Biological bacteriostatic agent, preparation method and application in prevention and control of postharvest diseases of fruits and vegetables
By using the CGMCC 9951 metabolite VOCs as biological antibacterial agents, the drug resistance and environmental pollution caused by chemical fungicides were solved, and effective prevention and control and preservation effects on sweet potatoes and other fruits and vegetables were achieved.
Patent Information
- Application Number
- CN202411541597.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-31
- Publication Date
- 2025-02-21
AI Technical Summary
In the prior art, chemical fungicides are often used when preventing and controlling black spot diseases caused by sweet potato saccharin bacteria, but their frequent use leads to increased resistance to pathogens, environmental pollution and human health threats, and the development of resistant varieties takes a long time. , generally lower resistance.
VOCs, the metabolite of Bacillus coagulis CGMCC 9951, was used as biological antibacterial agent, and cultured the activated Bacillus coagulis on LB medium for 48 hours, and prepared antibacterial agents and stored them with fruits and vegetables. The bacterial culture dishes were replaced regularly. .
It significantly reduces peroxidative damage to fruits and vegetables such as sweet potatoes, delays the deterioration of fruit quality, effectively reduces black spot lesions, has a good inhibitory effect on common molds, and is environmentally friendly.
Smart Images
Figure CN119488115A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of storage disease prevention and control and preservation of fruits and vegetables, and specifically relates to a biological antibacterial agent, a preparation method and application in preventing and controlling post-harvest diseases of fruits and vegetables. Background Art
[0002] sweet potato( Ipomoea batatas ) is the bottom line crop to ensure national food security. In recent years, global climate change has been abnormal, and plant pathogens such as fungi and bacteria have seriously affected sweet potato production and safe storage after harvest. Ceratocystis fimbriata ) occurs during the sweet potato seedling, growth and storage periods, which can cause seedling death and cellar rot, resulting in an annual sweet potato yield loss of about 5% to 10%, and up to 20% to 50% in severe cases, causing significant economic losses.
[0003] At present, the main ways to reduce the infection of sweet potato long-beaked shell fungus and overcome sweet potato black spot disease include the use of fungicides and the breeding of disease-resistant varieties. Chemical fungicides such as carbendazim, methyl thiophanate, mancozeb and ethionine are commonly used at home and abroad to inhibit sweet potato long-beaked shell fungus and prevent and control sweet potato black spot disease. However, the frequent use of chemical agents has increased the resistance of pathogens, and is accompanied by residues, causing environmental pollution and other problems, which in turn threatens human health and becomes a bottleneck for sustainable agricultural development. In addition, the development of resistant varieties takes a long time. Due to the easy mutation of pathogens and their adaptability to the environment, the resistance of new varieties is generally reduced after several years of planting. There is an urgent need to find safer and more effective ways in the production and preservation of sweet potatoes. Microorganisms have the advantages of short growth cycle, strong antibacterial ability and environmental friendliness, and have good application prospects in the preservation of sweet potatoes after harvest. Summary of the invention
[0004] To solve the above problems, the present invention provides a biological antibacterial agent, a preparation method and application in preventing and controlling post-harvest diseases of fruits and vegetables, which can significantly reduce peroxidation damage of sweet potatoes and other fruits and vegetables and delay deterioration of fruit quality.
[0005] The present invention is achieved through the following technical solutions: The invention provides a biological bacteriostatic agent, wherein the bacteriostatic agent is a metabolite VOCs of Bacillus coagulans.
[0006] Furthermore, the deposit number of the Bacillus coagulans is CGMCC NO.9951.
[0007] Furthermore, the antibacterial substances in the metabolite VOCs include but are not limited to benzaldehyde, acetic acid, isobutyric acid, and isovaleric acid.
[0008] The present invention also provides a method for preparing a biological antibacterial agent, which comprises diluting and coating the activated Bacillus coagulans on the surface of an LB culture medium and culturing the culture medium at 37° C. for 48 h.
[0009] Furthermore, the components of the LB medium include: 15.0 g / L glucose, 15.0 g / L tryptone, 15.0 g / L yeast powder, 5.0 g / L magnesium sulfate, 20.0 g / L agar, the pH is adjusted to 7.0, and the medium is sterilized at 121° C. for 20 min before use.
[0010] The present invention also provides the use of biological antibacterial agents in preventing and controlling post-harvest diseases of fruits and vegetables, wherein the pathogens include but are not limited to Fusarium, Mucor, Aspergillus, and Aspergillus niger.
[0011] Furthermore, the application method specifically comprises the following steps: S1. Pretreatment: Soak freshly harvested fruits and vegetables in 1-2% sodium hypochlorite for 1-2 min, then wash with water for later use; S2. Fresh-keeping treatment: Place the fruits and vegetables treated in S1 and the biological antibacterial agent in a carton and store at room temperature. Replace the culture dish with bacteria every 1 to 2 weeks.
[0012] The beneficial effects of the present invention are: 1. The present invention utilizes Bacillus coagulans CGMCC 9951 VOCs to show obvious biocontrol effects, effectively reducing black spot lesions of vegetables and fruits such as sweet potatoes during storage; 2. During use, the present invention can significantly reduce the peroxidation damage of sweet potatoes and other fruits and vegetables, delay the deterioration of fruit quality, and has potential application value in the storage and preservation of sweet potatoes and other fruits and vegetables; 3. The present invention clarifies the main antibacterial components of Bacillus coagulans CGMCC 9951 VOCs, including benzaldehyde, acetic acid, isobutyric acid, isovaleric acid and other components; 4. The biological antibacterial agent of the present invention has a good inhibitory effect on common molds that cause fruit and vegetable lesions, such as Fusarium oxysporum, Mucor, Verticillium serrata, mildew, and Fusarium solani; 5. The biological antibacterial agent and preservation method of the present invention can be used to prevent and control sweet potatoes and other fruits and vegetables from being infected by pathogenic bacteria such as Aspergillus niger and Fusarium spp., and has a wide range of applications. BRIEF DESCRIPTION OF THE DRAWINGS
[0013] Figure 1 This is a graph showing the inhibition of Bacillus coagulans on common sweet potato pathogens; Figure 2 (A) Figure 2 (B) are the comparison chart and bar chart of the effects of various volatile components of metabolites on the mycelial growth of sweet potato black spot disease; Figure 3 (A) Figure 3 (B) is a comparison chart of the effects of Bacillus coagulans VOCs on the storage quality of sweet potatoes. Figure 3 (C) Figure 3(D) Figure 3 (E) Figure 3 (F) Figure 3 (G) Figure 3 (H) are bar charts comparing the effects of Bacillus coagulans VOCs on various properties of sweet potatoes. DETAILED DESCRIPTION
[0014] The present invention provides a biological antibacterial agent, which is a VOCs metabolite of Bacillus coagulans. The deposit number of the Bacillus coagulans is CGMCC NO.9951, and the antibacterial substances in the metabolite VOCs include but are not limited to benzaldehyde, acetic acid, isobutyric acid, and isovaleric acid.
[0015] The present invention also provides a method for preparing a biological antibacterial agent, comprising: 8 cfu / mL) was spread on the surface of LB medium and cultured at 37℃ for 48 h. The components of LB medium include: 15.0 g / L glucose, 15.0 g / L tryptone, 15.0 g / L yeast powder, 5.0 g / L magnesium sulfate, 20.0 g / L agar, pH was adjusted to 7.0, and sterilized at 121℃ for 20 min before use.
[0016] The present invention also provides the application of biological antibacterial agents in preventing and controlling post-harvest diseases of fruits and vegetables, including but not limited to Fusarium, Mucor, Aspergillus, and Aspergillus. The application method specifically comprises the following steps: S1. Pretreatment: Soak freshly harvested fruits and vegetables in 1-2% sodium hypochlorite for 1-2 min, then wash with water for later use; S2. Fresh-keeping treatment: Place the fruits and vegetables treated in S1 and the biological antibacterial agent in a carton and store at room temperature. Replace the culture dish with bacteria every 1 to 2 weeks.
[0017] In order to make the purpose, technical solution and advantages of the embodiments of the present invention more clear, the technical solution in the embodiments of the present invention will be clearly and completely described below according to the accompanying drawings.
[0018] Embodiment 1: 1. Test Materials The experimental materials used were sweet potatoes (watermelon red) with basically uniform fruit size and color, and no damage or disease on the surface of the sweet potatoes.
[0019] 2. Test methods 2.1 Culture medium Culture medium for Bacillus coagulans: 15.0 g / L glucose, 15.0 g / L tryptone, 15.0 g / L yeast powder, 5.0 g / L magnesium sulfate, 20.0 g / L agar, adjust pH to 7.0, sterilize at 121°C for 20 min, and set aside.
[0020] 2.2 Source of Bacillus coagulans CGMCC 9951 The Bacillus coagulans CGMCC 9951 strain was independently screened and isolated by the applicant's research group in 2014 and has been made public. The strain has been deposited in the General Microbiology Center of China Culture Collection Administration (referred to as CGMCC, address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Postal Code: 100101), the deposit date is November 13, 2014, and the deposit number is CGMCC NO.9951.
[0021] 2.3 Preparation of Bacillus coagulans VOCs antibacterial agent The activated Bacillus coagulans CGMCC 9951 bacterial solution was diluted to 10 5 cfu / mL, 10 6 cfu / mL, 10 7 cfu / mL, 10 8 cfu / mL, 100 μL of bacterial solution was spread on the surface of Bacillus coagulans solid culture medium, and a blank culture dish without bacterial solution was used as a control, and cultured in a 37°C incubator for 2 days.
[0022] Figure 1 This is the inhibitory effect of Bacillus coagulans VOCs on common sweet potato pathogens. As shown in the figure, compared with the control group, the pathogens in the experimental group grew slowly, indicating that VOCs have a good prevention and control effect on sweet potato Mycelium, Mucor, and Verticillium serrata.
[0023] 2.4 Antibacterial spectrum of Bacillus coagulans VOCs Using the plate interlocking method, after inoculating pathogens (Schistolonifer, Mucor, Verticillium, Penicillium, Fusarium, etc.) on the surface of the PDA culture medium, the culture dish in 2.3 was interlocked with the PDA culture dish, sealed and inverted at 28℃ to observe the growth of the colonies.
[0024] 2.5 Analysis of main components of Bacillus coagulans VOCs The VOCs produced during the cultivation of Bacillus coagulans were collected, and the VOCs components were determined by HS-SPME / GC-MS.
[0025] Taking the sweet potato long beak fungus as an example, the main antibacterial components of VOCs were further clarified. Figure 2 Among them, benzaldehyde, acetic acid, isobutyric acid, isovaleric acid and other ingredients have the best inhibitory effect on sweet potato long-beaked shell fungus.
[0026] 2.6 Postharvest processing of sweet potatoes The sweet potatoes were washed and dried, soaked in 2% sodium hypochlorite for 2 min, washed twice with sterile water, and then dried. The sweet potato epidermis was damaged and 5 μL 10 6 spores / mL sweet potato long beaked shell spore liquid. The bacterial suspension of Bacillus coagulans CGMCC 9951 was inoculated on the solid medium of Bacillus coagulans and placed at the bottom of a sterile plastic box. The solid medium of Bacillus coagulans was placed at the bottom of a sterile plastic box, and sweet potatoes were placed on top. Sweet potatoes that were not inoculated with sweet potato long beaked shell spores were used as controls, and the culture was sealed and cultured at 28°C for 2 weeks.
[0027] 2.7 Analysis of sweet potato storage quality After 2 weeks of cultivation, the sweet potatoes were taken out and the storage quality indexes of the sweet potatoes were analyzed. The determination method was as follows: Measure the diameter and depth of sweet potato lesions and record them.
[0028] Total soluble solids: Cut 10 g of sweet potato tissue and place it in a mortar. Add appropriate amount of distilled water, grind it and filter it with 2 layers of gauze. Use a rubber-tipped dropper to absorb a drop of filtered liquid on the lens of a handheld sugar meter. The value of the blue and white dividing line is the soluble solid content.
[0029] Soluble sugar, MDA and total antioxidant capacity: operate according to the instructions of the kit.
[0030] from Figure 3 It can be seen that after VOCs treatment, the area and diameter of the lesions inoculated with E. coli on the surface of sweet potatoes were reduced, the soluble solids and soluble sugar content of sweet potatoes increased, and the antioxidant activity was enhanced.
[0031] Example 2 The difference from Example 1 is that the sweet potato is replaced by melon, and after VOCs treatment, the area and diameter of the lesions inoculated with Fusarium on the surface of the melon are reduced.
[0032] It should be noted that although the present invention is described by the above embodiments, the present invention may also have other various embodiments. Without departing from the spirit and scope of the present invention, it is obvious that those skilled in the art may make various corresponding changes and deformations to the present invention, but these changes and deformations should all fall within the scope of protection of the appended claims of the present invention and their equivalents.
Claims
1. A biological antibacterial agent, characterized in that: The antibacterial agent is VOCs, a metabolite of Bacillus coagulans.
2. A biological antibacterial agent according to claim 1, characterized in that: The deposit number of the Bacillus coagulans is CGMCC NO.9951.
3. A biological antibacterial agent according to claim 1, characterized in that: The antibacterial substances in the metabolite VOCs include but are not limited to benzaldehyde, acetic acid, isobutyric acid, and isovaleric acid.
4. A method for preparing a biological antibacterial agent according to any one of claims 1 to 3, characterized in that: The activated Bacillus coagulans was diluted and spread on the surface of LB medium and cultured at 37°C for 48 h.
5. The method for preparing a biological antibacterial agent according to claim 4, characterized in that: The ingredients of LB medium include: 15.0 g / L glucose, 15.0 g / L tryptone, 15.0 g / L yeast powder, 5.0 g / L magnesium sulfate, 20.0 g / L agar, adjust the pH to 7.0, sterilize at 121°C for 20 min and set aside.
6. The use of the biological antibacterial agent according to any one of claims 1 to 3 in preventing and controlling post-harvest diseases of fruits and vegetables, characterized in that: Pathogens include but are not limited to Fusarium, Mucor, Aspergillus, and Aspergillus.
7. The use of the biological antibacterial agent according to claim 6 in preventing and controlling post-harvest diseases of fruits and vegetables, characterized in that: The application method specifically comprises the following steps: S1. Pretreatment: Soak freshly harvested fruits and vegetables in 1-2% sodium hypochlorite for 1-2 min, then wash with water for later use; S2. Fresh-keeping treatment: Place the fruits and vegetables treated in S1 and the biological antibacterial agent in a carton and store at room temperature. Replace the culture dish with bacteria every 1 to 2 weeks.