Method for improving fruit quality

Abstract

This invention relates to methods for improving the flavor, color, texture, or any combination thereof of plant materials, including pre-harvest or post-harvest treatment with an effective amount of aromatic amino acids or their analogues.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 134,403, filed January 6, 2021, entitled "Methods for Improving Fruit Quality," and U.S. Provisional Patent Application No. 63 / 164,051, filed March 22, 2021, also entitled "Methods for Improving Fruit Quality." The contents of both applications are incorporated herein by reference in their entirety. Technical Field

[0003] In some embodiments of the present invention, methods for improving plant quality, such as flavor, color, wound healing, or combinations thereof, are involved. Background Technology

[0004] It is widely believed that the flavor quality of many fruits has declined significantly in recent decades. This decline appears to be at least partly attributable to early harvesting, damage, long-term storage, and biased selection for certain desired phenotypes, such as shelf life and firmness, which are negatively correlated with flavor, aroma, nutritional value, and color, at least in some cases. Flavor involves the integration of sugars, acids, and volatile organic compounds (VOCs).

[0005] There is a great need for methods to improve fruit quality parameters such as flavor, aroma, wound healing, color, or combinations thereof. Furthermore, how and when to select suitable fruits for quality improvement treatments has not yet been determined. Summary of the Invention

[0006] According to a first aspect, a method is provided for improving the flavor, color, damage healing, or combination thereof of plant material, leaves of a plant containing said plant material, or both, comprising treating the plant material with an effective amount of phenylalanine or an analogue thereof before or after harvest.

[0007] In some implementations, the improvement does not include an increase due to a reduction in any of the flavor, color, cure, or combinations thereof induced by plant pathogens.

[0008] In some implementations, improving flavor includes increasing the amount of anthocyanins, flavonoids, or both in the plant material or juice extracted from it.

[0009] In some implementations, improving flavor includes increasing the Brix value, decreasing the acidity value, increasing aroma, or both, in the plant material or juice extracted from it.

[0010] In some implementations, improving color includes inducing reddening in plant material, reducing the amount of chlorophyll, increasing the ratio of anthocyanins to chlorophyll, or any combination thereof.

[0011] In some implementations, the plant material includes either fruit or tuber.

[0012] In some implementations, the damage is caused by non-biological agents.

[0013] In some implementations, non-biological agents include physical or mechanical damage.

[0014] In some implementations, improved healing includes: increasing scratch healing, increasing cut healing, increasing scratch color healing in plant material, reducing weight loss percentage, increasing lignin production rate or cork resin production rate or both, or any combination thereof.

[0015] In some embodiments, the effective amount of phenylalanine or its analogues is greater than 2 mM.

[0016] In some embodiments, the effective amount of phenylalanine or its analogues is in the range of 4 mM to 20 mM.

[0017] In some embodiments, the method also includes the step of providing the plant material with a period of 1 to 30 days of pre-harvest abiotic stress.

[0018] In some implementations, abiotic stress conditions include light, radiation, temperature, lack of nutrients or water, or any combination thereof.

[0019] In some embodiments, the method further includes the step of selecting plant material that needs to be treated with the phenylalanine or its analogues.

[0020] In some embodiments, selection includes determining the amount of phytochemical in the plant material compared to a predetermined threshold, wherein plant material containing an amount of phytochemical greater than the predetermined threshold is suitable for pre-harvest or post-harvest treatment with an effective amount of phenylalanine or its analogues.

[0021] In some embodiments, the phytochemical is selected from flavonoids, anthocyanins, pigments, and any combination thereof.

[0022] In some implementations, the pigment includes chlorophyll.

[0023] In some embodiments, selections including determining the total soluble solids (TSS) % or dry matter, or TSS / acid in the fruit, wherein the fruit containing at least 5% TSS % or TSS / acid is suitable for pre-harvest or post-harvest treatment with an effective amount of phenylalanine or its analogues.

[0024] In some embodiments, the treatment includes: drenching, dipping, soaking, injection, spraying, coating, or any combination thereof.

[0025] In some implementations, the treatment takes place in an open area, a greenhouse, a storage facility, or any combination thereof.

[0026] In some implementations, the improvement is compared with control plant material.

[0027] In some implementations, plant materials include vegetables or fruits.

[0028] In some embodiments, vegetables include: stems, leaves, roots, tubers, or any combination thereof.

[0029] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While similar or equivalent methods and materials to those described and used herein may be used in the practice or testing of embodiments of the invention, exemplary methods and / or materials will be described below. In case of conflict, the patent specification, including its definitions, shall prevail. Furthermore, the materials, methods, and examples described are illustrative only and are not intended to be necessarily limiting.

[0030] Further embodiments and the full scope of the invention will become apparent from the detailed description given below. However, it should be understood that while indicating preferred embodiments of the invention, this detailed description and specific examples are given merely by way of illustration, as various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. Attached Figure Description

[0031] This document describes some embodiments of the invention by way of example only, with reference to the accompanying drawings. Referring now to the details of the drawings, it should be emphasized that the details shown are by way of example and are for the purpose of illustrative discussion of embodiments of the invention. In this regard, the description taken in conjunction with the drawings will enable those skilled in the art to understand how embodiments of the invention can be practiced.

[0032] Figure 1 This includes photomicrographs and graphs showing the improvement in flavor of mango fruit (“Shelly”) after harvesting, achieved by applying Phe during a 15-day storage period at 22°C in an experiment conducted in 2019.

[0033] Figure 2This includes photomicrographs and graphs showing the improvement in flavor of mango fruit (“Shelly”) after harvest by applying Phe during a 13-day storage period at 22°C in an experiment conducted in 2020.

[0034] Figure 3 This includes photomicrographs and graphs showing the improvement in flavor of mango fruit (“Tali”) after post-harvest application of Phe during a 12-day storage period at 22°C in an experiment conducted in 2020.

[0035] Figure 4 Includes representative images showing the induction of reddening (e.g., the appearance of red color) in mango fruits (“Shelly”) by pre-harvest application of Phe. Control – untreated; 1W Phe – phenylalanine treatment 1 week before harvest; 2W Phe – phenylalanine treatment 2 weeks before harvest; and 2W PDJ – prohydrojasmon treatment 2 weeks before harvest.

[0036] Figure 5 This includes vertical bar graphs showing the induction of reddening (e.g., red coloration) in mango fruits (“Shelly”) by pre-harvest application of Phe. T-0—harvest; CS—after refrigeration; SL—after shelf life. The red surface area was assessed as a percentage, and the red intensity was assessed using a scale of 0-5. For each treatment at different time points (harvest and after shelf life at 22°C), the percentage of red surface area on the fruit was assessed. Similarly, the red intensity of each mango fruit was assessed using a visual rating scale, where 0 = no red, 1 = faint red, and 5 = very intense red. Fifty fruits were assessed for each treatment. The skin color (hue and a*) of 15 mango fruits for each treatment was measured at the reddest point on the equator of each fruit using a CR-400 / 410 colorimeter (Konicka Minolta, Osaka, Japan).

[0037] Figure 6 This includes a vertical bar graph showing the induction of reddening (e.g., turning red) in mango fruits (“Shelly”) by pre-harvest application of Phe. T-0—harvest; CS—after refrigeration; SL—after shelf life. Chlorophyll, anthocyanin, and flavonoid contents were measured using a Multiplex III fluorescence detector (Force A, Orsay, France), which consists of 12 fluorescence signals. The ratios between these signals in different mathematical expressions are correlated with the fluorescence of major chemical groups, such as anthocyanins (FER_RG, the ratio of infrared emission excited by red or green light), flavonoids (FLAV), and chlorophyll (SER_R). Fifteen fruits for each treatment were evaluated on the red side.

[0038] Figure 7 This includes photomicrographs and graphs showing the improvement in flavor of mango fruit (“Shelly”) after shelf life following application of Phe before harvest, as demonstrated in an experiment conducted in 2020.

[0039] Figure 8 This includes photomicrographs and graphs showing improvements in the quality of juice extracted from mango fruit (“Shelly”) by applying Phe before harvest in an experiment conducted in 2020.

[0040] Figure 9 Includes representative images showing the induction of reddening (e.g., turning red) in mango fruits (“Kent”) by pre-harvest application of Phe. Control—untreated; 1W Phe—phenylalanine treatment 1 week before harvest; 2W Phe—phenylalanine treatment 2 weeks before harvest; and 2W PDJ—jasmone treatment 2 weeks before harvest.

[0041] Figure 10 This includes vertical bar graphs showing the induction of reddening (e.g., red coloration) in mango fruits (“Kent”) by pre-harvest application of Phe. T-0—harvest; CS—after refrigeration; SL—after shelf life. The red surface area was assessed as a percentage, and the red intensity was assessed using a scale of 0-5. For each treatment at different time points (harvest and after shelf life at 22°C), the percentage of red surface area on the fruit was assessed. Similarly, the red intensity of each mango fruit was assessed using a visual rating scale, where 0 = no red, 1 = faint red, and 5 = very intense red. Fifty fruits were assessed for each treatment. The skin color (hue and a*) of 15 mango fruits for each treatment was measured at the reddest point on the equator of each fruit using a CR-400 / 410 colorimeter (Konicka Minolta, Osaka, Japan).

[0042] Figure 11 This includes a vertical bar graph showing the induction of reddening (e.g., turning red) in mango fruits (“Kent”) by pre-harvest application of Phe. T-0—harvest; CS—after refrigeration; SL—after shelf life. Chlorophyll, anthocyanin, and flavonoid contents were measured using a Multiplex III fluorescence detector (Force A, Orsay, France), which consists of 12 fluorescence signals. The ratios between these signals in different mathematical expressions are correlated with the fluorescence of major chemical groups, such as anthocyanins (FER_RG, the ratio of infrared emission excited by red or green light), flavonoids (FLAV), and chlorophyll (SER_R). Fifteen fruits for each treatment were evaluated on the red side.

[0043] Figure 12 This includes photomicrographs and graphs showing the improvement in flavor of mango fruit (“Kent”) after shelf life following application of Phe before harvest in an experiment conducted in 2020.

[0044] Figure 13 This includes photomicrographs and graphs showing improvements in the quality of juice extracted from mango fruit (“Kent”) by applying Phe before harvest in an experiment conducted in 2020.

[0045] Figure 14 This includes charts showing the flavor improvement of white Muscat grapes after pre-harvest application of Phe, as demonstrated in an experiment conducted in 2020.

[0046] Figure 15 This includes charts showing improvements in the quality of juice extracted from Muscat Blanc grapes after pre-harvest application of Phe, as demonstrated in an experiment conducted in 2020.

[0047] Figure 16 This includes charts showing that, in an experiment conducted in 2020, the quality of juice extracted from Petit Verdot grapes was improved by applying Phe before harvest.

[0048] Figure 17 Includes representative images showing the induction of reddening (e.g., turning red) in apple fruits (“Jonathan”) after refrigeration via pre-harvest application of Phe. Control – untreated; 1W Phe – phenylalanine treatment 1 week before harvest; 2W Phe – phenylalanine treatment 2 weeks before harvest; and 2W PDJ – jasmone treatment 2 weeks before harvest.

[0049] Figure 18 This includes vertical bar graphs showing the induction of reddening (e.g., red coloration) in apple fruits (“Jonathan”) by pre-harvest application of Phe. The red surface area was assessed as a percentage, and red intensity was assessed using a scale of 0-5. For each treatment, the percentage of red surface area on the fruit was assessed at different time points (harvest, after refrigeration at 0°C, and after shelf life at 22°C). Similarly, the red intensity of each apple fruit was assessed using a visual rating scale, where 0 = no red, 1 = faint red, and 5 = very intense red. Fifty fruits were assessed for each treatment. The skin color (hue) of 30 mango fruits for each treatment was measured at the reddest point on the equator of each fruit using a CR-400 / 410 colorimeter (Konicka Minolta, Osaka, Japan).

[0050] Figure 19This includes a vertical bar graph showing the induction of reddening (e.g., turning red) in apple fruits (“Jonathan”) by pre-harvest application of Phe. Chlorophyll, anthocyanin, and flavonoid contents were measured using a Multiplex III fluorescence detector (Force A, Orsay, France), which consists of 12 fluorescence signals. The ratios between these signals in different mathematical expressions are correlated with the fluorescence of major chemical groups, such as anthocyanins (FER_RG, the ratio of infrared emission excited by red or green light), flavonoids (FLAV), and chlorophyll (SER_R). Thirty fruits from each treatment were evaluated on the red side.

[0051] Figure 20 Includes charts showing the improvement in flavor of apple fruit (“Jonathan”) after refrigeration after application of Phe before harvest.

[0052] Figure 21 Includes charts showing the flavor improvement of juice extracted from apple fruit (“Jonathan”) after pre-harvest application of Phe.

[0053] Figures 22A-22F Includes vertical bar graphs and photomicrographs showing the induction of scratch healing in potato tubers treated with phenylalanine at 7 and 14 days post-harvest. Scratch healing in Sifra potatoes (22A); weight loss in Sifra potatoes (22B); scratch healing in Memphis potatoes (22C); weight loss in Memphis potatoes (22D); and color healing in Memphis potatoes (22E). (22F) is a representative image of the skin analysis of Memphis potatoes treated with phenylalanine at 7 days post-scratch.

[0054] Figures 23A-23D Includes vertical bar graphs showing the induction of cut healing in potato tubers treated with phenylalanine at 7 and 14 days post-harvest. Sivra potato cut healing (23A); Sivra potato weight loss (23B); Memphis potato cut healing (23C); Memphis potato weight loss (23D).

[0055] Figures 24A-24EIncludes photomicrographs, fluorescence photomicrographs, and vertical bar graphs showing the induction of cut healing in potato tubers treated with phenylalanine after 7 days. Photomicrographs (24A-24B) show cut healing in Sivra potatoes from control (24A) and Phe-treated (24B) potatoes treated after 7 days. Fluorescence photomicrographs (24C-24D) show lignin stained with Calcofluor and fluorescence microscopy images of control (24C) and Phe-treated (24D) potatoes. A graph showing the fluorescence intensity of 10 replicates (24C-24D) measured by Image J.

[0056] Figure 25A-25F Includes photomicrographs showing the induction of cure in potato tubers treated with phenylalanine 14 days after harvest. (25A-25D) Sivra potatoes; and (25E-25F) Memphis potatoes. (25A, 25C, and 25E) controls; and (25B, 25D, and 25F) Phe treatments.

[0057] Figures 26A-26C This includes vertical bar graphs showing the effects of pre-harvest spraying treatments on reddening and TSS of mango (Shelly) fruit. Red surface area (%; 26A), red intensity (26B), and TSS (% Brix; 26C) were measured under pre-harvest treatment with 0.01%, 0.06%, 0.012%, or 0.24% phenylalanine (Phe). 0.01% (equivalent to ~0.5 mM) Phe was found to be ineffective in improving reddening and is shown as the negative control (26A–26B, rectangular boxes). Pre-harvest treatment with 0.01% Phe did not contribute to TSS, as in the negative control (26C).

[0058] Figures 27A-27D This includes vertical bar graphs showing the effects of pre-harvest spraying treatments on reddening and TSS of Starking Delicious apple fruit. Red surface (%; 27A), red intensity (27B), and TSS (% Brix; 27C–27D) were assessed under pre-harvest treatments with 0.01% and 0.012% phenylalanine (Phe). 0.12% (equivalent to ~6 mM) Phe was found to be more effective in improving reddening, with TSS comparable to the negative control at 0.01%. In 27D, TSS was assessed at time 0 (T0), after refrigeration (CS), and after time under shelf conditions (e.g., shelf life (SL)).

[0059] Figure 28Includes a vertical bar graph showing the effect of pre-harvest spraying on the reddening (indices 0-5) of Skarlota grapes. 0.12% (equivalent to ~6 mM) of Phe was more effective in improving the reddening index compared to 0.01%.

[0060] Figures 29A-29E Includes photomicrographs showing the effects of pre-harvest treatment on decay parameters of Starking (29A-29B) and Anna (29C-29E) apples. Enzymatic browning (i.e., oxidation) was very pronounced in the control fruits (29A and 29C). Treatment with 0.12% Phe two weeks before harvest (29B and 29D) or four weeks before harvest (29E) was found to effectively reduce enzymatic browning in the apple fruits.

[0061] Figure 30A-30G This includes a vertical bar graph showing the increase in aroma-related VOC levels in mango fruit after pre-harvest treatment with 8 mM phenylalanine. Mango (Tali) fruit was immersed in 8 mM phenylalanine and stored at 22°C for 11 days; the volatiles in the fruit sediment were then examined. The concentrations of the following volatile aroma compounds (expressed in μg / gFW) were determined: (30A) α-pinene; (30B) 3-carene; (30C) D-limonene; (30D) guargenene; (30E) α-terpinene; (30F) α-phellandrene; and (30G) caryophyllene.

[0062] Figures 31A-31C Includes fluorescence micrographs (31A) and vertical bar graphs showing that post-harvest treatment with 8 mM phenylalanine reduced the accumulation of reactive oxygen species (ROS) in the peel (31B) and pulp (31C) of the treated damaged fruit compared to the control (untreated) damaged fruit. This effect was also observed in fruit stored at 12°C and beyond shelf life (SL). (31A) Visualization 20 minutes after induced damage. (31B-31C) ROS quantification 7 days after induced damage. Detailed Implementation

[0063] In some embodiments, this invention relates to methods for improving the quality of plant materials. In some embodiments, the plant material includes vegetables. In some embodiments, the vegetables include stems, leaves, roots, tubers, stem-tubers, or any combination thereof. In some embodiments, the plant material includes fruits. In some embodiments, the method includes improving fruit quality parameters. In some embodiments, this invention relates to methods for improving the flavor of plant materials (e.g., fruits or juices extracted or derived therefrom). In some embodiments, this invention relates to methods for improving the quality of plant materials (e.g., fruits or juices extracted or derived therefrom). In some embodiments, the method includes treating the plant material, such as a fruit or vegetable, with an effective amount of aromatic amino acids or their analogues.

[0064] According to some aspects, the methods of the present invention relate to shortening the growth and / or maturity period of plant materials, as described herein. According to some aspects, the methods disclosed herein result in a shorter growth and / or maturity period for plant materials (e.g., vegetables), as described herein. According to some aspects, the methods disclosed herein relate to increasing the total annual crop yield of plant materials (e.g., vegetables) compared to a control. In some embodiments, plant materials (e.g., vegetables) treated according to the methods disclosed herein are suitable for harvesting within a shorter time period compared to a control, for example, in terms of quality, color, sweetness, phytochemical content, or any combination thereof. In some embodiments, plant materials (e.g., plants) treated according to the methods disclosed herein are harvested earlier than control plant materials (e.g., vegetables). In some embodiments, plant materials (e.g., vegetables) treated according to the methods disclosed herein reach the suitable quality, color, sweetness, phytochemical content, or any combination thereof described herein earlier than a control (e.g., fruit or plant material, such as vegetables).

[0065] According to another aspect, the methods disclosed herein relate to increasing the sweetness properties of plant materials (e.g., fruits or vegetables). In some embodiments, increasing the sweetness properties of plant materials includes polysaccharide decomposition or catabolism. In some embodiments, increasing the sweetness properties of plant materials includes reducing the amount of polysaccharides in the plant material. In some embodiments, according to the methods disclosed herein, the amount of polysaccharides is reduced in the cells of the treated plant material (e.g., fruit), in the exocarp, epicarp, cortex, or any equivalent thereof, or any combination thereof. In some embodiments, according to the methods disclosed herein, the polysaccharides in the treated plant material are converted, decomposed, metabolized, or any combination thereof and equivalent forms of monosaccharides, disaccharides, or combinations thereof. In some embodiments, increasing the sweetness properties of plant materials includes producing monosaccharides, disaccharides, or both in the plant material. In some embodiments, the produced monosaccharides, disaccharides, or both are obtained through the conversion, decomposition, metabolism, or any combination thereof and equivalent forms of polysaccharides.

[0066] In some implementations, the polysaccharide includes starch.

[0067] In some embodiments, the methods disclosed herein include increasing the sweetness properties of plant materials while maintaining or retaining the nutritional value of the plant materials. In some embodiments, the methods disclosed herein include increasing the sweetness properties of plant materials while maintaining or retaining the calorie value, calorie index, or both of the plant materials.

[0068] In some embodiments, the methods disclosed herein include increasing the sweetness characteristics of plant materials without increasing or decreasing their nutritional value. In some embodiments, the methods disclosed herein include increasing the sweetness characteristics of plant materials without increasing their calorie value, calorie index, or both.

[0069] As used herein, the term "aromatic amino acid (AAA)" refers to phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), or any analogue thereof. As used herein, any AAA analogue may be used, provided that the analogue retains the antipathogenicity and performance-enhancing activities disclosed herein.

[0070] According to some embodiments, a method for improving fruit flavor is provided, which includes a pre-harvest or post-harvest treatment of plant material by contacting it with an effective amount of phenylalanine or an analogue thereof, thereby improving the flavor of the plant material.

[0071] According to some embodiments, methods for reducing or inhibiting fruit decay are provided, which include pre-harvest or post-harvest treatment of plant material by contacting an effective amount of phenylalanine or its analogues, thereby reducing or inhibiting fruit decay.

[0072] In some embodiments, improving the quality of plant material includes reducing or inhibiting decay of the plant material. In some embodiments, the plant material includes or is composed of fruit. In some embodiments, improving the quality of plant material includes reducing or inhibiting fruit decay.

[0073] In some embodiments, fruit decay includes decay of the cuticle, epidermis, parenchyma, subcutaneous tissue, pericarp, cortex, or any combination thereof.

[0074] In some implementations, decay includes enzymatic browning, such as oxidation.

[0075] In some embodiments, the method includes reducing or inhibiting enzymatic browning of the fruit. In some embodiments, the reduction or inhibition of enzymatic browning is determined in sliced ​​or cut fruit.

[0076] In some embodiments, reducing or inhibiting enzymatic browning of fruit includes reducing the rate, volume, surface area, or any combination thereof of enzymatic browning in the fruit.

[0077] In some implementations, the treatment includes contact, application, or a combination thereof.

[0078] In some implementations, the method includes pre-harvest treatment, post-harvest treatment, or both.

[0079] In some embodiments, the method includes treating or contacting the plant material at least once. In some embodiments, the method includes treating or contacting the plant material at least 2, 3, 4, 5, 6, 8, or 10 times, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, the method includes treating or contacting the plant material 1-10 times, 2-10 times, 3-8 times, or 4-10 times. Each possibility represents a separate embodiment of the invention.

[0080] In some implementations, the method includes treating or contacting the plant material once or at least twice (e.g., multiple treatments).

[0081] In some embodiments, during multiple treatments or contacts of plant material, the intervals between each treatment or contact event are at least 5 days, at least 6 days, at least 7 days, at least 9 days, at least 10 days, at least 12 days, at least 15 days, at least 20 days, at least 25 days, at least 30 days, at least 40 days, or any value and range between these intervals. Each possibility represents a separate embodiment of the invention. In some embodiments, during multiple treatments or contacts of plant material, the intervals between each treatment or contact event are 5-20 days, 7-21 days, 7-28 days, 7-35 days, 5-30 days, or 6-30 days. Each possibility represents a separate embodiment of the invention.

[0082] In some embodiments, treating plant material with aromatic amino acids or their analogues includes contacting the plant material with aromatic amino acids or their analogues before harvest, contacting the plant material with aromatic amino acids or their analogues after harvest, or both.

[0083] In some implementations, pre-harvest treatment includes multiple pre-harvest treatments.

[0084] In some implementations, post-harvest contact includes multiple post-harvest contacts.

[0085] In some implementations, the term "multiple" includes any integer equal to or greater than 2.

[0086] In some implementations, improving flavor includes increasing the amount of anthocyanins, flavonoids, or both in the plant material or juice extracted from it.

[0087] In some implementations, improving flavor includes increasing the Brix value of the fruit or juice extracted from it, decreasing the acidity value, increasing aroma, or both.

[0088] In some embodiments, increasing aroma includes increasing the amount of at least one volatile organic compound (VOC) that is associated with or related to aroma.

[0089] The types of VOCs associated with or related to fruit aromas, and the methods for determining them, are readily apparent to those skilled in the art.

[0090] In some embodiments, improving fruit quality includes increasing the amount of anthocyanins, flavonoids, or both in the plant material or juice extracted therefrom.

[0091] In some implementations, improving fruit quality includes inducing reddening in the plant material, reducing the amount of chlorophyll, increasing the ratio of anthocyanins to chlorophyll, or any combination thereof.

[0092] In some implementations, improving fruit quality includes increasing the Brix value, decreasing the acidity value, increasing the aroma, or both, in the fruit or the juice extracted from it.

[0093] In some embodiments, increasing fruit quality or flavor includes increasing the amount, secretion level, or both of at least one compound or VOC selected from: α-pinene, 3-carene, D-limonene, guarene, α-terpinene, α-phellandrene, caryophyllene, or any combination thereof.

[0094] In some embodiments, increasing fruit quality or flavor includes increasing the amount, secretion level, or both of α-pinene, 3-carene, D-limonene, guarene, α-terpinene, α-phellandrene, and caryophyllene.

[0095] In some embodiments, the increase is an increase of at least 5%, at least 20%, at least 50%, at least 75%, at least 100%, at least 250%, at least 500%, or at least 1000%, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

[0096] In some embodiments, the increase is 5-200%, 20-300%, 50-400%, 75-500%, at least 100-1,200%, or 250-1,500%. Each possibility represents a separate embodiment of the invention.

[0097] In some embodiments, improving fruit quality includes reducing the rate, amount, or both of ROS accumulation in the fruit. In some embodiments, improving fruit quality includes reducing the rate, amount, or both of ROS accumulation in the fruit peel. In some embodiments, improving fruit quality includes reducing the rate, amount, or both of ROS accumulation in the fruit pulp. In some embodiments, improving fruit quality includes reducing the rate, amount, or both of ROS accumulation in both the fruit peel and pulp. In some embodiments, the method includes reducing the rate, amount, or both of ROS accumulation in the pulp, peel, or both of damaged fruit. In some embodiments, the damaged fruit is damaged before harvest, damaged after harvest, or both.

[0098] In some embodiments, improving fruit flavor includes reducing the rate, amount, or both of ROS accumulation in fruit treated according to the methods disclosed herein.

[0099] In some embodiments, improving fruit quality or parameters includes improving fruit firmness. In some embodiments, the method includes increasing fruit firmness. In some embodiments, fruit treated according to the methods disclosed herein is firmer than untreated control fruit.

[0100] Methods for determining fruit firmness are common and readily apparent to those skilled in the art.

[0101] In one implementation, hardness is expressed in Newtons (N).

[0102] Methods for determining anthocyanins, flavonoids, acidity, and blister sugar content are common and readily apparent to those skilled in the art. Non-limiting examples of these methods include, but are not limited to, pH testing, GC-MS, colorimetric examination, multiplex fluorescence detection, or others, some of which are illustrated herein.

[0103] According to some embodiments, a method for improving the color of plant material is provided, comprising pre-harvest treatment with an effective amount of phenylalanine or an analogue thereof, thereby improving the color of the plant material.

[0104] In some implementations, improving color includes inducing reddening and anthocyanin in the plant material, reducing the amount of chlorophyll, increasing the ratio of anthocyanins to chlorophyll, or any combination thereof.

[0105] In some embodiments, improving color includes increasing the surface area of ​​plant material that appears reddish. In some embodiments, improving color includes reducing the surface area of ​​plant material that appears green. In some embodiments, improving color includes increasing the ratio of the surface area of ​​plant material that appears reddish to the surface area of ​​plant material that appears green.

[0106] According to the method of the invention, in some embodiments, the culture duration of the fruit or plant material described herein is shorter compared to control fruit or plant material (e.g., not treated according to the method of the invention). In some embodiments, the fruit or plant material treated according to the method of the invention is harvested after a shorter culture period compared to control fruit or plant material (e.g., not treated according to the method of the invention). In some embodiments, the method of the invention provides at least one more harvest opportunity per year for each crop type (e.g., fruit or plant material) compared to the control described herein, as described herein.

[0107] In some embodiments, compared to a control, the method of the present invention can shorten the cycle from cultivation to harvesting of fruit and / or plant material, increase the annual harvest opportunities for a crop type (e.g., fruit or plant material), or both, as described herein.

[0108] In some implementations, the ratio includes a mole-to-mole (m:m) ratio. In some implementations, the ratio includes a weight-to-weight (w / w) ratio.

[0109] In some embodiments, "increase" is an increase of at least 5%, at least 15%, at least 30%, at least 50%, at least 75%, at least 100%, at least 200%, at least 350%, at least 500%, at least 750%, or at least 1000%, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

[0110] In some embodiments, "decrease" (decreasing) is a reduction of at least 5%, at least 15%, at least 30%, at least 50%, at least 75%, at least 100%, at least 200%, at least 350%, at least 500%, at least 750%, or at least 1000%, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

[0111] In some embodiments, the effective amount of phenylalanine or its analogues is at least 2 mM, at least 3 mM, at least 4 mM, at least 6 mM, at least 8 mM, at least 10 mM, at least 12 mM, at least 15 mM, at least 17 mM, or at least 20 mM, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

[0112] In some embodiments, the effective amount of phenylalanine or its analogues ranges from 2 mM to 20 mM, 4 mM to 20 mM, 6 mM to 20 mM, 8 mM to 20 mM, 10 mM to 20 mM, 12 mM to 20 mM, 4 mM to 8 mM, 4 mM to 10 mM, 4 mM to 12 mM, or 15 mM to 20 mM, or any values ​​and ranges therebetween. Each possibility represents a separate embodiment of the invention.

[0113] In some implementations, the method also includes the step of providing the fruit with pre-harvest abiotic stress conditions.

[0114] In some embodiments, the duration of application of abiotic stress conditions is 1-30 days, 2-30 days, 5-30 days, 7-30 days, 10-30 days, 14-30 days, 18-30 days, 23-30 days, 27-30 days, 1-20 days, 1-15 days, 5-30 days, 8-24 days, 10-20 days, or 7-21 days. Each possibility represents a separate embodiment of the invention.

[0115] In some implementations, abiotic stress conditions include light, radiation, temperature, lack of nutrients, lack of water, or any combination thereof.

[0116] In some embodiments, the method further includes the step of selecting plant material for the treatment disclosed herein.

[0117] In some implementations, the method includes determining the amount of phytochemicals in the plant material compared to a predetermined threshold.

[0118] In some embodiments, the selection includes determining the total soluble solids (TSS) percentage of the plant material. In some embodiments, the selection includes determining the TTS to acid ratio. In some embodiments, the selection includes determining the TTS% or TTS / acid ratio in the dry or wet matter of the plant material. In some embodiments, the selection includes determining the TTS% or TTS / acid ratio in a sample comprising dry or wet matter derived from or extracted from the plant material.

[0119] In some embodiments, fruit or plant material, such as vegetables, suitable for treatment according to the methods disclosed herein is characterized by a TSS% or TSS / acid or any equivalent thereof being at least 5%, at least 10%, at least 15%, at least 25%, or at least 30%, or any value and range therebetween, that is known to those skilled in the art as a minimum TSS% or TSS / acid or any equivalent thereof for harvesting fruit or plant material. Each possibility represents a separate embodiment of the invention.

[0120] In some implementations, selection, compared to a control, includes determining the weight % of TSS or the TSS / acid ratio in the fruit or plant material, as described herein.

[0121] In some embodiments, fruit or plant material (e.g., vegetables) containing at least 5% less, at least 10% less, at least 15% less, at least 20% less, at least 25% less, or at least 30% less TSS% or TSS / acid compared to a control is suitable for pre-harvest or post-harvest treatment with an effective amount of phenylalanine or its analogues.

[0122] In some embodiments, the term "control" includes fruit or plant material characterized by a minimum TSS weight % or TSS / acid or any equivalent standard thereof known to those skilled in the art as a minimum TSS weight % or TSS / acid or any equivalent standard thereof for harvesting.

[0123] In some embodiments, the method includes providing fruit and determining whether the fruit is characterized by a TSS% or TSS / acidity or any equivalent thereof being at least 5%, at least 10%, at least 15%, at least 25%, or at least 30% lower than the minimum TSS% or TSS / acidity or any equivalent thereof known to a person skilled in the art for harvesting.

[0124] In some embodiments, the method includes treatment with an effective amount of aromatic amino acids, such as phenylalanine, characterized in that the TSS% or TSS / acid or any equivalent thereof is at least 5%, at least 10%, at least 15%, at least 25%, or at least 30% lower than the minimum TSS% or TSS / acid or any equivalent thereof known to those skilled in the art.

[0125] In some embodiments, fruits unsuitable for treatment according to the methods disclosed herein are characterized by TSS% or TSS / acid or any equivalent thereof being at most 4% lower, equal to, or greater than the minimum TSS% or TSS / acid or any equivalent thereof known to those skilled in the art for harvesting fruits, or any value and range thereof. Each possibility represents a separate embodiment of the invention.

[0126] Table 1 below provides several examples of minimum threshold values ​​for soluble solids that indicate the suitability of the fruit to be harvested.

[0127]

[0128] In some embodiments, apples suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 10, at most 11, or at most 12, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, apples suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 12 or 13.

[0129] In some embodiments, apricot fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 6, at most 7, at most 8, or at most 9, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, apricot fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 10.

[0130] In some embodiments, Asian pear fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 7, at most 8, at most 9, or at most 10, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, Asian pear fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 11.

[0131] In some embodiments, avocado fruit suitable for treatment according to the methods disclosed herein is characterized by a TSS% value of at most 20, at most 21, at most 22, at most 23, at most 24, at most 25, or any value and range between these values. Each possibility represents a separate embodiment of the invention. In some embodiments, avocado fruit suitable for treatment according to the methods disclosed herein is characterized by a TSS% value of less than 24.5.

[0132] In some embodiments, banana fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 8, at most 9, at most 10, or at most 11, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, banana fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 12.

[0133] In some embodiments, cherry fruit suitable for treatment according to the methods disclosed herein is characterized by a TSS% value of at most 10, at most 11, at most 12, or at most 13, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, cherry fruit suitable for treatment according to the methods disclosed herein is characterized by a TSS% value of less than 14.

[0134] In some embodiments, date palms suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 35, at most 40, at most 45, or at most 49, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, date palms suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 50.

[0135] In some embodiments, grapes suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 17, at most 18, at most 19, or at most 20, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, grapes suitable for treatment according to the methods disclosed herein are characterized by a TSS% value below 18-21.

[0136] In some embodiments, grapefruits suitable for treatment according to the methods disclosed herein are characterized by a TSS / acid value of at most 2, at most 3, at most 4, or at most 5, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, grapefruits suitable for treatment according to the methods disclosed herein are characterized by a TSS / acid value below 5.5-6.

[0137] In some embodiments, kiwifruit suitable for treatment according to the methods disclosed herein is characterized by a TSS% value of at most 10, at most 11, at most 12, or at most 13, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, kiwifruit suitable for treatment according to the methods disclosed herein is characterized by a TSS% value of less than 14.

[0138] In some embodiments, litchi fruit suitable for treatment according to the methods disclosed herein is characterized by a TSS / acid value of at most 26, at most 27, at most 28, or at most 29, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, litchi fruit suitable for treatment according to the methods disclosed herein is characterized by a TSS / acid value below 30.

[0139] In some embodiments, citrus fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS / acid value of at most 3, at most 4, at most 5, or at most 6, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, citrus fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS / acid value below 6.5.

[0140] In some embodiments, mango fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 5, at most 6, at most 7, or at most 8, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, mango fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value below 7-8.

[0141] In some embodiments, mangosteen fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 14, at most 15, at most 16, or at most 8, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, mangosteen fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 17.

[0142] In some embodiments, peaches suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 6, at most 7, at most 8, or at most 9, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, mangoes suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 10.

[0143] In some embodiments, nectarines suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 7, at most 8, at most 9, or at most 8, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, nectarines suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 10.

[0144] In some embodiments, orange fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS / acid value of at most 4, at most 5, at most 6, or at most 7, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, orange fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS / acid value below 8.

[0145] In some embodiments, papaya fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 8, at most 9, at most 10, or at most 11, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, papaya fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 11.5.

[0146] In some embodiments, passion fruit suitable for processing according to the methods disclosed herein is characterized by a TSS% value of at most 10, at most 11, at most 12, or at most 13, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, passion fruit suitable for processing according to the methods disclosed herein is characterized by a TSS% value of less than 14.

[0147] In some embodiments, pear fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 6, at most 7, at most 8, or at most 9, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, pear fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 10.

[0148] In some embodiments, persimmon fruit suitable for treatment according to the methods disclosed herein is characterized by a TSS% value of up to 15, up to 16, up to 17, up to 18, up to 19, or up to 20, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, persimmon fruit suitable for treatment according to the methods disclosed herein is characterized by a TSS% value below 18-21.

[0149] In some embodiments, pineapple fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 8, at most 9, at most 10, or at most 11, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, pineapple fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 12.

[0150] In some embodiments, plum fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 6, at most 7, at most 8, or at most 9, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, plum fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 10.

[0151] In some embodiments, pomegranate fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 13, at most 14, at most 15, or at most 16, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, pomegranate fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 17.

[0152] In some embodiments, rambutan fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 12, at most 13, at most 14, or at most 15, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, rambutan fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 16.

[0153] In some embodiments, the fruit suitable for processing according to the methods disclosed herein is characterized by a TSS% value of at most 9, at most 10, at most 11, or at most 12, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, the fruit suitable for processing according to the methods disclosed herein is characterized by a TSS% value of less than 13.

[0154] In some embodiments, strawberry fruit suitable for treatment according to the methods disclosed herein is characterized by a TSS% value of at most 3, at most 4, at most 5, or at most 6, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, strawberry fruit suitable for treatment according to the methods disclosed herein is characterized by a TSS% value of less than 7.

[0155] In some embodiments, tree tomato fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 6, at most 7, at most 8, or at most 9, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, tree tomato fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 10.

[0156] In some embodiments, watermelon fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of at most 6, at most 7, at most 8, or at most 9, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, watermelon fruits suitable for treatment according to the methods disclosed herein are characterized by a TSS% value of less than 10.

[0157] In some embodiments, plant materials, such as fruits, containing phytochemicals in amounts greater than a predetermined threshold are suitable for treatment with an effective amount of phenylalanine or its analogues before, after, or both.

[0158] In some embodiments, plant materials, such as fruits, containing amounts of phytochemicals below a predetermined threshold are not suitable for treatment with an effective amount of phenylalanine or its analogues before or after harvest, or both.

[0159] In some embodiments, plant materials, such as fruits, containing amounts of phytochemicals below a predetermined threshold are suitable for treatment with an effective amount of phenylalanine or its analogues before, after, or both.

[0160] In some embodiments, plant materials, such as fruits, containing phytochemicals in amounts greater than a predetermined threshold are not suitable for treatment with an effective amount of phenylalanine or its analogues before or after harvest, or both.

[0161] In some implementations, "less than" or "greater than" compared to a predetermined threshold is at least 5% lower or greater.

[0162] In some embodiments, the phytochemical is selected from flavonoids, anthocyanins, pigments, or any combination thereof.

[0163] In some implementations, the pigment includes chlorophyll.

[0164] In some embodiments, the treatment includes wetting, immersion, soaking, injection, spraying, coating, or any combination thereof.

[0165] In some implementations, the treatment takes place in an open area, a greenhouse, a storage facility, or any combination thereof.

[0166] In some embodiments, the improvement is compared with a control plant material (e.g., fruit or vegetable) or a plant containing it. In some embodiments, the control includes a control tuber or a plant containing it.

[0167] In some embodiments, the AAA used herein, such as Phe, improves the flavor, color, or both of the fruit or the juice extracted or derived from it.

[0168] In some embodiments, a composition comprising AAA is provided for improving the flavor, color, quality parameters, or combinations thereof of a fruit or juice extracted or derived therefrom.

[0169] In one embodiment, the composition containing AAA is used to improve the flavor, color, quality parameters, or combinations thereof of plant material (e.g., fruit or juice extracted or derived from it), wherein the concentration of AAA (such as phenylalanine or its analogues) is from 2 mM to 20 mM. In another embodiment, the concentration of AAA (such as phenylalanine) from 2 mM to 20 mM is used to improve the flavor, color, quality parameters, or combinations thereof of plant material (e.g., fruit or juice extracted or derived from it).

[0170] As used herein, the term "juice" includes any portion or fraction of juice extracted or derived from a fruit, as disclosed herein.

[0171] In some implementations, the term "weight" refers to dry weight. In other implementations, weight refers to wet weight.

[0172] According to some embodiments of the present invention, the plant is a dicotyledonous plant. According to some embodiments of the present invention, the plant is a monocotyledonous plant.

[0173] According to one embodiment, the fruit is the fruit of a cultivated fruit tree. According to another embodiment, a cultivated fruit tree refers to a plant whose fruit has economic value. According to another embodiment, the cultivated fruit trees are selected from apples, apricots, Asian pears, avocados, bananas, bell peppers, chili peppers, cherries, corn, cucumbers, dates, eggplants, grapes, grapefruits, kiwis, lychees, citrus fruits, mangoes, mangosteens, melons, pumpkins, peaches, pears, nectarines, oranges, papayas, passion fruit, pears, persimmons, pineapples, plums, pomegranates, rambutans, olives, strawberries, tree tomatoes, tomatoes, watermelons, and lemons.

[0174] In one implementation, the cultivated fruit trees include any citrus fruits.

[0175] According to some implementation methods, a method for improving the quality of plant materials is provided.

[0176] In some implementations, improving the quality of plant material includes increasing scratch healing, increasing cut healing, increasing scratch color healing, reducing weight loss percentage, increasing lignin production rate or cork resin production rate or both, or any combination thereof.

[0177] According to some implementation methods, a method for improving the healing of tuber damage is provided.

[0178] In some embodiments, the method includes treating the tuber, the leaves of the plant containing the tuber, or both, with an effective amount of phenylalanine or an analogue before, after, or both of harvesting, thereby improving the healing of tuber damage.

[0179] In some embodiments, the damage is induced by a biological agent, a non-biological agent, or both. In some embodiments, the damage is biological damage. In some embodiments, the damage is non-biological damage (e.g., but not limited to physical or mechanical damage).

[0180] In some implementations, biological agents include plant pathogens or pests.

[0181] In some implementations, plant pathogens or pests include viruses, bacteria, nematodes, arthropods or any developmental stage thereof, or any combination thereof.

[0182] In some implementations, arthropods include insects or worms.

[0183] In some implementations, non-biological agents include physical or mechanical damage.

[0184] In some implementations, improved healing includes: increased scratch healing, increased cut healing, increased scratch color healing, reduced weight loss percentage, increased lignin or cork resin production rate, or any combination thereof.

[0185] In some implementations, the tuber includes stem tubers or root tubers.

[0186] In some embodiments, the tuber is a stem tuber. In some embodiments, the tuber is a root tuber.

[0187] The types of tubers are obvious to those skilled in the art.

[0188] In some embodiments, the tuber is selected from: potato, sweet potato (e.g., yam), cassava, carrot, radish, jicama, or any combination thereof.

[0189] In some embodiments, the tuber is a potato tuber.

[0190] As used herein, the term "tuber" includes plants that contain tubers. In some embodiments, the tuber is a potato plant.

[0191] As used herein, the term "phenylalanine" or "Phe" refers to the α-amino acid having the formula C9H11NO2. It can be considered as a benzyl-substituted methyl group of alanine, or a phenyl-substituted terminal hydrogen group of alanine. Due to the inertness and hydrophobicity of the benzyl side chain, this essential amino acid is classified as neutral and nonpolar. The L-isomer is used in the biochemical formation of proteins encoded by DNA. The codons for L-phenylalanine are UUC and UUU. Phenylalanine is a precursor to tyrosine; the monoamine neurotransmitters dopamine, norepinephrine, and epinephrine; and the skin pigment melanin.

[0192] As used herein, the term "tyrosine" or "Tyr" refers to the amino acid having the formula C9H11NO3. It can be considered as a tyrosine group replacing the methyl group of alanine. Due to the inertness and hydrophobicity of the tyrosine side chain, this non-essential amino acid is classified as neutral and nonpolar. The L-isomer is used in the biochemical formation of proteins encoded by DNA. The codons for L-tyrosine are UAC and UAU.

[0193] As used herein, the term "tryptophan" or "Trp" refers to the α-amino acid with the formula C11H12N2O2. Due to the inertness and hydrophobicity of its benzyl side chain, this essential amino acid is classified as neutral and nonpolar. The L-isomer is used in the biochemical formation of proteins encoded by DNA. The codon for tryptophan is UGG. Tryptophan is a precursor to the neurotransmitters serotonin, melatonin, and vitamin B3.

[0194] “Phenylalanine analogue” or “Phe analogue” means any naturally occurring or synthetically produced (chemical or biosynthetic) Phe analogue that improves fruit flavor, color, quality parameters or any combination thereof, as disclosed herein.

[0195] According to one embodiment, a Phe analog is a naturally occurring compound. In one embodiment, the use of the term "Phe" includes compositions comprising an effective amount of Phe or an analog of Phe. According to some embodiments, a Phe analog comprises or is an aromatic compound. According to one embodiment, a Phe analog is tyrosine or a synthetic analog of tyrosine, which can improve fruit flavor, color, quality parameters, or any combination thereof, as disclosed herein. Synthetic analogs are commercially available, for example from AnaSpec. A list of non-limiting examples is provided below. Measures should be taken to test for phytotoxicity before application to plants. Table 2 below lists some non-limiting examples of Phe and Tyr analogs.

[0196] Table 2. Phe and Tyr analogues

[0197]

[0198]

[0199]

[0200]

[0201]

[0202] Table 3. Non-limiting examples of Trp analogues

[0203] Boc-4-methyl-DL-tryptophan Boc-6-Fluoro-DL-Tryptophan Boc-6-methyl-DL-tryptophan Boc-DL-7-azine Fmoc-(R)-7-azatryptophan Fmoc-5-benzyloxy-DL-tryptophan Fmoc-5-bromo-DL-tryptophan Fmoc-5-chloro-DL-tryptophan Fmoc-5-hydroxy-L-tryptophan Fmoc-6-chloro-L-tryptophan Fmoc-6-methyl-DL-tryptophan Fmoc-7-methyl-DL-tryptophan Fmoc-DL-7-azatryptophan

[0204] According to one embodiment, the analog is aspartame. According to one embodiment, the analog is tyrosine.

[0205] Low concentrations (e.g., 0.01-10 mM, 0.01-5 mM, 0.01-1 mM, 0.1-10 mM, 0.1-5 mM, and 0.1-1 mM) of AAAs, such as Phe, Tyr, Trp, or their analogues, can be used, especially when used in combination with surfactants or when using a combination of AAAs and analogues (e.g., Phe, Tyr, and Trp).

[0206] According to one embodiment, AAA, such as Phe or its analogues, is applied at a concentration that will not cause precipitation on plants. According to one embodiment, Phe or its analogues are applied at a concentration that will not cause Phe precipitation on plants.

[0207] In some implementations, AAA, such as Phe or its analogues, is applied at concentrations that do not cause or induce fruit ripening and / or decay.

[0208] As used herein, “plant” means the whole plant, plant tissue, plant organ, fruit, vegetable, edible part of a plant, grafted plant, including seeds, buds, stems, roots (including tubers), rhizomes, scions, and plant cells, tissues, fruits, flowers, and organs. A plant can be in any form, including cuttings and harvested material (e.g., fruit).

[0209] Phe (or similar substances) can be applied to the fruit by spraying, sprinkling, coating, soaking, irrigating, immersing, or treating with active ingredients.

[0210] When referring to a specific stage, application may be limited to that stage or the stated stage plus another stage. For example, when referring to application during flowering, application may be carried out during flowering or flowering + post-flowering (i.e., fruiting), or it may be carried out before and during flowering, or before and after flowering.

[0211] According to one embodiment, phenylalanine or the like is formulated as a composition selected from extracts, sprays, or concentrates. According to one embodiment, it is applied near or on plant fruits. According to one embodiment, application is by irrigation, soaking, maceration, immersion, injection, coating, or spraying. According to one embodiment, application is in an open area. According to one embodiment, application is in a greenhouse. According to one embodiment, application is in a storage facility (e.g., a dark room, a cold storage). According to one embodiment, application is a single application. According to one embodiment, application is applied at least twice as needed and / or as described herein in any scheme or duration.

[0212] According to one implementation, application includes repeated application (two or more applications, e.g., weekly, seasonal, every two weeks, every two months, etc.). Repeated application is particularly suitable for field / greenhouse treatments.

[0213] According to one implementation, repeated application includes application during flowering, after flowering, before flowering, or any combination thereof, weekly, daily, monthly, or every two months. For example, proposed programs may include, but are not limited to, spraying plants in open fields and greenhouses, increasing irrigation for plants grown in open fields, greenhouses, and pots, immersing whole leaves in the solution after harvest, and adding it to a bottle of cut flowers before and / or after harvest and possibly before shipment.

[0214] According to one embodiment, an active ingredient (Phe and / or the like) is formulated into a composition wherein it is mixed with other active ingredients (e.g., fungicides) and / or agriculturally acceptable carriers.

[0215] According to one embodiment, such compositions of the present invention are shelf-stable. The term "shelf-stable" means that the compositions of the present invention retain their activity during a given storage period under recommended conditions (e.g., temperature) and optionally do not separate into separate phases or produce any unpleasant odors.

[0216] As used herein, the term "agriculturally acceptable carrier" refers to a material that facilitates the application of the compositions of the present invention to an intended target, such as plants, plant material, compost, soil, the environment, or equipment, or a material that facilitates storage, transport, or disposal. The carrier used in the compositions applied to plants and plant material is preferably non-phytotoxic or only mildly phytotoxic. Depending on the desired formulation, a suitable carrier may be solid, liquid, or gaseous. In one embodiment, the carrier comprises a polar liquid carrier, such as water, mineral oil, and vegetable oil. In one embodiment, the carrier enhances the stability of the active ingredient described herein.

[0217] Examples of liquid carriers include, but are not limited to, water; alcohols, especially butanol or glycol, and their ethers or esters, especially methyl glycol acetate; ketones, especially acetone, cyclohexanone, methyl ethyl ketone, methyl isobutyl ketone, or isophorone; petroleum fractions such as alkanes or aromatics, especially xylene or alkylnaphthalenes; mineral oils or vegetable oils; aliphatic chlorinated hydrocarbons, especially trichloroethane or dichloromethane; aromatic chlorinated hydrocarbons, especially chlorobenzene; water-soluble or strongly polar solvents such as dimethylformamide, dimethyl sulfoxide, or N-methylpyrrolidone; liquefied gases; and mixtures thereof.

[0218] Examples of solid carriers include, but are not limited to, fillers such as kaolin, bentonite, dolomite, calcium carbonate, talc, powdered magnesium oxide, fullerite, gypsum, diatomaceous earth, and Chinese clay. Carriers providing slow or delayed release of the compounds of the present invention (Phe or the like) may also be included in the compositions of the present invention (particularly for short-life-cycle pathogens).

[0219] In another embodiment, the composition of the present invention (or its active ingredient—Phe or the like) is applied in an amount capable of inhibiting bacterial spore germination or bacterial spread. According to one embodiment, the composition of the present invention (or its active ingredient—Phe or the like) is applied in an amount capable of reducing the standard concentration recommended by regulatory agencies (e.g., FDA, USDA) for commonly used agricultural technical formulations.

[0220] The application can be applied directly to the fruit, plant, or close enough to the surface of the fruit or plant to improve flavor, color, or parameters, as described herein. In one embodiment, "close enough" is within a distance of approximately 1 meter. In one embodiment, "close enough" is within a distance of approximately 0.7 meters. In one embodiment, "close enough" is within a distance of approximately 0.5 meters. In one embodiment, "close enough" is within a distance of approximately 0.2 meters.

[0221] Therefore, application can be to any target surface of a plant or plant organ such as a fruit, and the compounds or compositions of the present invention can be applied to such surface, for example, to a plant, plant material (including fruit, root, bulb, tuber, bulb, leaf, flower, seed, stem, callus, nut, grain, cutting, rhizome, scion), harvested crop (including root, bulb, tuber, bulb, leaf, flower, seed, stem, callus, nut, grain, fruit, cutting, rhizome, scion), or any surface that may come into contact with the harvested crop (including harvesting equipment, packaging equipment, and packaging materials).

[0222] The compound or composition of the present invention is applied to surfaces such as harvesting equipment, packaging equipment, and packaging materials before use of the harvesting equipment, packaging equipment, or packaging materials.

[0223] According to one embodiment, the compounds or compositions of the present invention are formulated as extracts, powders, sprays, or concentrates. According to one embodiment, the formulation includes a surfactant. According to one embodiment, the surfactant is a cationic surfactant, such as benzalkonium chloride or hexadecylpyridinium chloride. According to one embodiment, the surfactant is anionic, such as alkyl sulfates or alkyl ethoxylate sulfates. According to one embodiment, the surfactant is a nonionic surfactant, such as alkyl polyglycosides, Triton X-100, polyoxyethylene (20) sorbitan monooleate (Tween-80), or Silwett L-77. According to one embodiment, the surfactant is Tween-80 or Silwett L-77.

[0224] According to one embodiment, the concentration of the surfactant is at least 0.1%. In one embodiment, the composition of the present invention may further comprise at least one additional agricultural agent. In an alternative embodiment, the composition of the present invention may be delivered separately, simultaneously, or sequentially with at least one additional agricultural agent.

[0225] In some embodiments, AAA can be a composition having a coating agent, such as, but not limited to, polysaccharides.

[0226] According to one embodiment, the agricultural composition may contain phenylalanine and tyrosine for improving fruit flavor (including juice extracted or derived therefrom), color, quality parameters, or any combination thereof.

[0227] As used in this article, the term “about” means ±10%.

[0228] The terms “comprises”, “comprising”, “includes”, “having”, and their grammatical variations mean “including but not limited to”.

[0229] The term "composed of" means "including and limited to". The term "substantially composed of" means that a composition, method, or structure may include additional ingredients, steps, and / or portions, provided that the additional ingredients, steps, and / or portions do not materially alter the essential and novel characteristics of the claimed composition, method, or structure. As used herein, the singular forms "an", "an", and "the" include the plural forms unless the context clearly specifies otherwise. For example, the terms "a compound" or "at least one compound" may include multiple compounds, including mixtures thereof.

[0230] Throughout this application, various embodiments of the invention may be presented in a range format. It should be understood that the range format is merely for convenience and brevity and should not be construed as a rigid limitation on the scope of the invention. Therefore, the description of a range should be considered as specifically disclosing all possible subranges and individual numerical values ​​within that range. For example, a description of a range such as 1 to 6 should be considered as specifically disclosing subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., and individual numbers within that range, such as 1, 2, 3, 4, 5, and 6. This applies regardless of the width of the range.

[0231] Whenever a range of numbers is indicated herein, it means to include any numbers (fractions or integers) mentioned within the indicated range. The phrases “range between the first indicated number and the second indicated number” and “range from the first indicated quantity to the second indicated quantity” are used interchangeably herein and mean to include the first indicated quantity and the second indicated quantity, as well as all decimals and integers in between.

[0232] As used herein, the term "method" refers to the manner, means, technique, and procedure used to accomplish a given task, including but not limited to those manner, means, technique, and procedure known to practitioners in the fields of chemistry, pharmacology, biology, biochemistry, and medicine, or readily developed from known manner, means, technique, and procedure. As used herein, the term "treatment" includes eliminating, substantially inhibiting, slowing, or reversing the progression of a condition, substantially improving the clinical or aesthetic symptoms of the condition, or substantially preventing the occurrence of the clinical and aesthetic symptoms of the condition.

[0233] Other objects, advantages, and novel features of the invention will become apparent to those skilled in the art upon examination of the following embodiments, which are not intended to be limiting. Furthermore, each of the various embodiments and aspects of the invention as described above and claimed in the claims section has been experimentally supported in the following embodiments.

[0234] It should be understood that, for clarity, certain features of the invention described in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, for brevity, various features of the invention described in the context of a single embodiment may also be provided separately, or in any suitable sub-combination, or appropriately provided in any other described embodiment of the invention. Certain features described in the context of various embodiments should not be considered essential features of those embodiments unless the embodiment cannot be practiced without these elements.

[0235] Example

[0236] The following embodiments, together with the foregoing description, illustrate some implementations of the invention in a non-limiting manner.

[0237] Example 1

[0238] Improving fruit quality with post-harvest treatment using phenylalanine

[0239] Materials and methods

[0240] Improve fruit flavor

[0241] Mangoes – Unripe mangoes were harvested and immersed in 4-8 mM phenylalanine for 30 seconds. The fruits were stored at 22°C until ripe (days 11-15), during which sensory analysis was performed by a panel of 20 tasters. Additionally, the Brix content, acidity % (calibrated according to citric acid), and firmness (in Newtons) of five different fruit samples were determined.

[0242] Apples – The apple trees were treated with 8 mM phenylalanine one or two weeks before harvest; or with 0.2% jasmone (also known as “blushing”). The fruit was harvested from the outer region of the tree. After harvest, the fruit was stored at 0°C for three months, during which a panel of 20 tasters performed sensory analysis on fruit slices and juice extracted from them. Brix values ​​were also determined.

[0243] Grapes—Muscat and Petit Verdot grapes were treated with 8 mM phenylalanine for (i) one or two weeks; and (ii) one, two, or three weeks prior to harvest, as well as combinations thereof. A panel of 20 tasters performed sensory analysis on juices extracted from the fruit, and Brix and acidity % (calibrated to citric acid) values ​​were also determined. In the case of Muscat grapes, whole fruit was also analyzed (e.g., by the panel, Brix and acidity %).

[0244] Improve fruit color

[0245] Mango or apple trees were treated with 8 mM phenylalanine or 0.2% jasmone (“reddening”) one or two weeks before harvest. Fruit was harvested from the outer areas of the tree where it was exposed to sunlight (photon flux of 1500 ± 200 μmol·m⁻¹). -2 ·sec -1 After harvest: (a) store mangoes at 12°C for 3 weeks and then at 22°C for another week (e.g., shelf life); and (b) store apples at 0°C for 3 months and then at 22°C for another week (e.g., shelf life).

[0246] The red surface area was assessed as a percentage, and the red intensity was assessed using a scale of 0-5. For each treatment at different time points (harvest, after refrigeration at 12°C, and after shelf life at 22°C), the percentage of red surface area on each mango fruit was assessed. Similarly, the red intensity of each mango was assessed using a visual rating scale, where 0 = no red, 1 = faint red, and 5 = very intense red. Fifty fruits were assessed for each treatment. The hue of the skin of 15 mango fruits from each treatment was measured using a CR-400 / 410 colorimeter (Konicka Minolta, Osaka, Japan) at the reddest point on the equator of each fruit. Hue angles were used to measure color (120 for green; 60-70 for yellow; 30-40 for red).

[0247] Chlorophyll, anthocyanin, and flavonoid contents were measured using a Multiplex III fluorescence detector (Force A, Orsay, France), which consists of 12 fluorescence signals. The ratios between these signals in different mathematical expressions were correlated with the fluorescence of major chemical groups, such as anthocyanins (FER_RG, the ratio of infrared emission excited by red or green light), flavonoids (FLAV), and chlorophyll (SER_R). Fifteen fruits from each treatment were evaluated on the red side.

[0248] result

[0249] According to the sensory analysis of the tasting panel, the treated mangoes were found to be of better quality, for example, sweeter, less acidic, more fragrant, firmer, and more visually appealing. Figure 1-3 , 7 and 12). In addition, as in respectively Figure 1-3 7 and 12 and Figure 8 and 13 As can be seen, the Brix value measured in the treated mango fruit or the juice extracted or derived from it was greater than that in the control.

[0250] Furthermore, it was shown that treatment with phenylalanine can effectively induce the formation of redness in mango fruit. Figure 4-6 (and 9-11).

[0251] It was found that Muscat grapes and Petit Verdot grapes treated according to the methods disclosed herein and / or juices extracted or derived from them included increased Brix values ​​compared to controls. Figure 14-16 ).

[0252] Based on sensory analysis by a panel of tasters, the processed apples and / or juices extracted or derived from them were found to be of higher quality, for example, sweeter, lower in acidity, and more aromatic (see [link to relevant documentation]). Figure 20 and 21The team also determined that the treated apples had a better texture. Figure 20 Furthermore, the Brix values ​​determined in the treated apple fruit were higher than those in the control, such as in... Figure 20 visible.

[0253] Furthermore, it was shown that treatment with phenylalanine can effectively induce the reddening of apple fruits. Figure 17-19 ).

[0254] Furthermore, the inventors have demonstrated that the post-harvest treatment disclosed herein improves fruit aroma. The inventors soaked mango fruit and, after storing it at 22°C for 11 days, determined the levels of aroma-related VOCs. Indeed, post-harvest treatment with an effective amount of phenylalanine induced a significant increase in VOC levels in all tests (Figure 30). Specifically, compared to untreated fruit, the treated mango fruit showed significantly higher levels of α-pinene (30A), 3-carene (30B), D-limonene (30C), guarbenene (30D), α-terpinene (30E), α-phellandrene (30F), and caryophyllene (30G) in the pulp.

[0255] Furthermore, the inventors have demonstrated that the post-harvest treatment disclosed herein reduces the accumulation of ROS (reactive oxygen species) in post-harvest fruit damage (Figure 31). Specifically, the inventors have demonstrated that mango fruits post-harvest treated with phenylalanine according to the method disclosed herein accumulate significantly less ROS in the peel and pulp compared to untreated damaged fruits.

[0256] Example 2

[0257] Post-harvest treatment with phenylalanine induced potato tuber healing.

[0258] Red (Memphis) and white (Sivra) potato tubers were immersed in water or 8 mM Phe for 1 minute and then dried. The tubers were stored at 20°C for 2 days. Each tuber was scratched at 3 points (2 cm long, 1 cm wide). After one and two weeks, the healing of the scratches was assessed using a 0-3 scale, the recovery of the red tubers was assessed using a 0-3 scale, and weight loss was measured. Scratch healing was significantly improved in both types of potatoes at 7 and 14 days post-treatment. Figure 22A and 22C In red (Memphis) potatoes, color recovery was also significantly improved at 7 and 14 days post-treatment. Figure 22E-22F The weight loss of both types of potatoes was also reduced. Figure 22B and 22D (This improvement was found to be statistically significant in white potatoes).

[0259] Red (Memphis) and white (Sivra) potato tubers were immersed in water or 8 mM Phe for 1 minute and then dried. The tubers were stored at 20°C for 2 days. Each tuber was cut at 3 points (2 cm long, 2 cm deep, 2 mm wide). Wound healing was assessed using a 0-3 scale and weight loss was measured after one and two weeks. As early as 7 days post-treatment, wound healing was significantly improved in Sivra potatoes compared to the control group, and this improvement continued further at 14 days post-treatment. Figure 23A A similar trend was initially observed 7 days after Memphis potato treatment. Figure 23C Although the degree was smaller after 14 days. Furthermore, both types of potatoes showed reduced weight loss. Figure 23B and 23D The improvement was statistically significant 14 days after treatment.

[0260] White (Sivrac) potato tubers were immersed in water or 8 mM Phe for 1 minute and then dried. The tubers were stored at 20°C for 2 days. Each tuber was cut (2 cm long, 2 cm deep, 2 mm wide). Wound healing was recorded after one week. Lignin was stained white with Calcofluor and photographed under a fluorescence microscope; fluorescence intensity was measured in 10 replicates using ImageJ. Compared with the control group (… Figures 24A-24C Compared to the previous treatment, increased lignin production was observed in Phe-treated potatoes one week after treatment. Figure 24B and 24D -24E).

[0261] Soak red (Memphis) and white (Sivra) potato tubers in water or 8mM Phe for 1 minute, then dry. Store the tubers at 20°C for 2 days. Cut each tuber (2 cm long, 2 cm deep, 2 mm wide). After two weeks, record the surface and internal healing. Phe-treated Sivra potatoes ( Figure 25B and 25D ) and Memphis potatoes ( Figure 25F ) surface (e.g., exterior; Figure 25B ) or inside ( Figure 25D and 25F ) and control ( Figure 25A , 25C Both have been improved compared to 25E.

[0262] Example 3

[0263] Pre-harvest treatment with phenylalanine improves fruit quality.

[0264] The inventors further demonstrated that pre-harvest treatment with phenylalanine in different types of fruits, such as mangoes (Fig. 26) and apples (Fig. 27), significantly improved fruit color and TSS. Furthermore, improvements in fruit color were also observed in grapes. Figure 28 Furthermore, the desired results of improving this fruit quality characteristic were achieved when the phenylalanine concentration was 0.06% by weight (~3 mM) or higher, such as 0.012% and 0.24% (approximately 7 mM and 14.5 mM, respectively). Specifically, a concentration of 0.01% (equivalent to ~0.5 mM) of phenylalanine was shown to be ineffective in improving: (i) reddening in any of mangoes, apples, and grapes (Figs. 26–28); and (ii) TSS levels in mangoes or apples (Figs. 26–27). In fact, in most of these cases, 0.01% was comparable to or the same as the control group and therefore could not be considered relevant to achieving the sought technical effect.

[0265] Furthermore, the inventors have examined the effect of pre-harvest treatment with phenylalanine on enzymatic browning (i.e., oxidation) in fruit. The inventors demonstrated that enzymatic browning was significantly reduced in sliced ​​apples treated with 0.12% phenylalanine 2 or 4 weeks before harvest (Figure 29).

[0266] Therefore, the current data provides substantial scientific evidence that phenylalanine at concentrations between 0.06% and 0.24% (~3mM to 14.5mM) is suitable for improving fruit quality.

[0267] While certain features of the invention have been described herein, many modifications, substitutions, alterations, and equivalents will now occur to those skilled in the art. Therefore, it should be understood that the appended claims are intended to cover all such modifications and alterations that fall within the true spirit of the invention.

Claims

1. A method for improving the color of fruit of a cultivated fruit tree, the method comprising treating the cultivated fruit tree or its fruit with an effective amount of phenylalanine of 2 mM to 20 mM before or after harvest, wherein the improvement comprises inducing reddening of the fruit, and wherein the improvement comprises not an increase caused by a reduction in color induced by plant pathogens.

2. The method according to claim 1, wherein the effective amount of phenylalanine is greater than 2 mM.

3. The method according to claim 1, wherein the effective amount of phenylalanine is 4 mM to 20 mM.

4. The method of claim 1, further comprising the step of providing the cultivated fruit tree or its fruit with a period of 1 to 30 days of pre-harvest abiotic stress.

5. The method of claim 4, wherein the abiotic stress conditions include light, radiation, temperature, lack of nutrients or water, or any combination thereof.

6. The method of claim 1, further comprising the step of selecting cultivated fruit trees or their fruits that require color improvement using the phenylalanine.

7. The method of claim 6, wherein the selection comprises determining the amount of phytochemical in the cultivated fruit tree or its fruit compared to a predetermined threshold, wherein cultivated fruit trees or their fruit containing an amount of the phytochemical greater than the predetermined threshold are suitable for pre-harvest or post-harvest treatment with an effective amount of phenylalanine.

8. The method according to claim 7, wherein the phytochemical is selected from anthocyanins, pigments, and both.

9. The method of claim 8, wherein the pigment comprises chlorophyll.

10. The method of claim 1, wherein the processing comprises: Impregnation, soaking, immersion, injection, spraying, coating, or any combination thereof.

11. The method of claim 1, wherein the treatment is carried out in an open area, a greenhouse, a storage facility, or any combination thereof.

12. The method of claim 1, wherein the improvement is compared with control plant material.

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