Protein hydrolysate and its use

Abstract

The present invention relates to a protein hydrolysate, wherein at least 90 wt.% of the peptides in the protein hydrolysate have a molecular mass of at most 500 Dalton, and wherein between 35 and 55 wt.% of the peptides have a molecular mass between 204 and 300 Dalton.

Description

[0001] PROTEIN HYDROLYSATE AND ITS USE

[0002] TECHNICAL FIELD

[0003] The invention relates to a protein hydrolysate and use thereof.

[0004] PRIOR ART

[0005] Traditional seed treatments often have limitations regarding their effectiveness and sustainability. Many of these treatments use synthetic or non-biodegradable materials that do not align with the growing need for environmentally friendly agricultural practices. Furthermore, the interactions between plants and symbiotic nitrogen-fixing bacteria are often suboptimal, leading to reduced efficiency in nitrogen fixation and ultimately limiting plant growth. These challenges highlight the need for innovative solutions that not only promote plant growth but also support natural symbiotic processes and reduce dependence on chemical fertilizers.

[0006] SUMMARY OF THE INVENTION

[0007] In a first aspect, the invention relates to a protein hydrolysate according to claim 1. The protein hydrolysate has a beneficial influence on the growth rate of the plant and promotes the formation of root nodules in symbiosis with nitrogen-fixing bacteria, leading to improved nitrogen fixation and nutrient uptake. The protein hydrolysate is biodegradable and offers a sustainable solution for the agricultural sector, with potential for cost savings and a reduced dependence on chemical fertilizers through the promotion of natural symbiotic processes. Furthermore, the specific molecular mass distribution of the peptides leads to faster germination and more uniform growth of treated seeds, while the use of small amounts of protein hydrolysate per seed increases cost efficiency.

[0008] In a second aspect, the invention relates to a composition comprising the protein hydrolysate and an inoculum of bacteria. In a third aspect, the invention relates to the use of the protein hydrolysate.

[0009] DETAILED DESCRIPTION

[0010] The invention relates to a protein hydrolysate, as well as to a method for optimizing seeds.Unless defined otherwise, all terms used in the description of the invention, including technical and scientific terms, have the meaning as commonly understood by a person skilled in the technical field of the invention. For a better assessment of the description of the invention, the following terms are explicitly explained.

[0011] The term "wt.%" is synonymous with "m%" and refers to weight percent, a measure of the concentration of a particular ingredient in a mixture, expressed as a percentage of the total weight of the composition.

[0012] As used in this document, the articles "a", "an" and "the" refer to both the singular and the plural, unless the context clearly dictates otherwise. For example, "a segment" means one or more than one segment.

[0013] The terms "comprise", "comprising", "consist of", "consisting of", "provided with", "contain", "containing", "encompass", "encompassing", "include" and "including" are used herein as synonyms and are intended as inclusive or open terms indicating the presence of what follows, without excluding or preventing the presence of other components, characteristics, elements, members or steps known from or disclosed in the prior art.

[0014] The citation of numerical ranges by their end points includes all integers, fractions, and / or real numbers between the end points, including the end points themselves.

[0015] The term "protein hydrolysate" refers to a product obtained by breaking down proteinrich material via hydrolysis, wherein larger molecules are converted into smaller ones.

[0016] In the present invention, the term "animal protein hydrolysate" refers to a protein hydrolysate derived from animal proteins, such as those originating from poultry, cattle, sheep, pigs, pets, birds, fur-bearing animals, fish, insects, crustaceans and shellfish, or animal species such as deer or camels.

[0017] The term "thermal hydrolysis" refers to a method of breaking down chemical compounds using heat.

[0018] The term "enzymatic hydrolysis" refers to a process in which enzymes are used to break down chemical compounds, resulting in the formation of a hydrolysate.In the present invention, the term "peptides" refers to the short chains of amino acids that are formed by the cleavage of proteins during the hydrolysis process.

[0019] The term "molecular mass" in the present invention refers to the mass of a molecule, expressed in Dalton (Da). The molecular mass of peptides is measured using techniques such as gel permeation chromatography (GPC) or mass spectrometry, preferably according to a standardized method such as ISO 10927:2018 for the determination of the molecular mass of polymers.

[0020] In the present invention, the term "wt.%" or "weight percent" indicates the weight percentage of a certain component relative to the total weight of the peptides in the protein hydrolysate. This percentage is calculated by dividing the mass of the respective peptides by the total mass of all peptides and subsequently multiplying by 100.

[0021] In the present invention, the term "inoculum" refers to a material containing bacteria that is used to inoculate a substrate (such as, for example, agricultural soil). This inoculum may comprise nitrogen-fixing bacteria that live in symbiosis with plant roots and promote nitrogen fixation.

[0022] In the present invention, the term "treating seeds" refers to bringing a protein hydrolysate into contact with the seed surface, for example by means of a coating, or to simultaneously bringing seeds and protein hydrolysate into contact during the sowing process.

[0023] In a first aspect, the invention relates to a protein hydrolysate. The protein hydrolysate is suitable for agricultural applications, and preferably a protein hydrolysate for optimizing seeds. By optimizing seeds is meant the simultaneous, sequential or separate application of the seeds in combination with the protein hydrolysate to the soil.

[0024] In the context of the present invention, optimizing seeds refers to performing one or more operations on or to the seeds, prior to or during sowing, with the aim of favorably influencing the early development of the plant. Such operations may inter alia be aimed at improving germination, initial root and shoot development, early growth rate and / or the interaction of the young plant with soil microorganisms.

[0025] In a preferred embodiment, the protein hydrolysate comprises peptides with a lower molecular mass. In an embodiment, the protein hydrolysate comprises at least 90 wt.% peptides with a molecular mass of at most 500 Dalton. The inventors found that thislow molecular mass fraction provides for improved uptake of nutrients by the plants, in particular nitrogen, which contributes to accelerated and improved plant growth.

[0026] The low molecular mass fraction of the peptides in the protein hydrolysate ensures the effectiveness of the seed optimization. This distribution optimizes the symbiotic interactions of the plant with beneficial microorganisms, such as symbiotic nitrogenfixing bacteria, which are important for nitrogen fixation in some plants, such as legumes. By improving these interactions, the resilience of plants against diseases and pests is strengthened, resulting in healthier and more robust plants.

[0027] Besides the increased nutrient uptake and improved symbiotic interactions, the treatment offers a biological and environmentally friendly solution that aligns with sustainable agricultural practices. This can lead to a reduced dependence on chemical fertilizers, which both reduces the ecological footprint of agricultural practices and lowers economic costs for farmers.

[0028] The described hydrolysate is effective not only in increasing plant growth and the formation of root nodules, but also in promoting the overall health of the plant. This is achieved through a synergistic interaction between the Rhizobium bacteria and the biostimulant, resulting in improved nitrogen fixation and overall growth conditions. These improved growth conditions contribute to a higher yield and quality of the crops, which is of great importance to the agricultural sector.

[0029] In an embodiment, at least 80 wt.% of the peptides in the protein hydrolysate have a molecular mass of at most 500 Dalton. Preferably, the protein hydrolysate comprises at least 85 wt.% peptides, more preferably at least 90 wt.% peptides, even more preferably at least 95 wt.%, and even more preferably at least 98 wt.%, with a molecular mass of at most 500 Dalton.

[0030] In another or further embodiment, between 80 and 99 wt.% of the peptides have a molecular mass of at most 500 Dalton. Preferably, the protein hydrolysate comprises between 85 and 99 wt.% peptides, more preferably between 90 and 99 wt.% peptides, even more preferably between 95 and 99 wt.%, with a molecular mass of at most 500 Dalton.

[0031] In an embodiment, at most 10 wt.% of the peptides have a molecular mass of at least 500 Dalton. Preferably, the protein hydrolysate comprises at most 9 wt.%, more preferably at most 8 wt.%, even more preferably at most 7 wt.%, even more preferablyat most 6 wt.%, even more preferably at most 5 wt.% peptides with a molecular mass of at least 500 Dalton.

[0032] In another or further embodiment, between 0.1 and 10 wt.% of the peptides have a molecular mass between 500 and 1,000 Dalton. Preferably, the protein hydrolysate comprises between 1 and 9 wt.%, more preferably between 1 and 8 wt.%, even more preferably between 1 and 7 wt.%, even more preferably between 1 and 6 wt.%, even more preferably between 1 and 5 wt.% peptides with a molecular mass between 500 and 1,000 Dalton. These higher molecular mass peptides can play a supporting role in maintaining the bioactivity of the treatment, whereby the plant remains capable of efficiently taking up nutrients for a longer period of time.

[0033] In an embodiment, between 30 and 60 wt.% of the peptides have a molecular mass between 204 and 300 Dalton, preferably between 35 and 55 wt.%, or even between 40 and 50 wt.%.

[0034] In a preferred embodiment, at least 30 wt.% of the peptides in the protein hydrolysate have a molecular mass between 204 and 300 Dalton, preferably at least 35 wt.% of the peptides, more preferably at least 40 wt.% of the peptides, more preferably at least 45 wt.% of the peptides, more preferably at least 50 wt.% of the peptides. In another or further preferred embodiment, at most 99 wt.% of the peptides in the protein hydrolysate have a molecular mass between 204 and 300 Dalton, preferably at most 95 wt.% of the peptides, more preferably at most 90 wt.% of the peptides, more preferably at most 85 wt.% of the peptides, more preferably at most 80 wt.% of the peptides, more preferably at most 75 wt.% of the peptides, more preferably at most 70 wt.% of the peptides.

[0035] In a preferred embodiment, at least 90 wt.% of the peptides in the protein hydrolysate have a molecular mass of at most 500 Dalton, and between 35 and 55 wt.% of the peptides have a molecular mass between 204 and 300 Dalton.

[0036] In an embodiment, at least 70 wt.% of the peptides have a molecular mass of at most 400 Dalton. Preferably, the protein hydrolysate comprises at least 75 wt.% peptides, more preferably at least 80 wt.% peptides, even more preferably at least 85 wt.%, and even more preferably at least 90 wt.%, with a molecular mass of at most 400 Dalton.

[0037] In another or further embodiment, between 70 and 99 wt.% of the peptides have a molecular mass of at most 400 Dalton. Preferably, the protein hydrolysate comprisesbetween 75 and 99 wt.% peptides, more preferably between 80 and 99 wt.% peptides, even more preferably between 80 and 95 wt.%, with a molecular mass of at most 400 Dalton.

[0038] In another or further embodiment, at least 85 wt.% of the peptides have a molecular mass of at most 500 Dalton, and at least 80 wt.% of the peptides have a molecular mass of at most 400 Dalton. In another or further embodiment, at least 90 wt.% of the peptides have a molecular mass of at most 500 Dalton, and at least 80 wt.% of the peptides have a molecular mass of at most 400 Dalton.

[0039] In an embodiment, between 5 and 50 wt.% of the peptides have a molecular mass of at most 204 Dalton, preferably between 10 and 40 wt.%, even more preferably between 10 and 30 wt.%, even more preferably between 10 and 25 wt.%, even more preferably between 15 and 25 wt.%.

[0040] In an embodiment, between 5 and 40 wt.% of the peptides have a molecular mass between 300 and 400 Dalton, preferably between 10 and 30 wt.%, even more preferably between 10 and 25 wt.%, even more preferably between 15 and 25 wt.%.

[0041] In an embodiment, between 1 and 15 wt.% of the peptides have a molecular mass between 400 and 500 Dalton, preferably between 1 and 10 wt.%.

[0042] In a preferred embodiment, at least 90 wt.% of the peptides in the protein hydrolysate have a molecular mass of at most 500 Dalton,

[0043] wherein between 10 and 30 wt.% of the peptides have a molecular mass of at most 204 Dalton, preferably between 15 and 25 wt.%;

[0044] between 35 and 55 wt.% of the peptides have a molecular mass situated between 204 and 300 Dalton, preferably between 40 and 50 wt.%; between 10 and 30 wt.% of the peptides have a molecular mass situated between 300 and 400 Dalton, preferably between 15 and 25 wt.%; and between 1 and 15 wt.% of the peptides have a molecular mass situated between 400 and 500 Dalton, preferably between 1 and 10 wt.%.

[0045] This specific molecular mass distribution of peptides contributes to an improved symbiotic interaction with nitrogen-fixing bacteria, which can lead to an increased formation of root nodules. This effect can further improve the efficiency of nitrogen fixation, which optimizes the overall growth conditions of the plant. Such animprovement in symbiosis can result in a reduced dependence on chemical fertilizers, which contributes to more sustainable agricultural practices.

[0046] The protein hydrolysate of the present invention may be of animal, microbial or plant origin. Preferably, the protein hydrolysate is an animal protein hydrolysate. This origin can contribute to the biodegradability and environmental compatibility of the protein hydrolysate. These advantages make the hydrolysate particularly suitable for sustainable agricultural practices.

[0047] Examples of animal protein hydrolysates are protein hydrolysates originating from poultry, cattle, sheep, pigs, pets, birds, fur-bearing animals, fish, insects, crustaceans and shellfish, or animal species such as deer or camels.

[0048] The protein hydrolysate may also originate from animal slaughterhouse waste, such as slaughterhouse waste from poultry, pigs, cattle, fish, sheep, blood, viscera, hides, feathers, hooves, heads, or other waste material that is not suitable for human consumption. In a preferred embodiment, the protein hydrolysate originates from animal slaughterhouse waste, such as blood, hides, viscera, feathers, feet, and bones, preferably blood.

[0049] In an embodiment, the protein hydrolysate originates from animal blood.

[0050] In an embodiment, the protein hydrolysate originates from poultry slaughterhouse waste, such as blood, skins, viscera, feathers, feet, and / or bones of chickens, ducks, or turkeys.

[0051] In an embodiment, the protein hydrolysate originates from poultry blood.

[0052] In another aspect, the invention relates to a method for producing a protein hydrolysate as described herein. The protein hydrolysate can be obtained by means of enzymatic, thermal or chemical hydrolysis.

[0053] The term "thermal hydrolysis" refers to a method of breaking down chemical compounds using heat.

[0054] The term "enzymatic hydrolysis" refers to a process in which enzymes are used to break down chemical compounds, resulting in the formation of a hydrolysate.In an embodiment, the method comprises the steps of:

[0055] a. providing a protein-rich material and water;

[0056] b. thermally hydrolyzing the protein-rich material, whereby a protein hydrolysate is obtained.

[0057] In an embodiment, the obtained protein hydrolysate is treated with peroxides. This can deodorize the protein hydrolysate.

[0058] The protein-rich material can be, for example, slaughterhouse waste. This waste can consist of feathers, organs, bones, blood, hooves, and other parts of the animal that are not intended for human consumption.

[0059] In a further embodiment, the protein hydrolysate of protein-rich material can be further treated to increase the concentration of certain beneficial components, or to remove unwanted components. For example, the protein hydrolysate can be filtered to remove solid particles, or it can be centrifuged to separate heavier components. It can also be treated with enzymes to further promote the breakdown of proteins into peptides and amino acids, or it can be treated with acids or bases to adjust the pH.

[0060] In a further embodiment, enzymatic hydrolysis or chemical hydrolysis, preferably enzymatic hydrolysis, is also performed on the protein-rich material.

[0061] The enzymatic hydrolysis of protein-rich material can be performed using various types of enzymes, such as pepsin, trypsin, pancreatin, keratinase, and papain, wherein the choice of the enzyme may depend on the specific type of protein-rich material that is used. This combination of enzymatic and thermal hydrolysis can result in a higher yield of low molecular weight peptides.

[0062] In a second aspect, the invention relates to a composition comprising a protein hydrolysate as described above, and an inoculum comprising bacteria.

[0063] The protein hydrolysate and the inoculum can thereby be provided as separate formulations or as one combined composition, and can be applied simultaneously or sequentially.

[0064] The bacteria are preferably nitrogen-fixing bacteria, and even more preferably symbiotic or associative nitrogen-fixing bacteria.The symbiotic nitrogen-fixing bacteria can be selected from: Rhizobium spp., Bradyrhizobium spp., Sinorhizobium spp., Mesorhizobium spp., Azorhizobium spp. Frankia spp. (actinorhiza), Allorhizobium spp., Neorhizobium spp., preferably chosen from Rhizobium leguminosarum, Rhizobium phaseoli, Rhizobium trifolii, Rhizobium etli, Rhizobium galegae, Bradyrhizobium japonicum, Bradyrhizobium elkanii, Bradyrhizobium liaoningense, Bradyrhizobium yuanmingense, Sinorhizobium meliloti, Sinorhizobium fredii, Sinorhizobium saheli, Mesorhizobium loti, Mesorhizobium ciceri, Mesorhizobium mediterraneum, Azorhizobium caulinodans, Azorhizobium doebereinerae, Frankia alni, Frankia casuarinae, Frankia discariae, Frankia coriariae, Allorhizobium undicola, Neorhizobium galegae.

[0065] The associative nitrogen-fixing bacteria can be selected from: Azospirillum spp., Herbaspirillum spp., Gluconacetobacter spp., Azoarcus spp., Burkholderia spp., Enterobacter spp., Klebsiella spp., Pseudomonas spp., preferably selected from

[0066] Azospirillum brasilense, Azospirillum lipoferum, Azospirillum amazonense, Azospirillum halopraeferens, Herbaspirillum seropedicae, Herbaspirillum frisingense, Herbaspirillum rubrisubalbicans, Gluconacetobacter diazotrophicus, Gluconacetobacter johannae, Azoarcus indigens, Azoarcus communis, Azoarcus olearius, Burkholderia vietnamiensis, Burkholderia kururiensis, Burkholderia tropica, Enterobacter cloacae, Enterobacter asburiae, Klebsiella pneumoniae, Klebsiella oxytoca, Pseudomonas stutzeri, Pseudomonas fluorescens.

[0067] In a preferred embodiment, the composition comprises an inoculum comprising symbiotic nitrogen-fixing bacteria. Virtually all symbiotic nitrogen-fixing bacteria work with plants that form nitrogen root nodules (nodules). The root nodules are formed under the influence of the nitrogen-fixing bacteria from, among others, the genera Rhizobium spp., Bradyrhizobium spp., Sinorhizobium spp., Mesorhizobium spp., Azorhizobium spp. Frankia spp. (actinorhiza), Allorhizobium spp., and Neorhizobium spp., which live in these nodules in mutualistic symbiosis with the plant, the bacteria receive sugar from the plant.

[0068] With the aid of nitrogenase, they fix nitrogen (N2) from the air into ammonia (NH3) for these plants. The formed ammonia is further converted by other types of free-living soil bacteria, via so-called nitrification, into the nitrogen compound nitrate, which the plant, dissolved in soil moisture, can absorb via its roots. The nitrogen-fixing bacteria derive their energy from glucose that the plant assimilates during photosynthesis.In an embodiment, the composition comprises an inoculum comprising symbiotic nitrogen-fixing bacteria selected from Rhizobium spp., Bradyrhizobium spp., Sinorhizobium spp., Mesorhizobium spp., Azorhizobium spp. Frankia spp. (actinorhiza), Allorhizobium spp., Neorhizobium spp., preferably selected from Rhizobium spp., Bradyrhizobium spp., or a combination thereof, and most preferably Bradyrhizobium spp.

[0069] In another or further embodiment, the composition comprises an inoculum comprising symbiotic nitrogen-fixing bacteria selected from Rhizobium leguminosarum, Rhizobium phaseoli, Rhizobium trifolii, Rhizobium etli, Rhizobium galegae, Bradyrhizobium japonicum, Bradyrhizobium elkanii, Bradyrhizobium liaoningense, Bradyrhizobium yuanmingense, or a combination thereof.

[0070] In another or further embodiment, the composition comprises an inoculum comprising symbiotic nitrogen-fixing bacteria selected from Bradyrhizobium japonicum, Bradyrhizobium elkanii, Bradyrhizobium liaoningense, Bradyrhizobium yuanmingense, or a combination thereof.

[0071] In a third aspect, the protein hydrolysate or the composition as described above is used in agricultural applications. Agricultural applications include, among others, a seed treatment, seed coating, soil conditioning, and application in liquid fertilizers.

[0072] When sowing plants, farmers can treat the seeds with a protein hydrolysate or a composition comprising the protein hydrolysate. The use of this protein hydrolysate or this composition ensures that the seeds get an optimal start, which results in stronger root growth and better resistance to stress in the early growth phase. It has been found that the protein hydrolysate stimulates the early formation of nitrogen root nodules, which is beneficial for effective nitrogen fixation during the growing season, and which further results in increased above-ground plant mass. Seed coating is a specific method of seed treatment wherein a visible, often thicker layer is applied around the seed. In a preferred embodiment, the protein hydrolysate or the composition is a protein hydrolysate or a composition as described above.

[0073] A protein hydrolysate or a composition comprising the protein hydrolysate can be combined with liquid fertilizers and applied around the seeds directly during sowing. In this way, the protein hydrolysate or the composition provides a combination of directly available nutrients such as nitrogen, phosphate, and potassium, together with other benefits for the young plants. This approach ensures that the seedlings have sufficientnutrients and protection from the start, thereby maximizing growth chances, especially in soils where nutrient availability is limited.

[0074] A protein hydrolysate or a composition comprising the protein hydrolysate can also be used for improving soil quality prior to sowing. By adding the protein hydrolysate or the composition to the soil, the subsequently sown seeds get an optimal start, resulting in stronger root growth and better resistance to stress in the early growth phase. It has been found that the protein hydrolysate stimulates the early formation of nitrogen root nodules, which is beneficial for effective nitrogen fixation during the growing season, and which further results in increased above-ground plant mass.

[0075] In an embodiment, the protein hydrolysate or the composition is used in a seed treatment. The seed treatment can comprise a seed coating, an immersion of the seeds, a mist treatment, a slurry treatment, or a vacuum filtration.

[0076] In an embodiment, the protein hydrolysate or the composition is used in a seed coating. When coating seeds, the protein hydrolysate or a solution thereof can be used with a binder to be applied to the seed. The hydrolysate can be added to the seeds in a drum mixer. While the seeds rotate, a thin layer of liquid is applied evenly, optionally in combination with powders such as clay or minerals, which adhere to the liquid.

[0077] In an embodiment, the protein hydrolysate is used in an immersion. In immersion, the seeds are immersed in the protein hydrolysate for a certain period. The seeds are dried after immersion before they are sown.

[0078] In an embodiment, the protein hydrolysate is used in a mist treatment. In a mist treatment, seeds are treated by misting them with the protein hydrolysate or a solution thereof. The seeds are placed in a rotating drum or a special machine, while a fine mist of the liquid is sprayed over the seeds. This ensures an even treatment without the seeds becoming too wet.

[0079] In an embodiment, the protein hydrolysate is used in a slurry treatment. In a slurry treatment, the seeds are dipped in or passed through a thick liquid suspension (slurry) containing the protein hydrolysate or a solution thereof. This method offers good adhesion and ensures that the seed is well covered. After application of the slurry, the seeds can be dried to ensure that they are easier to handle and do not clump together.In an embodiment, the protein hydrolysate is used in a vacuum filtration. In vacuum infiltration, the seeds are placed in the protein hydrolysate or a solution thereof, and subsequently the air is removed from the seeds using a vacuum pump. This allows the protein hydrolysate or a solution thereof to penetrate deep into the seed tissues, which is especially useful for seeds with a hard coat. After the vacuum, the pressure is normalized again, whereby the protein hydrolysate or a solution thereof is effectively drawn into the seed.

[0080] In an embodiment, the protein hydrolysate or the composition is used separately, sequentially, or simultaneously with the sowing of seeds. Herein, the protein hydrolysate or the composition is applied to the soil prior to the sowing of the seeds, simultaneously with the sowing of the seeds, or after the sowing of the seeds.

[0081] In an embodiment, the protein hydrolysate can be added to a fertilizer or a soil conditioner and applied to the soil prior to the sowing of the seeds. Herein, the hydrolysate can be applied to the soil up to a maximum of 30 days before the sowing of the seeds, preferably at most 14 days.

[0082] In another or further embodiment, the protein hydrolysate is used separately, sequentially, or simultaneously with the sowing of seeds, wherein an inoculum comprising bacteria is also applied to or into the soil separately, sequentially, or simultaneously with the sowing of seeds. Herein, the protein hydrolysate is applied to the soil prior to the sowing of the seeds, simultaneously with the sowing of the seeds, or after the sowing of the seeds, and an inoculum comprising bacteria can also be applied to the soil prior to the sowing of the seeds, simultaneously with the sowing of the seeds, or after the sowing of the seeds. The inoculum can contain bacteria as in the embodiments described above.

[0083] In a preferred embodiment, the composition comprising the protein hydrolysate and the inoculum is used as a seed coating, wherein the protein hydrolysate is applied in an amount of at most 0.5 pL / seed, preferably at most 0.3 pL / seed, preferably even of at most 0.2 pL / seed. Preferably, the protein hydrolysate has a dry matter content between 10 and 20 wt.%, preferably between 12 and 18 wt.%.

[0084] In an embodiment, the protein hydrolysate or the composition is used for seeds of plants or crops that can have a symbiotic relationship with nitrogen-fixing bacteria. In a further embodiment, the protein hydrolysate or the composition is used for seeds of plants or crops selected from Actinorhizal plants, and legumes.Actinorhizal plants are a group of angiosperms that are characterized by their ability to enter into a symbiosis with the nitrogen-fixing bacterium Frankia. Actinorhizal plants comprise plants from the families Coriariaceae (coriaria family), Datiscaceae, Betulaceae (birch family), Casuarinaceae, Myricaceae (bayberry family), Elaeagnaceae (oleaster family), Rhamnaceae (buckthorn family), and Rosaceae (rose family).

[0085] Legumes comprise plants from the families Fabaceae (legume family), Polygalaceae (milkwort family), Quillajaceae (Quillaja family), and Surianaceae.

[0086] In an embodiment, the protein hydrolysate or the composition is used for seeds of plants or crops that form nitrogen root nodules. Preferably, the protein hydrolysate or the composition is used for plants or crops selected from the families Fabaceae (legume family), Coriariaceae (coriaria family), Datiscaceae, Betulaceae (birch family), Casuarinaceae, Myricaceae (bayberry family), Elaeagnaceae (oleaster family), Rhamnaceae (buckthorn family), and Rosaceae (rose family).

[0087] Specifically, plants or crops can be selected from the genera: Coriaria, Datisca, Alnus, Allocasuarina, Casuarina, Ceuthostoma, Gymnostoma, Comptonia, Myrica, Elaeagnus, Hippophae, Shepherdia, Adolphia, Colletia, Discaria, Kentrothamnus, Retanilla, Talguenea, Trevoa, Ochetophila, Ceanothus, Cercocarpus, Chamaebatia, Cowania, Dryas, Purshia, Caesalpinia, Cercis, Detarium, Dialium, Duparquetia, Faboideae, Polygala, Dakotanthus, Quillaja, Suriana., preferably Coriaria, Datisca, Alnus, Allocasuarina, Casuarina, Ceuthostoma, Gymnostoma, Comptonia, Myrica, Elaeagnus, Hippophae, Shepherdia, Colletia, Discaria, Ceanothus, Cercocarpus, Cowania, Purshia, Caesalpinia, Cercis, Detarium, Dialium, Duparquetia, Abrus, Acmispon, Acosmium, Adenocarpus, Adenodolichos, Adesmia, Aenictophyton, Aeschynomene, Afgekia, Aganope, Airyantha, Aldina, Alexa, Alhagi, Alistilus, Almaleea, Alysicarpus, Amburana, Amicia, Ammodendron, Ammopiptanthus, Ammothamnus, Amphiodon, Amorpha, Amphicarpaea, Amphimas, Amphithalea, Anagyris, Anarthrophyllum, Ancistrotropis, Andira, Angylocalyx, Antheroporum, Anthyllis, Antopetitia, Aotus, Aphyllodium, Apios, Apoplanesia, Apurimacia, Arachis, Argyrocytisus, Argyrolobium, Arthroclianthus, Aspalathus, Astragalus, Ateleia, Austrocallerya, Austrodolichos, Austrosteenisia, Baphia, Baphiastrum, Baphiopsis, Baptisia, Barbieria, Behaimia, Bionia, Bituminaria, Bobgunnia, Bocoa, Bolusafra, Bolusanthus, Bolusia, Bossiaea, Bowdichia, Bowringia, Brongniartia, Brya, Bryaspis, Burkilliodendron, Butea, Cadia, Cajanus, Calia, Calicotome, Callerya, Callistachys, Calobota, Calophaca, Calopogonium, Calpurnia, Camoensia, Camptosema, Campylotropis, Canavalia, Candolleodendron, Caragana, Carmichaelia, Carrissoa,Cascaronia, Castanospermum, Centrolobium, Centrosema, Chadsia, Chaetocalyx, Chamaecytisus, Chapmannia, Chesneya, Chorizema, Christia, Cicer, Cladrastis, Clathrotropis, Cleobulia, Clianthus, Clitoria, Clitoriopsis, Cochlianthus, Cochliasanthus, Codariocalyx, Collaea, Cologania, Colutea, Condylostylis, Cordyla, Coronilla, Coursetia, Craibia, Cranocarpus, Craspedolobium, Cratylia, Cristonia, Crotalaria, Cruddasia, Cullen, Cyamopsis, Cyathostegia, Cyclocarpa, Cyclolobium, Cyclopia, Cymbosema, Cytisophyllum, Cytisopsis, Cytisus, Dahlstedtia, Dalbergia, Dalbergiella, Dalea, Dalhousiea, Daprainia, Daviesia, Decorsea, Dendrolobium, Derris, Dermatophyllum, Desmodiastrum, Desmodium, Dewevrea, Dichilus, Dicraeopetalum, Dillwynia, Dioclea, Diphyllarium, Diphysa, Diplotropis, Dipogon, Dipteryx, Discolobium, Disynstemon, Dolichopsis, Dolichos, Dorycnium, Droogmansia, Dumasia, Dunbaria, Dussia, Dysolobium, Ebenus, Echinospartum, Eleiotis, Eminia, Endosamara, Eremosparton, Erichsenia, Erinacea, Eriosema, Errazurizia, Erythrina, Etaballia, Euchilopsis, Euchlora, Euchresta, Eutaxia, Eversmannia, Exostyles, Eysenhardtia, Ezoloba, Fairchildia, Fiebrigiella, Fissicalyx, Flemingia, Fordia, Galactia, Galega, Gastrolobium, Geissaspis, Genista, Genistidium, Geoffroea, Gliricidia, Glycine, Glycyrrhiza, Gompholobium, Gonocytisus, Goodia, Grazielodendron, Guianodendron, Gueldenstaedtia, Halimodendron, Hammatolobium, Haplormosia, Hardenbergia, Harleyodendron, Harpalyce, Hebestigma, Hedysarum, Helicotropis, Herpyza, Hesperolaburnum, Hippocrepis, Hoita, Holocalyx, Hosackia, Hovea, Huangtcia, Humularia, Hymenocarpos, Hymenolobium, Hypocalyptus, Indigastrum, Indigofera, Inocarpus, Isotropis, Jacksonia, Kanburia, Kennedia, Kotschya, Kummerowia, Lablab, Laburnocytisus, Laburnum, Lackeya, Ladeania, Lamprolobium, Lathyrus, Latrobea, Lebeckia, Lecointea, Lembotropis, Lennea, Lens, Leobordea, Leptoderris, Leptodesmia, Leptolobium, Leptosema, Leptospron, Lespedeza, Lessertia, Leucomphalos, Limadendron, Liparia, Listia, Lonchocarpus, Lotononis, Lotus, Luetzelburgia, Lupinus, Luzonia, Maackia, Machaerium, Macropsychanthus, Macroptilium, Macrotyloma, Maraniona, Margaritolobium, Marina, Mastersia, Mecopus, Medicago, Melilotus, Melliniella, Melolobium, Microcharis, Mildbraediodendron, Millettia, Mirbelia, Monopteryx, Mucuna, Muellera, Muelleranthus, Mundulea, Myrocarpus, Myrospermum, Mysanthus, Nanhaia, Neocollettia, Neoharmsia, Neonotonia, Neorautanenia, Neorudolphia, Nephrodesmus, Nesphostylis, Nissolia, Nogra, Oberholzeria, Olneya, Onobrychis, Ononis, Ophrestia, Orbexilum, Oreophysa, Ormocarpopsis, Ormocarpum, Ormosia, Orphanodendron, Ornithopus, Oryxis, Ostryocarpus, Otholobium, Otoptera, Ottleya, Oxylobium, Oxyrhynchus, Oxytropis, Pachyrhizus, Padbruggea, Panurea, Paracalyx, Paragoodia, Paramachaerium, Parochetus, Parryella, Pearsonia, Pediomelum, Pedleya, Periandra, Pericopsis, Petaladenium, Peteria, Petteria, Phaseolus, Phylacium, Phyllodium, Phyllota, Phylloxylon, Physostigma, Pickeringia, Pictetia, Piptanthus, Piscidia, Pisum,Plagiocarpus, Platycelyphium, Platycyamus, Platylobium, Platymiscium, Platypodium, Platysepalum, Podalyria, Podocytisus, Podolobium, Poecilanthe, Poiretia, Poitea, Polhillia, Polhillides, Pongamiopsis, Pseudarthria, Pseudeminia, Pseudoeriosema, Pseudovigna, Psophocarpus, Psoralea, Psorothamnus, Pterocarpus, Pterodon, Ptycholobium, Ptychosema, Pueraria, Pultenaea, Pullenia, Pycnospora, Pyranthus, Rafnia, Ramirezella, Ramorinoa, Retama, Rhodopis, Rhynchosia, Rhynchotropis, Riedeliella, Robinia, Robynsiophyton, Rothia, Rupertia, Sakoanala, Salweenia, Sarcodum, Sartoria, Schefflerodendron, Scorpiurus, Sellocharis, Sesbania, Shuteria, Sigmoidala, Sigmoidotropis, Sinodolichos, Smirnowia, Smithia, Soemmeringia, Sophora, Spartium, Spathionema, Spatholobus, Sphaerolobium, Sphaerophysa, Sphenostylis, Sphinctospermum, Spirotropis, Spongiocarpella, Stauracanthus, Staminodianthus, Steinbachiella, Stirtonanthus, Stonesiella, Streblorrhiza, Strongylodon, Strophostyles, Stylosanthes, Styphnolobium, Swainsona, Swartzia, Sweetia, Sylvichadsia, Syrmatium, Tabaroa, Tadehagi, Taralea, Taverniera, Templetonia, Tephrosia, Teramnus, Teyleria, Thermopsis, Thinicola, Tipuana, Trifidacanthus, Trifolium, Trigonella, Tripodion, Trischidium, Uleanthus, Ulex, Uraria, Uribea, Urodon, Vandasina, Vatairea, Vataireopsis, Vatovaea, Vavilovia, Vermifrux, Verdesmum, Vicia, Vigna, Viminaria, Virgilia, Vuralia, Wajira, Weberbauerella, Whitfordiodendron, Wiborgia, Wiborgiella, Wisteria, Wisteriopsis, Xanthocercis, Xiphotheca, Zollernia, Zornia, Zygocarpum.

[0088] In a preferred embodiment, the protein hydrolysate or the composition is used for seeds of plants or crops of the legume family Fabaceae), and more specifically the subfamily Faboideae, comprising the genera of: Abrus, Acmispon, Acosmium, Adenocarpus, Adenodolichos, Adesmia, Aenictophyton, Aeschynomene, Afgekia, Aganope, Airyantha, Aldina, Alexa, Alhagi, Alistilus, Almaleea, Alysicarpus, Amburana, Amicia, Ammodendron, Ammopiptanthus, Ammothamnus, Amphiodon, Amorpha, Amphicarpaea, Amphimas, Amphithalea, Anagyris, Anarthrophyllum, Ancistrotropis, Andira, Angylocalyx, Antheroporum, Anthyllis, Antopetitia, Aotus, Aphyllodium, Apios, Apoplanesia, Apurimacia, Arachis, Argyrocytisus, Argyrolobium, Arthroclianthus, Aspalathus, Astragalus, Ateleia, Austrocallerya, Austrodolichos, Austrosteenisia, Baphia, Baphiastrum, Baphiopsis, Baptisia, Barbieria, Behaimia, Bionia, Bituminaria, Bobgunnia, Bocoa, Bolusafra, Bolusanthus, Bolusia, Bossiaea, Bowdichia, Bowringia, Brongniartia, Brya, Bryaspis, Burkilliodendron, Butea, Cadia, Cajanus, Calia, Calicotome, Callerya, Callistachys, Calobota, Calophaca, Calopogonium, Calpurnia, Camoensia, Camptosema, Campylotropis, Canavalia, Candolleodendron, Caragana, Carmichaelia, Carrissoa, Cascaronia, Castanospermum, Centrolobium, Centrosema, Chadsia, Chaetocalyx, Chamaecytisus, Chapmannia, Chesneya, Chorizema, Christia, Cicer, Cladrastis,Clathrotropis, Cleobulia, Clianthus, Clitoria, Clitoriopsis, Cochlianthus, Cochliasanthus, Codariocalyx, Collaea, Cologania, Colutea, Condylostylis, Cordyla, Coronilla, Coursetia, Craibia, Cranocarpus, Craspedolobium, Cratylia, Cristonia, Crotalaria, Cruddasia, Cullen, Cyamopsis, Cyathostegia, Cyclocarpa, Cyclolobium, Cyclopia, Cymbosema, Cytisophyllum, Cytisopsis, Cytisus, Dahlstedtia, Dalbergia, Dalbergiella, Dalea, Dalhousiea, Daprainia, Daviesia, Decorsea, Dendrolobium, Derris, Dermatophyllum, Desmodiastrum, Desmodium, Dewevrea, Dichilus, Dicraeopetalum, Dillwynia, Dioclea, Diphyllarium, Diphysa, Diplotropis, Dipogon, Dipteryx, Discolobium, Disynstemon, Dolichopsis, Dolichos, Dorycnium, Droogmansia, Dumasia, Dunbaria, Dussia, Dysolobium, Ebenus, Echinospartum, Eleiotis, Eminia, Endosamara, Eremosparton, Erichsenia, Erinacea, Eriosema, Errazurizia, Erythrina, Etaballia, Euchilopsis, Euchlora, Euchresta, Eutaxia, Eversmannia, Exostyles, Eysenhardtia, Ezoloba, Fairchildia, Fiebrigiella, Fissicalyx, Flemingia, Fordia, Galactia, Galega, Gastrolobium, Geissaspis, Genista, Genistidium, Geoffroea, Gliricidia, Glycine, Glycyrrhiza, Gompholobium, Gonocytisus, Goodia, Grazielodendron, Guianodendron, Gueldenstaedtia, Halimodendron, Hammatolobium, Haplormosia, Hardenbergia, Harleyodendron, Harpalyce, Hebestigma, Hedysarum, Helicotropis, Herpyza, Hesperolaburnum, Hippocrepis, Hoita, Holocalyx, Hosackia, Hovea, Huangtcia, Humularia, Hymenocarpos, Hymenolobium, Hypocalyptus, Indigastrum, Indigofera, Inocarpus, Isotropis, Jacksonia, Kanburia, Kennedia, Kotschya, Kummerowia, Lablab, Laburnocytisus, Laburnum, Lackeya, Ladeania, Lamprolobium, Lathyrus, Latrobea, Lebeckia, Lecointea, Lembotropis, Lennea, Lens, Leobordea, Leptoderris, Leptodesmia, Leptolobium, Leptosema, Leptospron, Lespedeza, Lessertia, Leucomphalos, Limadendron, Liparia, Listia, Lonchocarpus, Lotononis, Lotus, Luetzelburgia, Lupinus, Luzonia, Maackia, Machaerium, Macropsychanthus, Macroptilium, Macrotyloma, Maraniona, Margaritolobium, Marina, Mastersia, Mecopus, Medicago, Melilotus, Melliniella, Melolobium, Microcharis, Mildbraediodendron, Millettia, Mirbelia, Monopteryx, Mucuna, Muellera, Muelleranthus, Mundulea, Myrocarpus, Myrospermum, Mysanthus, Nanhaia, Neocollettia, Neoharmsia, Neonotonia, Neorautanenia, Neorudolphia, Nephrodesmus, Nesphostylis, Nissolia, Nogra, Oberholzeria, Olneya, Onobrychis, Ononis, Ophrestia, Orbexilum, Oreophysa, Ormocarpopsis, Ormocarpum, Ormosia, Orphanodendron, Ornithopus, Oryxis, Ostryocarpus, Otholobium, Otoptera, Ottleya, Oxylobium, Oxyrhynchus, Oxytropis, Pachyrhizus, Padbruggea, Panurea, Paracalyx, Paragoodia, Paramachaerium, Parochetus, Parryella, Pearsonia, Pediomelum, Pedleya, Periandra, Pericopsis, Petaladenium, Peteria, Petteria, Phaseolus, Phylacium, Phyllodium, Phyllota, Phylloxylon, Physostigma, Pickeringia, Pictetia, Piptanthus, Piscidia, Pisum, Plagiocarpus, Platycelyphium, Platycyamus, Platylobium, Platymiscium, Platypodium, Platysepalum, Podalyria, Podocytisus, Podolobium, Poecilanthe, Poiretia, Poitea,Polhillia, Polhillides, Pongamiopsis, Pseudarthria, Pseudeminia, Pseudoeriosema, Pseudovigna, Psophocarpus, Psoralea, Psorothamnus, Pterocarpus, Pterodon, Ptycholobium, Ptychosema, Pueraria, Pultenaea, Pullenia, Pycnospora, Pyranthus, Rafnia, Ramirezella, Ramorinoa, Retama, Rhodopis, Rhynchosia, Rhynchotropis, Riedeliella, Robinia, Robynsiophyton, Rothia, Rupertia, Sakoanala, Salweenia, Sarcodum, Sartoria, Schefflerodendron, Scorpiurus, Sellocharis, Sesbania, Shuteria, Sigmoidala, Sigmoidotropis, Sinodolichos, Smirnowia, Smithia, Soemmeringia, Sophora, Spartium, Spathionema, Spatholobus, Sphaerolobium, Sphaerophysa, Sphenostylis, Sphinctospermum, Spirotropis, Spongiocarpella, Stauracanthus, Staminodianthus, Steinbachiella, Stirtonanthus, Stonesiella, Streblorrhiza, Strongylodon, Strophostyles, Stylosanthes, Styphnolobium, Swainsona, Swartzia, Sweetia, Sylvichadsia, Syrmatium, Tabaroa, Tadehagi, Taralea, Taverniera, Templetonia, Tephrosia, Teramnus, Teyleria, Thermopsis, Thinicola, Tipuana, Trifidacanthus, Trifolium, Trigonella, Tripodion, Trischidium, Uleanthus, Ulex, Uraria, Uribea, Urodon, Vandasina, Vatairea, Vataireopsis, Vatovaea, Vavilovia, Vermifrux, Verdesmum, Vicia, Vigna, Viminaria, Virgilia, Vuralia, Wajira, Weberbauerella, Whitfordiodendron, Wiborgia, Wiborgiella, Wisteria, Wisteriopsis, Xanthocercis, Xiphotheca, Zollernia, Zornia, Zygocarpum.

[0089] In an embodiment, the protein hydrolysate or the composition is used for seeds of plants or crops from the genus Glycine, such as Glycine albicans, Glycine aphyonotos, Glycine arenaria, Glycine argyrea, Glycine canescens, Glycine clandestina, Glycine curvata, Glycine cyrtoloba, Glycine dolichocarpa, Glycine falcata, Glycine gracei, Glycine hirticaulis, Glycine hirticaulis subsp. leptosa, Glycine koidzumii, Glycine lactovirens, Glycine latifolia, Glycine latrobeana, Glycine microphylla, Glycine montis-douglas, Glycine peratosa, Glycine pescadrensis, Glycine pindanica, Glycine pullenii, Glycine remota, Glycine rubiginosa, Glycine stenophita, Glycine syndetika, Glycine tabacina, Glycine tomentella, Glycine max, Glycine soja.

[0090] In an embodiment, the seeds are soybean seeds (Glycine max).

[0091] In an embodiment, the seeds are treated with protein hydrolysate in an amount between 0.01 and 100 pL per seed, preferably between 0.1 and 100 pL per seed, even more preferably between 0.1 and 75 pL per seed, more preferably between 0.1 and 50 pL per seed, even more preferably between 0.1 and 25 pL per seed, even more preferably between 0.1 and 20 pL per seed, and most preferably between 0.1 and 10 pL per seed. Preferably, the protein hydrolysate has a dry matter content between 10 and 20 wt.%, preferably between 12 and 18 wt.%.In a preferred embodiment, the protein hydrolysate is used as a seed treatment, preferably seed coating, wherein the protein hydrolysate is applied in an amount of at most 1 pL per seed, preferably of at most 0.9 pL per seed, more preferably of at most 0.8 pL per seed, even more preferably of at most 0.7 pL per seed, and even more preferably of at most 0.6 pL per seed, and most preferably of at most 0.5 pL per seed. In another or further preferred embodiment, the protein hydrolysate is used as a seed treatment, preferably seed coating, wherein the protein hydrolysate is applied in an amount between 0.01 and 1 pL per seed, preferably between 0.05 and 0.9 pL per seed, more preferably between 0.05 and 0.8 pL per seed, even more preferably between 0.05 and 0.7 pL per seed, and even more preferably between 0.05 and 0.6 pL per seed, and most preferably between 0.05 and 0.5 pL per seed. Preferably, the protein hydrolysate has a dry matter content between 10 and 20 wt.%, preferably between 12 and 18 wt.%.

[0092] An advantage of such low dosages per seed is that a biological effect can be achieved even with very small amounts of protein hydrolysate, which contributes to an efficient use of the product, limited material costs, and a simple industrial application in large-scale seed treatment.

[0093] In an embodiment, the seeds are treated with protein hydrolysate in an amount between 0.01 and 15 g dry matter per seed, preferably between 0.01 and 10 g dry matter per seed, even more preferably between 0.01 and 1 g dry matter per seed, more preferably between 0.1 and 1 g dry matter per seed, even more preferably between 0.1 and 0.5 g dry matter per seed.

[0094] In a preferred embodiment, the seeds are optimized with protein hydrolysate in an amount between 0.001 and 10 mg dry matter per seed, preferably between 0.005 and 5 mg, even more preferably between 0.01 and 1 mg, more preferably between 0.01 and 0.5 mg, and even more preferably between 0.01 and 0.1 mg dry matter per seed.

[0095] In a preferred embodiment, the protein hydrolysate is used as a seed treatment, preferably as a seed coating, wherein the protein hydrolysate is applied in a dosage expressed as volume per mass of seed. In particular, the protein hydrolysate is applied in an amount of at most 5 mL per kg seed, preferably at most 4 mL per kg seed, more preferably at most 3 mL per kg seed, even more preferably at most 2.5 mL per kg seed, and even more preferably at most 2 mL per kg seed, even more preferably at most 1.5 mL per kg seed.In a further preferred embodiment, the protein hydrolysate is applied in an amount between 0.1 and 5 mL per kg seed, preferably between 0.2 and 4.5 mL per kg seed, more preferably between 0.2 and 4 mL per kg seed, even more preferably between 0.2 and 3.5 mL per kg seed, even more preferably between 0.5 and 3 mL per kg seed, even more preferably between 0.5 and 2.5 mL per kg seed, even more preferably between 0.5 and 2 mL per kg seed, even more preferably between 0.5 and 1.5 mL per kg seed.

[0096] The bacteria present in the inoculum can vary, depending on the seeds that are to be optimized. Each plant has specific symbionts that work best for their root environment and nutritional requirements. Thus, soybeans, such as Glycine max, form a symbiosis with Bradyrhizobium japonicum, while peas (Pisum sativum) thrive better with Rhizobium leguminosarum. For lucerne (Medicago sativa), Sinorhizobium meliloti is the most suitable. Also alders (Alnus spp.) have a unique symbiosis, wherein Frankia alni helps in forming nitrogen root nodules.

[0097] It should be understood that the embodiments of the aspects described above all also relate to the aspects described hereinafter. The use aspects described above thus also relate to the methods for optimizing seeds described hereinafter, and vice versa.

[0098] In another aspect, the invention relates to a method for optimizing seeds. Optimizing seeds is intended to mean actions that are performed on the seeds or during the seed phase of the crops.

[0099] In an embodiment, the invention relates to a method for optimizing seeds with a protein hydrolysate. Herein, the method comprises the sequential, separate, or simultaneous application of a protein hydrolysate with the seeds. Herein, the protein hydrolysate is applied to the soil prior to the sowing of the seeds, simultaneously with the sowing of the seeds, or after the sowing of the seeds.

[0100] In an embodiment, the method comprises the sequential, separate, or simultaneous application of a protein hydrolysate or a composition as discussed above with the seeds.

[0101] The simultaneous application can comprise a seed treatment, wherein the seeds are treated with the protein hydrolysate or the composition prior to sowing; or the simultaneous application of the seeds and the protein hydrolysate or the composition separately to the soil.In an embodiment, the protein hydrolysate can be added to a fertilizer or a soil conditioner and applied to the soil prior to the sowing of the seeds. Herein, the hydrolysate can be applied to the soil up to a maximum of 30 days before the sowing of the seeds, preferably at most 14 days.

[0102] In an embodiment, the seeds are optimized with protein hydrolysate in an amount between 0.01 and 100 pL per seed, preferably between 0.1 and 100 pL per seed, even more preferably between 0.1 and 75 pL per seed, more preferably between 0.1 and 50 pL per seed, even more preferably between 0.1 and 25 pL per seed, even more preferably between 0.1 and 20 pL per seed, and most preferably between 0.1 and 10 pL per seed. Preferably, the protein hydrolysate has a dry matter content between 10 and 20 wt.%, preferably between 12 and 18 wt.%.

[0103] In a preferred embodiment, the protein hydrolysate is used as a seed treatment, preferably seed coating, wherein the protein hydrolysate is applied in an amount of at most 1 pL per seed, preferably of at most 0.9 pL per seed, more preferably of at most 0.8 pL per seed, even more preferably of at most 0.7 pL per seed, and even more preferably of at most 0.6 pL per seed, and most preferably of at most 0.5 pL per seed. In another or further preferred embodiment, the protein hydrolysate is used as a seed treatment, preferably seed coating, wherein the protein hydrolysate is applied in an amount between 0.01 and 1 pL per seed, preferably between 0.05 and 0.9 pL per seed, more preferably between 0.05 and 0.8 pL per seed, even more preferably between 0.05 and 0.7 pL per seed, and even more preferably between 0.05 and 0.6 pL per seed, and most preferably between 0.05 and 0.5 pL per seed. Preferably, the protein hydrolysate has a dry matter content between 10 and 20 wt.%, preferably between 12 and 18 wt.%.

[0104] In an embodiment, the seeds are optimized with protein hydrolysate in an amount between 0.01 and 15 g dry matter per seed, preferably between 0.01 and 10 g dry matter per seed, even more preferably between 0.01 and 1 g dry matter per seed, more preferably between 0.1 and 1 g dry matter per seed, even more preferably between 0.1 and 0.5 g dry matter per seed.

[0105] In a preferred embodiment, the seeds are optimized with protein hydrolysate in an amount between 0.001 and 10 mg dry matter per seed, preferably between 0.005 and 5 mg, even more preferably between 0.01 and 1 mg, more preferably between 0.01 and 0.5 mg, and even more preferably between 0.01 and 0.1 mg dry matter per seed.In another or further embodiment, the method comprises the separate, sequential, or simultaneous application of the protein hydrolysate and the seeds to the soil, wherein an inoculum comprising bacteria is also applied to or in the soil separately, sequentially, or simultaneously with the sowing of seeds. Herein, the protein hydrolysate is applied to the soil prior to the sowing of the seeds, simultaneously with the sowing of the seeds, or after the sowing of the seeds, and an inoculum comprising bacteria can also be applied to the soil prior to the sowing of the seeds, simultaneously with the sowing of the seeds, or after the sowing of the seeds. The inoculum can contain bacteria as in the embodiments described above.

[0106] In an embodiment, the method comprises optimizing seeds wherein the seeds originate from plants or crops that can have a symbiotic relationship with nitrogen-fixing bacteria. Preferably selected from Actinorhizal plants, and legumes as in embodiments described above.

[0107] In a preferred embodiment, if an inoculum comprising symbiotic nitrogen-fixing bacteria is applied simultaneously, the protein hydrolysate is applied in an amount of at most 0.3 pL / seed, preferably even of at most 0.2 pL / seed. Preferably, the protein hydrolysate has a dry matter content between 10 and 20 wt.%, preferably between 12 and 18 wt.%.

[0108] In an aspect, the invention relates to a method for increasing the nitrogen uptake in seeds and / or plants or crops. This can be achieved by embodiments as described above. In an embodiment, the seeds of the plants or crops, when these are sown, are treated with a protein hydrolysate as described above, preferably the seeds are coated with the protein hydrolysate.

[0109] In this method, an inoculum comprising symbiotic nitrogen-fixing bacteria can also be sown with the seeds.

[0110] In another or further aspect, the invention relates to a method for increasing the growth rate in plants or crops. In another or further aspect, the invention relates to a method for increasing the number of root nodules in plants or crops. In another or further aspect, the invention relates to a method for increasing the number of pods in plants or crops.

[0111] The invention can thus also be described with reference to the following embodiments: 1. A method for optimizing seeds with a protein hydrolysate, wherein the protein hydrolysate is derived from poultry, and wherein the protein hydrolysate is applied sequentially, separately, or simultaneously with the seeds.2. The method according to embodiment 1, wherein the protein hydrolysate is sown simultaneously with the seeds.

[0112] 3. The method according to embodiment 2, wherein the protein hydrolysate is applied to the seeds as a seed coating.

[0113] 4. The method according to any of the preceding embodiments, wherein the protein hydrolysate is applied in an amount between 0.1 and 100 pL per seed.

[0114] 5. The method according to embodiment 4, wherein the protein hydrolysate is sown in an amount between 0.1 and 10 pL per seed.

[0115] 6. The method according to any of the preceding embodiments, wherein further an inoculum of nitrogen-fixing bacteria is applied sequentially, separately, or simultaneously with the seeds and the protein hydrolysate.

[0116] 7. The method according to embodiment 6, wherein the nitrogen-fixing bacteria are selected from Rhizobium spp., Bradyrhizobium spp., Sinorhizobium spp., Mesorhizobium spp., Azorhizobium spp. Frankia spp. (actinorhiza), Allorhizobium spp., and Neorhizobium spp., preferably selected from Rhizobium spp., Bradyrhizobium spp., or a combination thereof.

[0117] 8. The method according to any of the preceding embodiments, wherein the protein hydrolysate is derived from slaughterhouse waste, such as blood, hides, entrails, feathers, feet, and bones.

[0118] 9. The method according to any of the preceding embodiments, wherein the seeds belong to the legume family (Fabaceae).

[0119] 10. The method according to embodiment 9, wherein the seeds belong to the Faboideae.

[0120] 11. The method according to embodiment 10, wherein the seeds are soybean seeds (Glycine max).

[0121] 12. The method according to any of the preceding embodiments, wherein at least 90 wt.% of the peptides in the protein hydrolysate have a molecular weight of at most 500 Dalton, and wherein between 35 and 55 wt.% of the peptides have a molecular mass between 204 and 300 Dalton.

[0122] 13. Use of a protein hydrolysate derived from poultry in the treatment of seeds.

[0123] 14. Use according to embodiment 13, wherein the seeds belong to the legume family (Fabaceae).

[0124] 15. Use according to embodiment 14, wherein the seeds are soybean seeds (Glycine max).

[0125] 16. Method for increasing the nitrogen uptake in plants or crops, wherein when the seeds of the plants or crops are sown, these seeds have been treated with a protein hydrolysate, wherein the protein hydrolysate is derived from poultry.17. The method according to embodiment 16, wherein the seeds are coated with the protein hydrolysate.

[0126] 18. The method according to embodiment 16 or 17, wherein an inoculum comprising symbiotic nitrogen-fixing bacteria is also applied with the seeds.

[0127] In the following, the invention is described by means of non-limiting examples illustrating the invention, and which are not intended or to be interpreted to limit the scope of the invention.

[0128] EXAMPLES

[0129] Example 1

[0130] In this experiment, it was investigated whether the seed treatment (A) with a protein hydrolysate according to an embodiment of the invention has an influence on the growth of soybeans (variety: Acardia) in comparison with an untreated control (C). For this study, use was made of an oligotrophic mineral substrate, in which eight replicate pots were set up, each with three soybean plants. The protein hydrolysate was obtained from chicken blood by clotting, boiling, and centrifuging the chicken blood, so that a serum was obtained. Subsequently, the serum was subjected to a thermal and enzymatic hydrolysis.

[0131] The seed treatment consisted of immersing each seed in 1 pL of protein hydrolysate with a dry matter content of 5 wt.%, while the control received no treatment. All seeds were subsequently planted in the same type of substrate to standardize the growth conditions. The plants were additionally inoculated with Bradyrhizobium japonicum, a bacterium known for its symbiotic relationship with soybeans and its ability to fix nitrogen.

[0132] Plant height

[0133] The seed treatment led to an increase in the growth rate of the shoots by 20%. This is also shown in Figure 1 wherein the plant height in cm is plotted as a function of the number of days after emergence of the seedlings.

[0134] Root length and mass

[0135] The seed treatment led to an increase in the root mass. This is also shown in Figures 2A and 2B wherein the root length and mass (g dry matter) for the control and the seed treatment are shown.

[0136] Number of podsThe seed treatment led to an increase in the number of pods on the plants. This is also shown in Figure 3 wherein the number of pods for the control and the seed treatment is shown. The dry weight of the pods was also higher in the plants that had undergone a seed treatment.

[0137] Number of root nodules

[0138] The seed treatment led to an increase in the number of root nodules by 66%. This is also shown in Figure 4A wherein the number of nodules for the control and the seed treatment is shown. Figures 4B and 4C show the nodules of the control group and the seed treatment, respectively. The dry weight of the nodules was also higher in the plants that had undergone a seed treatment.

[0139] Example 2

[0140] The experiment from Example 1 was repeated but with four different test setups: untreated (C), seed treatment (A), Bradyrhizobium japonicum inoculum (R), and a seed treatment in combination with a Bradyrhizobium japonicum inoculum (A+R).

[0141] Plant height

[0142] The seed treatment in combination with inoculum led to a significant increase in the growth rate of the shoots compared to the control as early as four weeks after sowing, but also compared to the Bradyrhizobium japonicum inoculum alone, whereas the latter only showed a significant growth response from about nine weeks after sowing. This is also shown in Figure 5 wherein the plant height in cm is plotted as a function of the number of weeks after sowing. The seed treatment in combination with inoculum thus resulted in an approximately five weeks faster growth response compared to the inoculum alone. The fact that the protein hydrolysate showed no detectable effect on the plant height in the absence of rhizobia confirms that the observed effect is rhizobium-mediated.

[0143] Number of root nodules

[0144] The seed treatment in combination with a Bradyrhizobium japonicum inoculum led to an increase in the number of root nodules compared to the Bradyrhizobium japonicum inoculum alone. This is also shown in Figure 6 wherein the number of nodules for the Bradyrhizobium japonicum inoculum alone and the seed treatment in combination with a Bradyrhizobium japonicum inoculum is shown. The dry weight of the nodules was also higher in the plants that had undergone a seed treatment.Number of pods

[0145] The seed treatment in combination with a Bradyrhizobium japonicum inoculum also led to an increase in the number of pods compared to the Bradyrhizobium japonicum inoculum alone. This is also shown in Figure 7 wherein the number of pods for the Bradyrhizobium japonicum inoculum alone and the seed treatment in combination with a Bradyrhizobium japonicum inoculum is shown. The dry weight of the pods was also higher in the plants that had undergone a seed treatment.

[0146] Example 3

[0147] A multi-location field trial was carried out to evaluate the effect of a protein hydrolysate as seed treatment (seed coating) on the early growth and yield of soybean under field conditions. The field trials were carried out at four different trial locations. The trial locations represented diverse soil types, pH values, and yield potentials, in order to evaluate the robustness of the effect across different agro-ecological conditions.

[0148] At each location, three seed treatments were compared, namely an untreated control, a low dose of protein hydrolysate (1 mL / kg seed) and a high dose of protein hydrolysate (5 mL / kg seed). Converted, the low dose corresponds to approximately 0.2 pL / seed, while the high dose corresponds to approximately 1 pL / seed.

[0149] The product used in this field trial was a protein hydrolysate derived from chicken blood-derived protein fractions, with a dry matter content of approximately 15% (w / w), a peptide molecular weight distribution, expressed as relative contributions to the total protein content, of approximately 20% below 200 Da, 45% between 200 and 300 Da, 17% between 300 and 400 Da, 6% between 400 and 500 Da, 3% between 500 and 600 Da, approximately 5% between 600 and 1,000 Da in total, and approximately 3% above 1,000 Da, and an amino acid composition, expressed as a percentage of the total protein content, consisting of approximately glutamic acid (2.5%), leucine (2.3%), lysine (2.3%), aspartic acid (2.0%), alanine (1.7%), tyrosine (1.5%), valine (1.5%), phenylalanine (1.4%), threonine (1.3%), histidine (1.1%), serine (1.1%), glycine (1.1%), isoleucine (1.0%), arginine (0.9%), proline (0.9%), tryptophan (0.4%), methionine (0.3%) and cysteine (0.3%).

[0150] The trials were set up according to a randomized block design with six replicates per treatment at each location. No separate bacterial inoculum was added, so that the effect of the protein hydrolysate could be evaluated exclusively in interaction with the naturally occurring soil microbiology. The results are shown below in comparison with the untreated control, UTC:Loca% effect of low % effect of Total yield Number of root tion dose on low dose on in UTC nodules per plant in biomass (V6) yield (tons / ha) UTC

[0151] 1 16.0 2.8 4.07 27.0

[0152] 2 5.5 15.4 3.14 42.0

[0153] 3 20.0 3.2 5.93 41.0

[0154] 4 15.0 6.9 4.27 41.0

[0155]

[0156] During the growing season, early crop development was monitored by measuring the above-ground biomass at the six-leaf stage. In addition, the yield was determined at the end of the growing season.

[0157] The results show that the low dose of protein hydrolysate led to a significant increase in the above-ground biomass at the six-leaf stage, with a model-estimated average increase of approximately 14% compared to the untreated control. This effect was consistent across the different trial locations.

[0158] Also at the level of the final yield, a statistically significant increase was observed for the low dose of protein hydrolysate. The model -estimated yield increase amounted to approximately 7% on average, which corresponds to approximately 0.28 tons / ha.

[0159] For the high dose of protein hydrolysate, no statistically significant effect on the early biomass was observed when using the protein hydrolysate as a seed coating. These results show that the protein hydrolysate exhibits a pronounced dose-dependent effect when applied as a seed coating, wherein even low doses suffice to achieve a significant positive effect on both early crop development and final yield, whereas higher doses provide no additional benefit.

Claims

CLAIMS1. A protein hydrolysate for optimizing seeds, characterized in that at least 90 wt.% of the peptides in the protein hydrolysate have a molecular mass of at most 500 Dalton, and that between 35 and 60 wt.% of the peptides have a molecular mass between 204 and 300 Dalton.

2. Protein hydrolysate according to claim 1, wherein at least 80 wt.% of the peptides have a molecular mass of at most 400 Dalton.

3. Protein hydrolysate according to any one of the preceding claims, wherein between 10 and 30 wt.% of the peptides have a molecular mass of at most 204 Dalton.

4. Protein hydrolysate according to claim 3, wherein between 10 and 25 wt.% of the peptides have a molecular mass of at most 204 Dalton.

5. Protein hydrolysate according to any one of the preceding claims, wherein between 10 and 30 wt.% of the peptides have a molecular mass between 300 and 400 Dalton.

6. Protein hydrolysate according to claim 5, wherein between 15 and 25 wt.% of the peptides have a molecular mass between 300 and 400 Dalton.

7. Protein hydrolysate according to any one of the preceding claims, wherein between 40 and 50 wt.% of the peptides have a molecular mass between 204 and 300 Dalton.

8. Protein hydrolysate according to any one of the preceding claims, wherein between 1 and 15 wt.% of the peptides have a molecular mass between 400 and 500 Dalton.

9. Protein hydrolysate according to claim 1, whereinbetween 10 and 30 wt.% of the peptides have a molecular mass of at most 204 Dalton;between 10 and 30 wt.% of the peptides have a molecular mass between 300 and 400 Dalton; andbetween 1 and 15 wt.% of the peptides have a molecular mass between 400 and 500 Dalton.

10. Protein hydrolysate according to any one of the preceding claims, wherein the protein hydrolysate is an animal protein hydrolysate.

11. Protein hydrolysate according to any one of the preceding claims, wherein the protein hydrolysate is derived from poultry.

12. Protein hydrolysate according to claim 11, wherein the protein hydrolysate is derived from poultry blood.

13. A composition comprising a hydrolysate according to any one of the preceding claims, and an inoculum comprising bacteria.

14. Composition according to claim 13, wherein the bacteria are nitrogen-fixing bacteria.

15. Composition according to claim 14, wherein the bacteria are selected from Rhizobium spp., Bradyrhizobium spp., Sinorhizobium spp., Mesorhizobium spp., Azorhizobium spp. Frankia spp. (actinorhiza), Allorhizobium spp., and Neorhizobium spp., preferably selected from Rhizobium spp., Bradyrhizobium spp., or a combination thereof.

16. Use of a protein hydrolysate according to any one of claims 1-12 during the treating or sowing of seeds, wherein the seeds belong to the legume family Fabaceae).

17. Use according to claim 16, wherein the seeds are soybean seeds (Glycine max).

18. Use according to claim 16 or 17, wherein the seeds are sown with protein hydrolysate in an amount between 0.1 and 100 pL per seed, wherein the protein hydrolysate has a dry matter content of 10-20 wt.%.

19. Use according to claim 18, wherein the seeds are treated with protein hydrolysate in an amount between 0.01 and 10 pL per seed, wherein the protein hydrolysate has a dry matter content of 10-20 wt.%.

20. Use according to claim 19, wherein the seeds are treated with protein hydrolysate in an amount between 0.01 and 0.5 pL per seed, wherein the protein hydrolysate has a dry matter content of 10-20 wt.%.

21. Method for increasing nitrogen uptake in plants or crops, wherein when the seeds of the plants or crops are sown, said seeds have been treated with a protein hydrolysate according to one of claims 1 to 12.

22. Method according to claim 21, wherein the seeds are coated with a protein hydrolysate according to one of claims 1 to 12.

23. Method according to claim 21 or 22, wherein an inoculum comprising symbiotic nitrogen-fixing bacteria is also sown with the seeds.

24. A method for optimizing seeds with a protein hydrolysate, wherein the protein hydrolysate is administered sequentially, separately or simultaneously with the seeds.

25. Method according to claim 24, wherein the protein hydrolysate is derived from poultry.

26. Method according to claim 25, wherein the protein hydrolysate is applied to the seeds as a seed coating.

27. Method according to claim 26, wherein the protein hydrolysate is applied in an amount between 0.05 and 0.5 pL per seed.

28. Method according to claim 26 or 27, wherein the seeds are soybean seeds.

29. Method according to claims 26, 27 or 28, wherein the protein hydrolysate is a protein hydrolysate according to one of claims 1-12.

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