Methods of weed control

Description

1 / 44 “WEED CONTROL METHODS” RELATED ORDERS

[0001] This application claims priority from U.S. Provisional Patent Application No. 63 / 459,717, filed April 17, 2023, which is incorporated herein by reference in its entirety. FIELD AND BACKGROUND OF THE INVENTION

[0002] The present invention, in some of its embodiments, relates to methods of weed control and, specifically, to the use of xenogeneic pollen in weed control.

[0003] Weeds have been the leading biotic cause of agricultural productivity losses since the dawn of agriculture. The potential for damage caused by weeds is estimated at an average of 34% loss in agricultural productivity worldwide [Oerke, EC., 2006]. In the US alone, the annual cost of crop losses due to weeds exceeds $26 billion [Pimentel D et al., 2000]. Furthermore, according to the Weed Science Society of America, weeds are estimated to cause over $40 billion in annual global losses [wssa(dot)net / wssa / weed / biological-control / ]. Weeds therefore represent a major threat to food security [Delye et al., 2013].

[0004] Herbicides are the most commonly used and effective weed control tools. Due to the intense selective pressure exerted by herbicides, herbicide resistance is constantly growing, and in 2016, there were more than 470 weed biotypes identified as resistant to one or more herbicides. Petition 870250094224, dated 10 / 15 / 2025, page 7 / 57 2 / 44 herbicides according to the International Survey of Herbicide-Resistant Weeds (weedscience(dot)org / ).

[0005] Weeds, like other plants, have various mechanisms of sexual reproduction: self-pollination, cross-pollination, or both. Self-pollination describes pollination using pollen from a flower that is transferred to the same or another flower of the same plant. Cross-pollination describes pollination using pollen from a flower of another plant. Weeds depend on wind or animals such as bees and other insects to pollinate them.

[0006] Since the 1940s, the use of sterile organisms has been reported to reduce pest populations, and the success of these methods has been demonstrated in many cases, such as the tsetse fly [Klassen & Curtis, 2005], the melon fly [Yosiakie et al. 2003] and the sweet potato weevil [Kohama et al., 2003].

[0007] Planting sterile pollen-producing plants in the field to produce infertile seeds was mentioned, but immediately rejected for practical, regulatory and economic reasons.

[0008] PCT Publication No. WO2017 / 203519 describes a method of weed control comprising the artificial pollination of a weed species of interest with pollen from the same species, which reduces the fitness of the weed species of interest, thereby blocking the next generation of viable weed seeds. Petition 870250094224, dated 10 / 15 / 2025, page 8 / 57 3 / 44

[0009] PCT Publication No. WO2020 / 084586 describes a method for inhibiting weed growth. The additional technique of background checks includes:

[0010] Gaines, TA et al. (2012) Interspecific hybridization transfers a previously unknown glyphosate resistance mechanism in Amaranthus species. Evolutionary Applications 5(1):29-38.

[0011] Trucco et al. 2007 Weed Science 55:119122 . SUMMARY OF THE INVENTION

[0012] According to one aspect of some embodiments of the present invention, a method of weed control is provided, the method comprising artificially pollinating a weed species of interest with irradiated pollen that reduces the fitness of the weed species of interest, the pollen being of a species different from that of the weed species of interest and capable of competing with the pollen of the weed species of interest, wherein the artificial pollination is carried out under conditions of competition with native pollen.

[0013] According to some embodiments of the invention, the pollen consists of pollen from a species different from that of the weed species of interest.

[0014] According to some embodiments of the invention, artificial pollination is carried out on a large scale of at least 0.1 acre.

[0015] According to some embodiments of the invention, the pollen is contained in a formulation that allows the distribution of pollen by artificial pollination. Petition 870250094224, dated 10 / 15 / 2025, p. 9 / 57 4 / 44

[0016] According to some embodiments of the invention, the pollen is irradiated by γ radiation.

[0017] According to some embodiments of the invention, the pollen is irradiated by X radiation.

[0018] According to some embodiments of the invention, the pollen is irradiated by alpha radiation.

[0019] According to some embodiments of the invention, the pollen is of the same genus as the weed species of interest.

[0020] According to some embodiments of the invention, the pollen and the weed species of interest are of different

[0021] genera. According to some embodiments of the In the invention, at least one of the aforementioned pollen and the aforementioned weed species of interest is a hybrid.

[0022] According to some embodiments of the invention, the pollen is non-genetically modified pollen.

[0023] According to some embodiments of the invention, the pollen is genetically modified pollen.

[0024] According to some embodiments of the invention, the pollen is from the Amaranthaceae family.

[0025] According to some embodiments of the invention, the pollen is of the genus Amaranthus.

[0026] According to some embodiments of the invention, the pollen is A. palmeri.

[0027] According to some embodiments of the invention, the pollen and weed species of interest are selected from the group consisting of A. palmeri and A. tuberculatus. Petition 870250094224, dated 10 / 15 / 2025, p. 10 / 57 5 / 44

[0028] According to some embodiments of the invention, the pollen is A. palmeri.

[0029] According to some embodiments of the invention, the pollen is a hybrid of A. palmeri and A. tuberculatus.

[0030] According to some embodiments of the invention, the weed species of interest is a hybrid of A. palmeri and A. tuberculatus.

[0031] According to some embodiments of the invention, the pollen is of the genus Lolium.

[0032] According to some embodiments of the invention, the pollen is of the genus Alopecurus.

[0033] According to some embodiments of the invention, the pollen and the weed species of interest are selected from the group of pairs shown in Table 1.

[0034] According to some embodiments of the invention, the ability to compete is achieved by fertilizing the weed species of interest.

[0035] Unless otherwise defined, all technical and / or scientific terms used in this invention have the same meaning commonly understood by a person skilled in the art to which the invention relates. Although methods and materials similar or equivalent to those described in this invention may be used in the practice or testing of embodiments of the invention, exemplary methods and / or materials are described below. In case of conflict, the descriptive report of the patent, including the definitions, shall prevail. Furthermore, materials, methods Petition 870250094224, dated 10 / 15 / 2025, p. 11 / 57 6 / 44 and examples are for illustrative purposes only and are not intended to be exhaustive.

[0036] Implementing the method and / or system of the embodiments of the invention may involve performing or completing selected tasks manually, automatically, or a combination thereof. Furthermore, depending on the current instrumentation and equipment of the embodiments of the method and / or system of the invention, various selected tasks may be implemented by hardware, software, firmware, or a combination thereof, using an operating system. DESCRIPTION OF SPECIFIC EMBODIMENTS OF THE INVENTION

[0037] The present invention, in some embodiments thereof, relates to methods of weed control.

[0038] Before explaining at least one embodiment of the invention in detail, it should be understood that the invention is not necessarily limited in its application to the details presented in the following description or exemplified by the Examples. The invention may be carried out in other embodiments or practiced or performed in various ways.

[0039] The present inventors have previously developed a technology for weed control using irradiated pollen from the same species as the target weed. This pollen is typically applied during the flowering period of the weeds by means of artificial pollination to prevent the generation of viable seeds of resistant weeds, while competing with wild pollen present in the field. Petition 870250094224, dated 10 / 15 / 2025, page 12 / 57 7 / 44

[0040] Upon further developing the technology, the present inventors have surprisingly discovered that pollen from a weed of a different species than the target weed can be used effectively in a weed control method. This is surprising, since irradiated pollen from another species of the target plant is not expected to fertilize and subsequently produce a similar set of seeds of a hybrid weed in a manner comparable to the application of irradiated pollen and even non-irradiated pollen (see Examples section) of the same species. In both cases, the resulting seeds are aborted and fail to germinate effectively (see, for example, Example 1). These results were reinforced in experiments conducted under competitive conditions, demonstrating the ability of irradiated pollen to successfully compete with non-irradiated pollen from the target plant (of the same species, see, for example, Example 2).Thus, the pollen of the present invention that is non-(i.e., xenogeneic) from the target weed species can be used as universal pollen for weed control.

[0041] Thus, according to one aspect of the invention, a method of weed control is provided. The method comprises artificially pollinating a weed species of interest with irradiated pollen that reduces the fitness of the weed species of interest, the pollen being of a different species from that of the weed species of interest and capable of competing with the pollen of the weed species of interest, wherein the pollination Petition 870250094224, dated 10 / 15 / 2025, page 13 / 57 8 / 44 artificial pollen is produced under conditions of competition with native pollen.

[0042] As used in the present invention, “competition” refers to the ability of pollen to reduce the production of viable seeds resulting from natural fertilization by target pollen. This can be done in numerous ways, such as physically occupying the position of the target pollen in the female organ and / or fertilizing the target plant. Thus, seed production may be approximately the same in terms of quantity, but the number of non-viable seeds in seed production (e.g., as manifested by seed weight and / or germination capacity) is greater than that of the control.

[0043] As used in the present invention, “control” refers to fertilization by non-irradiated pollen of the same species as the weed species of interest (also referred to as “target” or “target plant”).

[0044] As used in the present invention, the term “weed control” refers to the suppression of the growth and, optionally, the spread of a population of at least one weed species of interest or of a member or members of a genus or family thereof, and even to the reduction of the size of its population in a given growing area.

[0045] According to some embodiments of the invention, weed control is carried out in a growing area of ​​at least 0.1 acre and optionally not exceeding 100,000 acres, for example, 50,000 acres. Petition 870250094224, dated 10 / 15 / 2025, page 14 / 57 9 / 44

[0046] According to a specific modality, the growth is from 1 to 10,000 acres, for example, 1 to 5,000, 1 to 2,000, 1 to 1,000 acres.

[0047] According to some embodiments of the invention, weed control is carried out in a growing area of ​​at least 1 acre.

[0048] According to a specific modality, the growth area is an urban area, for example, golf courses, athletic fields, parks, cemeteries, roadsides, residential gardens / lawns and the like.

[0049] According to an additional or alternative modality, the growth area is a rural area.

[0050] According to an additional or alternative modality, the growth area is an agricultural growth area, for example, open field, greenhouse, plantation, vineyard, orchard, etc.

[0051] As used in the present invention, the term “weed species of interest” refers to a wild plant that grows where it is not wanted and that may be in competition with cultivated plants of interest (i.e., plants desirable for crops). Weeds are typically characterized by rapid growth and / or ease of germination and / or competition with crops for space, light, water, and nutrients.

[0052] According to some embodiments of the invention, the weed species of interest is not traditionally cultivated. The weed species of interest may also be referred to as the target weed. Petition 870250094224, dated 10 / 15 / 2025, page 15 / 57 10 / 44

[0053] According to another embodiment of the invention, the weed of interest or irradiated pollen is a perennial weed.

[0054] According to another embodiment of the invention, the weed of interest or irradiated pollen is a biennial weed.

[0055] According to another embodiment of the invention, the weed of interest or irradiated pollen is an annual weed.

[0056] According to another embodiment of the invention, the weed of interest or irradiated pollen is a therophyte.

[0057] According to one embodiment, the weed of interest or irradiated pollen is a parasitic plant.

[0058] According to one embodiment, the weed of interest or irradiated pollen reproduces sexually.

[0059] According to one embodiment, the weed A weed of interest, or irradiated pollen, is a broad-leaved weed.

[0060] According to one embodiment, the weed of interest or irradiated pollen is a grassy weed.

[0061] According to one embodiment, the weed of interest or irradiated pollen is a non-grass monocotyledonous weed.

[0062] According to one embodiment, the weed of interest or irradiated pollen is from a monoecious plant. Petition 870250094224, dated 10 / 15 / 2025, page 16 / 57 11 / 44

[0063] According to one embodiment, the weed of interest or irradiated pollen is from a dioecious plant.

[0064] According to one embodiment, the weed of interest or radiated pollen is from a protogynous plant, in which the flowering of male and female flowers is regulated spatially or temporally. For example, female flowers that mature before male flowers, ensuring a high level of cross-pollination (temporal regulation), are found in Bassia scoparia (kochia). The genus Amaranthus L. is a diverse group of plants with 70 to 80 species distributed throughout the temperate and tropical regions of the world [Raiyemo et al. BMC Biology (2023) 21:37]. Nine of these species [Amaranthus acanthochiton JD Sauer, Amaranthus Arenicola IM Johnson, Amaranthus australis (A. Gray) JD Sauer, Amaranthus cannabinus (L.) JD Sauer, Amaranthus floridanus (S. Watson) JD Sauer, Amaranthus tuberculatus (Moq.) JD Sauer, Amaranthus greggii S. Watson, Amaranthus watsonii Standley and Amaranthus palmeri s.Watson] are dioecious (i.e., separate individual male and female plants), native to North America and collectively grouped in the subgenus Acnida (L.) Aellen ex KR. In monoecious plants, which are also contemplated in the present invention, male and female flowers can be found on one plant.

[0065] As used in the present invention, "genus" refers to a plurality of species.

[0066] As used in the present invention, "family" refers to a plurality of genders. Family is Petition 870250094224, dated 10 / 15 / 2025, page 17 / 57 12 / 44 a basic taxonomic grouping, classified above the genus and below the order.

[0067] Typically, the classification follows the Linnaean classification system.

[0068] Exemplos de famílias de ervas daninhas incluem, mas não estão limitados a, Amaranthaceae, Anacardiaceae, Compositae, Apocynaceae, Curcurbitaceae, Poaceae (Gramínaceae), Cannabaceae, Chenopodiaceae, Leguminosae; Fabaceae, Lythraceae, Malvaceae, Asclepiadaceae, Labiatae, Convolvulaceae, Cruciferae, Solanaceae, Plantaginaceae, Portulacaceae, Rosaceae, Cyperaceae e Polygoneaceae.

[0069] Examples of weed species that may be targeted (mitigated) in accordance with these teachings include, but are not limited to, Amaranthus species - A. albus, A. blitoides, A. hybridus, A. palmeri, A. powellii, A. retroflexus, A. rudis, A. spinosus, A. tuberculatus, A. thunbergii, A. graecizans and A. viridis; Ambrosia species - A. trifida, A. artemisifolia; Lolium species - L. multiflorum, L. rigidium, L. perenne; Digitaria species - D. insularis, D. sanguinalis; Euphorbia species - E. heterofila; Kochia species - K. scoparia; Sorghum species - S. halepense; Conyza species - C. bonariensis, C. canadensis, C. sumatrensis; Cloris species - C. truncaris; Echinochola species - E. colona, ​​E. crus-galli; Eleusina species - E. indica; Poa species - P. annalis; Plantago species - P. lanceolata; Avena species - A. fatua; Chenopodium species - C. album; Setaria species - S.viridis, Abutilon theophrasti, Ipomoea species, Sesbania, Xanthium strumarium species, Cassia species,. Petition 870250094224, dated 15 / 10 / 2025, p. 18 / 57 13 / 44 Sida Species, Brachiaria Species, Sporobolus Species - S. pyramidalis, S. natalensis, S. jacquemontii, S. fertilis, S. africanus S. indicus, Solanum nigrum, Solanum carolinense and Solanum elaeagnifolium.

[0070] Additional weed species found in cultivated areas include Alopecurus myosuroides, Avena sterilis, Avena sterilis ludoviciana, Brachiaria plantaginea, Bromus diandrus, Bromus rigidus, Cynosurus echinatus, Digitaria ciliaris, Digitaria ischaemum, Digitaria sanguinalis, Echinochloa oryzicola, Echinochloa phyllopogon, Eriochloa punctata, Hordeum glaucum, Hordeum leporinum, Ischaemum rugosum, Leptochloa chinensis, Lolium persicum, Phalaris minor, Phalaris paradoxa, Rottboellia exalta, Setaria faberi, Setaria viridisvar, robusta-alba schreiber, Setaria viridisvar, robusta-purpurea, Snowdenia polystachea, Sorghum Sudanese, Alisma plantago-aquatica, Amaranthus lividus, Ammaniaauriculata, Ammania coccinea, Anthemis cotula, Apera spica-venti, Bacopa rotundifolia, Bidens pilosa, Bidens subalternans, Brassica tournefortii, Bromus tectorum, Camelina microcarpa, Chrysanthemum coronarium, Cuscuta campestris, Cyperus dijformis, Damasonium minus, Descurainia sophia,Diplotaxis tenuifolia, Echium plantagineum, Elatine triandravar, pedicellata, Euphorbia heterophylla, Fallopia convolvulus, Fimbristylis miliacea, Galeopsis tetrahit, Galium spurium, Helianthus annuus, Iva xanthifolia, Ixophorus sunisetus, Ipomoea indica, Ipomoea purpurea, Ipomoea sepiaria, Ipomoea aquatica, Ipomoea triloba, Lactuca serriola, Limnocharis flava, Limnophila erecta, Limnophila sessiliflora, Lindernia dubia, Lindernia dubiavar, major, Petition 870250094224, dated 15 / 10 / 2025, p. 19 / 57 14 / 44 Lindernia micrantha, Lindernia procumbens, Mesembryanthemum crystallinum, Monochoria korsakowii, Monochoria vaginalis, Neslia paniculata, Papaver rhoeas, Parthenium hysterophorus, Pentzia suj fruticosa, Phalaris minor, Raphanus raphanistrum, Raphanus sativus, Rapistrum rugosum, Rotalaindicavar, uliginosa, Sagittaria guyanensis, Sagittaria montevidensis, Sagittaria pygmaea, Salsola iberica, Scirpus juncoidesvar, ohwianus, Scirpus mucronatus, Setaria lutescens, Sida spinosa, Sinapis arvensis, Sisymbrium orientale, Sisymbrium thellungii, Solanum ptycanthum, Sonchus asper, Sonchus oleraceus, Sorghum bicolor, Stellaria media, Thlaspi arvense, Xanthium strumarium, Arctotheca calendula, Conyza sumatrensis, Crassocephalum crepidiodes, Cuphea carthagenenis, Epilobium adenocaulon, Erigeron philadelphicus, Landoltia punctata, Lepidium virginicum, Monochoria korsakowii, Solanum americanum, Solanum nigrum, Vulpia bromoides, Youngia j aponica, Hydrilla verticillata, Carduus nutans,Carduus pycnocephalus, Centaurea solstitialis, Cirsium arvense, Commelina diffusa, Convolvulus arvensis, Daucuscarota, Digitaria ischaemum, Echinochloa crus-pavonis, Fimbristylis miliacea, Galeopsis tetrahit, Galium spurium, Limnophila erecta, Matricaria perforada, Papaver rhoeas, Ranunculus acris, Soliva sessilis, Sphenoclea zeylanica, Stellaria media, Nassella trichotoma, Stipa neesiana, Agrostis stolonifera, Polygonum aviculare, Alopecurus j aponicus, Beckmannia syzigachne, Bromus tectorum, Chloris inflate, Echinochloa erecta, Portulaca oleracea and Senecio vulgaris. Examples of parasitic plants include, but are not limited to, Striga sp, Orobanche sp, Cuscuta sp, Visco., Petition 870250094224, dated 15 / 10 / 2025, p. 20 / 57 15 / 44

[0071] According to a specific embodiment, weed species are selected from or belong to the group consisting of Amaranthus: A. palmeri, A. tuberculatus, A. retroflexus, A. hybridus, A. powellii, A. spinosus, A. albus, A. blitoides, A. rudis, Lolium rigidum, Lolium multiflorum, Lolium perenne Ambrosia: A. trifida, A. artemisiifolia, Kochia scoparia, Conyza: C. canadensis, C. bonariensis, Echinochloa, Alopecurus myosuroides, Sorghum halepense, Digitaria insularis, Eleusine indica, Avena fatua, Euphorbia Heterophylla and Chenopodium album.

[0072] According to a specific modality, the pollen is from the Amaranthaceae family.

[0073] According to a specific modality, the pollen is of the genus Amaranthus.

[0074] According to a specific type, the pollen is from the Poaceae family.

[0075] According to a specific modality, the pollen is of the genus Lolium.

[0076] According to a specific modality, the pollen is of the genus Alopecurus.

[0077] According to a specific modality, the pollen is of the genus Ambrosia.

[0078] According to a specific modality, the pollen is of the genus Erigeron.

[0079] According to a specific modality, the pollen is from the Asteraceae Family.

[0080] According to a specific embodiment, at least one of the pollen and the weed species of interest is a hybrid. Thus, the pollen may be a hybrid, the weed species of interest may Petition 870250094224, dated 10 / 15 / 2025, p. 21 / 57 16 / 44 could be a hybrid, or both could be hybrids, although they are different hybrids.

[0081] According to a specific variant, the pollen in question is A. palmeri.

[0082] According to a specific embodiment, the pollen in question is a hybrid of A. palmeri and A. tuberculatus.

[0083] According to a specific embodiment, the aforementioned weed species of interest is a hybrid of A. palmeri and A. tuberculatus.

[0084] According to a specific embodiment, the said pollen and the said weed species of interest are selected from the group consisting of A. palmeri and A. tuberculatus. Thus, the pollen may be A. palmeri or a hybrid thereof and the weed species of interest A. tuberculatus or a hybrid thereof. The weed species of interest may be A. palmeri or a hybrid thereof and the pollen A. tuberculatus or a hybrid thereof.

[0085] Other examples of amaranth hybrids include, but are not limited to, A. palmeri x A. spinosus, A. palmeri x A. hybridus, A. tuberculatus x A. hybridus, A. tuberculatus x A. powellii, A. tuberculatus x retroflexus, and A. tuberculatus x A. spinosus.

[0086] According to a specific embodiment, the pollen or target is not a hybrid or, specifically, none of the hybrids mentioned in the present invention.

[0087] As mentioned, the pollen is not of the same species as the target weed, but rather of the same family as the target weed. The pollen may be of a different species. Petition 870250094224, dated 10 / 15 / 2025, page 22 / 57 17 / 44 of the target weed genus. Alternatively, the pollen may be of a different genus than that of the target weed, however, in some embodiments of the invention, the pollen is of the same family as the target weed. For example, C. album with A. palmeri.

[0088] According to a specific embodiment, the pollen is a mixture of pollen (e.g., 2, 3, 4, 5) from species or genera different from the target weeds in the treated field. For example, more than 40%, 50%, 60%, 70% of the weeds in the treated area are not of the same species as the pollen used in weed control.

[0089] According to a specific embodiment, the pollen consists of pollen from a species different from that of the weed species of interest (as mentioned, the target weed).

[0090] Different weeds can have different growth habits and therefore specific weeds usually characterize a particular crop under certain growing conditions.

[0091] According to a specific embodiment, the weed is a herbicide-resistant weed.

[0092] According to a specific modality, a weed is defined as herbicide-resistant when it meets the Weed Science Society of America (WSSA) definition of resistance.

[0093] Consequently, the WSSA defines herbicide resistance as “the inherited ability of a plant to survive and reproduce after exposure to a dose of herbicide that is normally lethal to the wild type.” Alternatively, herbicide resistance is defined as Petition 870250094224, dated 10 / 15 / 2025, page 23 / 57 18 / 44 the developed capacity of a previously herbicide-susceptible weed population to resist a herbicide and complete its life cycle when the herbicide is used at its normal rate in an agricultural situation (Source: Heap and Lebaron, 2001 in Herbicide Resistance and World Grains).

[0094] As mentioned, weed control, according to the present teachings, is carried out by reducing the fitness of at least one weed species of interest.

[0095] As used in the present invention, fitness refers to the relative ability of the weed species of interest to grow, reproduce or propagate and transmit its genes to the next generation.

[0096] As used in the present invention, relative means in comparison to a weed of the same species that has not been artificially pollinated with the pollen of the invention and grown under the same conditions.

[0097] It should be noted that the effect of pollen treatment, according to the present teachings, typically manifests itself in the first generation after fertilization. As demonstrated in the Examples section, fertilization with xeno-irradiated pollen of the present invention resulted in aborted seed production, manifested by significantly lower weight (results that were further corroborated by competition) and lower germination capacity compared to control seed production. As mentioned above, the control is non-irradiated pollen of the same species as the target weed; such control reflects natural competitive conditions. Petition 870250094224, dated 10 / 15 / 2025, page 24 / 57 19 / 44 In accordance with other methods, control is irradiated pollen from the same species as the target weed.

[0098] Fitness can be affected by reduced productivity, propagation, fertility, fecundity, biomass, tolerance to biotic stresses, tolerance to abiotic stresses and / or herbicide resistance.

[0099] As used in the present invention, “productivity” refers to the potential rate of incorporation or generation of energy or organic matter by an individual, population or trophic unit per unit of time per unit of area or volume; carbon fixation rate.

[0100] As used in the present invention, “fecundity” refers to the potential reproductive capacity of an organism or population, measured by the number of gametes.

[0101] According to a specific modality, pollen affects any stage of seed development or germination.

[0102] According to a specific modality, the reduction in productivity manifests itself through at least one of: (i) inability to develop an embryo; (ii) abortion of an embryo; (iii) seed unviability; (iv) seed that fails to develop completely; and / or (v) seed that fails to germinate. [010 3] According to a specific modality, the reduction in productivity manifests itself through at least one of the following: (i) inability to develop an embryo; (ii) abortion of an embryo; Petition 870250094224, dated 10 / 15 / 2025, p. 25 / 57 20 / 44 (iii) seed unviability; (iv) seed that fails to develop completely; (v) seed that fails to germinate; and / or (vi) reduced or non-existent seed production.

[0104] It is important to highlight that when pollen reduces the productivity, fertility, propagation capacity, or fecundity of the weed in the next generation, it may be termed sterile pollen by a qualified professional, even though it fertilizes the weed of interest. Therefore, sterile pollen, as used in this work, is still capable of fertilizing, but normally leads to interrupted seed development or seed abortion.

[0105] According to a specific embodiment, seed production is approximately similar in quantity to that obtained with the use of irradiated or non-irradiated pollen from the same target weed species. + / - 0.1 to 5 times (e.g., 0.4 to 5 times).

[0106] According to a specific modality, the reduction in fitness is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97% or even 100%, within the first generation after fertilization and optionally the second generation after fertilization and optionally the third generation after fertilization.

[0107] According to a specific modality, the reduction in fitness is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97% or even 100%, within the first generation after fertilization.

[0108] According to a specific modality, reduced fitness results from reduced tolerance to Petition 870250094224, dated 10 / 15 / 2025, page 26 / 57 21 / 44 biotic or abiotic conditions, for example, herbicide resistance.

[0109] Non-limiting examples of abiotic stress conditions include salinity, osmotic stress, drought, water deprivation, excess water (e.g., flooding, inundation), etiolation, low temperature (e.g., cold stress), high temperature, heavy metal toxicity, anaerobiosis, nutrient deficiency (e.g., nitrogen deficiency or limitation), excess nutrients, air pollution, herbicides, pesticides, and UV irradiation.

[0110] Biotic stress is stress that occurs as a result of damage caused to plants by other living organisms, such as bacteria, viruses, fungi, parasites, beneficial and harmful insects, weeds, and cultivated or native plants.

[0111] As used in the present invention, pollen refers to viable pollen that is capable of fertilizing the weed species of interest and therefore competes with native pollination.

[0112] According to some embodiments of the invention, the pollen is selected so that it is effective enough to compete with the pollen of the target weed. According to some embodiments, the term "effective enough" refers to obtaining a seed yield approximately similar in quantity to that obtained using irradiated or non-irradiated (native) pollen of the same target weed species, + / - 0.1 to 5 times (e.g., 0.4 to 5 times). Alternatively, when there is no competition for native pollen, or when pollen levels are very low. Petition 870250094224, dated 10 / 15 / 2025, p. 27 / 57 22 / 44 native species are present; pollination by the pollen used inhibits apomixis of weeds and thus also reduces their quantities [Ribeiro et al. 2012].

[0113] According to a specific modality, pollen exhibits susceptibility to a single growth condition, for example, herbicide and temperature.

[0114] Depending on the specific modality, pollen exhibits susceptibility to multiple growth conditions, for example, different herbicides.

[0115] According to a specific modality, the pollen is not genetically modified.

[0116] According to a specific modality, the pollen is genetically modified.

[0117] According to a specific embodiment, the pollen is of the same genus as the weed species of interest.

[0118] According to a specific modality, the pollen and the weed species of interest are from different genera.

[0119] For example, the genus Amaranthaceae comprises several species of red amaranth (Amaranthus retroflexus), smooth amaranth (Amaranthus hybridus), Powell amaranth (Amaranthus powellii), spiny amaranth (Amaranthus spinosus), collar amaranth (Amaranthus albus), prostrate amaranth (Amaranthus blitoides), common aquatic hemp (Amaranthus rudis), tall aquatic hemp (Amaranthus tuberculatus), pilgrim amaranth (Amaranthus palmeri), Amaranthus tuberculatus, Amaranthus thunbergii, Amaranthus graecizans or Amaranthus viridis. Petition 870250094224, dated 10 / 15 / 2025, page 28 / 57 23 / 44

[0120] According to a specific modality, the pollen is not from Amaranthus spinosus.

[0121] According to a specific modality, the pollen is not from Amaranthus spinosus when the female is Amaranthus palmeri.

[0122] An example of pollen that is not of the same species as the target weed can be found in the genus Amaranthus: female A. retroflexus x irradiated A. palmeri pollen.

[0123] Examples of pollination within different genera can be for the Amaranthaceae family: female of B. scoparia X pollen of irradiated A. palmeri, or C. album X pollen of irradiated A. palmeri.

[0124] Other examples are provided in Table 1 below. Table 1 - a non-exhaustive list of pollination pairs. Each pair (one row in the table) is considered an incorporation. # Female Pollen Classification 1 A. tuberculatus A. palmeri irradiated Genus Amaranthus 2 A. retroflexus A. palmeri irradiated Genus Amaranthus 3 A. albus A. palmeri irradiated Genus Amaranthus 4 A. blitoides A. palmeri irradiated Genus Amaranthus 5 A. hybridus A. palmeri irradiated Genus Amaranthus 6 A. powellii A. palmeri irradiated Genus Amaranthus 7 A. rudis A. palmeri irradiated Genus Amaranthus 9 A. palmeri A. retroflexus irradiated Genus Amaranthus 10 A. tuberculatus A. retroflexus irradiated Genus Amaranthus 11 A. albus A. retroflexus irradiated Genus Amaranthus Petition 870250094224, dated 10 / 15 / 2025, page 29 / 57 24 / 44 12 A. blitoides A. retroflexus irradiated Genus Amaranthus 13 A. hybridus A. retroflexus irradiated Genus Amaranthus 14 A. powellii A. retroflexus irradiated Genus Amaranthus 15 A. rudis A. retroflexus irradiated Genus Amaranthus 16 A. spinosus A. retroflexus irradiated Genus Amaranthus 17 A. viridis A. retroflexus irradiated Genus Amaranthus 18 A. palmeri A. tuberculatus irradiated Genus Amaranthus 19 A. retroflexus A. tuberculatus irradiated Genus Amaranthus 20 A. albus A. tuberculatus irradiated Genus Amaranthus 21 A. blitoides A. tuberculatus irradiated Genus Amaranthus 22 A. hybridus A. tuberculatus irradiated Genus Amaranthus 23 A. powellii A. tuberculatus irradiated Genus Amaranthus 24 A. rudis A. tuberculatus irradiated Genus Amaranthus 25 A. spinosus A. tuberculatus irradiated Genus Amaranthus 26 A. viridis A. tuberculatus irradiated Genus Amaranthus 27 B. scoparia A. palmeri irradiated Family Amaranthaceae 28 C. album A. palmeri irradiated Family Amaranthaceae 29 B.scoparia C. album irradiated Family Amaranthaceae 30 C. album B. scoparia irradiated Family Amaranthaceae 31 B. scoparia A. tuberculatus irradiated Family Amaranthaceae 32 C. album A. tuberculatus irradiated Family Amaranthaceae 33 L. multiflorum L. rigidum irradiated Genus Lolium 34 L. perenne L. rigidum irradiated Genus Lolium 35 L. temulentum L. rigidum irradiated Genus Lolium 36 L. canariense L. rigidum irradiated Genus Lolium 37 L. persicum L. rigidum irradiated Genus Lolium 38 L. rigidum L. perenne irradiated Genus Lolium 39 A. pratensis A. myosuroides irradiated Genus Alopecurus. Petition 870250094224, dated 15 / 10 / 2025, p. 30 / 57 25 / 44 40 A. japonicus A. myosuroides irradiated Genus Alopecurus 41 A. aequalis A. myosuroides irradiated Genus Alopecurus 42 A. myosuroides A. japonicus irradiated Genus Alopecurus 43 L. rigidum A. myosuroides irradiated Family Poaceae 44 A. myosuroides L. rigidum irradiated Family Poaceae 45 A. confertiflora A. artemisiifolia irradiated Genus Ambrosia 46 A. psilostachya A. artemisiifolia irradiated Genus Ambrosia 47 A. trifida A. artemisiifolia irradiated Genus Ambrosia 48 A. artemisiifolia A. trifida irradiated Genus Ambrosia 49 A. artemisiifolia A. contertiflora irradiated Genus Ambrosia 50 C. bonariensis C. canadensis irradiated Genus Erigeron 51 C. canadensis C. bonariensis irradiated Genus Erigeron 52 A. artemisiifolia C. canadensis irradiated Family Asteraceae 53 C. canadensis A. artemisiifolia irradiated Family Asteraceae

[0125] According to some modalities, the pollen is not A. hybridus when the target plant is A. tuberculatus.

[0126] According to some embodiments, the weed species of interest is selected so that the female flowers mature before the male flowers, ensuring a high level of cross-pollination. According to some embodiments, the weed species of interest is Bassia scoparia (Kochia).

[0127] The pollen used according to some embodiments of the invention is irradiated pollen which will reduce the fitness of the target weed, as explained above.

[0128] According to a specific embodiment, a method is provided for producing pollen that reduces fitness Petition 870250094224, dated 10 / 15 / 2025, page 31 / 57 26 / 44 of at least one weed species of interest, the method comprising treating the weed species of interest (e.g., seeds, seedlings, tissues / cells) or pollen thereof with an agent that reduces fitness.

[0129] According to a specific modality, the agent is radiation.

[0130] According to a specific embodiment, the method comprises collecting pollen from the weed species of interest after treatment with the fitness-reducing agent.

[0131] According to other specific modalities, the pollen can first be collected and then treated with the agent (i.e., radiation) that reduces the fitness of the weed species of interest.

[0132] According to some methods, the pollen used is mature pollen, for example, at least 20%, 50% or 80% of the pollen is mature.

[0133] According to some methods, mature pollen is collected after the natural process of anther dehiscence.

[0134] Depending on the specific modality, radiation is selected from the group consisting of X-ray radiation, gamma radiation, particle irradiation such as alpha, beta or other accelerated particles, and UV radiation. The qualified professional will know which agent to select.

[0135] According to a specific modality, radiation is X-ray radiation.

[0136] Depending on the specific modality, the radiation dose (X-ray) is 50 to 600 Gy, for example, 100 to 400 Gy, 150 to 400 Gy, 100 to 300 Gy, 150 to 250 Gy, 100 Petition 870250094224, dated 10 / 15 / 2025, p. 32 / 57 27 / 44 to 250 Gy, 150 to 300 Gy, for example, 150, 200, 250 or 300 Gy, as in the case of the genus Amaranthus (for example, A. palmeri).

[0137] According to a specific modality, the cultivation of the pollen-producing weed that reduces fitness is carried out on a large scale (e.g., hundreds to thousands of m2, 0.01 to 100 Acres).

[0138] According to some embodiments of the invention, the weed-producing pollen comprises only male plants.

[0139] Pollen collection is well known in the art. For example, by the use of paper bags. Another example is presented in document US 20060053686, which is incorporated into the present invention by reference in its entirety.

[0140] Once the pollen is obtained, it can be stored for future use. Examples of storage conditions include, but are not limited to, storage temperatures in degrees Celsius, for example, -196, -160, -130, -80, -20, -5, 0, 4, 20, 25, 30 or 35; percentage of relative humidity, for example, 0, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100. Humidity control can be achieved using a dehydrating agent or a humidity chamber, as known in the art. Additionally, the pollen can be stored in light or dark.

[0141] Alternatively, the pollen product of the present teachings is subjected to post-harvest treatment.

[0142] Thus, according to one aspect of the invention, a method for producing pollen for use in artificial pollination is provided, the method comprising: Petition 870250094224, dated 10 / 15 / 2025, page 33 / 57 28 / 44 (a) to obtain pollen that reduces the fitness of at least one weed species of interest, for example, as described in the present invention; and (b) to treat the pollen for use in artificial pollination.

[0143] Consequently, a composition of matter comprising weed pollen is provided which reduces the fitness of at least one weed species of interest, the pollen having been treated to improve its use in artificial pollination.

[0144] Examples of such treatments include, but are not limited to, coating, preparation, formulation, chemical inducers, physical inducers [for example, potential inducers include, but are not limited to, ethanol, hormones, steroids (e.g., dexamethasone, glucocorticoid, estrogen, estradiol), salicylic acid, pesticides and metals such as copper, antibiotics such as, but are not limited to, tetracycline, ecdysone, ACE inhibitors, benzothiadiazole and safener, tebufenozide or methoxyfenozide], solvent solubilization, drying, heating, cooling and irradiation (e.g., gamma, UV, X-rays, particles).

[0145] Additional ingredients and additives may be advantageously added to the pollen composition of the present invention, and may additionally contain sugar, potassium, calcium, boron, and nitrates. These additives may promote pollen tube growth after pollen distribution on flowering plants. Petition 870250094224, dated 10 / 15 / 2025, page 34 / 57 29 / 44

[0146] In some embodiments, the pollen composition of the present invention contains dehydrated or partially dehydrated pollen.

[0147] Thus, the pollen composition may comprise a surfactant, a stabilizer, a buffer, a preservative, an antioxidant, a diluent, a solvent, an emulsifier, an inverted emulsifier, a spreader, an adhesive, a penetrant, a foaming agent, an antifoaming agent, a thickener, a protective agent, a compatibility agent, a vegetable oil concentrate, a viscosity regulator, a binder, an adhesion agent, a drift control agent, a fertilizer, a controlled-release coating, a water-resistant coating, an antibiotic, a fungicide, a nematicide, a herbicide or a pesticide.

[0148] As used in the present invention, the term “about” refers to ± 10%.

[0149] The terms “compreende”, “compreendendo”, “inclui”, “incluindo”, “tendo” and their conjugates mean “including, but not limited to”.

[0150] The term “consisting of” means “including and limited to”.

[0151] The term “essentially consisting of” means that the composition, method or structure may include additional ingredients, steps and / or parts, but only if the additional ingredients, steps and / or parts do not materially alter the basic and innovative characteristics of the claimed composition, method or structure.

[0152] As used in the present invention, the singular forms “a”, “an” and “the” include references Petition 870250094224, dated 10 / 15 / 2025, page 35 / 57 30 / 44 in the plural, unless the context clearly indicates otherwise. For example, the term "a compound" or "at least one compound" can include a plurality of compounds, including mixtures thereof.

[0153] Throughout this application, various embodiments of this invention may be presented in range format. It should be understood that the range format description is merely for convenience and brevity and should not be interpreted as an inflexible limitation to the scope of the invention. Consequently, a range description should be considered as having specifically disclosed all possible subranges as well as individual numerical values ​​within that range. For example, a range description such as 1 to 6 should be considered as having specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the range width.

[0154] Whenever a numerical range is indicated in the present invention, it shall include any cited numeral (fractional or integer) within the indicated range. The expressions “ranging between” a first indicative number and a second indicative number and “ranging from” a first indicative number “to” a second indicative number are used interchangeably in the present invention and shall include the first and second indicated numbers and all fractional and integer numerals between them.

[0155] As used in the present invention, the term “method” refers to ways, means, techniques and Petition 870250094224, dated 10 / 15 / 2025, page 36 / 57 31 / 44 procedures for performing a given task, including, but not limited to, those ways, means, techniques and procedures known or easily developed from ways, means, techniques and procedures known to professionals in the chemical, pharmacological, biological, biochemical and medical fields.

[0156] It is recognized that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, several features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as appropriate in any other described embodiment of the invention. Certain features described in the context of several embodiments should not be considered essential features of those embodiments unless the embodiment is inoperable without those elements.

[0157] Several embodiments and aspects of the present invention, as outlined above and as claimed in the claims section below, find experimental support in the following examples. EXAMPLES

[0158] Reference is now made to the following examples, which, together with the descriptions above, illustrate some embodiments of the invention in a non-limiting manner. EXAMPLE 1 Genus Amaranthus: A. tuberculatus female X pollen of A. palmeri irradiated Petition 870250094224, dated 10 / 15 / 2025, p. 37 / 57 32 / 44 Experiment 98 Materials and Methods Plant Cultivation

[0159] The experiment was conducted between May and June in an indoor grow room in Rehovot, Israel. A. tuberculatus seeds were sown in germination trays. Three weeks after seedling emergence, they were transplanted into 5 L pots with a potting soil mix. When the plants began to flower, they were monitored daily to identify their sex at an early stage. Immediately after sex identification, females and males were separated and placed in different locations: male plants were grown in a greenhouse with netting and female plants were grown in an indoor grow room (35°C / 25°C conditions, 16 / 8 day / night photoperiod), where the pollination experiment was conducted. After maturation, 6 female A. tuberculatus plants were selected for the experiment. A. palmeri seeds were sown and cultivated using the same protocol.Immediately after sex identification, the males were separated and placed in a greenhouse with a net for pollen production. Pollen Collection and Irradiation

[0160] As soon as the male plants of A. tuberculatus and A. palmeri began to flower, the pollen was collected separately with a vacuum cleaner. The A. palmeri pollen was placed in 15 cm Petri dishes. The A. palmeri Petri dish was X-ray irradiated with a dose of 300 Gy. The A. tuberculatus pollen was not irradiated. Petition 870250094224, dated 10 / 15 / 2025, page 38 / 57 33 / 44 Pollination

[0161] To test the pollination potential of irradiated A. palmeri pollen on A. tuberculatus, each of the six female A. tuberculatus plants was artificially pollinated with irradiated A. palmeri pollen, while A. tuberculatus pollen served as a positive control. On each plant, a paper tube containing 10 mg of irradiated A. palmeri pollen (later referred to as the “Cross with Palmer” treatment) and another paper tube containing 10 mg of A. tuberculatus pollen (later referred to as the “Control”) were carefully placed on an inflorescence, leaving it to rest for 20 minutes before removal.Furthermore, two empty paper tubes, without pollen inside, were placed in an additional ear of corn before pollination (“blank,” to assess the level of natural pollination caused by airborne pollen) and in another ear at the end of the pollination process (“blank end,” pollination to assess the level of pollination unintentionally caused in non-target ears during artificial pollination), in order to evaluate the level of pollen contamination. Sixteen days after pollination, all treated ears were cut and dried. The seeds were harvested manually. The total seed count per ear was measured, the average weight of a single seed was calculated, and the seed morphology was examined. Results Seed Set

[0162] As can be seen in Table 2 below, pollination of female ears of A. tuberculatus with irradiated A. palmeri pollen produced at least the same Petition 870250094224, dated 10 / 15 / 2025, page 39 / 57 34 / 44 quantity of seeds that pollinate with pollen from A. tuberculatus and even more.

[0163] To evaluate the quality of the seeds obtained, the average weight of a seed obtained from the cross of A. tuberculatus x irradiated A. palmeri (“Cross with Palmer”) was calculated and compared to the average weight of a seed obtained from the cross of A. tuberculatus x A. tuberculatus (“Control”). The results showed that the average weight of a seed obtained from the cross with irradiated Palmer is almost 6 times lower than the average weight of the control seed, indicating that the seeds obtained from the cross with Palmeri treatment are probably aborted seeds, as expected (Table 3). Table 2: Average number of seeds

[0164] The values ​​refer to the average number of seeds from 6 ears of corn, each from a different female plant. Total number of seeds Blank tip Blank end Control Cross with Palmer Average 13.17 22.33 396.17 655.17 STE 6.50 10.07 110.46 151.85 Table 3: Average seed weight Average seed weight (mg) Control cross with Palmer Average 0.353 0.06 STE 0.025 0.005 P-value 1.97E-07 Germination Test

[0165] In general, the seeds obtained from the cross with pollen from A. palmeri and irradiated pollen were thin, partially empty and light brown in color, Petition 870250094224, dated 10 / 15 / 2025, pp. 40 / 57 35 / 44 while those obtained from the control pollen appeared fuller and dark brown / black in color. A germination assay was conducted to estimate the different germination levels between seeds obtained by artificial pollination with irradiated A. palmeri pollen versus those obtained by artificial pollination with regular A. tuberculatus pollen. The seeds were separated according to their visual appearance into two subgroups of each category. In the control, there were normal seeds versus seeds that appeared aborted, while in the cross-pollination with A. palmeri pollen, there were seeds suspected of being “normal” versus seeds that appeared aborted. Up to 50 seeds from each group (according to the quantity detected in each group) were taken from each of the samples, and each set of seeds was sown in small pots for the germination test.These pots were placed in a growth chamber under conditions of 34 / 25 °C, 16 / 8 hours day / night, for 14 days. After 14 days, the seedlings were counted and the germination rate was calculated for each sample. None of the seeds classified as aborted in either group germinated. Regarding the seeds classified as “normal”, while the average germination rate obtained with the control pollen was 44%, none of the seeds obtained with the irradiated Palmeri pollen germinated (Table 4). Table 4: Germination test Percentage of Germination Normal Control Cross with Palmer suspected of being “normal” Average 0.44 0 STE 0.12 0 Depth Value (compared to control) 0.0025 Petition 870250094224, dated 10 / 15 / 2025, page 41 / 57 36 / 44 (at the intersection) EXAMPLE 2 Interspecific pollen application: A. tuberculatus X A. palmeri with competition Experiment 720 Objectives of the Experiment

[0166] To test the pollination potential of irradiated A. palmeri pollen on A. tuberculatus when competing with non-irradiated A. tuberculatus pollen.

[0167] Compare the pollination potential of irradiated A. palmeri pollen on A. tuberculatus with that of irradiated A. tuberculatus pollen, either alone or in competition with non-irradiated A. tuberculatus pollen. Materials and Method Plant Cultivation

[0168] The experiment was conducted during the month of August in Rehovot, Israel. Seeds of A. tuberculatus were sown in germination trays and grown in indoor growing chambers. Three weeks after seedling emergence, they were transplanted into 5 L pots with a potting soil mix. When the plants began to flower, they were monitored daily to identify their sex at an early stage. Immediately after sexing, the females and males were separated and placed in different locations: the male plants were grown in a greenhouse with netting, and the female plants were grown outdoors, approximately 300 meters from the greenhouse with netting for the males, where the pollination experiment was conducted. After the Petition 870250094224, dated 10 / 15 / 2025, page 42 / 57 At 37 / 44 maturity, three female A. tuberculatus plants were selected for the experiment. Seeds of A. palmeri were sown and cultivated following the same protocol. Immediately after sexing, the males were separated and placed in a greenhouse with a separate net for pollen production. Pollen Collection and Irradiation

[0169] As soon as the male plants of A. tuberculatus and A. palmeri began to flower, pollen was collected separately for each species using a vacuum cleaner. The pollen from A. tuberculatus was divided into two batches originating from the same pollen mixture: one to be irradiated and the other to remain non-irradiated. Irradiation of the pollen from A. tuberculatus and A. Palmer was performed by placing the pollen in a 14 cm Petri dish (A. Palmer) or in a 1.5 mL Eppendorf tube (A. tuberculatus; because the quantity was less than that of the A. Palmer pollen) and irradiating by X-ray at a dose of 250 Gy. Pollination

[0170] Each of the three female A. tuberculatus plants was pollinated with the following pollen treatments. Two inflorescences ('spikes') were pollinated for each treatment on the same plant. 1. Non-irradiated A. tuberculatus 2. Irradiated palmer's arachnid 3. A. tuberculatus irradiated 4. Irradiated A. palmeri & non-irradiated A. tuberculatus (1:1 ratio) 5. Irradiated A. tuberculatus & Non-irradiated A. tuberculatus (1:1 ratio) Petition 870250094224, dated 10 / 15 / 2025, pp. 43 / 57 38 / 44

[0171] Pollinations were carried out by covering each ear with a 14 cm long paper tube containing 10 mg of pollen. Pollinations in which competition between irradiated and non-irradiated pollen was tested contained 10 mg of pollen from a premixed stock containing A. tuberculatus pollen and irradiated A. palmeri pollen or irradiated A. tuberculatus pollen in a 1:1 ratio. In addition, two empty paper tubes, without pollen inside, were placed in an additional ear before pollination (“blank”) and in another inflorescence at the end of the pollination process (“blank end”) to assess the level of pollen contamination.

[0172] Sixteen days after pollination, all treated ears were cut and dried. The seeds were harvested manually, the total seed count per ear was measured, and the average weight of a single seed was calculated. Results Seed Set

[0173] The total number of seeds (normal and defective) did not differ significantly between pollination treatments (ANOVA, p = 0.12; Table 1), indicating that the pollination potential of irradiated pollen, whether from A. palmeri or A. tuberculatus, does not decrease in relation to that of non-irradiated A. tuberculatus pollen. Seed Weight

[0174] The average seed weight differed significantly between treatments (ANOVA, p=0.001; Table 1), with irradiated A. palmeri pollen seeds weighing less (0.05 mg) and A. Petition 870250094224, dated 10 / 15 / 2025, pp. 44 / 57 39 / 44 non-irradiated tuberculatus pollen weighed more (0.19 mg). Since seed weight is an indicator of seed development, the low seed weight from irradiated pollen suggests that these seeds were aborted. The average seed weight from irradiated pollen competing with non-irradiated A. tuberculatus pollen was similar, regardless of whether the irradiated pollen was from A. palmeri or A. tuberculatus (average seed weight of 0.15 and 0.16 mg, respectively). These statistically significant results indicate that the ability of irradiated A. palmeri pollen to pollinate female A. tuberculatus flowers does not decrease compared to that of irradiated A. tuberculatus pollen when competing with non-irradiated pollen. Table 5: Total seed count (normal and aborted) and average seed weight, for inflorescences (“ears”) pollinated with non-irradiated pollen from A. tuberculatus, irradiated pollen from A. tuberculatus and A. palmeri, and mixed irradiated and non-irradiated pollen. Treatment Average total seed count* ± SEM Average individual seed weight (mg) ± SEM Blank 120 ± 33 0.08 ± 0.01 Blank end 80 ± 30 0.08 ± 0.02 Non-irradiated A. tuberculatus 1180±207 0.19 ± 0.01 Irradiated A. palmeri 617± 128 0.05 ± 0.00 Irradiated A. tuberculatus 935±236 0.09 ± 0.04 Irradiated A. palmeri & Non-irradiated A. tuberculatus (1:1 ratio) 1003 ±243 0.15 ± 0.03 Irradiated A. tuberculatus & Non-irradiated A. tuberculatus (1:1 ratio) 1206 ± 267 0.16 ± 0.01 Petition 870250094224, dated 10 / 15 / 2025, pp. 45 / 57 40 / 44 * The values ​​are calculated as an average for six ears of corn (2 ears for each of the three plants). EXAMPLE 3 Genus Amaranthus: Female A. retroflexus X pollen of irradiated A. palmeri

[0175] Produced as in Example 1 or Example 2 with only the pollen and target weed indicated. EXAMPLE 4 Amaranthaceae family: Female of B. scoparia X pollen of A. palmeri irradiated

[0176] Produced as in Example 1 or Example 2 with only the pollen and target weed indicated. EXAMPLE 5 Family Amaranthaceae: Chenopodi A. album female X pollen of A. palmeri irradiated Experiment 683 Objective of the Experiment

[0177] The objective of this experiment was to test the pollination potential of A. palmeri pollen irradiated on Chenopodium album, a monoecious and self-compatible plant of the Amaranthaceae family, but not belonging to the Amaranthus genus. Materials and Method Plant Cultivation

[0178] The experiment was conducted in June at the Rehovot farm, Israel. C. album seeds were sown in germination trays and grown in indoor growth chambers. Three weeks after seedling emergence, they were transplanted into 5 L pots with a Petition 870250094224, dated 10 / 15 / 2025, pp. 46 / 57 41 / 44 potting soil mix and grown in a greenhouse with netting until used in the experiment, one week later.

[0179] A. palmeri seeds were sown and cultivated using the same protocol. When the plants began to flower, they were monitored daily to identify their sex at an early stage. Immediately after sex identification, the males were separated and placed in a greenhouse with a separate net for pollen production. Pollen Collection and Irradiation

[0180] As soon as the male A. palmeri plants began to flower, the pollen was collected with a vacuum cleaner. Irradiation of A. palmeri pollen was performed by placing it in a 14 cm Petri dish and irradiating it with X-rays at a dose of 250 Gy. Pollination

[0181] Ten C. album plants were selected in total, of which five were artificially pollinated with irradiated A. palmeri pollen (treatment) and five were left untreated (control). Each of the five treated plants was artificially pollinated with irradiated A. palmeri pollen three times, four days apart. Artificial pollination was performed by “spraying” a mixture of irradiated pollen and talc powder (in a 2:1 ratio), using a Mini Duster (©Kiwi Pollen), directly onto all inflorescences. The five control C. album plants were not artificially pollinated but left to self-pollinate. The treatment and control plants were Petition 870250094224, dated 10 / 15 / 2025, pp. 47 / 57 42 / 44 placed in two separate areas of a single greenhouse-net, on the agricultural farm in Rehovot.

[0182] As a positive control for the quality of irradiated A. palmeri pollen, two inflorescences from a single female A. palmeri were artificially pollinated with irradiated pollen, as described above. Both inflorescences produced many aborted seeds (460 and 758), confirming the quality of the irradiated pollen. Seed harvesting and germination

[0183] The inflorescences of each plant were harvested 14 days after the last pollination. The seeds were harvested manually and separated according to appearance into “normal” seeds (dark brown and round) and “aborted” seeds (yellowish-brown and flat).

[0184] To evaluate the germination rate, “normal” seeds of each plant were sown in small pots of soil and kept in a growth chamber (16 hours of light at 35 °C and 8 hours of darkness at 25 °C). The seeds were kept refrigerated in water for one week before sowing to increase germination. Germination was monitored over a period of 5 months. Preliminary Results

[0185] The germination rate of seeds that appeared “normal” was significantly lower for plants artificially pollinated with irradiated pollen from A. palmeri compared with control plants: 22.7 ± 4.7% and 39.5 ± 7.8%, respectively; one-tailed t-test, p-value = 0.05. Petition 870250094224, dated 10 / 15 / 2025, pp. 48 / 57 43 / 44 EXAMPLE 6 Genus Lolium: L. multiflorum female X Irradiated L. rigidum pollen

[0186] Produced as in Example 1 or Example 2 with only the pollen and target weed indicated. EXAMPLE 7 Genus Lolium: L. perenne female X Irradiated L. rigidum pollen

[0187] Produced as in Example 1 or Example 2 with only the pollen and target weed indicated. EXAMPLE 8 Genus Alopecurus: Female A. japonicus X pollen of irradiated A. myosuroides

[0188] Produced as in Example 1 or Example 2 with only the pollen and target weed indicated. EXAMPLE 9 Poaceae family: Female L. rigidum X pollen of irradiated A. myosuroides

[0189] Produced as in Example 1 or Example 2 with only the pollen and target weed indicated. EXAMPLE 10 Genus Ambrosia: Female of A. trifida X Pollen radiated from A. artemisiifolia

[0190] Produced as in Example 1 or Example 2 with only the pollen and target weed indicated.

[0191] Although the invention has been described together with its specific embodiments, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Consequently, it is intended to cover all of these. Petition 870250094224, dated 10 / 15 / 2025, pp. 49 / 57 44 / 44 alternatives, modifications and variations that fit within the spirit and broad scope of the appended claims.

[0192] It is the Applicant(s)' intention that all publications, patents and patent applications mentioned in this descriptive report be incorporated in their entirety by reference to the descriptive report, as if each individual publication, patent or patent application were specifically and individually mentioned, when referenced, to be incorporated into the present invention by reference. Furthermore, the citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent that section headings are used, they shall not be construed as necessarily limiting. In addition, any prior document(s) of this application are incorporated into the present invention by reference in their entirety. Petition 870250094224, dated 10 / 15 / 2025, pp. 50 / 57

Claims

1 / 3 CLAIMS 1. Weed control method, the method characterized in that it comprises artificially pollinating a weed species of interest with irradiated pollen that reduces the fitness of said weed species of interest, said pollen being of a species different from that of said weed species of interest and capable of competing with the pollen of said weed species of interest, wherein said artificial pollination is carried out under conditions of competition with native pollen.

2. Method according to claim 1, characterized in that said pollen consists of pollen from a species different from that of said weed species of interest.

3. Method, according to claim 1 or 2, characterized in that said artificial pollination is carried out on a large scale of at least 0.1 acre.

4. A method, according to any one of claims 1 to 3, characterized in that said pollen is contained in a formulation that allows for the distribution of pollen by artificial pollination.

5. A method, according to any one of claims 1 to 4, characterized in that said pollen is irradiated by γ radiation.

6. Method, according to any one of claims 1 to 4, characterized in that said pollen is irradiated by X-ray radiation. Petition 870250094224, dated 10 / 15 / 2025, p. 51 / 57 2 / 3 7. A method, according to any one of claims 1 to 4, characterized in that said pollen is irradiated by alpha radiation.

8. A method according to any one of claims 1 to 7, characterized in that said pollen is of the same genus as said weed species of interest.

9. A method according to any one of claims 1 to 8, characterized in that at least one of said pollen and said weed species of interest is a hybrid.

10. Method, according to any one of claims 1 to 9, characterized in that said pollen is not genetically modified pollen.

11. Method, according to any one of claims 1 to 9, characterized in that said pollen is genetically modified pollen.

12. Method, according to any one of claims 1 to 11, characterized in that said pollen is from the Amaranthaceae family.

13. Method, according to any one of claims 1 to 12, characterized in that the pollen said is of the genus Amaranthus.

14. Method according to claim 12 or 13, characterized in that said pollen and said weed species of interest are selected from the group consisting of A. palmeri and A. tuberculatus. Petition 870250094224, dated 10 / 15 / 2025, pp. 52 / 57 3 / 3 15. Method, according to any one of claims 1 to 13, characterized in that the pollen said is A. palmeri.

16. Method according to claim 13, characterized in that said pollen is a hybrid of A. palmeri and A. tuberculatus.

17. Method according to claim 13, characterized in that the said weed species of interest is a hybrid of A. palmeri and A. tuberculatus.

18. Method, according to any one of claims 1 to 11, characterized in that said pollen and said weed species of interest are selected from the group of pairs shown in Table 1.

19. Method, according to any one of claims 1 to 18, characterized in that said ability to compete is given by fertilization of said weed species of interest. Petition 870250094224, dated 10 / 15 / 2025, pp. 53 / 57