Lactobacillus pentosus and application thereof in mango storage and mango e-commerce logistics
Patent Information
- Application Number
- CN202610221403.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2026-02-25
- Publication Date
- 2026-09-08
- Estimated Expiration
- 2046-02-25
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of biotechnology and relates to a strain of Lactobacillus pentosus and its application in mango storage and e-commerce logistics of mangoes. Background Technology
[0002] Mango anthracnose is a latent disease that occurs during the flowering period. Under high temperature and humidity, the incidence rate can reach 100%, severely impacting the development of the mango industry. In the context of "smart agriculture + e-commerce," mango e-commerce and delivery have become important channels for mango sales and logistics, possessing enormous development potential. However, improper handling can significantly affect mango quality, especially for latent diseases. Fruits that appear disease-free in the field often develop the disease during the ripening and storage periods, causing fruit rot and severely impacting post-harvest quality, resulting in substantial economic losses. Losses during storage and transportation are generally 30%-50%, and in severe cases, can reach 100%.
[0003] Biological control has developed rapidly in recent years, effectively inhibiting the growth and reproduction of pathogens. It is safe, environmentally friendly, and economical, making it a key factor driving the development of the fruit and vegetable industry. Microbial antagonistic preservation refers to the process by which one microorganism produces antibacterial metabolites during its life activities, inhibiting or killing another microorganism, or competing with harmful microorganisms for nutrients, thus achieving the purpose of preservation.
[0004] Antagonistic bacteria are applied to fruits mainly through spraying, soaking, coating, and molding to act on the surface of mangoes. However, different application methods have different adaptability to the same type of fruit. Summary of the Invention
[0005] The purpose of this invention is to provide a strain of Lactobacillus pentosus and its application in mango storage and e-commerce logistics of mangoes.
[0006] This invention provides Lactobacillus pentosolicus ( Lactobacillus pentosus Lactobacillus pentosus GHY-5 was deposited on October 22, 2025, at the China General Microbiological Culture Collection Center (CGMCC, address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences), with accession number CGMCC No. 36297.
[0007] Furthermore, the present invention provides a bacterial agent of Lactobacillus pentosus GHY-5.
[0008] Specifically, the bacterial agent is a liquid bacterial agent.
[0009] The active ingredient provided by the bacterial agent is Lactobacillus pentosolicus GHY-5.
[0010] The bacterial agent contains 1×10⁻⁶ Lactobacillus pentosolicus GHY-5. 8 CFU / ml or higher.
[0011] The bacterial agent contains 1×10⁻⁶ Lactobacillus pentosolicus GHY-5. 8 cfu / ml up to 1×10 12 cfu / ml.
[0012] The bacterial agent contains 1×10⁻⁶ Lactobacillus pentosolicus GHY-5. 10 cfu / ml.
[0013] Specifically, the preparation method of the bacterial agent is as follows: Lactobacillus pentosus GHY-5 is inoculated into MRS liquid medium and cultured at 37°C with shaking at 170 rpm until the bacterial concentration reaches 1×10⁻⁶. 8 CFU / ml or higher.
[0014] Specifically, the preparation method of the bacterial agent is as follows: Lactobacillus pentosus GHY-5 is inoculated into MRS liquid medium and cultured at 37°C with shaking at 170 rpm until the bacterial concentration reaches 1×10⁻⁶. 8 cfu / ml up to 1×10 12 cfu / ml.
[0015] Specifically, the preparation method of the bacterial agent is as follows: Lactobacillus pentosus GHY-5 is inoculated into MRS liquid medium and cultured at 37°C with shaking at 170 rpm until the bacterial concentration reaches 1×10⁻⁶. 10 cfu / ml.
[0016] This invention also protects a liquid reagent for improving the storage performance of mangoes, comprising chitosan and Lactobacillus pentosus GHY-5; wherein the concentration of chitosan in the liquid reagent is 0.3-3 g / 100 mL, and the concentration of Lactobacillus pentosus GHY-5 is 0.1 × 10⁻⁶ g / mL. 10 -10×10 10 cfu / mL.
[0017] In the liquid reagent, the concentration of chitosan is 0.5-1.5 g / 100 mL, and the concentration of Lactobacillus pentosus GHY-5 is (0.4-0.6) × 10⁻⁶. 10 cfu / mL.
[0018] In the liquid reagent, the concentration of chitosan is 1 g / 100 mL, and the concentration of Lactobacillus pentosus GHY-5 is 0.5 × 10⁻⁶. 10 cfu / mL.
[0019] The liquid reagent further includes glycerol. The concentration of glycerol in the liquid reagent is 0.4-0.6 mL / 100 mL. The concentration of glycerol in the liquid reagent is 0.5 mL / 100 mL.
[0020] The liquid reagent also includes glacial acetic acid.
[0021] The liquid reagent also includes water.
[0022] In one specific form, the liquid reagent consists of Lactobacillus pentosus GHY-5 bacterial culture, chitosan, glycerol, glacial acetic acid, and water.
[0023] The preparation method of Lactobacillus pentosus GHY-5 bacterial culture is as follows: Lactobacillus pentosus GHY-5 is inoculated into liquid MRS medium and cultured until the bacterial concentration reaches (0.1-10)×10⁻⁶. 10 cfu / mL indicates Lactobacillus pentosus GHY-5 bacterial culture.
[0024] The preparation method of Lactobacillus pentosus GHY-5 bacterial culture is as follows: Lactobacillus pentosus GHY-5 is inoculated into liquid MRS medium and cultured until the bacterial concentration reaches (0.8-1.2) × 10⁻⁶. 10 cfu / mL indicates Lactobacillus pentosus GHY-5 bacterial culture.
[0025] The preparation method of Lactobacillus pentosus GHY-5 bacterial culture is as follows: Lactobacillus pentosus GHY-5 is inoculated into liquid MRS medium and cultured until the bacterial concentration reaches 1×10⁻⁶. 10 cfu / mL indicates Lactobacillus pentosus GHY-5 bacterial culture.
[0026] The liquid reagent is prepared as follows: chitosan solution and Lactobacillus pentosus GHY-5 bacterial solution are mixed in equal volumes to obtain the liquid reagent.
[0027] The chitosan solution is prepared as follows: Dissolve 1-3g of chitosan and 0.8-1.2mL of glycerol in 0.5-1.5g / 100mL glacial acetic acid aqueous solution, and then bring the volume up to 100mL with 0.5-1.5g / 100mL glacial acetic acid aqueous solution.
[0028] The chitosan solution is prepared as follows: Dissolve 2g of chitosan and 1mL of glycerol in 1g / 100mL glacial acetic acid aqueous solution, and then bring the volume up to 100mL with 1g / 100mL glacial acetic acid aqueous solution.
[0029] The preparation method of chitosan solution is as follows: Dissolve 2g chitosan and 1mL glycerol in 1g / 100mL glacial acetic acid aqueous solution in a 40℃ water bath, then make up to 100mL with 1g / 100mL glacial acetic acid aqueous solution, and then perform ultrasonic treatment (ultrasonic parameters: 40℃, 300W, 20 minutes).
[0030] The present invention also provides a method for pretreating mangoes, comprising the following steps: taking a mango, coating its surface with the liquid reagent, and then drying it; the purpose of the pretreatment is to improve the storage performance of the mango.
[0031] The mangoes mentioned are harvested mango fruits.
[0032] Specifically, apply 1-3 ml of the liquid reagent to each mango.
[0033] Specifically, each mango is coated with 2 ml of the liquid reagent.
[0034] The present invention also provides the application of Lactobacillus pentosus GHY-5 or the bacterial agent or the liquid reagent, as described in any of the following (a1) to (a6): (a1) Application in mango preservation; (a2) Application in the preparation of mango preservatives; (a3) Application in mango storage; (a4) Application in the preparation of reagents for mango storage; (a5) Application in the pretreatment of mangoes; the purpose of the pretreatment is to improve the storage performance of mangoes; (a6) Application in the preparation of products that pre-treat mangoes; the purpose of which is to improve the storage performance of mangoes.
[0035] The mangoes mentioned are harvested mango fruits.
[0036] The improvement in the storage performance of mangoes is manifested in any of the following (b1) to (b15): (b1) Inhibits mango fruit rot; (b2) Inhibit anthracnose in mango fruit; (b3) Delay the ripening process of mangoes; (b4) Reduce the respiration rate of mango fruit; (b5) Reduce the ethylene release from mango fruit; (b6) Inhibits the decrease in titratable acidity of mango fruit during storage; (b7) Inhibits the increase of soluble solids in mango fruit during storage; (b8) Inhibits the increase in electrical conductivity of mango fruit during storage; (b9) Inhibits the increase in cell membrane permeability during mango fruit storage; (b10) Delays the decrease in firmness of mango fruit during storage; (b11) Delays the decline in the elasticity of mango fruit during storage; (b12) Delays the recovery decline of mango fruit during storage; (b13) Delays the decline in cohesion of mango fruit during storage; (b14) Delays the decline in the adhesiveness of mango fruit during storage; (b15) Delays the decline in chewiness of mango fruit during storage.
[0037] The present invention also provides the application of Lactobacillus pentosus GHY-5 or the bacterial agent or the liquid reagent, as described in any of the following (b1) to (b15): (b1) Inhibits mango fruit rot; (b2) Inhibit anthracnose in mango fruit; (b3) Delay the ripening process of mangoes; (b4) Reduce the respiration rate of mango fruit; (b5) Reduce the ethylene release from mango fruit; (b6) Inhibits the decrease in titratable acidity of mango fruit during storage; (b7) Inhibits the increase of soluble solids in mango fruit during storage; (b8) Inhibits the increase in electrical conductivity of mango fruit during storage; (b9) Inhibits the increase in cell membrane permeability during mango fruit storage; (b10) Delays the decrease in firmness of mango fruit during storage; (b11) Delays the decline in the elasticity of mango fruit during storage; (b12) Delays the recovery decline of mango fruit during storage; (b13) Delays the decline in cohesion of mango fruit during storage; (b14) Delays the decline in the adhesiveness of mango fruit during storage; (b15) Delays the decline in chewiness of mango fruit during storage.
[0038] The present invention also provides the application of Lactobacillus pentosus GHY-5 or the bacterial agent or the liquid reagent in the preparation of products.
[0039] The present invention also provides a product comprising Lactobacillus pentosus GHY-5 or the bacterial agent or the liquid reagent.
[0040] The use of any of the products described above is as described in any of the following (b1) to (b15): (b1) Inhibits mango fruit rot; (b2) Inhibit anthracnose in mango fruit; (b3) Delay the ripening process of mangoes; (b4) Reduce the respiration rate of mango fruit; (b5) Reduce the ethylene release from mango fruit; (b6) Inhibits the decrease in titratable acidity of mango fruit during storage; (b7) Inhibits the increase of soluble solids in mango fruit during storage; (b8) Inhibits the increase in electrical conductivity of mango fruit during storage; (b9) Inhibits the increase in cell membrane permeability during mango fruit storage; (b10) Delays the decrease in firmness of mango fruit during storage; (b11) Delays the decline in the elasticity of mango fruit during storage; (b12) Delays the recovery decline of mango fruit during storage; (b13) Delays the decline in cohesion of mango fruit during storage; (b14) Delays the decline in the adhesiveness of mango fruit during storage; (b15) Delays the decline in chewiness of mango fruit during storage.
[0041] Any of (a1) to (a15): The mango fruit mentioned above refers to the harvested mango fruit.
[0042] Specifically, any of the mangoes mentioned above are Taiwanese mangoes.
[0043] Specifically, any of the mangoes mentioned above are small Taiwan mangoes.
[0044] Specifically, any of the mangoes mentioned above refers to the Tainong No. 1 mango.
[0045] This invention provides a novel *Lactobacillus pentosaceus* strain. It also provides a liquid bacterial agent prepared using this strain. Pretreatment of mango fruit with the liquid formulation prepared according to this invention (especially pretreatment by coating) can significantly improve the storage performance of mango fruit, delay the ripening process, and inhibit or delay decay. This invention has application and promotion value for mango storage, especially in the current e-commerce environment.
[0046] Preservation Instructions Strain name: Lactobacillus pentosus Latin name: Lactobacillus pentosus Classification and nomenclature: Lactobacillus pentosus ( Lactobacillus pentosus ) Strain number: GHY-5 Preservation Institution: China General Microbiological Culture Collection Center, China Microbiological Culture Collection Committee Abbreviation of depositary institution: CGMCC Address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing Deposit date: October 22, 2025 CGMCC Registration Number: CGMCC No. 36297 Preservation Instructions Strain name: Lactobacillus plantarum Latin name: Lactobacillus plantarum Classification and nomenclature: Lactobacillus plantarum ( Lactobacillus plantarum ) Strain number: GHY-1 Preservation Institution: China General Microbiological Culture Collection Center, China Microbiological Culture Collection Committee Abbreviation of depositary institution: CGMCC Address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing Deposit date: April 1, 2024 CGMCC Registration Number: 30237 Attached Figure Description
[0047] Figure 1 This is a photograph showing the morphological characteristics of strain GHY-5.
[0048] Figure 2 Examples of external and internal cross-sectional photographs of mango fruits during storage in Example 2.
[0049] Figure 3 The anthrax incidence rate, anthrax lesion diameter, and anthrax disease index during the storage process in Example 2 are shown.
[0050] Figure 4 The data represents the ethylene release and respiration rate during storage in Example 2.
[0051] Figure 5 The titratable acid content and soluble solids content during the storage process in Example 2 are given.
[0052] Figure 6 The relative conductivity during the storage process in Example 2.
[0053] Figure 7 The textural indicators are those used during the storage process in Example 2.
[0054] Figure 8 This is a photograph showing the morphological characteristics of strain GHY-1.
[0055] Figure 9 Examples of external and internal cross-sectional photographs of mango fruits during storage in Example 4.
[0056] Figure 10 The anthrax disease severity index during the storage process of Example 4. Detailed Implementation
[0057] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.
[0058] Unless otherwise specified, the experimental methods in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available. F-950 Portable Ethylene Analyzer: Beijing Sunshine Yishida Technology Co., Ltd. Rapid-TA Texture Analyzer: Shanghai Tengba Instrument Technology Co., Ltd. DDS-301 Conductivity Meter: Shanghai Instrument & Electronics Scientific Instrument Co., Ltd. PAL-1 Digital Saccharimeter: Shanghai Instrument & Electronics Scientific Instrument Co., Ltd. Unless otherwise specified, the quantitative experiments in the following examples were all performed in triplicate, and the results were averaged. Microsoft Excel 2019 and Origin 2018 software were used to process and analyze the data, calculate the mean and standard deviation, and plot the data. SPSS 26.0 software was used to perform significance and correlation analysis on the experimental data; p<0.05 indicated a significant difference. Chitosan (CAS No.: 9012-76-4): Sinopharm Chemical Reagent Co., Ltd., Biochemical Reagent BR (Shanghai Testing). The small Tainong mango used in the examples is Tainong No. 1 (Mangifera indica L.'Tainong No.1'), a cultivated variety of mango belonging to the Anacardiaceae family and the Mango genus.
[0059] Liquid MRS medium (pH 6.2±0.2): Add 10.0g peptone, 5.0g beef extract, 4.0g yeast extract, 20.0g glucose, 1.0ml Tween-80, 2.0g dipotassium hydrogen phosphate, 5.0g sodium acetate, 2.0g triammonium citrate, 15.0g agar, 0.2g MgSO4•7H2O, 0.05g MnSO4•4H2O per liter, with the remainder being distilled water.
[0060] Example 1: Isolation, Identification and Preservation of Strains The strain GHY-5 was obtained by screening traditional Gansu slurry.
[0061] Morphological characteristics of strain GHY-5: A straight, round-ended bacterium, approximately 0.8 μm × 3.6 μm, single in number; surface colonies approximately 2.2 mm in diameter, raised, round, smooth, dense, and white. See photo. Figure 1 .
[0062] Physiological and biochemical characteristics of strain GHY-5: Gram-positive, lactic acid produced.
[0063] Genomic DNA was extracted from strain GHY-5 and amplified by PCR using primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-TACGGCTACCTTGTTACGACTT-3'). The amplified products were then recovered and sequenced. Primers 27F and 1492R are universal primers targeting the coding gene of 16S rRNA. The sequencing results are shown in SEQ ID NO: 1. Homology alignment of the sequencing results was performed using NCBI.
[0064] Based on the above morphological characteristics, physiological and biochemical characteristics, and molecular identification results, strain GHY-5 belongs to Lactobacillus pentosus (… Lactobacillus pentosus It was named Lactobacillus pentosolicus GHY-5.
[0065] Lactobacillus pentosus ( Lactobacillus pentosus GHY-5 was deposited on October 22, 2025, at the China General Microbiological Culture Collection Center (CGMCC, address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences), with accession number CGMCC No. 36297.
[0066] Example 2: Application of Lactobacillus pentosolicus GHY-5 I. Preparation of GHY-5 bacterial culture 1. Inoculate the activated Lactobacillus pentosus GHY-5 into liquid MRS medium and incubate at 37°C and 170 rpm for 12 h.
[0067] 2. Inoculate the bacterial culture obtained in step 1 into liquid MRS medium and incubate at 37°C and 170 rpm until the bacterial concentration reaches 1×10⁻⁶. 10 cfu / mL is the concentration of GHY-5 bacterial culture.
[0068] II. Preparation of Compound Solution and Composite Membrane Preparation method of chitosan solution: Dissolve 2g of chitosan and 1mL of glycerol in 1g / 100mL glacial acetic acid aqueous solution in a 40℃ water bath, then make up to 100mL with 1g / 100mL glacial acetic acid aqueous solution, and then perform ultrasonic treatment (the purpose of ultrasonic treatment is to promote the dissolution of chitosan; the ultrasonic parameters are: 40℃, 300W, 20 minutes) to obtain chitosan solution.
[0069] Mix the chitosan solution with an equal volume of the GHY-5 bacterial solution prepared in step one to obtain the compound solution.
[0070] 2 ml of the compound solution was applied to the upper surface of a PE film (25 cm long and 25 cm wide) and air-dried at room temperature for 6 hours to obtain the composite film.
[0071] III. Group processing of mangoes In late May 2024, mangoes from Xiaotai Nong were harvested in Baise City, Guangxi Zhuang Autonomous Region. Diseased, rotten, or color- and ripe fruits with large differences were removed, and mangoes weighing 90-110g were retained.
[0072] Within 12 hours of harvesting, the mangoes were pre-cooled (placed in a temperature-controlled environment at 8-10℃ for 12 hours), and then randomly divided into five groups, with the following treatment methods: Coating group: Take a mango fruit, dip a brush into the compound solution, and evenly coat the entire surface (use 2ml of compound solution for each mango), and then let it dry.
[0073] Immersion group: Take mango fruit, immerse it completely in the compound solution for 1 minute, then take it out and let it air dry.
[0074] Spraying group: Take mango fruit, use a small spray bottle to evenly spray the compound solution onto the entire surface (use 2ml of compound solution for each mango), and then let it dry.
[0075] Molding assembly: Each mango fruit is completely wrapped with a composite film (the surface coated with the compound solution faces the mango).
[0076] Blank control group (CK): Mango fruits were taken and no treatment was performed.
[0077] IV. Transportation and Storage After completing step three, the mangoes are packed and transported to the storage site (Yangzhen Scientific Research Base, Beijing). The transportation method is land transport (natural conditions), which takes 72 hours. 96 hours after harvesting, the mangoes are placed in compartmentalized cardboard boxes at the storage site, with one mango in each compartment, and stored for 12 days (storage conditions: humidity 85%±0.5%, temperature 30±1℃).
[0078] V. Anthrax Disease Severity Index Statistics Disease index reflects the disease status of fruits and vegetables during post-harvest storage and is an important indicator for measuring fruit quality. The main cause of rotting in Xiaotai Nong mangoes during storage is anthracnose. Anthracnose lesions are characterized by: initially, small, pinhead-sized brown spots appear on the fruit peel, gradually expanding into large, irregular, sunken, dark brown spots, eventually leading to the entire fruit turning black and rotting.
[0079] Five disease severity levels are defined: Level 0 is no lesions; Level 1 is lesion area ≤ 10%; Level 2 is lesion area > 10% and ≤ 20%; Level 3 is lesion area ratio > 20% and ≤ 50%; and Level 4 is lesion area ratio > 50%. The lesion area ratio refers to the percentage of anthracnose lesion area to the fruit surface area.
[0080] Anthrax Disease Severity Index (DI) calculation formula: ; m represents the number of fruits at this disease level, n represents the disease level (0, 1, 2, 3, or 4), z represents the total number of fruits, and s represents the highest disease level that occurred.
[0081] Step 4: Exemplary exterior and interior photos of mangoes during storage. Figure 2 (0d refers to before being placed in the cardboard box, 4d refers to 4 days after being placed in the cardboard box, 8d refers to 8 days after being placed in the cardboard box, and 12d refers to 12 days after being placed in the cardboard box). It can be seen that as the storage time increased, all groups of mangoes began to show varying degrees of disease and color change. Compared with the control group at the same time point, the color change and disease severity of mangoes in the coating group, immersion group, spraying group, and molding group were significantly reduced.
[0082] Step four involves recording the anthracnose incidence rate, anthracnose lesion diameter, and anthracnose disease index during storage. At each time point, 12 mango fruits from each group are randomly selected for statistical analysis. The results are shown below. Figure 3 (0d refers to before placement in the cardboard box, 4d refers to 4 days after placement in the cardboard box, 8d refers to 8 days after placement in the cardboard box, and 12d refers to 12 days after placement in the cardboard box). With prolonged storage time, the anthrax disease index, anthrax incidence rate, and anthrax lesion diameter all showed an increasing trend. After 12 days of storage, compared with the control group, there was no significant difference in the anthrax disease index between the spraying group and the modeling group. p >0.05), and the disease indicators in both the coating and immersion groups were significantly reduced. This indicates that coating and immersion treatments can effectively inhibit the increase in the disease index, incidence, and lesion diameter of mango postharvest anthracnose during storage. Coating is more effective than immersion, effectively delaying fruit senescence, inhibiting bacterial growth, and promoting fruit self-protection.
[0083] VI. Determination of Physicochemical Indicators Step 4: Samples are taken every 4 days during storage. 0d refers to before being placed in the cardboard box, 4d refers to 4 days after being placed in the cardboard box, 8d refers to 8 days after being placed in the cardboard box, and 12d refers to 12 days after being placed in the cardboard box.
[0084] 1. Measurement of maturation physiological indicators Respiration intensity is an important indicator reflecting changes in the life activities of fruits and vegetables after harvest. It can be used to detect the rate of nutrient degradation and senescence of fruits and vegetables during storage. At each time point, three mango fruits from each group were collected, placed in a gas cylinder, and sealed for 30 minutes. The ethylene and CO2 contents in the cylinder were then measured using an F-950 portable ethylene analyzer. The ethylene release and respiration intensity (expressed as CO2) per unit mass (i.e., per kg fresh weight) of fruit per unit time (i.e., 1 h) were calculated. Ethylene release is expressed in mg / (kg·h), and respiration intensity in mg / (kg·h). Results are shown below. Figure 4 The respiration rate during storage showed a trend of first increasing and then decreasing. On day 8, the respiration rates of the coating group, impregnation group, spraying group, molding group, and CK group were 0.13 mg / (kg·h), 0.15 mg / (kg·h), 0.17 mg / (kg·h), 0.18 mg / (kg·h), and 0.17 mg / (kg·h), respectively. The respiration rates of the coating group and impregnation group were significantly lower than those of the CK group on day 8. p <0.05%, which was 22.69% and 13.36% lower than that of the CK group, respectively. On day 8, there was no significant difference in respiratory intensity between the modeling group and the spraying group and the CK group. p >0.05%. During storage, the ethylene release of the CK group and the modeling group showed a trend of first increasing and then decreasing, with no significant difference within 4-8 days. p >0.05). During storage, the ethylene release of the coating group, spraying group, and impregnation group showed an increasing trend. Within 4-12 days, the ethylene release of the coating group and impregnation group was significantly lower than that of the CK group (<0.05). p< On day 8, the ethylene release in the coated group and the impregnated group were 0.046 mg / (kg·h) and 0.058 mg / (kg·h), respectively, which were 56.29% and 44.83% lower than those in the control group. In conclusion, coating can effectively reduce the respiration intensity and ethylene release of mangoes during storage, thus delaying fruit ripening.
[0085] Fruits contain a large amount of organic acids. The content of organic acids mainly affects the taste, storability, and processing properties of fruit pulp. Soluble solids mainly refer to soluble sugars, which can be used to reflect the ripeness of the fruit. At each time point, three mango fruits were taken from each group, peeled and pitted, and the pulp was ground to obtain a homogenate. 5g of the homogenate was transferred to a 100mL volumetric flask, distilled water was added to the mark, mixed well, and allowed to stand for 30 minutes. The mixture was then filtered through qualitative filter paper, and the filtrate was collected. 600µL of the filtrate was taken and measured using a PAL-1 digital display saccharimeter. The titratable acid content was expressed as malic acid (%), and the soluble solids content was expressed as soluble sugar (%). The results are shown in […]. Figure 5During storage, the titratable acid content showed a decreasing trend. At 8 days of storage, the titratable acid content in the CK group, molding group, immersion group, spraying group, and coating group decreased by 75.95%, 56.38%, 46.27%, 69.32%, and 38.31% respectively compared to day 0. On day 12, the titratable acid content in the coating group and immersion group was significantly higher than that in the CK group (p<0.05), by 44.82% and 43.44% respectively, while the spraying group and molding group showed no significant difference from the CK group (p>0.05). The results indicate that coating and immersion treatments can effectively inhibit the decrease in titratable acid content in mangoes during storage, with coating treatment showing better results, thus effectively inhibiting fruit ripening. During the storage period, the soluble solids content in the immersion group, spraying group, molding group, and control group all showed a trend of first increasing and then decreasing, with peak values appearing on day 8, at 12.58%, 13.40%, 13.10%, and 14.17%, respectively. This is because some starch in mangoes is converted into sugar in the early stage of storage, leading to an increase in soluble solids content. In the later stage of storage, some sugar is consumed as a substrate for respiration, resulting in a decrease in the soluble solids content of mangoes. Within 12 days of storage, the soluble solids content in the coating group showed a continuous upward trend. On day 8, the soluble solids content was significantly lower than that in the control group (p<0.05), 21.65% lower than the control group, and significantly lower than the control group, molding group, and spraying group within 4-8 days (p<0.05). This indicates that coating treatment can effectively inhibit the increase of soluble solids in fruit, increase the storage resistance of mangoes, and slow down post-ripening.
[0086] 2. Cell membrane permeability Cell membrane permeability was expressed as conductivity. 3g of mango pulp was cut into 1mm thick slices, placed in a test tube, and deionized water was added to a volume of 25mL. The tube was shaken at 110rpm for 30min, and the conductivity was measured as P1. Then, the slices were boiled for 10min, cooled to room temperature, and deionized water was added to a volume of 25mL. The conductivity was measured as P2. The conductivity of the deionized water was measured as P0. Three mango fruits were tested at each time point for each group.
[0087] Relative conductivity (P): .
[0088] See results Figure 6As storage time increased, the relative conductivity of mangoes in each group continuously increased. On day 12, the conductivity of the sprayed group and the molded group was not significantly different from that of the control group (p>0.05), while the conductivity of the coating and immersion treatments was significantly lower than that of the control group (p<0.05). On day 12, the conductivity of the control group, the immersion group, and the coating group were 0.92%, 0.66%, and 0.53%, respectively, with the immersion group and the coating group being 27.92% and 41.94% lower than that of the control group. The results indicate that coating treatment can significantly inhibit the increase in the relative conductivity of mangoes. This may be because coating treatment maintains better cell membrane function. At the same time, the slowdown in metabolic rate also reduces the generation of free radicals, thus maintaining better cell structure and inhibiting the increase in relative conductivity.
[0089] 3. Texture determination Hardness reflects the ripeness and softening of fruit. Transpiration, respiration, and cell wall maturation lead to softening during storage. Elasticity refers to the percentage of a sample that returns to its original height or volume after compression and removal of external force. A larger range of elasticity changes in the pulp indicates active internal metabolism and rapid nutrient consumption. Resilience is an important indicator of a fruit's ability to quickly return to its original shape after compression. If the fruit tissue is severely damaged, its resilience will approach zero. Cohesion refers to a sample's relative resistance to a second compression after the first compression deformation. It reflects the sample's ability to resist external forces and maintain its structural integrity, and is also a manifestation of cell binding ability. Adhesion refers to the force that binds food together internally, and can be used to describe the texture of food. Chewability refers to the energy required to chew solid food, comprehensively reflecting the sustained resistance that food can provide during chewing. Higher chewability results in a better "bite" sensation in the texture of the food.
[0090] Mango fruits were collected, and pieces were cut from a 1cm thick section near the equator. Hardness was measured using a texture analyzer (P / 10 cylindrical probe, TPA mode, ambient temperature 25℃, detection speed 1mm / s, compression degree 20%, trigger load 0.981N). Hardness was measured in N. Texture characteristic curves were obtained, and hardness, elasticity, chewiness, adhesiveness, cohesiveness, and resilience were analyzed using Texture Lab Pro software. Twelve mango fruits from each group were tested at each time point. Results are shown below. Figure 7 .
[0091] With prolonged storage, the firmness of the fruit decreased. After 8 days of storage, the rate of decrease in mango fruit firmness slowed down. On the 8th day, the firmness of the coated and soaked groups was significantly higher than that of the control group (CK). p <0.05), there was no significant difference between the molding group, the spraying group, and the CK group. p>0.05). On day 8, the hardness of the coated group and the impregnated group were 67.81% and 41.89% higher than that of the control group, respectively. The results show that coating treatment can effectively maintain the hardness of mango fruits after harvest.
[0092] As storage time increased, the elasticity of the fruit gradually decreased. On day 12, the elasticity of the coated group, immersion group, and sprayed group was significantly different from that of the control group (CK). p <0.05). On day 12, the elasticity of the spraying group and the molding group was 0.61N and 0.55N, respectively, while the elasticity of the coating group was 0.68N. The results indicate that coating treatment can maintain the elasticity of mangoes during storage.
[0093] On day 4, the recovery rates of the coating and immersion groups were significantly different from those of the control group. p <0.05), there was no significant difference between the molding group and the spraying group and the CK group ( p >0.05). On day 8, the reactivity of all treatment groups showed a significant difference compared to the CK group ( p <0.05). The recovery rates of the coating group and the impregnation group were 51.00% and 17.39% higher than those of the control group, respectively, while those of the molding group and the spraying group were 36.81% and 10.61% lower than those of the control group, respectively. The results indicate that coating can effectively slow down the decrease in the recovery rate of mangoes during storage.
[0094] The cohesiveness of mangoes in the coated group was significantly higher than that in the CK group from 4 to 12 days. p <0.05). After 8-12 days, the cohesiveness of the fruits in the spraying group and the molding group showed little change, and the difference from the CK group was not significant. p >0.05). The results showed that the coated fruit still maintained good cell binding force and had good edibility after 12 days of storage.
[0095] After 4 days of storage, the adhesiveness of the coated mangoes was significantly higher than that of the control group. p <0.05), and there were no significant differences between the other three groups and the CK group. p >0.05). On day 8 of storage, all treatment groups showed significant differences compared to the CK group ( p <0.05), among which the coating and immersion treatments significantly increased the adhesiveness of mango fruits compared to the other three groups ( p <0.05), which were 129.41% and 81.37% higher than those in the control group, respectively. The results indicate that coating and impregnation treatments can effectively slow down the decrease in the adhesiveness of mangoes, with coating treatment being more effective than impregnation treatment.
[0096] The chewiness of mangoes decreased continuously during storage. At 12 days, the coated group showed significant differences compared to other groups. p<0.05), the coating group was 66.27% lower than the CK group on day 12, and the chewiness was 2184.52 N. There were no significant differences between the impregnation group, spraying group, and molding group and the CK group. p >0.05). This indicates that the coating treatment can maintain the chewiness of mangoes and preserve their taste during the later stages of storage.
[0097] Example 3: Isolation, Identification and Preservation of Strains The strain GHY-1 was obtained by screening traditional Gansu slurry.
[0098] Morphological characteristics of strain GHY-1: A straight, round-terminated bacterium, approximately 0.9 μm × 4.0 μm, single in number; surface colonies approximately 2.5 mm in diameter, raised, round, smooth, dense, and white. See photo. Figure 8 .
[0099] Physiological and biochemical characteristics of strain GHY-1: Gram-positive, lactic acid produced.
[0100] Genomic DNA was extracted from strain GHY-1 and amplified by PCR using primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-TACGGCTACCTTGTTACGACTT-3'). The amplified products were then recovered and sequenced. Primers 27F and 1492R are universal primers targeting the coding gene of 16S rRNA. The sequencing results are shown in SEQ ID NO: 2. Homology alignment of the sequencing results was performed using NCBI.
[0101] Based on the above morphological characteristics, physiological and biochemical characteristics and molecular identification results, strain GHY-1 belongs to Lactobacillus plantarum, and is named Lactobacillus plantarum GHY-1.
[0102] Lactobacillus plantarum ( Lactobacillus plantarum GHY-1 was deposited on April 1, 2024, at the China General Microbiological Culture Collection Center (CGMCC, address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences), with accession number CGMCC No. 30237.
[0103] Example 4: Evaluation of the effects of Lactobacillus plantarum GHY-1 and Lactobacillus pentosus GHY-5 on the storage and preservation of mangoes. I. Preparation of GHY-5 bacterial culture 1. Inoculate the activated Lactobacillus pentosus GHY-5 into liquid MRS medium and incubate at 37°C and 170 rpm for 12 h.
[0104] 2. Inoculate the bacterial culture obtained in step 1 into liquid MRS medium and incubate at 37°C and 170 rpm until the bacterial concentration reaches 1×10⁻⁶. 10 cfu / mL is the concentration of GHY-5 bacterial culture.
[0105] II. Preparation of GHY-1 bacterial culture 1. Inoculate the activated Lactobacillus plantarum GHY-1 into liquid MRS medium and incubate at 37℃ and 170rpm for 12h.
[0106] 2. Inoculate the bacterial culture obtained in step 1 into liquid MRS medium and incubate at 37°C and 170 rpm until the bacterial concentration reaches 1×10⁻⁶. 10 cfu / mL is the GHY-1 bacterial culture.
[0107] III. Preparation of Compound Solution and Composite Membrane Preparation method of chitosan solution: Same as the chitosan solution in step two of Example 2.
[0108] Mix the chitosan solution with the GHY-5 bacterial solution prepared in step one in equal volume to obtain the GHY-5 compound solution.
[0109] The chitosan solution was mixed with an equal volume of the GHY-1 bacterial culture prepared in step two to obtain the GHY-1 compound solution.
[0110] IV. Group processing of mangoes In early June 2024, Xiaotai Nong mangoes were harvested from Baise City, Guangxi Zhuang Autonomous Region. Diseased, rotten, or color- and ripe fruits with large differences were removed, and mangoes weighing 90-110g were retained.
[0111] Within 12 hours of harvesting, the mangoes were pre-cooled (placed in a temperature-controlled environment at 8-10℃ for 12 hours), and then randomly divided into three groups, with the following treatment methods: Blank control group (CK): Mango fruits were taken and no treatment was performed.
[0112] GHY-1 coating group (experimental group 1): Take mango fruit, dip a brush into GHY-1 compound solution, and evenly coat the entire surface (use 2ml of compound solution for each mango), and then let it dry.
[0113] GHY-5 coating group (experimental group 2): Take mango fruit, dip a brush into GHY-5 compound solution, and evenly coat the entire surface (use 2ml of compound solution for each mango), and then let it dry.
[0114] V. Transportation and Storage Same as step four in Example 2.
[0115] VI. Anthrax Disease Severity Index Statistics Same as step five in Example 2.
[0116] Step 5: Exemplary exterior and interior photos of mangoes during storage. Figure 9 (0d refers to before placement in the cardboard box, 4d refers to 4 days after placement in the cardboard box, 8d refers to 8 days after placement in the cardboard box, and 12d refers to 12 days after placement in the cardboard box). The mango peel turned yellow with prolonged storage. Compared to the control group (CK), the yellowing effect on the mango peel in the experimental group was not significant, and it had a significant inhibitory effect on mango disease. On the 12th day after treatment in the experimental group, the fruit peel and flesh showed no obvious disease, while the fruit in the control group showed numerous lesions on the 8th day, indicating that the treatment in the experimental group effectively inhibited fruit disease.
[0117] Step 5: Anthracnose disease severity index was calculated during storage. At each time point, 12 mango fruits from each group were randomly selected for analysis. The results are shown below. Figure 10 (0d refers to before placement in the cardboard box, 4d refers to 4 days after placement in the cardboard box, 8d refers to 8 days after placement in the cardboard box, and 12d refers to 12 days after placement in the cardboard box). With prolonged storage time, the disease index of the fruit in all treatment groups showed an increasing trend. During the storage period, the disease index of the second experimental group was inhibited compared to other groups. Therefore, the second experimental group treatment has an inhibitory effect on the disease development of mangoes during their shelf life.
[0118] The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some of the essential features can be applied within the scope of the following appended claims.
Claims
1. A method for pretreating mangoes, comprising the following steps: taking a mango, coating its surface with a liquid reagent, and then drying it; the purpose of the pretreating is to improve the storage performance of the mangoes; The improvement in the storage performance of mangoes is manifested in the following ways: (b1) and (b2) and (b3) and (b4) and (b5) and (b6) and (b7) and (b8) and (b9) and (b10) and (b11) and (b12) and (b13) and (b14) and (b15): (b1) Inhibits mango fruit rot; (b2) Inhibit anthracnose in mango fruit; (b3) Delay the ripening process of mangoes; (b4) Reduce the respiration rate of mango fruit; (b5) Reduce the ethylene release from mango fruit; (b6) Inhibits the decrease in titratable acidity of mango fruit during storage; (b7) Inhibits the increase of soluble solids in mango fruit during storage; (b8) Inhibits the increase in electrical conductivity of mango fruit during storage; (b9) Inhibits the increase in cell membrane permeability during mango fruit storage; (b10) Delays the decrease in firmness of mango fruit during storage; (b11) Delays the decline in the elasticity of mango fruit during storage; (b12) Delays the recovery decline of mango fruit during storage; (b13) Delays the decline in cohesion of mango fruit during storage; (b14) Delays the decline in the adhesiveness of mango fruit during storage; (b15) Delays the decline in chewiness of mango fruit during storage; The liquid reagent comprises chitosan and lactobacillus pentosus; in the liquid reagent, the concentration of chitosan is 1 g / 100 mL, and the concentration of lactobacillus pentosus GHY-5 is 0.5×10 10 cfu / mL; the lactobacillus pentosus is lactobacillus pentosus (Lactobacillus pentosus) Lactobacillus pentosus ) GHY-5 with the preservation accession number of CGMCC No.36297.
2. The application of liquid reagents is as follows (a5) or (a6): (a5) Application in the pretreatment of mangoes; the purpose of the pretreatment is to improve the storage performance of mangoes; (a6) Application in the preparation of products that pre-treat mangoes; the purpose of said pre-treatment is to improve the storage performance of mangoes; The improvement in the storage performance of mangoes is manifested in the following ways: (b1) and (b2) and (b3) and (b4) and (b5) and (b6) and (b7) and (b8) and (b9) and (b10) and (b11) and (b12) and (b13) and (b14) and (b15): (b1) Inhibits mango fruit rot; (b2) Inhibit anthracnose in mango fruit; (b3) Delay the ripening process of mangoes; (b4) Reduce the respiration rate of mango fruit; (b5) Reduce the ethylene release from mango fruit; (b6) Inhibits the decrease in titratable acidity of mango fruit during storage; (b7) Inhibits the increase of soluble solids in mango fruit during storage; (b8) Inhibits the increase in electrical conductivity of mango fruit during storage; (b9) Inhibits the increase in cell membrane permeability during mango fruit storage; (b10) Delays the decrease in firmness of mango fruit during storage; (b11) Delays the decline in the elasticity of mango fruit during storage; (b12) Delays the recovery decline of mango fruit during storage; (b13) Delays the decline in cohesion of mango fruit during storage; (b14) Delays the decline in the adhesiveness of mango fruit during storage; (b15) Delays the decline in chewiness of mango fruit during storage; The liquid reagent comprises chitosan and Lactobacillus pentosus; in the liquid reagent, the concentration of chitosan is 1 g / 100 mL, and the concentration of Lactobacillus pentosus GHY-5 is 0.5 × 10⁻⁶. 10 cfu / mL; the Lactobacillus pentosus is Lactobacillus pentosus (cfu / mL); Lactobacillus pentosus GHY-5, its accession number is CGMCC No.36297.
3. Applications of liquid reagents are as follows: (b1) and (b2) and (b3) and (b4) and (b5) and (b6) and (b7) and (b8) and (b9) and (b10) and (b11) and (b12) and (b13) and (b14) and (b15): (b1) Inhibits mango fruit rot; (b2) Inhibit anthracnose in mango fruit; (b3) Delay the ripening process of mangoes; (b4) Reduce the respiration rate of mango fruit; (b5) Reduce the ethylene release from mango fruit; (b6) Inhibits the decrease in titratable acidity of mango fruit during storage; (b7) Inhibits the increase of soluble solids in mango fruit during storage; (b8) Inhibits the increase in electrical conductivity of mango fruit during storage; (b9) Inhibits the increase in cell membrane permeability during mango fruit storage; (b10) Delays the decrease in firmness of mango fruit during storage; (b11) Delays the decline in the elasticity of mango fruit during storage; (b12) Delays the recovery decline of mango fruit during storage; (b13) Delays the decline in cohesion of mango fruit during storage; (b14) Delays the decline in the adhesiveness of mango fruit during storage; (b15) Delays the decline in chewiness of mango fruit during storage; The liquid reagent comprises chitosan and Lactobacillus pentosus; in the liquid reagent, the concentration of chitosan is 1 g / 100 mL, and the concentration of Lactobacillus pentosus GHY-5 is 0.5 × 10⁻⁶. 10 cfu / mL; the Lactobacillus pentosus is Lactobacillus pentosus (cfu / mL); Lactobacillus pentosus GHY-5, its accession number is CGMCC No.36297.
4. Application of liquid reagents in the preparation of products; the uses of said products are as follows (b1) and (b2) and (b3) and (b4) and (b5) and (b6) and (b7) and (b8) and (b9) and (b10) and (b11) and (b12) and (b13) and (b14) and (b15): (b1) Inhibits mango fruit rot; (b2) Inhibit anthracnose in mango fruit; (b3) Delay the ripening process of mangoes; (b4) Reduce the respiration rate of mango fruit; (b5) Reduce the ethylene release from mango fruit; (b6) Inhibits the decrease in titratable acidity of mango fruit during storage; (b7) Inhibits the increase of soluble solids in mango fruit during storage; (b8) Inhibits the increase in electrical conductivity of mango fruit during storage; (b9) Inhibits the increase in cell membrane permeability during mango fruit storage; (b10) Delays the decrease in firmness of mango fruit during storage; (b11) Delays the decline in the elasticity of mango fruit during storage; (b12) Delays the recovery decline of mango fruit during storage; (b13) Delays the decline in cohesion of mango fruit during storage; (b14) Delays the decline in the adhesiveness of mango fruit during storage; (b15) Delays the decline in chewiness of mango fruit during storage; The liquid reagent comprises chitosan and Lactobacillus pentosus; in the liquid reagent, the concentration of chitosan is 1 g / 100 mL, and the concentration of Lactobacillus pentosus GHY-5 is 0.5 × 10⁻⁶. 10 cfu / mL; the Lactobacillus pentosus is Lactobacillus pentosus (cfu / mL); Lactobacillus pentosus GHY-5, its accession number is CGMCC No.36297.