nematode population when the secondary plant is of the genus Brachiaria is achieved when the number of nematodes is less than 50, 40, 30, 20, 10, or 5, about 50, 40, 30, 20, 10, or 5, or about 50, 40, 30, 20, 10, or 5 per gram of root. In further embodiments, suppression of nematode populations when the secondary plant is of the genus Brachiaria is achieved when the number of nematodes per gram root is at or about 5-60, 10-60, 20-60, 5-50, 10-50, 20-50, 5-40, 5-30, 5-20, 10-40, 10-30, 10-20, 20-40, or 20-30 nematodes per gram root.

[0047] In some embodiments, suppression of nematode populations or maintaining suppression of nematode populations when the secondary plant is corn is achieved when the number of nematodes is less than 300, about 300, or about 300 per gram of root. In further embodiments, suppression of nematode populations when the secondary plant is corn is achieved when the number of nematodes is less than 250, 200, 150, 100, 50, 20, 10, or 5, about 250, 200, 150, 100, 50, 20, 10, or 5, or less than about 250, 200, 150, 100, 50, 20, 10, or 5 per gram of root. In further embodiments, suppression of nematode populations when the secondary plant is maize may be achieved by reducing the number of nematodes per gram of root to between 5-300, 5-250, 5-200, 5-150, 5-100, 5-50, 10-300, 10-250, 10-200, 10-150, 10-100, 10-50, 50-300, 50-250, 50-200, This is achieved when the number of cells is 50 to 150, or 50 to 100, or about 5 to 300, 5 to 250, 5 to 200, 5 to 150, 5 to 100, 5 to 50, 10 to 300, 10 to 250, 10 to 200, 10 to 150, 10 to 100, 10 to 50, 50 to 300, 50 to 250, 50 to 200, 50 to 150, or 50 to 100.

[0048] In some embodiments, suppression of nematode population or maintaining suppression of nematode population when the secondary plant is cotton is achieved when the number of nematodes is less than 60, about 60, or about 60 per gram of root. In further embodiments, suppression of nematode population when the secondary plant is cotton is achieved when the number of nematodes is less than 50, 40, 30, 20, 10, or 5, about 50, 40, 30, 20, 10, or 5, or about 50, 40, 30, 20, 10, or 5 per gram of root. In a further embodiment, suppression of nematode population when the secondary plant is cotton is achieved when the number of nematodes is at or about 5-60, 10-60, 20-60, 5-50, 10-50, 20-50, 5-40, 5-30, 5-20, 10-40, 10-30, 10-20, 20-40, or 20-30 nematodes per gram of root.

[0049] In some embodiments, suppression of nematode populations or maintaining suppression of nematode populations when the secondary plant is sorghum is achieved when the number of nematodes is less than 250, about 250, or about 250 per gram root. In further embodiments, suppression of nematode populations when the secondary plant is sorghum is achieved when the number of nematodes is less than 200, 150, 100, 50, 20, 10, or 5, about 200, 150, 100, 50, 20, 10, or 5, or about 200, 150, 100, 50, 20, 10, or 5 per gram root. In further embodiments, suppression of nematode populations when the secondary plant is sorghum is achieved when the number of nematodes per gram root is at or about 5-250, 5-200, 5-150, 5-100, 5-50, 10-250, 10-200, 10-150, 10-100, 10-50, 50-250, 50-200, 50-150, or 50-100 nematodes per gram root.

[0050] In some embodiments, suppression of nematode populations or maintaining suppression of nematode populations in the location where the secondary plants are grown is achieved when there is a 5% reduction, about a 5% reduction, or at least about a 5% reduction in the number of nematodes per gram of roots compared to the number of nematodes per gram of roots in a location comparable to the location where the secondary plants are grown. In further embodiments, suppression of nematode populations in a locus (e.g., a locus where a secondary plant is grown) is achieved when there is a 5%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, or 80% reduction, about a 5%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, or 80% reduction in the number of nematodes per gram of roots, or at least about a 5%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, or 80% reduction in the number of nematodes per gram of roots, compared to the number of nematodes per gram of roots in a comparable locus where no nematode management measures, including the cultivation of nematode-resistant plants, were taken. In some embodiments, the comparable locus is a locus where no nematode-resistant plants have been previously grown. In some embodiments, the comparable location is a location where the nematode-resistant plants were grown prior to but not during the current crop rotation cycle. In any of the aspects or embodiments of the present disclosure, the determination of the number of nematodes per gram of roots is performed by one skilled in the art.

[0051] The present disclosure relates to a method of suppressing nematode populations, the method comprising: growing a nematode-resistant plant at a locus in a first growing season, where growing the nematode-resistant plant results in suppression of the nematode population at the locus, and growing a nematode-susceptible plant at the same locus in the same or a subsequent growing season (e.g., a second, third, fourth, or fifth growing season), where the nematode-susceptible plant exhibits less nematode damage than would be observed if a nematode-resistant plant had not been grown at the locus in the first growing season. In some embodiments, the nematode population exhibits little resurgence during the subsequent growing season.

[0052] In some embodiments, the method includes growing a nematode-resistant plant at the location in a first growing season, growing a first nematode-susceptible plant at the location in a subsequent growing season (e.g., in a second, third, fourth, or other growing season after growing the nematode-resistant plant at the location), and growing a second nematode-susceptible plant at the location after growing the first nematode-susceptible plant. A third, fourth, fifth, etc. nematode-susceptible plant can be grown at the location in a subsequent growing season.

[0053] In some embodiments, a nematode-resistant plant is grown at the locus before growing the first nematode-susceptible plant. Those skilled in the art will appreciate that once nematode suppression is achieved at the locus, no particular order of planting is required.

[0054] In some embodiments, the nematode resistant plant is a monocotyledonous plant. In some embodiments, the nematode resistant plant is a dicotyledonous plant. In some embodiments, the nematode resistant plant is an annual plant. In some embodiments, the nematode resistant plant is a perennial plant.

[0055] In some embodiments, the nematode resistant plant is a soybean plant, a corn plant, a cotton plant, a canola plant, a sugarcane plant, a sugar beet plant, a potato plant, a wheat plant, a rice plant, an alfalfa plant, a barley plant, a sorghum plant, an oat plant, a rye plant, a cassava plant, a sweet potato plant, a sunflower plant, a vegetable plant, a fruit plant, an ornamental plant, an orchard plant (e.g., a fruit or nut-bearing tree), or a grape vine.

[0056] In some embodiments, the nematode resistant plant is a soybean plant, which is grown at a location in a first growing season, the nematode susceptible plant is a soybean plant, a corn plant, a cotton plant, a canola plant, a sugarcane plant, a sugar beet plant, a potato plant, a wheat plant, a rice plant, an alfalfa plant, a barley plant, a sorghum plant, an oat plant, a rye plant, a cassava plant, a sweet potato plant, a sunflower plant, a vegetable plant, a fruit plant, an orchard plant (e.g., a fruit or nut bearing tree), an ornamental plant, or a vine grown in the same field or at another location in a second season, and the nematode resistant soybean plant is grown in a third growing season.

[0057] In some embodiments, the nematode resistant plant is a soybean plant, and the soybean plant is grown at the location in a first growing season, and the nematode susceptible plant is a soybean plant, a corn plant, a cotton plant, a canola plant, a sugarcane plant, a sugar beet plant, a potato plant, a wheat plant, a rice plant, an alfalfa plant, a barley plant, a sorghum plant, an oat plant, a rye plant, a cassava plant, a sweet potato plant, a sunflower plant, a vegetable plant, a fruit plant, an orchard plant (e.g., a fruit or nut-bearing tree), an ornamental plant, or a vine, and is the same or a second In one embodiment, the nematode-resistant plant is grown at the same location for two growing seasons, and in another embodiment, the nematode-susceptible plant is grown at the same location for a third growing season, the nematode-susceptible plant being a soybean plant, a corn plant, a cotton plant, a canola plant, a sugarcane plant, a sugar beet plant, a potato plant, a wheat plant, a rice plant, an alfalfa plant, a barley plant, a sorghum plant, an oat plant, a rye plant, a cassava plant, a sweet potato plant, a sunflower plant, a vegetable plant, a fruit plant, an orchard plant (e.g., a fruit or nut-bearing tree), an ornamental plant, or a grapevine. For annuals, the seasonal order of planting is not important, as long as the nematode-resistant plant is grown at the location occasionally prior to or at the same time as the planting of the annuals, and as long as the nematode population at the location is sufficiently suppressed or remains suppressed to benefit the annuals.

[0058] In some embodiments, nematode-susceptible plants are grown at a locus with nematode-resistant plants during the same growing season or at the same time. In some embodiments, the nematode-susceptible plants can be perennial plants, including but not limited to vegetable plants, fruit plants, orchard plants (e.g., fruit trees or nut trees), ornamental plants, or grapevines. Thus, nematode-resistant plants can be used to protect perennial plants from nematode damage. In some embodiments, nematode-resistant plants are grown at a locus before planting perennial plants to suppress nematode populations at the locus before planting the perennial plants. In some embodiments, nematode-resistant plants are grown before the perennial plants are planted and grown simultaneously with the perennial plants or grown with the perennial plants, for example, on an annual basis if the nematode-resistant plants are annual plants. In some embodiments, nematode-resistant plants are grown at a locus that already contains perennial plants, and the perennial plants benefit from the addition of the nematode-resistant plants to the locus. For perennials, the seasonal sequence of planting is not important so long as the nematode-resistant plants are grown at the locus prior to, concurrently with, or after the planting of the perennials, and so long as the nematode population at the locus is sufficiently suppressed or remains suppressed to benefit the perennials.

[0059] Also provided is a method of protecting nematode susceptible plants from nematode damage or injury, comprising growing a nematode resistant plant at a locus for at least one growing season prior to growing the nematode susceptible plant; and growing a nematode susceptible plant at the locus for at least one growing season the same as or following the growing season of the nematode resistant plant. As used herein, the term "nematode injury" refers to physical damage or destruction caused to plants by nematodes. As used herein, the term "nematode damage" refers to financial loss caused to marketable commodities by nematodes.

[0060] Also provided is a method of increasing the yield of a nematode-susceptible plant. The method includes growing a nematode-resistant plant in a field during a first growing season; and growing a nematode-susceptible plant in the field during the same growing season or a subsequent growing season. Growing a nematode-resistant plant in the same growing season or a subsequent growing season of the nematode-susceptible plant increases the yield of the nematode-susceptible plant compared to the yield of a nematode-susceptible plant grown in the same or comparable location in the absence of nematode management measures, including growing a nematode-resistant plant. In some embodiments, the increase in the yield of the nematode-susceptible plant is compared to the yield of a nematode-susceptible plant grown in the same or comparable location in which a nematode-resistant crop was not grown during the most recent crop rotation cycle. In some embodiments, the comparable location is a location where a nematode-resistant plant has not been grown before. In some embodiments, the comparable location is a location where a nematode-resistant plant was grown before but not during the current crop rotation cycle.

[0061] As used herein, the term "yield" of a plant refers to the quality and / or quantity of biomass produced by the plant. By "biomass" is intended any plant product that can be measured. An increase in biomass production is any improvement in the yield of a measured plant product. Increasing plant yield has several commercial applications. For example, increasing plant leaf biomass can increase the yield of leafy vegetables for human or animal consumption. In addition, increasing leaf biomass can be used to increase the production of pharmaceutical or industrial products derived from the plant. In some embodiments, growing a nematode-susceptible plant after growing a nematode-resistant plant increases the yield of the nematode-susceptible plant by at least 1% (or at least 2%, at least 3%, at least 4%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%) compared to a nematode-susceptible plant grown in the same or comparable location when no nematode management measures, including growing a nematode-resistant plant, were taken. In some embodiments, the comparable location is a location where a nematode-resistant plant has not previously been grown. In some embodiments, the comparable location is a location where a nematode-resistant plant has been grown before but not during the current crop rotation cycle.

[0062] In various embodiments, the cultivation of nematode-susceptible plants simultaneously with the cultivation of nematode-resistant plants or in subsequent cultivation seasons provides the following benefits for nematode-susceptible plants, for example: improved root development (e.g., improved root or root hair growth); improved yield; faster emergence; improved plant stress management, including increased stress tolerance and / or improved stress recovery; increased mechanical strength; improved drought tolerance; reduced fungal, bacterial, and / or viral disease infection; and / or improved plant health, compared to nematode-susceptible plants grown in the same or comparable locations when no nematode management measures, including cultivation of nematode-resistant plants, are taken. In some embodiments, the comparable locations are locations where nematode-resistant plants have not been grown before. In some embodiments, the comparable locations are locations where nematode-resistant plants have been grown before, but not during, the current crop rotation cycle. Any combination of these benefits can also be obtained.

[0063] In various embodiments, growing plants in a growing season subsequent to or concurrent with the growing of nematode resistant plants in a particular location can allow for the growing of plants that are not likely to be nematode resistant due to technical, agronomic, and / or regulatory reasons. Non-limiting examples of plants that can be used in the present invention include fruit or nut trees, ornamentals, grape vines, or wheat in Australia.

[0064] The present invention can be used with both migratory and sedentary nematode species. Furthermore, nematode species in both the migratory and sedentary categories have been identified that may also be controllable by the present invention. In some embodiments, the nematode is from a nematode population, particularly a Pratylenchus spp. nematode population, such as Pratylenchus brachyurus, Meloidogyne spp., Heterodera spp., such as Heterodera glycines, Globodera spp., Rotylenchulus reniformis, Helicotylenchus spp., such as Helicotylenchus dihystera, Scutellonema brachyurus, Tubixaba tuxaua, or Aphelencoides besseyi.

[0065] Also contemplated are loci, such as fields or plots, in which nematode population density (e.g., nematodes per volume of soil) has been suppressed or nematode numbers (e.g., nematodes per gram of root) have been reduced (wherein the suppression of nematode population density or reduction of nematode numbers is achieved by the methods described herein). Measurements of suppression of nematode population density or reduction of nematode numbers at a locus are interchangeable in a practical sense, since both are indicators of nematode presence at a locus, the only difference being the method used to measure nematode presence. Suppression of nematode populations can be measured in a number of ways, none of which are limitations to the present invention. In various embodiments, the suppression of nematode population density is less than about 250, 200, 150, 100, 50, 20, or 10 nematodes per gram (g) of root.

[0066] Nematicidal nucleic acids, nematicidal proteins, and nematode-resistant plants The methods described herein describe the use of plants that express nematicidal nucleic acids. In some embodiments, the methods described herein include the use of plants that have been engineered by molecular biology techniques through the transformation of organisms or the use of organisms that contain heterologous nucleotide sequences encoding nematicidal proteins. There are several molecular biology techniques that can be used to enable plants to express nematicidal nucleic acids, and the technical approach used to obtain nematode-resistant plants is not a limitation for the methods described herein.

[0067] As used herein, the terms "nematicidal nucleic acid" and "nematicidal protein" refer to nematodes that are active against root-knot nematodes (Meloidogyne species), Rotylenchulus reniformis, Helicotylenchus dihystera, Scutellonema brachyurus, Tubixaba tuxaua, Aphelencoides besseyi, and Pratylenchus species (Pratylenchus alleni, Pratylenchus brachyurus, Pratylenchus coffeae, Pratylenchus crenatus, Pratylenchus dulscus, Pratylenchus fallax, Pratylenchus flakkensi, Pratylenchus goodeyi, Pratylenchus hexincisus, Pratylenchus loosi, Pratylenchus minutus, Pratylenchus mulchandi, Pratylenchus musicola, Pratylenchus neglectus, Pratylenchus penetrans, Pratylenchus pratensis, Pratylenchus reniformia, Pratylenchus scribneri, Pratylenchus solnai"Nematode" refers to a toxin that is active against one or more nematode pests, including, but not limited to, Pratylenchus thornei, Pratylenchus vulnus, and Pratylenchus zeae.

[0068] In some embodiments, the nematicidal protein is a Cry protein. Cry proteins are well known to those skilled in the art. The nematicidal activity of Cry proteins is described, for example, in WO 2010 / 027805, WO 2010 / 027809, WO 2010 / 027804, WO 2010 / 027799, WO 2010 / 027808, and WO 2007 / 147029. In some embodiments, the nematicidal protein comprises a Cry14 protein (see, for example, WO 2018 / 119336 and U.S. Provisional Patent Application No. 62 / 112,832, filed November 12, 2020, each of which is incorporated herein by reference in its entirety). In various embodiments, the Cry14 protein is Cry14Aa1 (GENBANK Accession No. AAA21516) or Cry14Ab1 (also known as Cry14Ab-1; GENBANK Accession No. KC156652). In some embodiments, the Cry14Ab-1 protein (SEQ ID NO:1 (amino acid sequence) and SEQ ID NO:2 (nucleotide sequence)) is as described in WO 2018 / 119361 and WO 2018 / 119364, as well as variants and fragments thereof. In various embodiments, the nematode-resistant plant expresses a nematicidal Cryl4Ab protein that has 95, 96, 97, 98, or 99%, about 95, 96, 97, 98, or 99%, at least 95, 96, 97, 98, or 99%, or at least about 95, 96, 97, 98, or 99% sequence identity to SEQ ID NO:1.

[0069] Some nucleic acids with nematocidal activity are well known to those skilled in the art, as well as molecular biology techniques that produce plants with nematode resistance.In some embodiments, the nucleic acid expressed by the nematode-resistant plant is produced by the sequence described in, for example, WO2011 / 82217, WO2013 / 078153, WO2020 / 243365, WO2018 / 005491, and WO2021 / 016098.

[0070] Molecular biology techniques used to generate nematode-resistant plants include, but are not limited to, genome editing to generate nucleic acids that generate nematocidal activity; expressing RNA molecules that generate nematocidal activity; and / or expressing heterologous or transgenic nematocidal proteins. Other techniques, such as mutagenesis, can also be used to generate nematode-resistant plants. In some embodiments, the nematode-resistant plant expresses nematode-resistant nucleic acids by Cry genes. In some embodiments, the Cry gene is a Cry14 gene. In some embodiments, the Cry14 gene is a Cry14Aa gene or a Cry14Ab gene.

[0071] In addition to the nucleic acid that provides nematocidal activity, the nematode-resistant plant may express one or more additional nucleic acids introduced by molecular biology techniques, including, but not limited to, herbicide resistance, resistance to coleopteran pests, resistance to lepidopteran pests, resistance to other pests, including other insects; and / or disease resistance. Such other nucleic acids may result from molecular biology techniques, such as, but not limited to, genome editing activity, expression of RNA, or expression of heterologous proteins. In addition, the nematode-resistant plant may express other foreign nucleic acids resulting from other techniques, such as introduction of mutations, introgression of traits through breeding, and / or other techniques known to those skilled in the art. The presence of such other nucleic acids and / or traits is not a limitation for the methods herein.

[0072] In some embodiments, the nematode-resistant plant expresses one or more additional exogenous nucleic acids in addition to the nucleic acid providing nematicidal activity. In some embodiments, the nucleic acid providing nematicidal activity is selected from the group consisting of glyphosate-based, glufosinate-based, HPPD inhibitor-based, sulfonylurea or imidazolinone-based, AHAS or ALS-inhibiting, and / or auxin-type (e.g., dicamba, 2,4-D) herbicides, such as Event EE-GM3 (also known as FG-072, MST-FG072-3, described in WO 2011 / 063411, USDA-APHIS Petition 09-328-01p), Event SYHT0H2 (also known as 0H2, SYN-000H2-5, described in WO 2012 / 082548 and 12-215-01p), Event DAS-68416-4 (also known as Enlist Soybean, described in WO 2011 / 066384 and WO 2011 / 066360, and USDA-APHIS Petition 09-349-01p), Event DAS-44406-6 (also known as Enlist E3, DAS-44406-6, described in WO 2012 / 075426 and USDA-APHIS 11-234-Olp), Event MON87708 (Dicamba Tolerance Event in Roundup Ready 2 Xtend Soybeans, described in WO 2011 / 034704 and USDA-APHIS Petition 10-188-Olp, and MON-87708-9), Event MON89788 (also known as Genuity Roundup Ready 2 Yield, described in WO 2006 / 130436 and USDA-APHIS Petition 06-178-01p), Event 40-3-2 (also known as Roundup Ready, GTS 40-3-2, MON-04032-6, described in USDA-APHIS Petition 93-258-01), Event A2704-12 (also known as LL27, ACS-GM005-3, described in WO 2006 / 108674 and USDA-APHIS Petition 96-068-Olp), Event127 (also known as BPS-CV127-9, described in WO 2010 / 080829), Event A5547-127 (also known as LL55, ACS-GM006-4, described in WO 2006108675 and USDA-APHIS Petition 96-068-01p), Event MON87705 (MON-87705-6, Vistive Gold, published in WO 2010 / 037016 and USDA-APHIS Petition 09-201-01p), Soybean Event HB4 (OECD Unique Identifier IND-00410-05, USDA-APHIS Petition 17-223-01p), or Event DP305423 (aka DP-305423-1, published in WO 2008 / 054747, USDA-APHIS Petition 06-354-01p), or nucleic acids providing nematicidal activity are combined with the following event combinations: Event MON98788 x MON87708 (aka Roundup Ready 2 Xtend Soybeans, MON-87708-9 x MON-89788-1), Event HOS x Event 40-3-2 (aka Plenish High Oleic Soybeans x Roundup Ready Soybeans), Event EE-GM3 x EE-GM2 (also known as FG-072 x LL55, described in the pamphlet of International Publication No. 2011 / 063413), Event MON 87701 x MON 89788 (also known as Intacta RR2 Pro Soybean, MON-87701-2 x MON-89788-1), DAS-81419-2 x DAS-44406-6 (also known as Conkesta (trademark) Enlist E3 (trademark) Soybean, DAS-81419-2 x DAS-44406-6), Event DAS-68416-4 x Event MON 89788 (also known as Enlist (trademark) RoundUp Ready (registered trademark) 2 Soybean, DAS-68416-4 x MON-89788-1), Event MON-87769-7 x EventMON-89788-1 (aka Omega-3 x Genuity Roundup Ready 2 Yield Soybeans), Event MON 87705 x Event MON 89788 (aka Vistive Gold, MON-87705-6 x MON-89788-1), or Event MON87769 x Event MON89788 (aka Omega-3 x Genuity Roundup Ready 2 Yield Soybeans, MON-87769-7 x MON-89788-1). In some embodiments, any of the above traits are modified using mutation introduction, genome editing, or other molecular biology techniques and combined, alone or in any combination, with a nematode resistance trait.

[0073] In some embodiments, the nematode-resistant plant is a soybean plant containing the EE-GM4 event described in WO 2018 / 119361, or a soybean plant containing the EE-GM5 event (also known as the GMB151 event) described in WO 2018 / 119364. The disclosures of WO 2018 / 119361 and WO 2018 / 119364 are incorporated herein by reference in their entireties.

[0074] In some embodiments, the nematode-resistant plant contains one or more native traits that confer resistance to nematodes. Such native traits are well known to those skilled in the art (see, for example, Fosu-Nyarko, J. and MGK Jones. 2015. Application of biotechnology for nematode control in crop plants. Pages 339-376 in: Advances in Botanical Research Vol. 73, Plant Nematode Interactions: A View on Compatible Interrelationships. Chapter 14. C. Escobar and C. Fenoll, eds. Elsevier, Oxford). The native traits may be complementary to the nematode-suppressing effect of the nematode-resistant plant, and in some embodiments, one or more such native traits are present in the germplasm of the nematode-resistant plant.

[0075] Thus, provided herein is a nematode pest population, such as, for example, a Pratylenchus spp. nematode population, such as Pratylenchus brachyurus, Meloidogyne spp., Heterodera spp., such as Heterodera glycines, Globodera spp., Rotylenchulus reniformis, Helicotylenchus spp., such as Helicotylenchus dihystera, Scutellonema brachyurus, Tubixaba tusauer, or the like, via a nematode resistant plant described herein. In certain embodiments, the nematicidal protein comprises a Cry14 protein as set forth in WO 2018 / 119361, WO 2018 / 119364, and U.S. Provisional Patent Application No. 62 / 112,832, and variants and fragments thereof.

[0076] system In another aspect, described herein is a system for increasing the efficiency of growing nematode-susceptible plants, the system comprising: growing a nematode-resistant plant at a locus during a first growing season; and growing a nematode-susceptible plant at the locus during the same or a subsequent growing season. In some embodiments, a second nematode-susceptible plant is grown at the locus after the growing of the first nematode-susceptible plant. In some embodiments, increased efficiency in growing nematode susceptible plants may be indicated or measured by one or more of the following: a. use of the locus for at least one additional growing season each year; b. reduced cultivation of the locus; c. reduced treatment of nematode susceptible crop seeds with nematicides; d. reduced treatment area of ​​the locus with nematicides prior to or during the growing season of the nematode susceptible plants; e. reduced rate of nematicides applied to nematode susceptible plants and / or locus prior to or during the growing season of the nematode susceptible plants; f. reduced number of applications of nematicides made to nematode susceptible plants and / or locus during the growing season; increased utility of the locus; increased value of the locus; improved sustainable agricultural practices; and / or g. increased yield of nematode susceptible crops.

[0077] In another aspect, described herein is a system for increasing use of a locus, the system comprising: growing a nematode resistant plant at the locus in a first growing season; and growing a first nematode susceptible plant at the locus in the same or a subsequent growing season. In some embodiments, the nematode resistant plant and the first nematode susceptible plant are grown in the same or successive growing seasons. In other embodiments, the nematode resistant plant and the first nematode susceptible plant are grown in non-successive growing seasons. In some embodiments, the locus is not left fallow for the subsequent growing season. In some embodiments, a cover crop is not planted for the subsequent growing season. In some embodiments, the locus is not tilled.

[0078] In another aspect, described herein is a system for increasing the use of a locus, such as, but not limited to, a field or plot, comprising: growing a nematode resistant plant at the locus in a first growing season; growing a first nematode susceptible plant at the locus in the same or a subsequent growing season; and growing a second nematode susceptible crop at the locus after growing the first nematode susceptible crop at the locus. In some embodiments, the nematode resistant plant, the first nematode susceptible plant, and the second nematode susceptible plant are grown in consecutive growing seasons. In other embodiments, the nematode resistant plant, the first nematode susceptible plant, and the second nematode susceptible plant are grown in non-consecutive growing seasons, or only two of the three are grown in consecutive or non-consecutive growing seasons. In some embodiments, the locus is not left fallow over subsequent growing seasons. In some embodiments, a cover crop is not planted over subsequent growing seasons. In some embodiments, the locus is not tilled.

[0079] The term "crop rotation system" or "cropping system" as used herein refers to the practice of following a crop rotation cycle in which one crop planted in a location, e.g., a field, is followed within a period of time by one or more successive crops. The period encompassed by a single crop rotation cycle does not necessarily coincide with a calendar year, nor is it necessarily a 12-month cycle. In some cases, a single crop rotation cycle may span a period of 2-3 years, or even longer. In some embodiments, sugarcane is on an approximately 5-year cycle (e.g., 4-5 years of sugarcane followed by one season of non-sugarcane). The exact timing of planting, duration of the cycle, and the crops to be planted are determined by the environment and are location-specific. Crop rotation systems are used for several reasons, such as reducing nutritional requirement issues, disease and / or pest pressure of each crop, and diversifying the business. As an example, a typical Brazilian soybean cropping system produces two crops that are planted in succession in a calendar year. The first growing season (called "safra") begins with planting a first crop, such as soybeans, between September and December. Harvesting of the safra season crop occurs between January and March. Immediately after harvesting the safra season crop, a second season crop, called "safrinha," is planted. The safrinha crop is then harvested between May and August. See FIG. 8. In the United States, crop rotation cycles vary depending on the environment and the farmer's objectives and are not necessarily tied to a calendar year. In one example, the first growing season begins with planting a first crop between February and May, with harvest occurring between August and October. A second crop is then planted between September and November, or February and May of the following year. Other locations or geographies support growing crops for a three growing season rotation cycle, and still others support growing crops for a four or more growing season rotation cycle.

[0080] In one aspect, described herein is a method of suppressing nematode populations at a locus, comprising growing a nematode resistant plant at the locus, wherein growing the nematode resistant plant suppresses nematode populations at the locus or maintains suppression of nematode populations at the locus during and beyond the growing of the nematode resistant plant. In some embodiments, the nematode resistant plant is grown during the safra season. In some embodiments, the method further comprises growing a secondary plant at the locus during the safrinha season immediately following the safra season. In some embodiments, the secondary plant is a nematode susceptible plant. In some embodiments, the nematode resistant plant and the secondary plant are both indigenous value crop plants. In some embodiments, the nematode resistant plant and the secondary plant are both commercial crop plants. In some embodiments, the nematode resistant plant is a soybean plant and the secondary plant is selected from the group consisting of corn, cotton, sorghum, wheat, and sugarcane.

[0081] In some aspects, described herein are methods of improving a crop rotation system, comprising: growing a nematode-resistant plant at a location during a first growing season; and growing a nematode-susceptible plant at the location during the same or a subsequent growing season, wherein growing the nematode-resistant plant at the location during the first growing season results in suppression of nematode populations at the location that allows or enhances growing the nematode-susceptible plant in the same or a subsequent growing season. In some embodiments, a second nematode-susceptible plant is grown at the location after growing the first nematode-susceptible plant.

[0082] In some embodiments, an improvement in a crop rotation system may be indicated or measured by one or more of the following: a. use of the locus for at least one additional growing season each year; b. reduced cultivation of the locus; c. reduced treatment of nematode-susceptible crop seeds with nematicides; d. reduced treatment area of ​​the locus with nematicides prior to or during the growing season of nematode-susceptible plants; e. reduced rate of nematicides applied to nematode-susceptible plants and / or locus prior to or during the growing season of nematode-susceptible plants; f. reduced number of applications of nematicides made to nematode-susceptible plants and / or locus during the growing season; increased utility of the locus; increased value of the locus; improved sustainable agricultural practices; and / or g. increased yield of nematode-susceptible crops.

[0083] It should be understood that the present invention is not limited to the particular methodology, protocols, plant species or genera, constructs, and reagents described per se. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention, which will be limited only by the appended claims. It should be noted that the singular forms "a," "an," and "the," as used herein and in the appended claims, include plural references unless the context clearly dictates otherwise. Thus, for example, a reference to a "vector" is a reference to one or more vectors, and includes equivalents thereof known to those of skill in the art, and so forth.

[0084] marketing "Marketing" as used herein is any communication by or on behalf of an entity for the purpose of informing the public / potential customers of a product and promoting its acceptance and / or use. Print marketing / materials may include, but are not limited to, booklets, brochures, flyers, catalogs, business cards, signs, posters, billboards, trade literature, seed bags, seed bag tags, labels, or other advertisements. Other types of marketing / materials may include, but are not limited to, any type of digital information, including websites, emails, texts, information about any company-sponsored communications or events that promote / disclose the product, or any social media platforms. Marketing also includes any rebates / incentives to farmers for the purchase of one or more products, the use of which during crop rotation cycles is enhanced by the use of nematode-resistant plants according to the methods provided herein.

[0085] In some embodiments, the disclosure also provides a method of marketing a crop rotation system, comprising: promoting the use of nematode resistant plants at a locus during a first growing season; and promoting the use of nematode susceptible plants at the locus during the same or a subsequent growing season, where growing nematode resistant plants at the locus during the first growing season results in suppression of nematode populations at the locus that allows or enhances growing nematode susceptible plants in the subsequent growing season. In further embodiments, a method of marketing a crop rotation system for suppressing nematode populations at a locus, comprising: promoting the use of nematode resistant plants at the locus during a first growing season; and promoting the use of nematode susceptible plants at the locus during the same or a subsequent growing season, where growing nematode resistant plants at the locus during the first growing season results in suppression of nematode populations at the locus that allows or enhances growing nematode susceptible plants in the subsequent growing season.

[0086] In a further aspect, the disclosure provides marketing materials related to a system of growing nematode resistant plants at a locus during a first growing season in conjunction with or followed by growing nematode susceptible plants at the locus during a subsequent growing season, where growing nematode resistant plants at the locus during the first growing season results in suppression of nematode populations at the locus that enables or enhances growing nematode susceptible plants in the subsequent growing season. In some embodiments, the marketing materials relate to promoting a system of growing nematode resistant plants at a locus during a first growing season in conjunction with or followed by growing nematode susceptible plants at the locus during a subsequent growing season, where growing nematode resistant plants at the locus during the first growing season results in suppression of nematode populations at the locus that enables or enhances growing nematode susceptible plants in the subsequent growing season.

[0087] In some embodiments, the present disclosure provides methods of improving marketing of a crop rotation system to reduce nematode populations, reduce damage to crops from nematodes, and / or increase crop yield through suppression of nematode populations, including, for example and without limitation, one or more features of a rebate program, marketing statement, connected offer, or integrated solution, or any sales offer related to the purchase of one or more products, the use of which during a crop rotation cycle is improved by the use of nematode resistant plants according to the methods provided herein.

[0088] In a further aspect, the disclosure provides a kit comprising marketing materials (e.g., related to marketing a crop rotation system) as described herein, and / or a nematode-resistant soybean plant, or a cell, part, seed, or progeny thereof, each as described herein. In some embodiments, the kit further comprises a secondary plant, or a cell, part, seed, or progeny thereof. In some embodiments, the kit further comprises any product useful for growing the plant, or a cell, part, seed, or progeny thereof, including, but not limited to, herbicides, fungicides, insecticides, nematicides, biologicals, fertilizers, seed treatments, inocula, decision-making or remote sensing tools, crop scouting services, crop diagnostic tools and / or services. In any of the aspects or embodiments of the disclosure, the kit is any package or material that includes marketing of a system or method for suppressing or maintaining a nematode population at a locus such that the resulting nematode population at the locus is about 250 nematodes or less per gram (g) of roots. In various embodiments, a kit is any package or material that includes marketing of a system or method for suppressing or maintaining a nematode population at a locus, where the resulting nematode population at the locus is about 250, 200, 150, 100, 50, 20, or 10 nematodes per gram (g) of roots, or less than about 250, 200, 150, 100, 50, 20, or 10 nematodes per gram (g) of roots.

[0089] The disclosed products, systems, and methods described herein also realize increased value and sustainability by enabling improved agricultural practices (e.g., through carbon credit opportunities). For example, sustainability opportunities created by embodiments of the present disclosure may include, but are not limited to, (i) improving a site to require less tillage; (ii) eliminating the need to grow cover crops or leave a site fallow for at least one growing season each year; and / or (iii) enabling the growth of plants with healthier and larger root systems that can capture more carbon than plants with smaller root systems or roots that are damaged by nematodes. A related additional benefit realized by the present disclosure relates to improved nitrogen fixation. Nematodes reduce nitrogen fixation in soybeans, thereby increasing the need for chemical nitrogen for crops (e.g., corn crops) planted the following season. In any of the aspects or embodiments of the present disclosure, the nematode-resistant plants described herein suppress nematode populations to enable improved or complete nitrogen fixation in soybeans, which in turn reduces the need for chemical nitrogen for corn crops the following season. In addition, chemical nitrogen poses environmental problems and is of concern (see, for example, Crews et al., Agriculture, Ecosystems and Environment 102 (2004) 279-297).

[0090] Embodiments: 1. A method for controlling nematode populations at a locus comprising: 1. A method comprising growing a nematode resistant plant at the locus, wherein growing the nematode resistant plant suppresses a nematode population at the locus or maintains suppression of a nematode population at the locus during and beyond the growing of the nematode resistant plant.

[0091] 2. The method of embodiment 1, wherein the period extends for one or more growing seasons after the growing season in which the nematode-resistant plants were grown.

[0092] 3. The method of embodiment 1 or 2, further comprising growing a secondary plant at the locus after growing the nematode resistant plant.

[0093] 4. The method of embodiment 1 or 2, further comprising growing a secondary plant at the locus prior to growing the nematode-resistant plant.

[0094] 5. The method of embodiment 1 or 2, further comprising growing the nematode resistant plant simultaneously with the secondary plant at the locus.

[0095] 6. The method of any one of embodiments 3 to 5, wherein the secondary plant is a nematode-susceptible plant.

[0096] 7. The method of any one of embodiments 1-6, wherein the nematode population is suppressed or maintained below detection limits.

[0097] 8. The method of any one of embodiments 3-6, wherein the secondary plant is of the genus Brachiaria and suppression of the nematode population is achieved when the number of nematodes is less than 60, about 60, or about 60 nematodes per gram of root.

[0098] 9. The method of any one of embodiments 3-6, wherein the secondary plant is corn and suppression of the nematode population is achieved when the number of nematodes is less than 300, about 300, or about 300 nematodes per gram of root.

[0099] 10. The method of any one of embodiments 3-6, wherein the secondary plant is cotton and suppression of the nematode population is achieved when the number of nematodes is less than 60, about 60, or about 60 nematodes per gram of root.

[0100] 11. The method of any one of embodiments 3-6, wherein the secondary plant is sorghum and suppression of the nematode population is achieved when the number of nematodes is less than 250, about 250, or about 250 nematodes per gram of root.

[0101] 12. The method of any one of embodiments 3-11, wherein suppression of the nematode population in the location where the secondary plant is grown is achieved when there is a 5% reduction, about a 5% reduction, or at least about a 5% reduction in the number of nematodes per gram of roots compared to the number of nematodes per gram of roots in a location comparable to the location where the secondary plant is grown.

[0102] 13. A method for protecting a plant from nematode damage or injury, comprising: growing the nematode-resistant plants at the locus for at least one growing season prior to planting the nematode-susceptible plants; growing nematode-susceptible plants at the locus for at least one growing season after growing the nematode-resistant plants; The method includes:

[0103] 14. The method of embodiment 13, wherein protecting nematode-susceptible plants from nematode damage comprises increasing the yield of harvested nematode-susceptible plant material and / or the revenue derived from its sale.

[0104] 15. A method for increasing the yield of a nematode-susceptible plant, comprising: growing the nematode-resistant plants at the locus for at least one growing season prior to planting the nematode-susceptible plants; growing nematode-susceptible plants at the locus for at least one growing season after growing the nematode-resistant plants; The method includes:

[0105] 16. The embodiment of embodiment 15, wherein the increase in yield of the nematode-susceptible plant is an increase compared to the yield of a nematode-susceptible plant grown in the same or a comparable location that did not previously grow a nematode-resistant crop, or the increase in yield of the nematode-susceptible plant is an increase compared to the yield of a nematode-susceptible plant grown in the same or a comparable location prior to growing the nematode-resistant plant.

[0106] 17. A method of nematode management for a locus, comprising: growing nematode resistant plants at the location; growing nematode-susceptible plants at the locus either simultaneously with or at a later time than the growing of the nematode-resistant plants; The method includes:

[0107] 18. The method of embodiments 6-17, wherein the nematode-susceptible plant is a perennial or annual plant.

[0108] 19. The method of any one of embodiments 6 to 18, wherein the nematode-susceptible plant is a soybean plant, a corn plant, a cotton plant, a canola plant, a sugarcane plant, a potato plant, a wheat plant, a vegetable plant, a rice plant, an alfalfa plant, a barley plant, a sorghum plant, an oat plant, a rye plant, a cassava plant, a sweet potato plant, a sunflower plant, a fruit plant, an orchard plant (e.g., a fruit or nut-bearing tree), an ornamental plant, or a grapevine.

[0109] 20. The method of any one of embodiments 1-19, wherein the nematode-resistant plant expresses a Cry protein, such as but not limited to a nematocidal Cry protein.

[0110] 21. The method of any one of embodiments 1-20, wherein the nematode resistant plant is a soybean plant, a corn plant, a cotton plant, a canola plant, a sugarcane plant, a sugar beet plant, a potato plant, a wheat plant, a rice plant, an alfalfa plant, a barley plant, a sorghum plant, an oat plant, a rye plant, a cassava plant, a sweet potato plant, a sunflower plant, a vegetable plant, a fruit plant, an ornamental plant, an orchard plant (e.g., a fruit or nut-bearing tree), or a grapevine.

[0111] 22. The nematode may be a Pratylenchus species nematode population, such as Pratylenchus brachyurus, Meloidogyne species, Heterodera species, such as Heterodera glycines, Globodera species, Rotylenchulus reniformis, Helicotylenchus species, such as Helicotylenchus dihystera, Scutellonema brachyurus, Tubixaba tuxaua, or Aphelencoides 22. The method of any one of the preceding embodiments, wherein the nematode species is selected from the group consisting of:

[0112] 23. The method of embodiment 22, wherein the Pratylenchus species is Pratylenchus brachyurus.

[0113] 24. A method of nematode management for a locus, comprising: Growing nematode-resistant plants in the same location as nematode-susceptible plants. The method includes:

[0114] 25. A method of nematode management for a locus, comprising: planting a nematode-susceptible plant at the locus; growing nematode-resistant plants at the locus after planting the nematode-susceptible plants; The method includes:

[0115] 26. A locus in which nematode population density is suppressed, wherein the suppression of nematode population density is achieved by the method of any one of embodiments 1 to 25.

[0116] 27. The locus of embodiment 26, which provides one or more of the following benefits: a. the locus does not need to lie fallow every year or for one growing season per rotation cycle; b. the locus does not need to be cultivated for one growing season per rotation cycle; and c. the locus does not need to be planted with a cover crop.

[0117] 28. The locus of embodiment 26 or embodiment 27, wherein the suppression of nematode population density is less than about 250, 200, 150, 100, 50, 20, or 10 nematodes per gram (g) of root, or less than about 250, 200, 150, 100, 50, 20, or 10 nematodes per gram (g).

[0118] 29. The location of embodiment 26, wherein the land need not lie fallow for one growing season per rotation cycle.

[0119] 30. A nematode-resistant plant grown in the location of embodiment 26.

[0120] 31. Plant material harvested from the plant of embodiment 30.

[0121] 32. Seeds produced by the plant of embodiment 31.

[0122] 33. A system for augmenting location usage comprising: growing nematode resistant plants at the location during a first growing season; growing nematode-susceptible plants at the location during the subsequent growing season; A system containing.

[0123] 34. The system of embodiment 33, wherein the locus is not left fallow during the subsequent growing season.

[0124] 35. The system of embodiment 33, wherein the cover plant or cover crop is not grown during the subsequent growing season.

[0125] 36. The system of embodiment 33, wherein the nematode-resistant and nematode-susceptible plants are grown in successive growing seasons.

[0126] 37. The system of any one of embodiments 33-36, wherein the nematode-resistant plant and the nematode-susceptible plant are each unique value crop plants.

[0127] 38. A method for improving a crop rotation system, comprising: growing a nematode resistant plant at the locus during a first growing season; growing nematode-susceptible plants at the location during a subsequent growing season; wherein growing nematode-resistant plants at the locus during a first growing season results in suppression of nematode populations at the locus that enables or enhances the growing of nematode-susceptible plants in a subsequent growing season.

[0128] 39. Improvement of crop rotation systems includes: Use of the location for at least one additional growing season each year; b. Reduction in cultivation at the location; c. Reduction in the treatment of nematode-susceptible crop seeds with nematicides; d. Reducing the area of ​​a site treated with a nematicide prior to or during the growing season of nematode-susceptible plants; e. A reduction in the rate of nematicides applied to nematode-susceptible plants and / or loci prior to or during the nematode-susceptible plant growing season; f. Reducing the number of nematicide applications made to nematode-susceptible plants and / or loci during the growing season; g. Increased location availability; h. Increase in value of location; i. Improving sustainable agricultural practices; and / or j. Increasing yields of nematode-susceptible crops The method of embodiment 38, which may further include one or more of the following:

[0129] 40. A method of nematode management for a locus, comprising: Growing nematode resistant plants at the location during a growing season; Cultivating perennial grasses at the location prior to, during, and / or after the growing season; wherein growing a nematode resistant plant at the locus results in suppression of nematode populations at the locus that enables or enhances the growth of perennial plants.

[0130] 41. The method of embodiment 40, wherein the perennial plant is a nematode-susceptible plant.

[0131] 42. The method of embodiment 40, wherein the perennial plant is a vegetable plant, a fruit plant, an orchard plant, an ornamental plant, or a vine.

[0132] 43. A method of nematode management for a locus, comprising: growing a nematode resistant plant at the locus for a first growing season, wherein the nematode resistant plant suppresses a nematode population at the locus or maintains suppression of a nematode population at the locus; Growing nematode-susceptible plants at the location during the same or a subsequent growing season; Achieving improved health and / or yield in nematode-susceptible plants compared to the health and / or yield that would be expected if the nematode population were not suppressed. The method includes:

[0133] 44. The method of embodiment 43, wherein the improved health of the nematode-susceptible plant includes one or more of the following: improved root development (e.g., improved root or root hair growth); improved yield; faster emergence; improved plant stress management, including increased stress tolerance and / or improved recovery from stress; increased mechanical strength; improved drought tolerance; reduced fungal, bacterial, and / or viral disease infection; or any combination thereof.

[0134] 45. Any of embodiments 1-44, wherein the nematode-resistant plant expresses a nematicidal Cryl4Ab protein having at least 95, 96, 97, 98, or at least 99% sequence identity to SEQ ID NO:1.

[0135] 46. ​​Any of embodiments 1-44, wherein the nematode-resistant plant comprises the elite event EE-GM5.

[0136] 47. Any of embodiments 1-44, wherein the nematode-resistant plant comprises the elite event EE-GM4.

[0137] 48. Any of embodiments 1-44, wherein the nematode-resistant plant expresses the Bacillus thuringiensis toxin Cry14Ab-1, or a functional truncation or variant thereof.

[0138] 49. A method of nematode management for a locus, comprising: growing nematode resistant plants at a locus, the nematode resistant plants being grown during the safra season in a South American country, such as Brazil or Paraguay; and growing the nematode-resistant plants and then growing nematode-susceptible plants at the location, the nematode-susceptible plants being grown during the safrinha season in a South American country, such as Brazil or Paraguay. The method includes:

[0139] 50. The method of embodiment 49, wherein the nematode-resistant plant expresses the Bacillus thuringiensis toxin Cry14Ab-1, or a functional truncation or variant thereof, and the nematode-susceptible plant is selected from the group consisting of corn, cotton, sorghum, wheat, and sugarcane.

[0140] The following examples are offered by way of illustration and not by way of limitation. EXAMPLES

[0141] Example 1 - Generation of nematode-resistant plants Soybean lines containing the GMB151 transgenic event (i.e., EE-GM5 as described in WO 2018 / 119364) were generated from the BC2:F2 selected plants. The soybean lines differed in their zygosity for the GMB151 event. One line homozygous for the GMB151 event was generated, while a second line nullzygous for the GMB151 event was also selected. These two lines shared approximately 87.5% genetic identity with the common backcross parent soybean line. Thus, the lines were agronomically similar except for the presence of the GMB151 event.

[0142] Example 2 - Control of the root-lesion nematode Pratylenchus brachyurus (Godfrey) by GMB151 transgenic soybean expressing the Bacillus thuringiensis toxin Cry14Ab-1 The ability of Cry14Ab-1 to control P. brachyurus in fields was investigated, specifically, the Cry14Ab-1 expression event GMB151 could reduce P. brachyurus reproduction in soybean growing fields, and this reduction in P. brachyurus reproduction could provide significant yield protection.

[0143] Materials and Methods: Field trials were conducted in the Brazilian states of Parana, Goias, and Mato Grosso during the 2018 / 19 soybean growing season. Field trial locations are listed below in Table 1:

[0144] [Table 1]

[0145] The GMB151 event was introgressed into a soybean maturity group IX background, which is adapted for production in Brazil. Introgression was accomplished by utilizing a maturity group IX line as the recurrent parent. BC2: In the F2 generation, plants homozygous and nullzygous for the GMB151 event were selected to produce two soybean lines that differed in the presence or absence of the GMB151 event but were otherwise approximately 87.5% genetically related to the recurrent parent.

[0146] Soybean trials were established at six study sites across a gradient of P. brachyurus population densities, as shown in Table 1. Three study sites contained P. brachyurus at population densities predicted to reduce soybean yield, while three sites were not predicted to have P. brachyurus at population densities high enough to significantly reduce soybean yield. Predictions of P. brachyurus pest pressure were made based on 2017 / 18 soybean crop samples or communications with farm managers.

[0147] The experimental design and test treatment differences varied between the predicted high and low pressure sites. The predicted low pressure test sites were run as randomized complete block design trials with five replications. Each field plot consisted of four 5 m long soybean rows spaced 0.5 m apart. Two treatments were included in each trial: GMB151 homozygous lines (homozygous) and GMB151 nullzygous lines (nullzygous).

[0148] At the three high pressure sites, two changes were made within the trial compared to the low pressure sites. First, the GMB151 event was evaluated in combination with a nematicidal seed treatment. Three seed treatments were included in the trial. The seed treatment treatment factor was fully crossed with the zygosity (i.e., homozygous or nullzygous) treatment factor to create six treatments. The six treatments were arranged in a split-plot design with zygosity as the whole-plot treatment and seed treatment as the subplot treatment. The subplots remained the same size as the other sites, four 5m long soybean rows spaced 76.2cm apart. The second change made to the trial at the high pressure sites was to reduce the number of replicates from five to three. The number of replicates was reduced due to the inclusion of seed treatment subplots and the limited space available for field trials.

[0149] Data collection: The efficacy of GMB151 against P. brachyurus was evaluated by measuring the population density of P. brachyurus in soybean root systems at 90 days after planting (dap). The 90 dap time point was chosen because it was close to the sampling time of a previous P. brachyurus study (Lima et al. 2015). At 90 dap, soybean root systems of 10 plants per plot were removed from the field. Five root systems were sampled from the first and fourth furrows of each plot. These furrows were chosen because they were not harvested and P. brachyurus sampling would not affect yield estimates taken at the end of the season. Root samples were transported to the laboratory, where they were weighed and homogenized in a blender. Nematodes were then extracted from the root segments using the method of Jenkins (1964). The extracted nematodes were then suspended in water and the number of P. brachyurus in 1 ml subsamples was counted under a microscope, and the number of P. brachyurus per gram of root was calculated for each sample.

[0150] Soybean yield was estimated for each plot at physiological maturity. The central 2 tiers of each 4-tier plot were harvested. Grain weight and moisture were measured for each plot and yield was standardized to 13% moisture.

[0151] Data Analysis: Pratylenchus brachyurus population density data were analyzed separately for the three high pressure and three low pressure locations. The number of P. brachyurus per gram (g) of root was natural log transformed prior to analysis to reduce heteroscadacity. Due to the wide variety of P. brachyurus population densities present across the study trial locations (affecting the amount of damage caused by P. brachyurus), yield data for each trial was analyzed separately. Yield data were analyzed as bushels per acre.

[0152] Data were analyzed to estimate the effect of GMB151 zygosity on P. brachyurus population density and soybean yield. Data were fitted to a mixed-effects model ANOVA to estimate the difference between homozygous and nullzygous lines with 95% confidence intervals. Trials that did not include seed treatment subplots utilized a model with replicates as a random effect and zygosity as a fixed effect. The interaction of replicates and zygosity was considered as an error term. Trials that included seed treatment subplots included zygosity, seed treatment, and the interaction of zygosity by seed treatment as fixed effects. Replicates and all other interactions between factors were considered as random effects. Interactions of replicates and zygosity were considered as whole plot error terms, while interactions of replicates, zygosity, and seed treatment were considered as subplot error terms. The effect of seed treatments on P. brachyurus, soybean yield, and their interactions with GMB151 traits are beyond the scope of the study reported here. Therefore, data were summarized only by zygosity across seed treatments.

[0153] result: Pratylenchus brachyurus population densities at two high pressure (Rio Verde and Sinop1) and one low pressure location (Sinop2) exceeded population densities observed to cause aboveground disease symptoms by Lima et al. (2015). The high pressure Lupinopolis location averaged P. brachyurus population densities within the range observed by Lima et al. (2015) outside symptomatic field areas, and therefore was expected to result in less yield reduction, if any. The final two low pressure locations (Ibipora and Trinidade) should not have affected soybean yield, as P. brachyurus populations were at the limit of detection.

[0154] Homozygous soybean lines reduced P. brachyurus population density by 96% across the three high-pressure locations (P < 0.0001). Homozygous lines also significantly reduced P. brachyurus populations by 90% at the predicted low-pressure location, Sinop2 (P < 0.01). Average P. brachyurus reductions ranged from 90% to 97% at the individual test locations (Figure 1).

[0155] Soybean yields were adversely affected by poor maturity group fit to the region at both the Lupinopolis and Ibipora sites. Poor fit resulted in low absolute yields across all test treatments. Treatment yields ranged from 12.4 bu / acre to 16.9 bu / acre at these two test sites. Treatment yields at the other four locations ranged from 28.0 to 57.1 bu / acre.

[0156] Yield differences between homozygous and nullzygous lines were dependent on the population density of P. brachyurus at the test location (Figure 2). At any of the three locations with the lowest P. brachyurus population densities, there was no significant yield difference between the two soybean lines. At each of the three sites with the highest P. brachyurus population densities, there was a significant yield difference between homozygous and nullzygous lines. The average yield improvement of homozygous lines compared to nullzygous lines at each location ranged from 6.2 to 7.8 bu / acre or 14 to 25%.

[0157] Consideration: The six study sites selected had Pratylenchus brachyurus population densities ranging from below the limit of detection to nearly 1,000 individuals per gram of root tissue. The six sites allowed us to evaluate the efficacy of the GMB151 transgenic soybean event against P. brachyurus and its ability to protect soybean yield. The GMB151 event was highly effective against P. brachyurus. Three months into the growing season, P. brachyurus populations were, on average, >90% lower in soybean lines homozygous for the GMB151 event compared to nullzygous lines. This level of control far exceeds that achieved by current management practices, which include crop rotation, fallow, and chemical control (Lima et al. 2015, Ribeiro et al. 2014, Rodrigues et al. 2014).

[0158] The benefit of the GMB151 event on soybean yield protection was dependent on P. brachyurus population density. Under low P. brachyurus pressure, soybean yield was not affected by the GMB151 trait. This result indicates that in the absence of P. brachyurus, GMB151 does not have any adverse effect on soybean yield. In each of the three test locations with the highest P. brachyurus population density, soybean yield was significantly greater in soybean lines homozygous for the GMB151 trait. Soybean yield improvements ranged from 14 to 25% at these sites. Thus, our results are consistent with previous studies that estimated yield damage from P. brachyurus to be 10–30% (Dias et al. 2007, Franchini et al. 2007).

[0159] The field trial results reported here show that the Cry14Ab1-expressing soybean trait GMB151 conferred significant efficacy against P. brachyurus resulting in improved soybean yield. These results were obtained under field production conditions with P. brachyurus population densities within the range reported for commercial production fields in Brazil (Franchini et al. 2007, Lima et al. 2015). Thus, the GMB151 trait represents a potentially unique new management tool to control an economically important crop pest with few viable management options today.

[0160] References for Example 2: Barbosa,BFF,JMdos Santos,JCBarbosa,PLMSoares,ARRuas,and RBde Carvalho.2013.Aggressiveness of Pratylenchus brachyurus to the sugarcane, compared with key nematode P.zeae.Nematropica 43:119-130. Dias,WP,NRRibeiro,IONLopes,A.Garcia,GESCarneiro,and JFVSilva.2007.Management of nematodes in soybean crops.In:Brazilian Congress of Nematology;12 August;Goiana,Brazil.Goiana:Brazilian Society of Nematology.p 26-30. Franchini,JC,OFSaraiva,H.Debiase,and SLGoncalves.2007.Contribution of soil management system for sustainable soybean production.Circular Tecnica 46:1-4. Inomoto,MM,AMCGoulart,ACZMachado,and ARMonteiro.2001.Effect of population densities of Pratylenchus brachyurus on the growth of cotton plants.Fitopatologia Brasileira 26:192-196. Jenkins,WR1964.A rapid centrifugal-flotation technique for separating nematodes from soil.Plant Disease Reporter 48:692. Lima,FSDO,GRDSantos,SRNogueira,PRRDSantos,and VRCorrea.2015.Population dynamics of the root lesion nematode,Pratylenchus brachyurus,in soybean fields in Tocantins State and its effect to soybean yield.Nematropica 45:170-177. Lima,FSO,VRCorrea,SRNogueira,and PRRSantos.2017.Nematodes affecting soybean and sustainable practices for their management.Intech doi:10.5772 / 67030 Ribeiro,NR,WPDias,and JMSantos.2010.Distribution of phytonematodes in soybean producing regions in the state of Mato Grosso.Soybean Research Bulletin.Fundacao MT,Rondonopolis,MT,289-296. Ribeiro,LM,HDCampos,CRDias-Arieira,DLNeves,and GCRibeiro.2014.Effect of soybean seed treatment on the population dynamics of Pratylenchus brachyurus under water stress conditions.Bioscience Journal 30:616-622. Rios,ADF,MRRocha,ASMachado,KAGBAvila,RATeixeira,LCSantos,and LRSRabelo.2016.Host suitability of soybean and corn genotypes to the lesion caused by nematode under natural infestation conditions.Ciencia Rural.46:580-584. Rodrigues,DB,CRDias-Arieira,MVVVedoveto,M.Roldi,HFDMolin,and VHFAbe.2014.Crop rotation for Pratylenchus brachyurus control in soybean.Nematropica 44:146-151. Wei, J., K. Hale, L. Carta, E. Platzer, C. Wong, S. Fang, and FVAroian.2003. Bacillus thuringiensis crystal proteins that target nematodes. Proceedings of the National Academy of Sciences 100:2760-2765.

[0161] Example 3 - GMB151 transgenic soybean expressing the Bacillus thuringiensis toxin Cry14Ab-1 provides nematicidal protection in safrinha ("second crop") crops The Brazilian soybean cropping system can produce two crops per calendar year. The first growing season, called "safra," begins with soybean planting from September to December. Harvesting of the safra season soybean crop occurs between January and March. Immediately following soybean harvest, a second season crop, called "safrinha," is planted. The safrinha crop is then harvested from May to August. In the soybean cropping system, the most common safrinha crops in Brazil are corn and cotton. In Brazil, soybean, corn, and cotton are all susceptible to the plant parasitic nematode Pratylenchus brachyurus. Therefore, GMB151 soybean was tested to determine whether planting in the safra season could confer nematode suppression to the subsequent nematode-susceptible safrinha season crop.

[0162] Two soybean lines (one homozygous and one null for the GMB151 trait) were planted in production fields infested with P. brachyurus. Three fields infested with P. brachyurus were available for the study in 2018 and 2019. The soybean field trial consisted of three (2018) or five (2019) replicates arranged in a randomized complete block design, with individual plots measuring 6 meters in length with 12 soybean row width units. Immediately after the soybean trial harvest in April / May 2019 and 2020, a safrinha crop was planted on top of the same ground. In 2019, signal grass was planted as the safrinha crop. Signal grass plots were planted parallel to the harvested soybean rows at each location. Three replicates of signal grass were planted on top of the soybean field trial ground. Thus, the plots were 6 meters long with 12 row width units. In April 2020, maize and cotton were planted as safrinha crops after soybean. Three replicates of maize plots and three replicates of cotton plots were planted on top of the soybean trial ground. The maize and cotton rows were planted perpendicular to the harvested soybean rows. The cotton plots were planted on top of the first 3 meters of replicates 1, 2, and replicate 3. The maize plots were planted on top of the last 3 meters of replicates 5, 4, and replicate 3. Thus, the cotton and maize plots were 9 meters long, with two replicates being 8 rows wide and the third replicate of each crop being 4 rows wide (Figure 3).

[0163] Data were collected to determine whether protection from P. brachyurus was conferred to safrinha crops planted after GMB151 soybean. Protection to safrinha crops was evaluated by measuring P. brachyurus population density and crop yield. Pratylenchus brachyurus population density was measured as the number of nematodes per gram of root tissue. P. brachyurus population density was measured in safrinha crops at 30 days after planting (dap), 60 dap, and 90 dap. The root systems of 10 plants per plot were collected for each P. brachyurus measurement. Five roots were collected from the first and last furrows of each plot. The middle furrow was not sampled. Grain yields were collected from the center rows of the cotton and corn safrinha plots in August 2020. Yields were not taken from the signal grass plots in 2019 because root samples are destructive and would have interfered with the collection of yield data from the center row of each plot. Yields were collected from the center rows of the cotton and corn safrinha plots in August 2020. Yields were not taken from the signal grass plots in 2019 because signal grass is grown as a pasture crop.

[0164] Data were analyzed to assess whether P. brachyurus population density and grain yield differed between plots following GMB151 soybean compared to conventional soybean. Data were analyzed separately for each safrinha crop; maize, cotton, and signal grass. All data were analyzed using a mixed-effects ANOVA model. The model included fixed effects of location, GMB151 zygosity, and their interaction. Experimental replicates were considered as random effects.

[0165] Consideration: Pratylenchus brachyurus population densities were significantly reduced in all three safrinha crops by prior planting with GMB151 soybean during the safra season. Depending on the safrinha crop, test location, and sampling time, P. brachyurus population densities were reduced by 28-100%, with an average reduction of 87%. There was no clear indication that P. brachyurus population suppression declined over the safrinha season in any of the three crops, or that there was a resurgence of nematode populations. Thus, planting with GMB151 soybean reduced P. brachyurus populations throughout the entire cropping system, beginning with the safra soybean crop and extending through the safrinha crop harvest (see, e.g., Figure 8). GMB151 soybean reduced P. brachyurus populations by 99% at the end of the soybean season, which led to significant P. brachyurus suppression throughout the safrinha crop season from planting to harvest in the corn safrinha crop (Figures 4 and 7) and cotton safrinha crop (Figure 7).

[0166] Safrinha crop protection resulted in substantial yield improvements for corn production in Rio Verde, Mato Grosso. For example, as shown in Figure 5, P. brachyurus control achieved by GMB151 soybeans resulted in numerically higher soybean yields and significantly greater corn yields in the safrinha planting season. The yield improvements achieved by GMB151 from P. brachyurus control were worth approximately $90 USD / acre in increased gross revenue to farmers. Gross revenue estimates are based on average Brazilian grain prices of $9.00 per bushel for soybeans and $6.30 per bushel for corn.

[0167] The data provided in this example demonstrates that the incorporation of a highly effective transgenic nematicidal protein into a host crop can provide protection not only to the transformed crop, but also to susceptible crops that are rotated with the transformed crop or subsequently planted in the same location. This provides an effective way to achieve nematode control throughout the entire farm production system. Transforming crops to provide protection throughout the production system limits the development and regulatory costs associated with the production and registration of transgenic crops. It also provides a cost-effective way to provide nematode resistance to crops where native resistance is not available, and where a particularly common problem is migratory plant parasitic nematodes, such as Pratylenchus spp. This approach also provides an opportunity to provide transgenic nematode protection to crops such as wheat, where end-user acceptance of transgenic grain can be a barrier, but where Pratylenchus spp. can be a significant yield-reducing pathogen with few effective management options available.

[0168] In one embodiment, transgenic GMB151 soybeans provide significant yield and crop system management benefits to growers in Brazil. The high efficacy of GMB151 against P. brachyurus provides growers with safra soybean protection, as well as protection for many safrinha crop species. The benefit to safrinha crops is also to provide growers with freedom and flexibility in their production systems to select the most profitable and agronomically advantageous crop rotations, regardless of crop susceptibility to P. brachyurus.

[0169] References for Example 3: Barker, KR, and SRKoenning. 1998. Developing sustainable systems for nematode management. Annual Review of Phytopathology 36:165-205. Cook,R.2004.Genetic resistance to nematodes: where is it useful? Australasian Plant Pathology 33:139-150. Inagaki,H.,and M.Tsutsumi.1971.Survival of the soybean cyst nematode Heterodera glycines Ichinohe(Tylenchida:Heteroderidae)under certain storage conditions.Applied Entomology and Zoology 6:156-162. IRAC.2018.Nematicide Resistance Risk Statement.https: / / irac-online.org / teams / nematodes. Nicol, JM, SJ Turner, DLCoyne, L. den Nijs, S. Hockland, and ZTMaafi.2011 Interaction,Springer,Germany,pp.21-43. Wei, J., K. Hale, L. Carta, E. Platzer, C. Wong, S. Fang, and FVAroian.2003.Crystal proteins of Bacillus thuringiensis that target nematodes.

[0170] Example 4 - Control of the root-lesion nematode Pratylenchus brachyurus (Godfrey) by GMB151 transgenic soybean expressing the Bacillus thuringiensis toxin Cry14Ab-1 The ability of Cry14Ab-1 to control P. brachyurus in fields was further explored by propagation in the experiments described in Example 2. Specifically, it was believed that the Cry14Ab-1 expression event GMB151 could reduce P. brachyurus reproduction in soybean growing fields, and that this reduction in P. brachyurus reproduction could provide significant yield protection.

[0171] Materials and Methods: Field trials were conducted in the Brazilian states of Bahia, Goias, Mato Gross, Parana, and Sao Paulo during the 2018 / 19, 2019 / 20, and 2020 / 21 soybean growing seasons. In total, 49 soybean trials were planted across the three safra seasons.

[0172] The GMB151 event was introgressed into multiple soybean backgrounds ranging from MG VI to IX, each of which is adapted for production in Brazil. Introgression was achieved by utilizing the Brazilian-adapted backgrounds as the recurrent parent. For each background, plants homozygous and nullzygous for the GMB151 event were selected in the BC2:F2 or BC3:F2 generation to generate two soybean lines that differed in the presence or absence of the GMB151 event but were otherwise approximately 87.5% (BC2:F2) or 93.75% (BC3:F2) genetically related to the recurrent parent.

[0173] Soybean trials were established at study sites infested with a wide range of population densities of P. brachyurus.

[0174] Variations in experimental design and test treatments varied among the test sites. Each of the 49 test sites contained two main plot treatments with 5–6 replicates replicated in a randomized complete block design. The two main plot treatments included per test were (1) BC2:F2 or BC3:F2 strains homozygous for the GMB151 event, and (2) BC2:F2 or BC3:F2 strains, respectively, nullzygous for the GMB151 trait. Additional split-plot treatments were included at a subset of locations. Nine locations received split-plot treatments of chemical nematicide treatments. At these nine locations, each soybean line was treated with (1) a base fungicide / insecticide seed treatment, (2) a base fungicide / insecticide seed treatment and a nematicide seed treatment, and (3) a base fungicide / insecticide seed treatment and an in-furrow application of a liquid nematicide. The other three locations utilized slightly different trial designs where the primary plot treatment was level of tillage, conventional tillage, or no-tillage production. The split-plot treatments at these three sites were soybean lines (homozygous or nullzygous for the GMB151 trait).

[0175] At all locations, plots were 5 m long and 4 rows wide.

[0176] Data collection: The efficacy of GMB151, chemical nematicides, and tillage against P. brachyurus was evaluated by measuring the population density of P. brachyurus in soybean root systems at 90 days after planting (dap). The 90 dap time point was chosen because it was close to the sampling time of a previous P. brachyurus study (Lima et al. 2015). At 90 dap, soybean root systems of 10 plants per plot were removed from the field. Three to five root systems were sampled from the first and fourth furrows of each plot. These furrows were chosen because they were not harvested and P. brachyurus sampling would not affect yield estimates taken at the end of the season. Root samples were transported to the lab, where they were weighed and homogenized in a blender. Nematodes were then extracted from the roots using the method of Jenkins (1964). The extracted nematodes were then suspended in water and the number of P. brachyurus in 1 ml subsamples was counted under a microscope. The number of P. brachyurus per gram of root was calculated for each sample.

[0177] Soybean yield was estimated for each plot at physiological maturity. The central 2 tiers of each 4-tier plot were harvested. Grain weight and moisture were measured for each plot and yield was standardized to 13% moisture.

[0178] Data Analysis: Pratylenchus brachyurus population density and yield data were first analyzed across all locations to estimate the effect of GMB151 on P. brachyurus population density. Numbers of P. brachyurus per gram (g) of root were natural log transformed prior to analysis to reduce variance heterogeneity. Yield data were analyzed as bushels per acre.

[0179] Pratylenchus brachyurus population density and yield data were then analyzed separately for nine trials evaluating chemical nematicides in combination with GMB 151. A third analysis was then completed to evaluate three trial locations comparing tillage practices and GMB 151 traits for Pratylenchus brachyurus control and soybean yield protection.

[0180] result: In 33 of 49 trials, Pratylenchus brachyurus population densities on GMB151 null strains exceeded the population density observed to cause aboveground disease symptoms by Lima et al. (2015).

[0181] Homozygous soybean lines reduced P. brachyurus population densities by 91% (P<0.0001) across 49 trials (Figure 9). The GMB151 trait also significantly improved soybean yield by an average of 4.2 bu / acre or 9% (Figure 10). Homozygous lines provided significantly better control of P. brachyurus than seed treatments or in-furrow nematicide applications (Figure 11). Homozygous lines also provided significantly greater control than conventional tillage (Figure 12).

[0182] Yield differences between homozygous and nullzygous lines were dependent on the population density of P. brachyurus at the trial location, with larger yield benefits generally observed in trials with higher P. brachyurus population densities. On average, homozygous lines had soybean yields of 4.2 bu / acre, or 9.4% greater than null lines (Figure 2).

[0183] Consideration: Forty-nine laboratory trials were conducted with Pratylenchus brachyurus population densities ranging from near the limit of detection to nearly 1,000 individuals per gram of root tissue. The trials allowed for the evaluation of the efficacy of the GMB151 transgenic soybean event against P. brachyurus and its ability to protect soybean yield. The GMB151 event was highly effective against P. brachyurus. Three months into the growing season, P. brachyurus populations were, on average, >90% lower in soybean lines homozygous for the GMB151 event compared to nullzygous lines. This level of control far exceeds that achieved by current management practices, which include crop rotation, fallow, and chemical control (Lima et al. 2015, Ribeiro et al. 2014, Rodrigues et al. 2014).

[0184] The field trial results reported here show that the Cry14Ab1-expressing soybean trait GMB151 conferred significant efficacy against P. brachyurus resulting in improved soybean yield. These results were obtained under field production conditions with P. brachyurus population densities within the range reported for commercial production fields in Brazil (Franchini et al. 2007, Lima et al. 2015). Thus, the GMB151 trait represents a potentially unique new management tool to control an economically important crop pest with few viable management options today.

[0185] References for Example 4: Barbosa,BFF,JMdos Santos,JCBarbosa,PLMSoares,ARRuas,and RBde Carvalho.2013.Aggressiveness of Pratylenchus brachyurus to the sugarcane, compared with key nematode P.zeae.Nematropica 43:119-130. Dias,WP,NRRibeiro,IONLopes,A.Garcia,GESCarneiro,and JFVSilva.2007.Management of nematodes in soybean crops.In:Brazilian Congress of Nematology;12 August;Goiana,Brazil.Goiana:Brazilian Society of Nematology.p 26-30. Franchini,JC,OFSaraiva,H.Debiase,and SLGoncalves.2007.Contribution of soil management system for sustainable soybean production.Circular Tecnica 46:1-4. Inomoto,MM,AMCGoulart,ACZMachado,and ARMonteiro.2001.Effect of population densities of Pratylenchus brachyurus on the growth of cotton plants.Fitopatologia Brasileira 26:192-196. Jenkins,WR1964.A rapid centrifugal-flotation technique for separating nematodes from soil.Plant Disease Reporter 48:692. Lima,FSDO,GRDSantos,SRNogueira,PRRDSantos,and VRCorrea.2015.Population dynamics of the root lesion nematode,Pratylenchus brachyurus,in soybean fields in Tocantins State and its effect to soybean yield.Nematropica 45:170-177. Lima,FSO,VRCorrea,SRNogueira,and PRRSantos.2017.Nematodes affecting soybean and sustainable practices for their management.Intech doi:10.5772 / 67030. Ribeiro,NR,WPDias,and JMSantos.2010.Distribution of phytonematodes in soybean producing regions in the state of Mato Grosso.Soybean Research Bulletin.Fundacao MT,Rondonopolis,MT,289-296. Ribeiro,LM,HDCampos,CRDias-Arieira,DLNeves,and GCRibeiro.2014.Effect of soybean seed treatment on the population dynamics of Pratylenchus brachyurus under water stress conditions.Bioscience Journal 30:616-622. Rios,ADF,MRRocha,ASMachado,KAGBAvila,RATeixeira,LCSantos,and LRSRabelo.2016.Host suitability of soybean and corn genotypes to the lesion caused by nematode under natural infestation conditions.Ciencia Rural.46:580-584. Rodrigues,DB,CRDias-Arieira,MVVVedoveto,M.Roldi,HFDMolin,and VHFAbe.2014.Crop rotation for Pratylenchus brachyurus control in soybean.Nematropica 44:146-151. Wei, J., K. Hale, L. Carta, E. Platzer, C. Wong, S. Fang, and FVAroian.2003. Bacillus thuringiensis crystal proteins that target nematodes. Proceedings of the National Academy of Sciences 100:2760-2765.

[0186] Example 5 - GMB151 transgenic soybean expressing the Bacillus thuringiensis toxin Cry14Ab-1 provides nematicidal protection in safrinha ("second crop") crops Building on and expanding on the experiments described in Example 3, this example sought to further determine whether planting GMB151 soybeans in the safra season could confer nematode suppression to a subsequent nematode-susceptible safrinha season crop.

[0187] Two soybean lines (one homozygous and one null for the GMB151 trait) were planted in production fields during the safra production season in fields infested with P. brachyurus. Conventional safrinha crops were planted on top of the same ground immediately following the soybean trial harvest in April / May 2019, 2020, and 2021. Each safrinha crop treatment trial consisted of three replicates arranged in a randomized complete block design. In 2019, Brachiaria spp. (signal grass) was planted as the safrinha crop. Signal grass plots were planted in parallel with the harvested soybean rows at each location. Plots were 5 meters long with 12 row width units. In April 2020 and 2021, corn and cotton were planted as safrinha crops after soybean. Three replicates of corn plots and three replicates of cotton plots were planted on top of the soybean trial ground. The corn and cotton furrows were planted perpendicular to the harvested soybean furrows. The cotton plots were planted on top of the first 3 m of replicates 1, 2, and replicate 3. The corn plots were planted on top of the last 3 m of replicates 5, 4, and replicate 3. Thus, the cotton and corn plots were 9 m long, with two replicates 8 furrows wide and the third replicate of each crop 4 furrows wide (Figure 3). In 2021, the cotton safrinha trial was replaced by a sorghum safrinha trial in three locations.

[0188] Data were collected to determine whether protection from P. brachyurus was conferred to a susceptible safrinha crop planted after GMB151 soybean. Protection against the safrinha crop was evaluated by measuring P. brachyurus population density and crop yield. Pratylenchus brachyurus population density was measured as the number of nematodes per gram of root tissue. P. brachyurus population density was measured in the safrinha crop at 30 days after planting (dap), 60 dap, and 90 dap. The root systems of 10 plants per plot were collected for each P. brachyurus measurement. Three to five roots were collected from the first and last furrows of each plot. The middle furrows were not sampled. Grain yields were collected from the center furrow of the cotton, maize, and sorghum safrinha plots in August 2020 and 2021. Yields were not taken from the signal grass plots in 2019 because root samples are destructive and would have interfered with the collection of yield data from the center furrow of each plot.

[0189] Data were analyzed to assess whether P. brachyurus population density and grain yield differed between plots following GMB151 soybean compared to conventional soybean. Data were analyzed separately for each safrinha crop; maize, cotton, sorghum, and Brachyaria. All data were analyzed using a mixed-effects ANOVA model. The model included fixed effects of location, GMB151 zygosity, and their interaction. Experimental replicates were considered as random effects.

[0190] Consideration: Pratylenchus brachyurus population densities were significantly reduced in all four safrinha crops by prior planting with GMB151 soybean during the safra season (Figure 13). Depending on the safrinha crop, brachyurus population densities were reduced by an average of 66-93% 90 days after planting, with an average reduction of 80%. There was no clear indication that P. brachyurus population suppression declined over the safrinha season in any of the three crops or that there was a resurgence of nematode populations. Thus, planting with GMB151 soybean reduced P. brachyurus populations throughout the entire cropping system, beginning with the safra soybean crop and extending through the safrinha crop harvest (see, for example, Figure 8). This led to improved yields in both the safra soybean crop (Figure 10) and the subsequent safrinha crop (Figure 7). In this exemplary location, GMB151 soybean reduced P. brachyurus populations by 99% at the end of the soybean season, which led to significant P. brachyurus suppression throughout the safrinha crop season from planting to harvest in the corn safrinha crop (Figures 4 and 6) and cotton safrinha crop (Figure 6).

[0191] Safrinha crop protection resulted in substantial yield improvements for corn production in Rio Verde, Mato Grosso. For example, as shown in Figure 5, P. brachyurus control achieved by GMB151 soybeans resulted in numerically higher soybean yields and significantly greater corn yields in the safrinha planting season. The yield improvements achieved by GMB151 from P. brachyurus control were worth approximately $90 USD / acre in increased gross revenue to farmers. Gross revenue estimates are based on average Brazilian grain prices of $9.00 per bushel for soybeans and $6.30 per bushel for corn.

[0192] The data provided in Examples 3 and 5 demonstrate that the incorporation of a highly effective transgenic nematicidal protein into a host crop can provide protection not only to the transformed crop, but also to susceptible crops that are rotated with the transformed crop or subsequently planted in the same location. This provides an effective way to achieve nematode control throughout the entire farm production system. Transforming a single crop to provide protection throughout the entire production system limits the development and regulatory costs associated with the production and registration of transgenic crops. It also provides a cost-effective way to provide nematode resistance to crops where native resistance is not available, and where a particularly common problem is migratory plant parasitic nematodes, such as Pratylenchus spp. This approach also provides an opportunity to provide transgenic nematode protection to crops such as wheat, where end-user acceptance of transgenic grain can be a barrier, but where Pratylenchus spp. can be a significant yield-reducing pathogen with few effective management options available.

[0193] In one embodiment, transgenic GMB151 soybeans provide significant yield and crop system management benefits to growers in Brazil. The high efficacy of GMB151 against P. brachyurus provides growers with safra soybean protection, as well as protection for many safrinha crop species. The benefit to safrinha crops is also to provide growers with freedom and flexibility in their production systems to select the most profitable and agronomically advantageous crop rotations, regardless of crop susceptibility to P. brachyurus.

[0194] It is also expected that the effects shown herein for one nematode susceptible plant will extend to hosts with the same susceptibility to the nematode. Below is a table showing the susceptibility of some common crops:

[0195] [Table 2]

[0196] For planted acreage forecasts, see also Camila Thaiana Rueda da Silva et al., Agriculture 2020, 10, 13; doi:10.3390 / agriculture10010013; Renato Lara De Assis et al., Expl Agric.:page 1 of 20 (Copyright) Cambridge University Press 2017 doi:10.1017 / S0014479717000333; and USDepartment of Agriculture, “World Agricultural Production”, Foreign Agriculture Service, Global Market Analysis, Circular Series WAP 2-22, February 2022.

[0197] References for Example 5: Barker, KR, and SRKoenning. 1998. Developing sustainable systems for nematode management. Annual Review of Phytopathology 36:165-205. Cook, R. 2004. Genetic resistance to nematodes: where is it useful? Australasian Plant Pathology 33:139-150. Inagaki, H., and M. Tsutsumi.1971.Survival of the soybean cyst nematode Heterodera glycines Ichinohe(Tylenchida:Heteroderidae)under certain storage conditions.Applied Entomology and Zoology 6:156-162. IRAC.2018.Nematicide resistance risk statement.https: / / irac-online.org / teams / nematodes. Nicol, JM, SJTurner, DLCoyne, L. den Nijs, S. Hockland, and ZTMaafi. 2011. Current nematode threats to world agriculture, in: J. Jones, G. Gheysen, C. Fenoll (Eds.), Genomics and Molecular Genetics of Plant-Nematode Interaction, Springer, Germany, pp. 21-43. Wei, J., K. Hale, L. Carta, E. Platzer, C. Wong, S. Fang, and FVAroian.2003. Bacillus thuringiensis crystal proteins that target nematodes. Proceedings of the National Academy of Sciences 100:2760-2765.

[0198] All publications and patent applications mentioned in this specification are indicative of the level of skill of those skilled in the art to which this invention pertains. All publications and patent applications are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

[0199] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be apparent that certain changes and modifications can be practiced within the scope of the appended claims.

Claims

1. 1. A method for controlling a nematode population at a locus comprising: Growing nematode-resistant plants at the location wherein growing the nematode resistant plant suppresses a nematode population at the locus or maintains suppression of the nematode population at the locus during and / or beyond the growing of the nematode resistant plant.

2. 2. The method of claim 1, wherein the period extends for one or more growing seasons after the growing season in which the nematode resistant plants were grown.

3. 10. The method of claim 1, further comprising growing a secondary plant at the locus after growing the nematode resistant plant.

4. 10. The method of claim 1, further comprising growing a secondary plant at said locus prior to growing said nematode resistant plant at said locus.

5. 10. The method of claim 1, further comprising growing the nematode resistant plant simultaneously with a secondary plant.

6. The method according to any one of claims 3 to 5, wherein the secondary plant is a nematode-susceptible plant.

7. 2. The method of claim 1, wherein the nematode population is suppressed or maintained below detection limits.

8. 6. The method of any one of claims 3 to 5, wherein the secondary plant is of the genus Brachiaria and the suppression of the nematode population is achieved when the number of nematodes is less than 60, about 60, or about 60 nematodes per gram of root.

9. 6. The method of any one of claims 3 to 5, wherein the secondary plant is maize and the suppression of the nematode population is achieved when the number of nematodes is less than 300, about 300, or about 300 nematodes per gram of root.

10. 6. The method of any one of claims 3 to 5, wherein the secondary plant is cotton and the suppression of the nematode population is achieved when the number of nematodes is less than 60, about 60, or about 60 nematodes per gram of root.

11. 6. The method of any one of claims 3 to 5, wherein the secondary plant is sorghum and the suppression of the nematode population is achieved when the number of nematodes is less than 250, about 250, or about 250 nematodes per gram of root.

12. 6. The method of any one of claims 3 to 5, wherein the suppression of the nematode population in the location where the secondary plants are grown is achieved when there is a 5% reduction, about a 5% reduction, or at least about a 5% reduction in the number of nematodes per gram of roots compared to the number of nematodes per gram of roots in a location comparable to the location where the secondary plants are grown.

13. 1. A method for protecting a nematode-susceptible plant from nematode damage or injury, comprising: growing a nematode-resistant plant at a locus for at least one growing season prior to planting said nematode-susceptible plant; growing nematode-susceptible plants at the locus for at least one growing season after growing the nematode-resistant plants; The method includes:

14. 1. A method for increasing yield of a nematode-susceptible plant, comprising: growing a nematode-resistant plant at a locus for at least one growing season prior to planting said nematode-susceptible plant; growing nematode-susceptible plants at said locus for said at least one growing season after growing said nematode-resistant plants; The method includes:

15. 15. The method of claim 14, wherein the increase in yield of the nematode susceptible plant is compared to the yield of the nematode susceptible plant grown in the same or a comparable location where the nematode resistant crop was not grown during the most recent crop rotation cycle.

16. The method according to any one of claims 3 to 5, wherein the nematode-susceptible plant is a perennial or annual plant.

17. 6. The method of any one of claims 3 to 5, wherein the nematode susceptible plant is a soybean plant, a corn plant, a cotton plant, a canola plant, a sugarcane plant, a potato plant, a wheat plant, a vegetable plant, a rice plant, an alfalfa plant, a barley plant, a sorghum plant, an oat plant, a rye plant, a cassava plant, a sweet potato plant, a sunflower plant, a fruit plant, an orchard plant (e.g. a fruit or nut bearing tree), an ornamental plant, or a grape vine.

18. The method of claim 1 , wherein the nematode-resistant plant expresses a nematocidal Cry protein.

19. 2. The method of claim 1, wherein the nematode resistant plant is a soybean plant, a corn plant, a cotton plant, a canola plant, a sugarcane plant, a sugar beet plant, a potato plant, a wheat plant, a rice plant, an alfalfa plant, a barley plant, a sorghum plant, an oat plant, a rye plant, a cassava plant, a sweet potato plant, a sunflower plant, a vegetable plant, a fruit plant, an ornamental plant, an orchard plant (e.g., a fruit or nut-bearing tree), or a grapevine.

20. The nematode may be a Pratylenchus spp. nematode population, e.g., Pratylenchus brachyurus, Meloidogyne spp., Heterodera spp., e.g., Heterodera glycines, Globodera spp., Rotylenchus reniformis, Helicotylenchus spp., e.g., Helicotylenchus dihystera, Scutellonema brachyurus, Scutellonema spp., Helicotylenchus ...

2. The method of claim 1, wherein the nematode species is selected from the group consisting of P. brachyurus, Tubixaba tuxaua, and Aphelencoides besseyi.

21. 21. The method of claim 20, wherein the Pratylenchus species is Pratylenchus brachyurus.

22. 13. A locus in which nematode population density has been suppressed, said suppression of nematode population density being achieved by the method of claim 1.

23. Benefits of: a. the locus does not have to lie fallow for one growing season per rotation cycle; b. the locus does not need to be cultivated for one growing season per crop rotation cycle; and c. The locus does not need to be planted with a cover crop.

23. The location of claim 22, providing one or more of the following:

24. 23. The locus of claim 22, wherein the suppression of nematode population density is about 250, 200, 150, 100, 50, 20, or 10 nematodes per gram (g) of root, or less than about 250, 200, 150, 100, 50, 20, or 10 nematodes per gram (g).

25. 23. A nematode resistant plant grown at the locus of claim 22.

26. 26. Plant material harvested from the plant of claim 25.

27. 26. A seed produced by the plant of claim 25.

28. 1. A system for augmenting location usage, comprising: growing nematode resistant plants at the location during a first growing season; growing nematode-susceptible plants at said location during a subsequent growing season. A system containing.

29. 30. The system of claim 28, wherein the locus is not left fallow between subsequent growing seasons.

30. 30. The system of claim 28, wherein no cover plants or cover crops are grown during the subsequent growing season.

31. 30. The system of claim 28, wherein the nematode-resistant plants and the nematode-susceptible plants are grown in successive growing seasons.

32. 30. The system of claim 28, wherein the nematode-resistant plant and the nematode-susceptible plant are each unique value crop plants.

33. 1. A method for improving a crop rotation system comprising: growing a nematode resistant plant at the locus during a first growing season; growing nematode-susceptible plants at said locus during a subsequent growing season; wherein growing the nematode resistant plants at the locus during the first growing season results in suppression of the nematode population at the locus that enables or enhances growing the nematode susceptible plants in the subsequent growing season.

34. The improvements in the crop rotation system include: a. use of said location for at least one additional growing season each year; b. A reduction in cultivation of said site; c. Reduced treatment of nematode-susceptible crop seeds with nematicides; d. reducing the area of ​​the locus treated with a nematicide prior to or during the growing season of the nematode-susceptible plants; e. reducing the rate at which nematicides are applied to the nematode-susceptible plants and / or the locus prior to or during the growing season of the nematode-susceptible plants; f. reducing the number of nematicide applications made to the nematode-susceptible plants and / or the locus during the growing season; g. increasing the availability of said location; h) an increase in the value of said location; i. Improving sustainable agricultural practices; and / or j. Increasing the yield of said nematode-susceptible crops The method of claim 33, further comprising one or more of the following:

35. 1. A method of nematode management for a locus comprising: growing a nematode resistant plant at a locus for a first growing season, the nematode resistant plant suppressing a nematode population at the locus or maintaining suppression of the nematode population at the locus; growing nematode-susceptible plants at said location during the same or a subsequent growing season; achieving improved health and / or yield of said nematode susceptible plants compared to the health and / or yield expected if said nematode population was not controlled; The method includes:

36. 36. The method of claim 35, wherein the improved health of the nematode-susceptible plant comprises one or more of the following: improved root development (e.g., improved root or root hair growth); improved yield; faster emergence; improved plant stress management, including increased stress tolerance and / or improved recovery from stress; increased mechanical strength; improved drought tolerance; reduced fungal, bacterial, and / or viral disease infection; or any combination thereof.

37. 1. A method of marketing a crop rotation system comprising: Promoting the use of nematode resistant plants during the first growing season; Promoting the use of nematode-susceptible plants in the location during the same or subsequent growing seasons; wherein growing the nematode resistant plants at the locus during the first growing season results in suppression of nematode populations at the locus that enables or enhances growing the nematode susceptible plants in a subsequent growing season.

38. 38. The method of claim 37, wherein the subsequent growing season is immediately adjacent to the first growing season.

39. 1. Marketing material relating to a crop rotation system in which a nematode resistant plant is grown during a first growing season in conjunction with or followed by growing a nematode susceptible plant at a locus during a subsequent growing season, wherein growing the nematode resistant plant at the locus during the first growing season enables or enhances the growing of the nematode susceptible plant, resulting in suppression of the nematode population at the locus.

40. 40. The marketing material of claim 39, wherein said marketing material relates to promoting said crop rotation system.

41. 1. A method of nematode management for a locus comprising: growing a nematode resistant plant at the locus prior to, simultaneously with, or after growing a secondary crop at said locus.

42. 42. The method of any one of claims 1, 33, 35, 37 and 41, wherein the nematode-resistant plant expresses a nematocidal Cryl4Ab protein having at least 95, 96, 97, 98, or at least 99% sequence identity to SEQ ID NO:

1.

43. 42. The method of any one of claims 1, 33, 35, 37, 40 and 41, wherein the nematode resistant plant comprises an elite event EE-GM5.

44. 42. The method of any one of claims 1, 33, 35, 37, 40 and 41, wherein the nematode resistant plant comprises an elite event EE-GM4.

45. 41. The locus of any one of claims 22 to 24, the nematode resistant plant of claim 25, the plant material of claim 26, the seed of claim 27, the line of any one of claims 28 to 32, or the marketing material of claim 39 or 40, wherein the nematode resistant plant expresses the Bacillus thuringiensis toxin Cry14Ab-1.

46. 41. The location of any one of claims 22 to 24, the nematode resistant plant of claim 25, the plant material of claim 26, the seed of claim 27, the line of any one of claims 28 to 32, or the marketing material of claim 39 or 40, wherein the nematode resistant plant comprises elite event EE-GM5.

47. 41. The location of any one of claims 22 to 24, the nematode resistant plant of claim 25, the plant material of claim 26, the seed of claim 27, the line of any one of claims 28 to 32, or the marketing material of claim 39 or 40, wherein the nematode resistant plant comprises elite event EE-GM4.

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