Algae-based biostimulant

Abstract

The present application pertains to the field of agriculture and the circular economy, in particular, the technology of the present application consists of a novel method for producing a biostimulant that can be used to promote plant growth and development, with a very low heavy metal content, formulated from widely available algae, as well as a biostimulant with said characteristics, uses of same, and methods for optimising crops using said biostimulant.

Description

[0001] ALGAE-BASED BIOSTIMULANT

[0002] FIELD OF TECHNOLOGY

[0003]

[0001] This application falls within the field of agriculture and the circular economy. Specifically, the technology relates to an organic biostimulant based on algae, its production method, application, and uses.

[0004] BACKGROUND OF THE TECHNOLOGY

[0005]

[0002] One of the main challenges of the current agricultural sector is to optimize the production of fruits and vegetables, minimizing the environmental impact, in which context it is desirable to have biostimulants that allow to promote and improve the growth and development of plants, as well as the transplanting of seedlings, both in conventional agricultural systems and in hydroponic systems.

[0006]

[0003] In particular, it is desirable that biostimulants provide a balanced supply of nutrients according to the requirements of the plantation in which they will be used, so that, for example, the quantity of fruit produced by the plant is improved without affecting its weight.

[0007]

[0004] Furthermore, the industry increasingly requires alternatives to chemically synthesized products, making organic biostimulants, formulated primarily from natural products, desirable. In this context, given the wide availability of different types of seaweed, many of which are byproducts of other industries, it is desirable to use or reuse these algae to formulate biostimulants capable of optimizing agricultural production. The use of this type of algae would promote a circular economy. However, the ongoing challenge remains in developing production methods that allow for the production of algae-based biostimulants.

[0008]

[0005] To a large extent, the fact that algae-based biostimulants are not yet a viable alternative in the industry is precisely because there are no methods that allow for their production on a commercial or industrial scale. Among other things, the challenges in designing methods for the production of algae-based biostimulants relate to the need for these processes to allow for the proper purification of the algae according to its intended use, using energy-efficient processes.

[0006] Another problem faced by organic biostimulants formulated from raw materials such as algae is that they are not usually able to provide all the nutrients required by the crops for which they are intended, especially minerals. In this sense, the challenge arises of designing formulations that include the correct addition of additional elements, which largely depends on the specific profile of the organic matrix used.

[0009]

[0007] Finally, it is also worth highlighting the challenge of ensuring that organic biostimulants meet the profiles required by local regulations and have low levels of heavy metals, such as lead (Pb), cadmium (Cd), arsenic (As), and mercury (Me). Therefore, it is desirable that biostimulant production methods reduce the presence of the aforementioned metals to below detectable limits, i.e., less than 1 mg / kg.

[0010]

[0008] In this context, while prior art documents exist that disclose processes for the production of algae-based biostimulants, there is still considerable room for optimization. One example is patent application 201401464, filed by Patagonia Biotecnología with the National Institute of Industrial Property in Chile, which describes a process for formulating an algae-based biostimulant. However, this patent presents several shortcomings. For example, among many others, it does not describe the formulation of a combined algae matrix that would provide the resulting product with adequate nutritional properties; it does not include specific processes for different types of algae that would maximize the extraction of the desired products or their purification; and it does not address the supplementation of the organic matrix with additional elements.

[0011]

[0009] In this context, the technology of the present application consists of a novel method for the production of a biostimulant useful for promoting the growth and development of plants, with a very low content of heavy metals, formulated from widely available algae, as well as a biostimulant with said characteristics, uses and methods for the optimization of crops from said biostimulant.

[0012] BRIEF DESCRIPTION OF THE TECHNOLOGY

[0010] This application falls within the field of agriculture and the circular economy. Specifically, the technology relates to an organic biostimulant based on algae and its production method.

[0013]

[0011] More specifically, the present invention comprises, on the one hand, a method for producing a biostimulant useful for promoting the growth and development of plants, based on algae, comprising at least the following steps:

[0014] I. A seaweed chopping stage, comprising at least the following steps:

[0015] a. Have at least the following types of algae available:

[0016] i. algae rich in ulvane (AU),

[0017] i. algae rich in carrageenan (AC), and

[0018] iii. tucano-rich algae (AF);

[0019] b. Place each type of seaweed in a separate container;

[0020] c. Carry out the following sub-steps for each type of algae:

[0021] i. Wash the seaweed;

[0022] i. Dry the washed seaweed at room temperature for 21 to 27 hours, preferably for 24 hours;

[0023] iii. Dry the algae again at a temperature between 40 and 60°C for up to 48 hours, preferably at 50°C; and

[0024] iv. Chop the dried seaweed until a particle size of no more than 1 cm is achieved, obtaining chopped seaweed for each of the types of seaweed.

[0025] II. A depigmentation stage, consisting of depigmenting each type of algae, by means of at least the following steps:

[0026] a. Immerse each type of previously chopped seaweed in ethanol at a ratio of approximately 2 L of ethanol per 100 g of chopped seaweed, forming a mixture (M1);

[0027] b. Stir the mixture (M1) for a period of 24 to 72 h;

[0028] c. Filter to obtain depigmented algae;

[0029] d. Dry the depigmented algae at room temperature for up to 48 hours.

[0030] III. An ulvane extraction stage, which in turn comprises:

[0031] a. Immerse the depigmented ulvano-rich algae in water, in a ratio of approximately 100 g of algae per 2 L of water; b. Stir constantly at a temperature between 70 and 90°C for approximately 5 hours;

[0032] c. Filter until an aqueous extract rich in ulvane is obtained;

[0033] d. Concentrate the ulvane-rich extract;

[0034] e. Precipitate in ethanol at between 80 and 95%, obtaining ulvane; f. Dry the ulvane at a temperature between 40 and 60°C, until dry ulvane is obtained;

[0035] g. Grind the dry ulvane until you obtain ulvane powder (UP).

[0036] IV. A tucano extraction stage, which in turn comprises:

[0037] a. Immerse the depigmented tucano-rich algae in an approximately 2% calcium chloride solution (S1), in an approximate ratio of 100 g of algae per 500 mL of the approximately 2% calcium chloride solution (S1);

[0038] b. Stir at a temperature between 70 and 100°C for approximately 5 hours;

[0039] c. Filter until you obtain an aqueous extract rich in tucano;

[0040] d. Concentrate the tucano-rich extract;

[0041] e. Precipitate in approximately 95% ethanol, obtaining a precipitated tucano extract;

[0042] f. Dry the precipitated tucano extract at a temperature between 40 and 60°C, until dry tucano is obtained;

[0043] g. Add to the dried tucano a solution of approximately 1 M hydrochloric acid (HCl) (S2) and subsequently add sodium hydroxide (NaOH) of approximately 1 M until the pH is adjusted to approximately 7; h. Concentrate and precipitate in ethanol, obtaining a concentrated extract of tucano;

[0044] i. Dry the concentrated tucano extract at a temperature between 40 and 60°C, obtaining tucano;

[0045] j. Grind the dried tucano, obtaining tucano powder (FP).

[0046] V. A carrageenan extraction stage, which in turn comprises:

[0047] a. Soak the depigmented carrageenan-rich algae in water at a ratio of approximately 100 g of algae per 2 L of water; b. Stir constantly at a temperature between 70 and 80°C for approximately 5 hours; c. Filter to obtain an aqueous extract rich in carrageenan;

[0048] d. Concentrate the aqueous extract rich in carrageenan;

[0049] e. Precipitate the aqueous extract rich in carrageenan in ethanol at between 80 and 95%, obtaining a precipitated carrageenan;

[0050] f. Dry the precipitated carrageenan at a temperature between 40 and 60°C; g. Grind the dried carrageenan, obtaining carrageenan powder (CP). VI. A step of obtaining an organic matrix, comprising mixing in a proportion of approximately 2:2:1 the ulvane powder (UP), carrageenan powder (CP) and tucano powder (FP), respectively, forming the organic matrix (MO).

[0051] VII. A stage of obtaining the biostimulant that comprises adding a mineral matrix (MM) to the organic matrix (MO).

[0052] Where stages III, IV and V are performed simultaneously or successively in any order.

[0053]

[0012] For its part, the biostimulant useful for promoting the growth and development of plants in this application comprises at least the following elements:

[0054] a. Approximately 10% of an organic matrix, which in turn comprises approximately:

[0055] - 40% Ulvano powder (UP);

[0056] - 40% Carrageenan powder (CP); and

[0057] - 20% Fucano powder (FP).

[0058] b. Approximately 90% of a mineral matrix, which in turn comprises:

[0059] iron; potassium sulfate; magnesium sulfate; manganese sulfate; boric acid; copper sulfate; zinc sulfate; and ethylenediaminetetraacetic acid (EDTA).

[0060]

[0013] The present invention also includes the use of the aforementioned biostimulant for the preparation of a solution useful for promoting the growth and development of plants and for the preparation of a solution useful for optimizing the transplanting of seedlings.

[0061]

[0014] The present invention also comprises a method for optimizing the growth and development of plants by applying the aforementioned biostimulant, comprising applying the biostimulant by foliar application or irrigation in a proportion of 1.5 to 3 L / ha.

[0062]

[0015] The present invention also comprises a method for optimizing the transplanting of seedlings based on the biostimulant of the present application, comprising at least the following steps:

[0063] - Prepare a solution comprising 6 mL of the biostimulant per liter of water;

[0064] - Immerse the seedlings to be transplanted in the solution containing the biostimulant and keep them for at least 3 minutes; and

[0065] - Plant the seedlings.

[0066]

[0016] In this context, the technologies in this application exhibit a number of advantages.

[0067]

[0017] First, the inventors determined that the technology of the present application allows for obtaining a biostimulant that improves plant growth and development. Specifically, and as evidenced in Example No. 3, it was even demonstrated that the present technology allowed for the formulation of a biostimulant that substantially increased the number of fruits in tomato plants compared to other commercially available biostimulants.

[0068]

[0018] Now, one of the greatest advantages of the technology is that the inventors managed to optimize the production of an organic biostimulant from widely available algae, supplemented only with a mineral matrix, without harmful elements. Thus, the biostimulant produced according to this application leaves no residues of artificial products harmful to the environment in the soil. Therefore, the biostimulant of this application is environmentally friendly and does not contaminate agricultural crops.

[0069]

[0019] Another advantage of the technology in this application is that the biostimulant is produced using low-value commercial algae as raw material, since these can be obtained as surplus or waste from industrial algae processing plants. Therefore, the technology in this application allows for obtaining a product with all the properties described above, but from waste that would otherwise have no secondary use, thus promoting a circular economy.

[0070]

[0020] Finally, the method of the present application has the advantage that it allows obtaining a biostimulant with lead (Pb), cadmium (Cd), arsenic (As) and mercury (Me) below detectable limits. DESCRIPTION OF THE FIGURES

[0071]

[0021] A brief description of the figures in this application is inserted below, which form a separate part of this descriptive report.

[0072]

[0022] Figure No. 1 corresponds to a photograph that shows a pilot trial in organic tomatoes, through which a plurality of treatments were evaluated.

[0073]

[0023] Figure No. 2 corresponds to a graph that shows the effects of different treatments on the height of organic tomato plants.

[0074]

[0024] Figure No. 3 corresponds to a graph that shows the effects of different treatments on the number of fruits of organic tomato plants.

[0075]

[0025] Figure No. 4 corresponds to a graph that shows the effects of different treatments on the weight of the fruits of organic tomato plants.

[0076] DETAILED DESCRIPTION OF THE TECHNOLOGY

[0077]

[0026] This application falls within the field of agriculture and the circular economy. Specifically, the technology relates to an organic biostimulant based on algae, its production method, application, and uses.

[0078]

[0027] Different preferred embodiments of the invention will be detailed below. However, unless otherwise stated, the specific configurations, such as certain steps, raw materials, and other aspects of the invention, which constitute the respective preferred embodiments, are to be interpreted as illustrative only and not as limiting the scope of the present technology, unless otherwise stated.

[0079]

[0028] Specifically, the invention consists of a method for producing a biostimulant useful for promoting plant growth and development, based on algal polysaccharides, comprising at least the following steps:

[0080] I. A seaweed chopping stage, comprising at least the following steps:

[0081] a. Have at least the following types of algae available:

[0082] i. algae rich in ulvane (AU),

[0083] i. algae rich in carrageenan (AC), and

[0084] iii. tucano-rich algae (AF);

[0085] b. Place each type of algae in a separate container; c. Perform the following sub-steps for each type of algae:

[0086] i. Wash the seaweed;

[0087] i. Dry the washed seaweed at room temperature for 21 to 27 hours, preferably for 24 hours;

[0088] iii. Dry the algae again at a temperature between 40 and 60°C for up to 48 hours, preferably at 50°C; and

[0089] iv. Chop the dried seaweed until a particle size of no more than 1 cm is achieved, obtaining chopped seaweed for each of the types of seaweed.

[0090] II. A depigmentation stage, consisting of depigmenting each type of algae, by means of at least the following steps:

[0091] a. Immerse each type of previously chopped seaweed in ethanol at a ratio of approximately 2 L of ethanol per 100 g of chopped seaweed, forming a mixture (M1);

[0092] b. Stir the mixture (M1) for a period of 24 to 72 h;

[0093] c. Filter to obtain depigmented algae;

[0094] d. Dry the depigmented algae at room temperature for up to 48 hours.

[0095] III. An ulvane extraction stage, which in turn comprises:

[0096] a. Soak the depigmented ulvano-rich algae in water, in a ratio of approximately 100 g of algae per 2 L of water; b. Stir constantly at a temperature between 70 and 90°C for approximately 5 hours;

[0097] c. Filter until an aqueous extract rich in ulvane is obtained;

[0098] d. Concentrate the ulvane-rich extract;

[0099] e. Precipitate in ethanol at between 80 and 95%, obtaining ulvane; f. Dry the ulvane at a temperature between 40 and 60°C, until dry ulvane is obtained;

[0100] g. Grind the dry ulvane until you obtain ulvane powder (UP).

[0101] IV. A tucano extraction stage, which in turn comprises:

[0102] a. Immerse the depigmented tucano-rich algae in an approximately 2% calcium chloride solution (S1), in an approximate ratio of 100 g of algae per 500 mL of the approximately 2% calcium chloride solution (S1); b. Stir at a temperature between 70 and 100°C for approximately 5 hours; c. Filter to obtain an aqueous extract rich in tucano;

[0103] d. Concentrate the tucano-rich extract;

[0104] e. Precipitate in approximately 95% ethanol, obtaining a precipitated tucano extract;

[0105] f. Dry the precipitated tucano extract at a temperature between 40 and 60°C, until dry tucano is obtained;

[0106] g. Add to the dried tucano a solution of approximately 1 M hydrochloric acid (HCl) (S2) and subsequently add sodium hydroxide (NaOH) of approximately 1 M until the pH is adjusted to approximately 7; h. Concentrate and precipitate in ethanol, obtaining a concentrated extract of tucano;

[0107] i. Dry the concentrated tucano extract at a temperature between 40 and 60°C, obtaining tucano;

[0108] j. Grind the dried tucano, obtaining tucano powder (FP).

[0109] V. A carrageenan extraction stage, which in turn comprises:

[0110] a. Soak the depigmented carrageenan-rich algae in water at a ratio of approximately 100 g of algae per 2 L of water; b. Stir constantly at a temperature between 70 and 80°C for approximately 5 hours;

[0111] c. Filter until you obtain an aqueous extract rich in carrageenan;

[0112] d. Concentrate the aqueous extract rich in carrageenan;

[0113] e. Precipitate the aqueous extract rich in carrageenan in ethanol at between 80 and 95%, obtaining a precipitated carrageenan;

[0114] f. Dry the precipitated carrageenan at a temperature between 40 and 60°C; g. Grind the dried carrageenan, obtaining carrageenan powder (CP). VI. A step of obtaining an organic matrix, comprising mixing in a proportion of approximately 2:2:1 the ulvane powder (UP), carrageenan powder (CP) and tucano powder (FP), respectively, forming the organic matrix (MO).

[0115] VII. A stage of obtaining the biostimulant that comprises adding a mineral matrix (MM) to the organic matrix (MO).

[0116] Where stages III, IV and V are performed simultaneously or successively in any order.

[0029] The method of the present invention has a number of advantages, among which the previously mentioned advantages stand out, which are derived precisely from its particular configuration, as well as those detailed below.

[0117]

[0030] The inventors preliminarily determined that, to begin with the method, it was necessary to have algae rich in ulvan (AU), carrageenan (AC), and tucano (AF). In short, the reason is that the inventors determined that these polysaccharides have properties useful for agricultural cultivation and that this was the most suitable combination. For the purposes of this application, algae rich in ulvan (AU) are understood to be, for example, green algae and, preferably, Ulva spp.; algae rich in carrageenan (AC) are preferably Chondracanthus spp.; and algae rich in tucano (AF) are preferably Macrocystis spp.

[0118]

[0031] The inventors determined that it was essential to begin by pulverizing each type of algae separately, as this offered several advantages. For example, they determined that the parameters of the subsequent extraction stage of ulvane, carrageenan, and tucano, respectively, depended substantially on the properties of each algae or product to be extracted. Therefore, while an expert might have been tempted to perform all the washing, chopping, pulverizing, and extraction processes together to simplify the method, the inventors determined that it was advisable to perform them separately anyway, in order to optimize the properties of the resulting biostimulant. Thus, the processes described below depend on the type of algae being processed.

[0119]

[0032] Regarding the specific operational aspects of the seaweed chopping stage, these were also specifically defined to optimize the properties of the resulting product.

[0120]

[0033] For example, washing the algae is relevant because it is the first step in the process designed to reduce unwanted impurities and purify the extracts to be added. If this step is not carried out, for example, the resulting biostimulant has a very high salt content that generates undesirable effects in the plants.

[0121]

[0034] Regarding the drying process, the inventors determined that it was convenient to do it in 2 sub-stages: the first at room temperature and the second in an oven.

[0122]

[0035] Regarding drying at room temperature, the inventors determined that it was advisable to carry it out for 21 to 27 hours, depending on the environmental conditions. Thus, typically, in a temperate climate (such as that of the coast of the Coquimbo Region in Chile), drying at room temperature should be carried out for approximately 24 hours. This drying stage has the advantage of reducing energy consumption and, consequently, the costs associated with the process, making it more environmentally sustainable. Additionally, the inventors determined that, in a preferred modality, it was advisable for this drying to be carried out in the shade, since a slow drying rate allows for more uniform water removal. Furthermore, the inventors determined that carrying out this drying in the shade was advisable to preserve its bioactive compounds, minimizing the risk of modifying or decomposing structurally complex compounds.

[0123]

[0036] The subsequent oven drying step, lasting up to 48 hours at a temperature between 40 and 60°C, is relevant because it allows the algae to reach a moisture content of no more than 10%, which the inventors determined to be optimal for proceeding to the next stage. Thus, the specific time and temperature of the oven drying will depend on the moisture content at the end of the ambient temperature drying process. However, the inventors determined that, preferably, this process should be carried out for more than 24 hours. This oven drying stage offers the advantage of removing as much water as possible from the algae, allowing for the precise determination of the dry-basis yield of polysaccharides during the final stage.

[0124]

[0037] Finally, regarding the seaweed chopping stage, reducing the particle size to 500 µm µm µm is relevant because it facilitates obtaining a higher yield in the extraction of the polysaccharide and simplifies the filtration process by separating the seaweed from the polysaccharide obtained.

[0125]

[0038] Regarding the depigmentation stage, it is also carried out independently for each type of algae because, as previously explained, this allows for a specific extraction stage for each type of extract. Furthermore, the inventors determined that this stage should be after, not before, the chopping stage, as this reduces the time and resources required.

[0126]

[0039] As for the specific operational aspects of the depigmentation stage, these were also specifically defined to optimize the properties of the resulting product.

[0127]

[0040] Immersing the chopped algae in ethanol at a ratio of approximately 2 L of ethanol per 100 g of algae to form mixture (M1) also contributes to the purification of the extract used and, therefore, the purity of the biostimulant. Furthermore, the inventors determined that the aforementioned concentrations made the process efficient, based on the previously described steps. For the purposes of this paragraph, this represents a difference of approximately ±50 mL per 100 g of algae. Alternatively, this step can be carried out using an 80% ethanol solution at 70°C to further facilitate the depigmentation process.

[0128]

[0041] For its part, keeping the aforementioned mixture in constant agitation is relevant because it allows for improved contact of the algae with the solvent.

[0129]

[0042] The subsequent filtration step is relevant to remove excess liquid before continuing with the process. In one embodiment of the technology, filtration is carried out using meshes smaller than 500 µm, which allows for the efficient separation of the algae from the solvent used.

[0130]

[0043] Finally, regarding depigmentation, drying the filtered product is essential for proper subsequent extraction. The inventors noted that one of the advantages of using ethanol for depigmentation is that it reduces the drying time, which can be as short as 5 hours, depending on environmental conditions. In one embodiment of the present invention, drying is carried out until the powder's moisture content is no more than 10%.

[0131]

[0044] Subsequently, the respective extraction stages for each of the extracts are carried out, starting from the depigmented algae. In particular, the specific operational parameters of each of the processes were adjusted to improve the extraction, depending on the type of extract.

[0132]

[0045] Regarding the ulvane extraction stages, the process begins by immersing the depigmented ulvane-rich algae in water, at a ratio of approximately 100 g of algae per 2 L of water. This represents a margin of error of approximately 10 g of algae per 2 liters of water. Constant agitation is preferably carried out at 80°C.

[0133]

[0046] As for the concentration sub-step, this can be carried out, for example, with a rotary evaporator at 60 °C, reducing the volume to approximately one-third of the initial volume.

[0047] As for the ethanol precipitation, an approximate ratio of 1:3 of extract to ethanol solution is preferably used (for these purposes, this represents a difference of approximately 0.1 parts of extract). In short, the extraction step allows for obtaining a substantially more purified ulvan powder and manufacturing a more effective biostimulant.

[0134]

[0048] Regarding the tucano extraction stage, it differs from the ulvano extraction, for example, because it involves immersing the depigmented powder in a calcium chloride solution of approximately 2% (where approximately represents a difference of plus or minus 0.1%). Specifically, the inventors determined that the optimal ratio of depigmented algae to calcium chloride solution was approximately 100 g of algae powder per 500 mL of the 2% calcium chloride solution, where approximately represents a difference of plus or minus 10 g of algae.

[0135]

[0049] Regarding the agitation for approximately 5 hours in the tucano extraction stage, the term approximately implies a difference of plus / minus 10 minutes.

[0136]

[0050] Regarding the Tucano concentration sub-stage, this can be carried out, for example, in a rotary evaporator at a temperature between 50 and 60°C. The concentration in this stage is preferably carried out until the product is concentrated to a volume of approximately one-third of the initial volume.

[0137]

[0051] As for the precipitate in ethane at approximately 95%, it represents a difference of plus or minus 2%.

[0138]

[0052] Another particularly distinctive aspect of the tucano extraction stage compared to that of ulvano is the step of adding to the dry tucano an approximately 1 M hydrochloric acid (HCl) solution (S2) and subsequently adding approximately 1 M sodium hydroxide (NaOH) until the pH is adjusted to approximately 7. In this respect, the term approximately, both in relation to the molarity of the solutions and to the pH, implies a difference of plus / minus 0.1.

[0139]

[0053] Furthermore, the extraction of the tucano stands out because, after the step described above, a new concentration and precipitation process is carried out, this time in ethanol.

[0054] In both drying steps of the tucano extraction stage, the temperature is preferably 50°C.

[0140]

[0055] Finally, in relation to the extraction processes, the one used to obtain carrageenan also has its own particularities.

[0141]

[0056] For example, regarding the step of immersing the depigmented algae in water, at a ratio of approximately 100 g of algae per 2 L of water, the term "approximately" implies a difference of plus or minus 10 g of algae. Regarding the step of constantly stirring at a temperature between 70 and 80°C for approximately 5 hours, the term "approximately" implies a difference of plus or minus 10 minutes.

[0142]

[0057] After extracting the three extracts and preparing the corresponding powders, the organic matrix (OM) is obtained by mixing them. The inventors determined that achieving a mixture with an approximate ratio of ulvano powder (UP): carrageenan powder (CP): tucano powder (FP) 2:2:1 is key at this stage, as they determined that this ratio allowed for the formulation of an optimal biostimulant. For the purposes of this paragraph, this approximate ratio represents a difference of 0.1 for each component.

[0143]

[0058] The mixing of the aforementioned extracts can be carried out by means of different techniques available in the state of the art for such purposes.

[0144]

[0059] Finally, the corresponding step of adding the mineral matrix (MM) is also fundamental, as it allows the biostimulant formulation to be completed. In a preferred embodiment of the invention, the matrix (MM) comprises the following elements: iron; potassium sulfate; magnesium sulfate; manganese sulfate; boric acid; copper sulfate; zinc sulfate; and ethylenediaminetetraacetic acid (EDTA). In a preferred embodiment, the aforementioned elements are in an approximate proportion as indicated below:

[0145] - 36.5% iron;

[0146] - 21% potassium sulfate;

[0147] - 8% magnesium sulfate;

[0148] - 0.3% manganese sulfate;

[0149] - 0.08% boric acid;

[0150] - 0.06% copper sulfate;

[0151] - 0.06% zinc sulfate; and - 34% ethylenediaminetetraacetic acid (EDTA).

[0152]

[0060] For the purposes of this formulation, approximately means a difference of 5% from the value. For example, 32% means a range of 32% plus or minus 1.6%. When the term “approximately” is used in relation to the formulation below, the interpretation of the concept should be the same.

[0153]

[0061] In another embodiment, the invention also comprises a biostimulant, preferably produced according to the method described above, which comprises at least the following elements:

[0154] a. Approximately 10% of an organic matrix, which in turn comprises approximately:

[0155] - 40% Ulvano powder (UP);

[0156] - 40% Carrageenan powder (CP); and

[0157] - 20% Fucano powder (FP).

[0158] b. Approximately 90% of a mineral matrix, which in turn comprises: iron;

[0159] potassium sulfate; magnesium sulfate; manganese sulfate; boric acid; copper sulfate; zinc sulfate; and ethylenediaminetetraacetic acid (EDTA).

[0160]

[0062] In a preferred embodiment of the invention, the mineral matrix (MM) comprises approximately the following percentages of each of the indicated elements:

[0161] - 36.5% iron;

[0162] - 21% potassium sulfate;

[0163] - 8% magnesium sulfate;

[0164] - 0.3% manganese sulfate;

[0165] - 0.08% boric acid;

[0166] - 0.06% copper sulfate;

[0167] - 0.06% zinc sulfate; and

[0168] - 34% Ethylenediaminetetraacetic acid (EDTA).

[0169]

[0063] The inventors determined that supplementing the organic matrix (OM) with the mineral matrix (MM) allowed for the optimization of the biostimulant, by adjusting it to the requirements of the target crops, for example, tomatoes.

[0064] In another embodiment, the present invention comprises the use of the aforementioned biostimulant for the preparation of a solution useful for promoting plant growth and development. Indeed, the biostimulant can be dissolved for administration via different routes. A particularly efficient use, as indicated above, is in tomatoes, as shown in Example No. 3.

[0170]

[0065] The inventors also determined that, in another embodiment, the biostimulant of this application is useful for preparing a solution to optimize the transplanting of seedlings. More specifically, in a preferred embodiment, the use of the biostimulant in seedlings involves preparing a solution comprising 6 mL of biostimulant per liter of water and, even more preferably, immersing the seedlings in this solution for 3 minutes before transplanting.

[0171]

[0066] In another modality, it comprises a method to optimize the growth and development of plants by applying the biostimulant described above, which comprises applying the biostimulant via foliar application or irrigation in a proportion of 1.5 to 3 L / ha.

[0172]

[0067] Finally, in another embodiment, the invention comprises a method for transplanting seedlings in agricultural crops by applying the biostimulant described above, comprising at least the following steps:

[0173] - Prepare a solution comprising 6 mL of the biostimulant per liter of water;

[0174] - Immerse the seedlings to be transplanted in the solution containing the biostimulant and keep them for at least 3 minutes; and

[0175] - Plant the seedlings.

[0176] EXAMPLES

[0177]

[0068] The method will be better understood by means of the following examples, which are merely illustrative and do not limit the scope of this application. Several changes and modifications to the described modalities would be evident to those skilled in the art, and such changes can be made without departing from the spirit of the technology and the scope of the appended claims.

[0069] It should be noted beforehand that, for the purpose of designing the method of the present invention, the inventors carried out a rigorous research and development process, which is shown in the examples below.

[0178] EXAMPLE No. 1: Preferred embodiment of the method of the invention.

[0179]

[0070] For the development of the biostimulant referred to in this application, the inventors carried out an arduous process of research, development, trial and error by means of which they systematically evaluated different modifications to the method, in order to identify a process with optimal ranges that would allow them to obtain a product with the desired properties.

[0180]

[0071] During the development process, they carried out the process described below:

[0181] I. They carried out a seaweed chopping stage, which comprised the following steps:

[0182] a. The following algae were available: Ulva, Chondracanthus, and Macrocystis.

[0183] b. Each type of algae was placed in a different container.

[0184] c. In each of the containers, the following steps were performed:

[0185] i. The seaweed was washed;

[0186] i. The washed algae were dried at low ambient temperature for 24 hours; ii. Drying was continued in an oven at a temperature of approximately 50°C; and

[0187] iv. The dried algae were chopped to a particle size ranging from approximately 500 µm to 1 cm. This yielded chopped algae (PA) for each type of algae.

[0188] II. Then they depigmented the algae (separately), using the following steps:

[0189] a. Each type of previously chopped algae was immersed in ethanol at a ratio of approximately 2 L of ethanol per 100 g of chopped algae, forming a mixture (M1);

[0190] b. The mixture (M1) was stirred for approximately 2 days;

[0191] c. It was filtered to obtain depigmented algae; d. the depigmented algae were dried at room temperature for approximately 48 hours.

[0192] III. They then carried out an ulvane extraction stage, which comprised the following steps:

[0193] a. The depigmented Ulva algae were submerged in water, in a proportion of approximately 100 g of algae per 2 L of water; b. They were constantly stirred at a temperature ranging between 70 and 80°C for approximately 5 hours;

[0194] c. They filtered until they obtained an aqueous extract rich in ulvane;

[0195] d. They concentrated the ulvane-rich extract;

[0196] e. They precipitated in ethanol at between 95% and ethanol, obtaining ulvane;

[0197] f. They dried the ulvane at a temperature of 50°C, until they obtained dry ulvane;

[0198] g. They ground the dried ulvane until they obtained ulvane powder (UP).

[0199] IV. In parallel, they carried out a tucano extraction stage, which included:

[0200] a. Immerse the depigmented Macrocystis algae in a 2% calcium chloride solution (S1), in an approximate ratio of 100 g of algae per 500 mL of the 2% calcium chloride solution (S1);

[0201] b. Stir at a temperature between 70 and 100°C for approximately 5 hours;

[0202] c. Filter until you obtain an aqueous extract rich in tucano;

[0203] d. Concentrate the tucano-rich extract;

[0204] e. Precipitate in 95% ethanol to obtain a precipitated tucano extract; f. Dry the precipitated tucano extract at a temperature of approximately 50°C, until dry tucano is obtained;

[0205] g. Add to the dried tucano a solution of approximately 1 M hydrochloric acid (HCl) (S2) and subsequently add sodium hydroxide (NaOH) approximately 1 M until the pH is adjusted to approximately 7;

[0206] h. Concentrate and precipitate in ethanol, until a concentrated extract of tucano was obtained; i. Dry the concentrated extract of tucano at a temperature between 40 and 60°C, until tucano was obtained;

[0207] j. Grind the tucano, obtaining tucano powder (FP).

[0208] V. They also carried out a carrageenan extraction stage in parallel, which included:

[0209] a. Soak the depigmented Chondracanthus algae in water at a ratio of approximately 100 g of algae per 2 L of water;

[0210] b. Stir constantly at a temperature between 70 and 80°C for approximately 5 hours;

[0211] c. Filter until you obtain an aqueous extract rich in carrageenan;

[0212] d. Concentrate the aqueous extract rich in carrageenan;

[0213] e. Precipitate the aqueous extract rich in carrageenan in 95% ethanol, until precipitated carrageenan is obtained;

[0214] f. Dry the precipitated carrageenan at a temperature between 40 and 60°C;

[0215] g. Grind the dry carrageenan until they obtained carrageenan powder (CP).

[0216] VI. They then carried out a stage of obtaining an organic matrix, which consisted of mixing the ulvane powder (UP), carrageenan powder (CP) and tucano powder (FP) in a proportion of approximately 2:2:1, respectively, forming the organic matrix (MO).

[0217] - Finally, they added a mineral matrix (MM) to the organic matrix (OM), which had been previously prepared with the following: 32% iron;

[0218] - 20% potassium sulfate;

[0219] - 7% magnesium sulfate;

[0220] - 0.2% manganese sulfate;

[0221] - 0.02% boric acid;

[0222] - 0.01% copper sulfate;

[0223] - 0.01% zinc sulfate; and

[0224] - 30.7% ethylenediaminetetraacetic acid (EDTA).

[0072] The inventors determined that the process described above was optimal for the production of the biostimulant.

[0225] EXAMPLE #2: Evaluation of the presence of heavy metals in the organic biostimulant obtained according to the process described in Example #1.

[0226]

[0073] One of the main challenges associated with the development of a biostimulant is ensuring that the resulting product has acceptable levels of heavy metals. In this case, the inventors' objective is that the biostimulant of the present invention does not contain lead (Pb), cadmium (Cd), arsenic (As), or mercury (Me) within detectable ranges. That is to say, if it does contain these metals, their concentration should be less than 1 mg / kg.

[0227]

[0074] In this context, and to evaluate whether the biostimulant produced according to the method described in Example No. 1 met these characteristics, an analysis was performed to assess the presence of minerals in it. Specifically, the analysis was carried out by Activation Labs. The results obtained indicate that BIOALG is composed of N (27 mg / L), Fe (215 mg / L), Zn (0.36 mg / L), B (5.3 mg / L), Mn (11.5 mg / L), Cu (2.92 mg / L), Ca (21.0 mg / L), Mg (58 mg / L), Na (136 mg / L), and K (738 mg / L).

[0228]

[0075] In this way, the inventors were able to verify that the production method had made it possible to eliminate heavy metals, so that the biostimulant had no detectable levels of lead (Pb), cadmium (Cd), arsenic (As) and mercury (Me).

[0229]

[0076] Likewise, the aforementioned analysis made it possible to verify that the biostimulant had a pH between 6.5 and 7.0.

[0230] EXAMPLE #3: Evaluation in organic tomatoes at pilot level.

[0231]

[0077] Following the characterization of the formulated biostimulant according to the method described in Example No. 1 (referred to as “Bio A” in this example), the inventors conducted a series of trials on vegetables, including the organic tomato experiment described below. The objective of this trial was to evaluate some agronomic variables of commercial interest by comparing different treatments on organic tomatoes.

[0232]

[0078] Experimental design of tomato trials.

[0079] For the present trial, 25 organic tomato plants were used, arranged in 5 rows of 5 plants each, as shown in Figure No. 1. A drip irrigation system was installed that allowed all plants to be watered under equal conditions.

[0233]

[0080] From the 25 plants, 5 groups of tomatoes were formed for the random application of 5 different treatments, which are detailed in Table No. 1 inserted below.

[0234] Table No. 1: Treatments applied to organic tomatoes

[0235]

[0236]

[0081] Where Stimplex® corresponds to a commercial biostimulant manufactured by the company ANASAC that is derived from Ascophyllum nodosum, which was used as a positive control in this trial.

[0237]

[0082] The fertilization of each of the tomato groups with the treatments indicated in Table No. 1 was carried out via foliar application every 21 days from the date of transplanting, applying 15 mL of solution per plant, each formulated according to the dose indicated in Table No. 1.

[0238]

[0083] Results of trials in tomatoes.

[0239]

[0084] After the trial was completed, plant height, fruit quantity, and fruit weight were evaluated according to treatment. The main results are as follows:

[0240]

[0085] Regarding plant height, although the differences were not statistically significant, it was observed that the plants treated with T2 were taller. Furthermore, it was found that the formulations based on the Bio A biostimulant of this application exhibited a similar effect to the positive control, even at lower doses. The results are shown in Figure 2.

[0086] Regarding the number of fruits, the results were even more surprising. Treatment T2, that is, the one formulated with the Bio A biostimulant of this application, consistently yielded a greater number of fruits than the positive control (T1).While the number of fruits resulting from treatments T3 and T4 was lower than that of the positive control (T1), a positive effect was observed (T1) compared to the negative control (T5), even though the dose was substantially lower. Therefore, all treatments based on the Bio A biostimulant in this application showed positive effects in terms of the number of fruits. The results are shown in Figure 3.

[0241]

[0087] Finally, regarding the average fruit weight, the results were also surprising, demonstrating that, although the differences were not statistically significant, treatment T2, namely the one formulated with the Bio A biostimulant from this application, resulted in a higher average weight than the positive control (T1). Thus, despite the increased number of fruits, T2 maintained a weight similar to that of the positive control and substantially higher than that of the negative control. The results are shown in Figure 4.

Claims

LIST OF CLAIMS 1. A method for producing a biostimulant useful for promoting plant growth and development, based on algae, CHARACTERIZED in that it comprises at least the following steps: I. A seaweed chopping stage, comprising at least the following steps: a. Have at least the following types of algae available: i. algae rich in ulvane (AU), i. algae rich in carrageenan (AC), and iii. tucano-rich algae (AF); b. Place each type of seaweed in a separate container; c. Carry out the following sub-steps for each type of algae: i. Wash the seaweed; i. Dry the washed seaweed at room temperature for 21 to 27 hours, preferably for 24 hours; iii. Dry the algae again at a temperature between 40 and 60°C for up to 48 hours, preferably at 50°C; and iv. Chop the dried algae until a particle size of no more than 1 cm is achieved until chopped algae is obtained for each of the types of algae. II. A depigmentation stage, consisting of depigmenting each type of algae, by means of at least the following steps: a. Immerse each type of previously chopped seaweed in ethanol at a ratio of approximately 2 L of ethanol per 100 g of chopped seaweed, forming a mixture (M1); b. Stir the mixture (M1) for a period of 24 to 72 h; c. Filter to obtain depigmented algae; d. Dry the depigmented algae at room temperature for up to 48 hours. III. An ulvane extraction stage, which in turn comprises: a. Immerse the depigmented ulvano-rich algae in water, in a ratio of approximately 100 g of algae per 2 L of water; b. Stir constantly at a temperature between 70 and 90°C for approximately 5 hours; c. Filter until an aqueous extract rich in ulvane is obtained; d. Concentrate the ulvane-rich extract; e. Precipitate in ethanol at between 80 and 95%, obtaining ulvane; f. Dry the ulvane at a temperature between 40 and 60°C, until dry ulvane is obtained; g. Grind the dried ulvano to obtain ulvano powder (UP). IV. A tucano extraction stage, which in turn comprises: a. Immerse the depigmented tucano-rich algae in an approximately 2% calcium chloride solution (S1), in an approximate ratio of 100 g of algae per 500 mL of the approximately 2% calcium chloride solution (S1); b. Stir at a temperature between 70 and 100°C for approximately 5 hours; c. Filter until an aqueous extract rich in tucano is obtained; d. Concentrate the extract rich in tucano; e. Precipitate in approximately 95% ethanol, obtaining a precipitated tucano extract; f. Dry the precipitated tucano extract at a temperature between 40 and 60°C, until dry tucano is obtained; g. Add to the dried tucano a solution of approximately 1 M hydrochloric acid (HCl) (S2) and subsequently add sodium hydroxide (NaOH) approximately 1 M until the pH is adjusted to approximately 7; h. Concentrate and precipitate in ethanol, obtaining a concentrated extract of tucano; i. Dry the concentrated tucano extract at a temperature between 40 and 60°C, obtaining tucano; j. Grinding the dried tucano, obtaining tucano powder (FP). V. A carrageenan extraction stage, which in turn comprises: a. Soak the depigmented carrageenan-rich algae in water at a ratio of approximately 100 g of algae per 2 L of water; b. Stir constantly at a temperature between 70 and 80°C for approximately 5 hours; c. Filter until an aqueous extract rich in carrageenan is obtained; d. Concentrate the aqueous extract rich in carrageenan; e. Precipitate the aqueous extract rich in carrageenan in ethanol at between 80 and 95%, obtaining a precipitated carrageenan; f. Dry the precipitated carrageenan at a temperature between 40 and 60°C; g. Grind the dry carrageenan, obtaining carrageenan powder (CP). VI. A step of obtaining an organic matrix, comprising mixing in a proportion of approximately 2:2:1 the ulvane powder (UP), carrageenan powder (CP) and tucano powder (FP), respectively, forming the organic matrix (MO). VII. A stage of obtaining the biostimulant that comprises adding a mineral matrix (MM) to the organic matrix (MO). Where stages III, IV and V are performed simultaneously or successively in any order.

2. The method according to claim 1, CHARACTERIZED in that the algae rich in ulvane (AU) are Ulva spp.

3. The method according to claim 1, CHARACTERIZED in that the carrageenan (AC) rich algae are Chondracanthus spp.

4. The method according to claim 1, CHARACTERIZED in that the tucano-rich algae (AF) are Macrocystis spp.

5. The method according to claim 1, CHARACTERIZED in that the mineral matrix (MM) comprises the following elements: iron; potassium sulfate; magnesium sulfate; manganese sulfate; boric acid; copper sulfate; zinc sulfate; and ethylenediaminetetraacetic acid (EDTA).

6. The method according to claim 5, CHARACTERIZED in that the mineral matrix comprises the elements in approximately the following proportions: - 36.5% iron; - 21% potassium sulfate; - 8% magnesium sulfate; - 0.3% manganese sulfate; - 0.08% boric acid; - 0.06% copper sulfate; - 0.06% zinc sulfate; and - 34% Ethylenediaminetetraacetic acid (EDTA).

7. A biostimulant useful for promoting plant growth and development, CHARACTERIZED in that it comprises at least the following elements: a. Approximately 10% of an organic matrix, which in turn comprises approximately: - 40% Ulvano powder (UP); - 40% Carrageenan powder (CP); and - 20% Fucano powder (FP). b. Approximately 90% of a mineral matrix, which in turn comprises: iron; potassium sulfate; magnesium sulfate; manganese sulfate; boric acid; copper sulfate; zinc sulfate; and ethylenediaminetetraacetic acid (EDTA).

8. The biostimulant according to claim 7, CHARACTERIZED in that the mineral matrix comprises the elements in approximately the following proportions: - 36.5% iron; - 21% potassium sulfate; - 8% magnesium sulfate; - 0.3% manganese sulfate; - 0.08% boric acid; - 0.06% copper sulfate; - 0.06% zinc sulfate; and - 34% of Ethylenediaminetetraacetic acid (EDTA).

9. Use of a biostimulant produced according to claim 7, CHARACTERIZED in that it serves for the preparation of a solution useful for promoting the growth and development of plants.

10. Use of a biostimulant produced according to claim 7, CHARACTERIZED in that it serves for the preparation of a solution useful for optimizing the transplanting of seedlings.

11. Use according to claim 11, CHARACTERIZED in that the solution is prepared so as to comprise 6 mL of biostimulant per liter of water.

12. Use according to claim 12, CHARACTERIZED in that the solution is used in such a way that the seedlings are immersed in it for 3 minutes before being transplanted.

13. A method for optimizing plant growth and development by applying a biostimulant according to claim 7, CHARACTERIZED in that it comprises applying the biostimulant by foliar application or irrigation in a proportion of 1.5 to 3 L / ha.

14. A method for optimizing the transplanting of seedlings in agricultural crops based on the biostimulant of claim 7, CHARACTERIZED in that it comprises at least the following steps: - Prepare a solution comprising 6 mL of the biostimulant per liter of water; - Immerse the seedlings to be transplanted in the solution containing the biostimulant and keep them for at least 3 minutes; and - Plant the seedlings.

Images