Formulation and method for preventing cracking or splitting in fruit

Abstract

The present invention relates to a formulation and a method for preventing or reducing cracking (splitting) in fruit. The formulation includes at least rhamnolipids, citric acid, boric acid, calcium chloride, gibberellic acid, chitosan, castor oil and water. Applying this formulation to fruit significantly decreases the occurrence of cracking, thus providing an effective and efficient solution to prevent or reduce damage of this type on the surface of fruit.

Description

[0001] FORMULATION AND METHOD TO PREVENT SPLITTING OR CRACKLING IN FRUIT TECHNICAL FIELD

[0002] The present invention relates to the technical field of agriculture and the agri-food industry. In particular, it relates to a new formulation to prevent splitting or cracking in fruits, preferably stone fruits, berries, and others.

[0003] BACKGROUND

[0004] One of the factors that significantly affects the production, quality, and marketing of cherries, especially in the south-central regions of Chile, is the appearance of cracks or splitting in the fruit's skin. This physiological phenomenon occurs due to water entering the fruit in various ways, but primarily through the cuticle. This process is exacerbated by rainfall near harvest time or by the condensation of water vapor when the fruit is packaged in modified atmospheres for export. This problem can have a considerable economic impact on producers, as some varieties can experience losses of up to 90% of production, representing millions of dollars in losses and thousands of tons discarded.This problem is intensified by current climate change-related issues that cause late spring rains and also negatively affect the national export industry, as cherries lose their commercial value in the fresh export market.

[0005] To reduce cracking in cherries, various strategies have been implemented, such as foliar application of calcium solutions to decrease osmotic potential, use of fixed or mobile plastic covers to protect the fruit from rain, application of phytohormones, or application of hydrophobic compounds and waxes by spraying. However, the effectiveness of these methods has been inconsistent: some have achieved only a minimal reduction in cracking, while the most effective methods involve high implementation costs. The massive losses caused by the decrease in cherry quality due to adverse weather conditions, such as late spring rains, result in low final productivity, which has led to the development of new strategies and practices to reduce cracking and, consequently, economic losses.

[0006] One strategy that promises high effectiveness is the use of plastic covers to completely protect cherry crops and prevent direct contact between rainwater and the fruit. While effective, its current cost ranges from $25,000 to $30,000 per hectare, with a lifespan of 3-5 years. This high initial investment makes it an alternative that few producers can afford, thus limiting its widespread use. Furthermore, studies have linked this strategy to problems with fruit firmness, potentially leading to quality losses and hindering marketing and export [Simon, G. (2006). Review on rain induced fruit cracking of sweet cherries (Prunus avium L), its causes and the possibilities of prevention. International Journal of Horticultural Science, 12(3), 27-35].

[0007] Another widely used methodology is the application of mineral salts, which aims to reduce water absorption by the fruit through osmosis. This creates a higher mineral concentration outside the cherry, preventing water from entering. Among the most commonly used mineral salts are those derived from calcium, such as calcium chloride (CaCl₂) and calcium hydroxide (Ca(OH)₂) [Demirsoy, LK, & Bilgener, S. (1998). The effects of preharvest calcium hydroxide applications on cracking in 'Ziraat', 'Lambert' and 'Van' sweet cherries. In III International Cherry Symposium 468, pp. 657-662; Lang, G., Guimond, C., Southwick, S., Kappel, F., Flore, JA, Facteau, T., & Azarenko, A. (1997). Performance of calcium / sprinkler-based strategies to reduce sweet cherry raincracking. In III International Cherry Symposium 468, pp.649-656], in addition to playing a fundamental role in maintaining the structural integrity and firmness of the fruit cell wall due to decreased permeability, thus reducing the rate of water absorption [Christensen, JV (1972). Cracking in cherries: IV. Physiological studies of the mechanism of cracking. Acta Agriculturae Scandinavica, 22(3), 153-162; Sekse, L. (1995). Cuticular fracturing in fruits of sweet cherry (Prunus avium L.) resulting from changing soil water contents. Journal of Horticultural Science, 70(4), 631-635]. This methodology, despite being economical and accessible, has a completely insignificant effect against large quantities of water, so in most cases it needs to be complemented with other methods such as plastic covers for its effectiveness, thus increasing its total investment value for crop protection.Furthermore, there are studies that have linked its use to effects of decreasing fruit size [Facteau, TJ, Rowe, KE, & Chestnut, NE (1987). Response of Bing and Lambert sweet cherry fruit to preharvest calcium chloride applications. HortScience, 22(2): 271-273].

[0008] On the other hand, there are commercial products to prevent cracking such as Premio® (Syngenta Biologicals), Parka® (Cultiva LLC), Crackguard® (Nutriprove SA), Fartum® Anticracking (Patagonia Biotecnología SA), RainGard® (Syngenta), among others, which are currently used to prevent cracking by generating a hydrophobic film through compounds that limit both the absorption and loss of water, reducing transpiration in the fruit [Correia, S., Schouten, R., Silva, AP, & Gongalves, B. (2018). Sweet cherry fruit cracking mechanisms and prevention strategies: A review. Scientia Horticulturae, 240, 369-377]. Premio® is a product of plant origin that increases the elasticity and permeability of the cell wall; it also contains mineral macronutrients that promote increased fruit size, uniformity of caliber, and coloration.Parka® (US patent 8,752,328 B2), for its part, forms a phospholipid biofilm on the cherry cuticle that prevents external damage caused by rain, improving the commercial yield and shelf life of the cherries. RainGard® is a product based on neutralized fatty acids that generate a transparent film that physically protects the cherries from water penetration, while also balancing the cherry tree's physiological processes in response to environmental demands. Fartum® Anticracking is an algae-based fertilizer and biostimulant that strengthens the cell wall, providing greater mechanical resistance to the cuticle, while also increasing the presence of minerals within the fruit, allowing for more soluble solids and reducing water content to prevent cracking.Crackguard® is a hydrophobic biofilm to prevent or control cracking in cherries, table grapes and blueberries composed of a mixture of phospholipids and natural plant organic acids.

[0009] There are also other technologies aimed at solving the same technical problem, such as the one described in US patent 11,737,454 B2, which presents a formulation and methods for forming an exogenous flexible film on a plant, particularly on the fruit, where this formulation includes a concentrated solution comprising: a) a solvent comprising 58 to 70% by weight of a concentrated solution, where the solvent includes water; b) a film-forming matrix component comprising 0.8 to 2.2% by weight of the concentrated solution, selected from compounds including chitosan; c) a hydrophobic barrier component comprising 12 to 25% by weight of the concentrated solution, selected from various oils or phenols; d) a plasticizing component comprising 4 to 15% by weight of the concentrated solution;(e) a film-enhancing component comprising between 8 and 18% by weight of the concentrated solution.;

[0010] The invention patent CN 103766474 B discloses a biological preservative for cherry and tomato fruits and its use, where each liter of preservative formulation contains 10 10 -10 12 Rhodotorula glutinis strain ZJU11 cells and between 400-600 mg of rhamnolipid containing 70-90% glycolipids.

[0011] US patent application 2006 / 0084577 A1 presents a method and a compound for preventing cracking of fruits and vegetables by means of a formulation containing an indole derivative that may include: tryptophan, auxin, an auxin derivative or naphthalene, and may be applied to any fruit or vegetable plant including tomatoes, drupes, pome fruits, cherries or grapes.

[0012] Patent application WO 2014 / 153210 A1 presents compositions comprising a cellulose nanomaterial and an inorganic salt component, which are useful for forming edible coatings or films on plants or parts thereof.

[0013] Despite existing commercial technologies and state-of-the-art developments, there remains a need for formulations to prevent cracking or splitting in fruit, particularly cherries. Therefore, formulations are required that are more efficient, more affordable, offer greater durability, and do not negatively affect other fruit characteristics such as size, color, etc.

[0014] SUMMARY OF THE INVENTION

[0015] A first object of the present invention relates to a formulation for preventing or reducing splitting or cracking in fruits comprising a solution of rhamnolipids, citric acid, boric acid, calcium chloride, gibberellic acid, a chitosan solution, castor oil and water.

[0016] In one embodiment of the present invention, the formulation is an emulsion comprising a rhamnolipid solution at a concentration between 0.1% and 10% v / v; citric acid at a concentration between 0.01% and 3% w / v; boric acid at a concentration between 0.1% and 5% w / v; calcium chloride at a concentration between 1% and 10% w / v; gibberellic acid at a concentration between 0.01% and 0.8% w / v; a 4% w / v chitosan solution at a concentration between 0.1% and 5% v / v; castor oil at a concentration between 0.1% and 5% v / v; and water q.s.

[0017] In one embodiment of the present invention, chitosan has a molecular weight range between 150,000 and 300,000 g / mol, and a degree of deacetylation greater than or equal to 85%.

[0018] Optionally, the formulation of the present invention comprises phosphoric acid in a concentration between 0.1% and 5% w / v.

[0019] In another embodiment of the present invention, the formulation is diluted between 0.1 - 5% v / v for use or application on the fruit.

[0020] A second object of the present invention relates to a method for preventing or reducing splitting or cracking in fruit, comprising a first step of diluting a formulation as described above in any of its forms, and applying the diluted formulation to the fruit. Preferably, the formulation is diluted to a concentration between 0.1 and 5% v / v before being applied to the fruit.

[0021] In one embodiment of the present invention, the formulation is applied to the fruit by spraying or immersion. In another embodiment of the present invention, the formulation is applied to drupe-type fruits (stone fruits) or berries. In a preferred embodiment of the invention, the fruits are cherries, grapes, plums, peaches, sour cherries, tomatoes, and blueberries.

[0022] A third object of the present invention relates to a method of preparing the formulation according to any of the embodiments described above, comprising the steps of: i) dissolving calcium chloride, boric acid, and citric acid in water to obtain a solution; ii) diluting gibberellic acid in water and adding it to the solution obtained in step i; and iii) adding to the solution obtained in step i) a solution of chitosan, castor oil, and a rhamnolipid solution and mixing to obtain the formulation; wherein all steps are carried out under constant stirring.

[0023] BRIEF DESCRIPTION OF THE FIGURES FIG. 1: Graph with the reported post-harvest results of productivity recorded in kilograms versus the total percentage of cracking observed with the treatments with each formulation (1, 2, 3, 4 and 5), control (without treatment) and commercial product (Crackguard®) for the early variety of cherries Santina.

[0024] FIG. 2: Graph showing the results of crack induction (% cracking) for each of the formulations (1, 2, 3, 4, and 5) and control (no treatment) using the water immersion test for the early cherry variety Santina over a period of 20 hours.

[0025] FIG. 3: Graph showing the results of crack induction (% cracking) with the treatment with each of the formulations (1, 2, 3, 4 and 5), control (without treatment) and commercial product (Crackguard®) by means of the water immersion test for the early variety of Lapins cherries during a period of 20 hours.

[0026] FIG. 4: Photographs of the splitting results after the water immersion test for a period of 20 hours for the Lapins (first and third photograph) and Santina (second photograph) cherry varieties.

[0027] FIG. 5: Graph showing the reported post-harvest results of productivity in kilograms recorded versus the total percentage of cracking (% splitting) observed in all cherries of the early variety Santina. Formulation 5, Control (no treatment), Commercial Product: Crackguard®.

[0028] FIG. 6: Photographs of the post-harvest states of the early variety Santina cherries for the 3 trial conditions. FIG. 6A: Application of Formulation 5, FIG. 6B: Control (no treatment), FIG. 6C: Application of Commercial Product (Crackguard®).

[0029] FIG. 7: Photographs of the post-harvest states of the Lapins late-ripening cherry variety for the 3 trial conditions. FIG. 7A: Application of Formulation 5, FIG.

[0030] 7B: Control (without treatment), FIG. 7C: Application of Commercial Product (Crackguard®).

[0031] FIG. 8: Graph showing the results of crack induction (% cracking) using the water immersion test for the Santina cherry variety over a period of 20 hours. Formulation 5, Control (no treatment).

[0032] FIG. 9: Graph showing the results of crack induction (% cracking) using the water immersion test for the Lapins cherry variety over a period of 20 hours. Formulation 5, Control (no treatment), Commercial Product: Crackguard®.

[0033] DESCRIPTION OF THE INVENTION

[0034] The present invention relates to a formulation, a method for preparing said formulation, and a method for preventing or reducing splitting or cracking in fruit.

[0035] The formulation of the present invention protects fruit by generating a hydrophobic layer that prevents or reduces water penetration. Furthermore, the formulation of the present invention improves the flexibility and resistance of the fruit cuticle, reduces physiological disturbances at the molecular level caused by water penetration, and promotes the self-repair of micro-cracks.

[0036] In one embodiment of the present invention, the formulation and method for preventing or reducing splitting in fruits were designed for stone fruits or drupes such as cherries, sour cherries, plums, peaches or apricots, apricots or basil, without being limited to those mentioned herein, and berries, berries being understood as fleshy fruits, generally with thin skin and multiple seeds such as grapes, blueberries, tomatoes, among others, without being limited to those mentioned herein.

[0037] The formulation of the present invention comprises at least the following components: a rhamnolipid solution, citric acid, boric acid, calcium chloride, gibberellic acid, a chitosan solution, castor oil, and water. Optionally, the formulation may also include phosphoric acid.

[0038] The components of this formulation were carefully selected to provide specific beneficial properties to both the cuticle and the fruit. First, rhamnolipids were incorporated to promote emulsion formation and improve the formulation's stability on the fruit surface. Citric acid interacts with the ester bonds of the cuticle, enhancing its flexibility and elasticity. Boric acid, in addition to being an essential micronutrient for plant growth and development, helps reduce the incidence of certain physiological disorders such as fruit cracking. Calcium chloride, besides its ability to reduce osmotic potential, promotes the activity of transcription factors that counteract the effect of melic acid, a compound produced by fruit that intensifies cracking. Gibberellic acid, for its part, counteracts the accumulation of melic acid and increases cuticle elasticity.Chitosan increases the resistance of the fruit cuticle and improves its ability to retain moisture. Castor oil provides octadecanoic fatty acids, which are an important component of the fruit cuticle. Optionally, some formulations include phosphoric acid to promote the repair of cracks through ester bonds. While the beneficial properties of these components are known individually, to date, no formulation combining them all to prevent or reduce fruit cracking is known.

[0039] In a preferred embodiment of the present invention, the formulation comprises at least a rhamnolipid solution between 0.1% and 10% v / v; citric acid between 0.01% and 3% w / v; boric acid between 0.1% and 5% w / v; calcium chloride between 1% and 10% w / v; gibberellic acid between 0.01% and 0.8% w / v (equivalent to between 10 and 800 ppm); a 4% w / v chitosan solution between 0.1% and 5% v / v; castor oil between 0.1% and 5% v / v; and water q.s., preferably distilled water. Optionally, the formulation includes phosphoric acid between 0.1% and 5% w / v.

[0040] The rhamnolipids of a preferred embodiment of the present invention are a mixture of mono- and di-rhamnolipids, CAS Registry Number 869062-42-0. Preferably, the rhamnolipid solution used as raw material for preparing the formulation is in an aqueous solution containing 40% or more rhamnolipids.

[0041] In one embodiment of the present invention, the chitosan used as a raw material for preparing the formulation is a 4% w / v aqueous solution. Preferably, the chitosan has a molecular weight between 150,000 and 300,000 g / mol, and a degree of deacetylation greater than or equal to 85%.

[0042] In one embodiment of the present invention, the castor oil used as a raw material for preparing the formulation has a composition that includes: between 85 and 92% ricinoleic acid, up to 2% C16 palmitic acid, up to 2.5% C18 stearic acid, between 2.5 and 6% C18:1 oleic acid, between 2.5 and 7% C18:2 linoleic acid, up to 1% C18:3 linolenic acid, up to 0.25% water, and up to 1% other components.

[0043] The formulation of the present invention, in any of its embodiments, is used in a method to prevent or reduce splitting or cracking in fruit. This method comprises diluting the formulation between 0, 1, and 5% v / v to obtain a diluted formulation and applying the diluted formulation to the fruit. The formulation is preferably diluted in water at a concentration appropriate to the type of fruit to which it will be applied. For example, for cherries, the formulation is preferably diluted to 1% v / v in water.

[0044] In one embodiment, the application of the diluted formulation to the fruit can be done by spraying or immersion, without being limited to these methods mentioned here, preferably by spraying.

[0045] Another object of the present invention relates to the process of preparing the formulation, which comprises the following stages:

[0046] i. Dissolve calcium chloride, boric acid, and citric acid in water to obtain a solution; ii. Dilute gibberellic acid in water and add it to the solution; and

[0047] iii. Add to the solution a chitosan solution, castor oil and a rhamnolipid solution and mix until the formulation is obtained; where all steps are carried out under constant stirring.

[0048] In one embodiment of the present invention, calcium chloride, boric acid, and citric acid are dissolved in a container with 50% of the final volume of water, preferably distilled water, and then diluted gibberellic acid is added. Subsequently, chitosan solution, castor oil, and rhamnolipid solution are added to the solution, and the volume is brought up to the mark with water, preferably distilled water.

[0049] In one embodiment of the present invention, the chitosan solution added in step i¡¡ is diluted to 4% w / v.

[0050] The final formulation can be stored at room temperature, preferably avoiding direct sunlight.

[0051] Table 1 below shows a comparison of different attributes of technologies used to prevent fruit cracking. Table 1: Comparison of different attributes of anti-cracking technologies Attributes Coatings Products Chemical plastic formulations of the invention Efficiency Reduce cracking in a variable way. Effectiveness is related to the maximum amounts of water that can withstand it. Prevents water from entering the fruit through the peduncle, but does not prevent excess water from entering due to rainfall greater than 5-10 mm. Prevents water from entering the fruit through the peduncle, but does not prevent excess water from entering due to root damage. Generally, they are limited to rainfall not greater than 5-10 mm.

[0052]

[0053] (physiological and self-repair of micro-cracks) Cost Between 25 - 30 Between 200 to 300 Market value million pesos thousand Chilean pesos low per

[0054] per hectare. per application. application compared to the competition.

[0055] Around 150,000 - 200,000 Chilean pesos per application.

[0056] Implementation Complexes Requires many Requires 2

[0057] Install applications, deploy applications (between 5 and 6), and when storing and caring for the color change, have trained personnel for straw yellow and the last season, use it in emergencies, of the fruit. In addition, it should only be diluted for a maximum of 3 to 5 seasons. It is required for unexpected rains, and therefore facilitates its annual maintenance. Implementation is required on occasions under climatic conditions.

[0058] It increases its cost. Adverse.

[0059]

[0060] Currently available chemical products should be used in conjunction with physical prevention methods, such as plastic covers. However, this strategy is designed to withstand rainfall of approximately 5-10 millimeters at the peduncle level. The low water tolerance, combined with the number of applications recommended by the manufacturer, renders this protection strategy ineffective and fails to contain the problem. In contrast, one formulation of the present invention is able to reduce cracking in Santina cherry varieties from 56.25% (control) to 8.75% (formulation), according to an experimental model in which the fruit was submerged in distilled water for 20 hours.It is worth noting that, under real-world conditions, cherries will not be subjected to the same volume of water or exposure time as those used in that experimental model, demonstrating that it is a highly effective formulation and could work independently, without the need for physical methods of crack prevention.

[0061] The following examples are intended to illustrate the invention and its preferred embodiments, but in no circumstances should they be considered to restrict the scope of the invention, which will be defined by the terms of the claims attached hereto.

[0062] APPLICATION EXAMPLES

[0063] Example 1: Preparation of formulations

[0064] Five formulations were prepared to determine the best combination for protection against splitting. Table 2 shows the components used and their respective concentrations.

[0065] Table 2. Percentage composition of the various components that make up each of the 5 formulations that will be used for the effectiveness tests against fruit splitting.

[0066] Formulation Component

[0067] Chemicals 1 2 3 4 5 Solution of 1.75% 3% 4% 4% 4% Rhamnolipids v / vv / vv / vv / vv / v 2% 2% 2% 1% 1% Citric Acid

[0068] p / vp / vp / vp / vp / v 1.15% 1.15% 1.15% 1.15%

[0069] Phosphoric Acid - p / vp / vp / vp / v

[0070]

[0071] 3% 3% 3% 1.5% 1.5% Boric Acid

[0072] p / vp / vp / vp / vp / v 6% 6% 6% 6% 6% Calcium Chloride

[0073] p / vp / vp / vp / vp / v 400 400 400 400 Gibberellic Acid 400 ppm

[0074] ppm PPm PPm PPm Solution of 1.8% 1.8% 2.5% 2.5% 2.5% Chitosan 4% w / vv / vv / vv / vv / vv / v 1.5% 2.5% 3% 3% 3% Castor Oil

[0075] v / vv / vv / vv / vv / v Distilled water cspcspcspcspcsp

[0076]

[0077] CSP: sufficient quantity to complete the final volume of the formulation.

[0078] The rhamnolipid product used was Rhamnolipid-RLH (Jiangyin Suoteng Biotechnology Co., Ltd. Wuxi, Jiangsu, China), which has the following physicochemical properties: appearance: yellow to brown, clear liquid; rhamnolipids: > 40.0%; water: < 60.0%; pH: 6.5 - 7.5.

[0079] The chitosan used had a molecular weight between 150,000 - 300,000 g / mol, a degree of deacetylation > 85%, a viscosity (1% HAc) between 50 ~ 20 cps, pH between 7.0 ~ 9.0, which was in aqueous solution at 4% w / v.

[0080] The castor oil used had the composition shown in Table 3 (information provided by the manufacturer).

[0081] Table 3. Composition of castor oil

[0082] Parameters Results Range Fatty acid composition (GC-FLD)

[0083] Ricinoleic acid 87.41 85-92%

[0084] Palmitic acid (C16) 1.24 Max. 2

[0085] Stearic acid (C18) 1.41 Max. 2.5

[0086] Oleic acid (C18:1) 3.36 2.5-6%

[0087] Linoleic acid (C18:2) 5.03 2.5-7%

[0088] Linolenic acid (C18:3) 0.44 Max. 1

[0089]

[0090] Water 0.13 Max. 0.25

[0091] Other 0.67 Max. 1

[0092]

[0093] Note: The information contained in this table represents a true copy of the analysis received from the supplier.

[0094] Prior to preparing these formulations, the components were readily available, and the different formulations were prepared in a container with constant stirring. The formulation preparation procedure was carried out as follows:

[0095] - The solid components such as calcium chloride, boric acid and citric acid were placed in concentrations according to each formulation shown in Table 2, in a container with 50% of the final volume of distilled water;

[0096] - once dissolved, phosphoric acid was added to the solution under a fume hood (except in formulation 5);

[0097] - Gibberellic acid, previously diluted in a sufficient amount of water, was added to the solution;

[0098] - The 4% chitosan solution, castor oil, and rhamnolipid solution were added in the amounts corresponding to each formulation in Table 2; and

[0099] - the volume was completed with distilled water.

[0100] The resulting formulations were stored at room temperature, avoiding direct contact with the sun.

[0101] The formulations obtained are of the emulsion type, therefore they require agitation prior to use to ensure homogeneous application on the fruit.

[0102] All formulations were validated on cherries, both in the laboratory, where cracking was induced using the distilled water immersion method, and in real-world fields located in Romeral, Chile. The formulations were diluted to 1% for use on cherries. Example 2: Field Trial

[0103] Once these five formulations were obtained, and in order to evaluate the emulsion's efficiency in reducing cracking levels on a real-world scale, a field trial was conducted at the Valle Frío farm in Romeral, Maule, Chile (34°57'49.5"S 71°03'59.2"W). After the fruit was ready for harvest and the experimental trial was completed, the percentage of cracking in all the fruit was determined.

[0104] It was decided to conduct the trials on 2 of the most produced cherry species in Chile, Santina and Lapins, since together they represent about 70% of the total Chilean production according to the latest Global Cherry Summit congress of cherry producers.

[0105] The experimental design was as follows: the five formulations were evaluated through two applications at different stages of fruit ripening. The first application was made when the fruit changed color from green to straw yellow in mid-November, and the second application when the fruit reached the reddish color stage in early December. References based on days after full bloom (DDFB) were not used, as this measurement is affected by annual climatic conditions and varies among species.

[0106] The trial was conducted on a total of 14 trees, selected based on their similar fruit load and size, and their proximity to one another. Seven trees of each variety, Lapins and Santina, were used. Five of these were treated with the formulations described in Table 2, one was designated as a control (no product was applied to protect against cracking), and one was treated with the commercial product Crackguard® (Nutriprove SA). Two applications of the formulations in Table 2 were made by spraying (using an agricultural sprayer).

[0107] In the Cuneó area, where the town of Romeral is located in the Maule Region of Chile, 39.8 mm of rainfall was recorded during November 2023, distributed over 5 days: November 9 (6.8 mm), November 10 (22.6 mm), November 11 (3.2 mm), November 16 (1 mm), and November 17 (6.2 mm). This data was obtained from the Chilean Meteorological Directorate - Climate Services.

[0108] Once the testing and maturation phases were completed, and after harvesting, the corresponding analyses were performed to determine productivity and total splitting. In this way, the efficiency percentage was obtained based on all the evaluated treatments, formulations, control, and commercial product, as shown in Figure 1.

[0109] All formulations significantly reduced the percentage of splitting or cracking reported post-harvest compared to the control and commercial product, in addition to reporting a higher final productivity in kilograms.

[0110] In detail, the control group (without any treatment) showed an 11.94% splitting rate with a yield of 5.04 kilograms. This splitting rate could be related to the fruit's late ripening, which reduces the incidence of cracking. The commercial product had the highest splitting rate at 25% of the total cherries, but its yield increased slightly to 6.88 kilograms. All the formulations showed a decrease in the percentage of split cherries, with formulation 5 exhibiting the lowest percentage at only 1.43% of the total cherries split, along with a yield of 9.16 kilograms. Thus, formulation 5 showed 29.4% higher yield than the commercial product and significantly reduced the percentage of split cherries.

[0111] Formulation 1 resulted in 5.25% of the total cherries being cracked and a yield of 5.93 kilograms harvested. Formulation 2 resulted in 8.09% of the total cherries being cracked and a yield of 7.85 kilograms harvested. Formulation 3 resulted in 7.16% of the total cherries being cracked and a yield of 8.79 kilograms harvested. Finally, formulation 4 resulted in 4.42% of the total cherries being cracked and a yield of 12.5 kilograms harvested.

[0112] The early-ripening Santina variety of cherries was affected by the November rains, which allowed for accurate results regarding the percentage of fruit splitting. In contrast, the later-ripening Lapins variety did not show significant changes in splitting percentages, as the rains did not coincide with the fruit's most susceptible stage of water absorption. Therefore, splitting results for Lapins cherries are not shown.

[0113] Example 3: Laboratory test using Christensen methodology Using the harvested cherries in good condition for each of the two cherry varieties (Santina and Lapins), a laboratory test was carried out following the Christensen methodology (Christensen, JV (1972). Cracking in cherries: IV. Physiological studies of the mechanism of cracking. Acta Agriculturae Scandinavica, 22(3), 153-162), where the aim is to induce the cracking of the cherries by immersing them in distilled water in order to simulate the behavior of the fruit in the rain.

[0114] The experimental design to carry out this methodology was as follows: 20 fruits were taken for each experimental group and, to ensure the validity of the results, the experiment was carried out in quadruplicate (a total of 80 cherries per treatment per variety) for immersion in distilled water.

[0115] Before proceeding with the water immersion of the fruit, each experimental group was sprayed with one of the formulations (1, 2, 3, 4, and 5) or the commercial product (Crackguard®), while one group received no treatment (control). The fruit was then fully submerged in water to cover its entire surface and left in this state for 20 hours to simulate prolonged exposure to rain without mechanical drying. Visual measurements of cracking signs were taken at 1, 2, 3, 4, 5, 6, 10, and 20 hours after the start of the methodology.

[0116] The results are shown in Figures 2 and 3 for the Santina and Lapins cherry varieties, respectively. The graphs also display the standard deviation bars for each value obtained in each measurement, based on the average of the four replicates.

[0117] Based on these results, it was observed in Santina that all formulations showed better results than the control treatment in the prolonged immersion test in distilled water. The control group finished the test with 56.25% of the cherries split, while formulation 1 had only 20% split cherries, formulation 2 25% split cherries, formulation 3 15% split cherries, formulation 4 17.5% split cherries, and the best result was obtained with formulation 5, with only 8.75% split cherries.

[0118] An analysis of variance (ANOVA) was performed on the results shown in Figure 2 to determine if there were significant differences between the samples. The following results were obtained: [F-statistic: 22.29] and [p-value: 3.92e-07]. This indicated that there were statistically significant differences between the treatments in terms of the percentage of split cherries. The relatively high F-value further supports the evidence that the observed differences between treatments were not due to chance.

[0119] The results for the Lapins variety showed that, as with the Santina variety, all groups of cherries treated with the formulations exhibited a lower percentage of splitting over the 20 hours of immersion in distilled water, both compared to the control treatment and the treatment with the commercial product. Unlike the experimental trial conducted with the Santina variety, the laboratory trials with the Lapins variety included a comparison with the commercial product due to the lack of real-world results (field trials).

[0120] Figure 3 shows that both the control treatment and the commercial product resulted in nearly identical percentages of total splitting, at 76.25% and 71.25% respectively, while all formulations had lower percentages of splitting. Cherries treated with formulation 3 had a splitting percentage of 52.5%. Cherries treated with formulations 1, 2, and 4 had similar percentages of splitting, with 40%, 41.25%, and 43.75% of split cherries, respectively. Finally, as in the immersion test of the Santina variety, formulation 5 showed the best results with only 27.5% split cherries, significantly reducing the percentage of split cherries compared to the other treatments.

[0121] An ANOVA analysis of variance was also performed on the results in FIG. 3 to determine if there were significant differences between the formulations versus the control, where the following values ​​were obtained [F statistic: 8.13] and [p value: 0.00036], and for the comparison between formulations versus the commercial product, the following values ​​were obtained [F statistic: 5.99] and [p value: 0.0019]. These results, like the previous analysis, have a p < 0.05, so the differences are statistically significant between the treatments in terms of the percentage of split cherries, and since the F values ​​are relatively high, the evidence is reinforced that the differences observed between the treatments are not due to chance.

[0122] Figure 4 shows photographs of cherries that split after being submerged in distilled water for 20 hours. The first and third photographs show Lapins cherries, and the second photograph shows a Santina cherry. The cracking caused by water absorption, which leads to swelling and subsequent splitting of the fruit, is clearly visible.

[0123] Example 4. Formulation with the best results obtained

[0124] Figure 5 shows the real-scale results (field trials) obtained after two applications of formulation 5 to the early variety Santina in November-December (2023), compared to the results with the commercial product (Crackguard®) and the control (no treatment). The high levels of cracking observed during the evaluation period are related to the altered climatic conditions caused by the El Niño phenomenon.

[0125] Cherries treated with formulation 5 showed only 1.43% post-harvest splitting compared to 25% splitting in cherries treated with the commercial product. The control group, on the other hand, showed 11.94% splitting, a lower percentage than that reported with the commercial product. The absence of any agrochemicals in the control group may have influenced slower fruit ripening, delaying it and consequently reducing water uptake by the fruit during its most susceptible stage to splitting. Figures 6A, 6B, and 60 show photographs of the post-harvest results for Santina cherries, where it can be observed that there is no significant difference in size and color between the cherries treated with formulation 5 (Figure 6A) and the commercial product (Figure 60). However, differences are observed when compared to the control group without agrochemicals, possibly associated with the delayed fruit ripening.

[0126] Furthermore, FIGS. 7A, 7B and 70 present photographs with the results in post-harvest Lapins cherries, where no differences in size and color were observed between the 3 conditions under study.

[0127] At the laboratory scale, the Christensen methodology explained in the previous example was used. Figure 8 shows the comparative results of the percentage of splitting in Santina cherries, using the treatment with formulation 5 versus the control (no treatment). At the end of the 20-hour immersion period in water, 56.25% splitting was observed in the cherries of the control group, while only 8.75% splitting was observed in the cherries treated with formulation 5.

[0128] In the laboratory-scale trial with Lapins variety cherries (FIG.

[0129] 9) Comparisons were made between formulation 5, a control group (no treatment), and a group treated with the commercial product Crackguard®. It was observed that, at the end of the 20-hour immersion period of the cherries in water, the control group (no treatment) had 76.25% of cherries with cracks, and the group treated with the commercial product had 71.25% of cherries with cracks, while in the group treated with formulation 5, only 27.5% ended up with cracks.

[0130] These results allow us to conclude that the formulations described here increase the fruit's tolerance to water penetration, which helps to prevent or significantly reduce cracking. Thus, it is demonstrated that applying any of these formulations represents an effective and efficient strategy for decreasing the fruit's vulnerability to climatic conditions that promote cracking.

Claims

CLAIMS 1. A formulation to prevent or reduce splitting or cracking in fruits, CHARACTERIZED in that it comprises a solution of rhamnolipids, citric acid, boric acid, calcium chloride, gibberellic acid, a chitosan solution, castor oil and water.

2. The formulation according to claim 1, CHARACTERIZED in that it is an emulsion comprising a rhamnolipid solution in a concentration between 0.1% and 10% v / v; citric acid in a concentration between 0.01% and 3% w / v; boric acid in a concentration between 0.1% and 5% w / v; calcium chloride in a concentration between 1% and 10% w / v; gibberellic acid in a concentration between 0.01% and 0.8% w / v; a 4% w / v chitosan solution in a concentration between 0.1% and 5% v / v; castor oil in a concentration between 0.1% and 5% v / v; and water q.s.

3. The formulation according to any of claims 1 or 2, CHARACTERIZED in that the chitosan has a molecular weight range between 150,000 and 300,000 g / mol, and a degree of deacetylation greater than or equal to 85%.

4. The formulation according to any of claims 1 to 3, CHARACTERIZED in that it further comprises phosphoric acid in a concentration between 0.1% and 5% w / v.

5. The formulation according to any of claims 1 to 4, CHARACTERIZED in that it is diluted between 0.1 and 5% v / v for application to the fruit.

6. A method for preventing or reducing splitting or cracking in fruit, CHARACTERIZED in that it comprises the following steps: - diluting a formulation according to any of claims 1 to 4 between 0.1% and 5% v / v to obtain a diluted formulation; and - Apply the diluted formulation to the fruit.

7. The method according to claim 6, CHARACTERIZED in that it comprises applying the diluted formulation to the fruit by spraying.

8. The method according to any of claims 6 or 7, CHARACTERIZED in that the fruits are stone fruits or berries.

9. The method according to claim 8, CHARACTERIZED in that the stone fruits are selected from the group consisting of cherries, sour cherries, plums, peaches, and apricots, and the berries are selected from the group consisting of grapes, blueberries, and tomatoes.

10. A method for preparing a formulation to prevent or reduce splitting or cracking in fruit, CHARACTERIZED in that it comprises the steps of: i. Dissolve calcium chloride, boric acid, and citric acid in water until a solution is obtained; i. Dilute gibberellic acid in water and add it to the solution; and iii. Add to the solution a chitosan solution, castor oil and a rhamnolipid solution and mix until the formulation is obtained; where all steps are carried out under constant stirring.

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