Application of cinnamyl alcohol in prevention and treatment of sweet potato soft rot

Abstract

The invention discloses application of cinnamyl alcohol in prevention and treatment of sweet potato soft rot, and belongs to the technical field of biology. Experiments find that low-concentration cinnamyl alcohol (200-400 [mu] g.ml <-1 >) can significantly inhibit soft rot pathogenic bacteria, can help eliminate or relieve sweet potato soft rot, provides a new way for safe and efficient prevention and treatment of sweet potato fungal diseases, provides a theoretical basis for cinnamyl alcohol development of a green biological fresh-keeping agent for sweet potato postharvest storage, and has broad application prospects. The method has important application value for reducing dependence on chemical pesticides and guaranteeing food safety in sweet potato production.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to the application of cinnamyl alcohol in the prevention and control of sweet potato soft rot. Background Technology

[0002] Sweet potatoes are high-yielding, highly adaptable to different environments, and contain a large amount of starch. They can be used not only as an important food source but also as animal feed, industrial raw material, and a new energy source. Sweet potatoes are an excellent source of nutrients, including vitamins, potassium, iron, calcium, and minerals. They also have medicinal value due to their anti-cancer, anti-diabetic, and anti-inflammatory activities, making them very popular.

[0003] Soft rot is a common disease of sweet potatoes during storage, mainly caused by Rhizopus stolonifera. This fungus invades the plant from wounds on the tubers and root pores, and secretes a variety of decomposing enzymes into the plant tissues. It quickly decomposes the mucilage and other components in the tuber cells, causing the sweet potato tissues to rot and decay. It spreads rapidly, often causing the entire cellar of sweet potatoes to rot, resulting in serious economic losses.

[0004] Currently, the main agents used to control sweet potato soft rot are chemical pesticides. However, with the extensive use of chemical pesticides causing serious environmental pollution and the increasing resistance of pathogens to drugs, traditional agents can no longer meet the requirements of safety and high efficiency. On the other hand, most pathogenic bacteria exhibit rapid physiological variation, which necessitates a highly efficient, safe, and broad-spectrum fungicide for effective control of pathogens. Summary of the Invention

[0005] The purpose of this invention is to provide the application of cinnamyl alcohol in the prevention and control of sweet potato soft rot, so as to solve the problems existing in the prior art. Through experiments, it was found that low concentrations of cinnamyl alcohol can effectively inhibit the pathogen of sweet potato soft rot, Rhizopus spp., which provides a scientific basis for the development of sweet potato storage bactericides and preservatives and long-term post-harvest storage of sweet potatoes.

[0006] To achieve the above objectives, the present invention provides the following solution:

[0007] This invention provides the application of cinnamyl alcohol in the prevention and control of sweet potato soft rot.

[0008] This invention also provides the application of cinnamyl alcohol in the control of creeping rhizopus.

[0009] This invention also provides the application of cinnamyl alcohol in the preparation of postharvest storage preservatives for sweet potatoes.

[0010] This invention also provides the application of cinnamyl alcohol in the preparation of biological agents for the prevention and control of sweet potato soft rot or Rhizopus stolonifer.

[0011] This invention also provides a postharvest storage preservative for sweet potatoes, comprising 200-400 μg·ml -1It consists of cinnamyl alcohol and a solvent used to dissolve cinnamyl alcohol.

[0012] Preferably, the solvent includes dimethyl sulfoxide.

[0013] This invention also provides a biological agent for preventing and controlling sweet potato soft rot or Rhizopus stolonifer, in a formulation of 200-400 μg·ml. -1 It consists of cinnamyl alcohol and a solvent used to dissolve cinnamyl alcohol.

[0014] Preferably, the solvent includes dimethyl sulfoxide.

[0015] The present invention discloses the following technical effects:

[0016] This invention discloses the use of cinnamyl alcohol in the control of sweet potato soft rot, thus broadening the application scope of cinnamyl alcohol. This invention, through experiments, has discovered that low concentrations of cinnamyl alcohol (200-400 μg / ml) can effectively control sweet potato soft rot. -1 It can significantly inhibit the pathogen of soft rot, help eliminate or alleviate sweet potato soft rot, provide a new way for safe and efficient prevention and control of fungal diseases in sweet potatoes, and provide a theoretical basis for the development of cinnamyl alcohol as a green biological preservative for post-harvest storage of sweet potatoes. It has important application value for reducing dependence on chemical pesticides and ensuring food safety in sweet potato production. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 The growth status of soft rot pathogens under different concentrations of cinnamyl alcohol treatment; DMSO treatment served as a control.

[0019] Figure 2 Line graph showing fungal growth under cinnamyl alcohol treatment;

[0020] Figure 3 The 24-hour inhibition rate of cinnamyl alcohol at various treatment concentrations is given. Detailed Implementation

[0021] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0022] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0023] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0024] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0025] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0026] This invention uses metabolomics analysis to screen 20 endogenous metabolites that are significantly associated with disease resistance (Nongdabai) and susceptible (Shangshu 19) sweet potato varieties, and further identifies 8 key antibacterial substances, one of which is cinnamyl alcohol. This invention further measures the antibacterial activity of cinnamyl alcohol through experiments, and specific examples are described below.

[0027] The soft rot pathogen used in this embodiment of the invention is Rhizopus stolonifer, provided by the Key Laboratory of Sweet Potato Biology and Biotechnology, China Agricultural University.

[0028] The antibacterial substance involved in the embodiments of this invention is cinnamyl alcohol, 98%, purchased from Beijing Solarbio Technology Co., Ltd.

[0029] Potato Dextrose Broth (PDB) was prepared by adding 20 g of agarose per liter, mixing thoroughly, and then autoclaving at 121°C for 30 mins. PDB was purchased from Shaanxi Puyinte Biotechnology Co., Ltd.

[0030] Example 1

[0031] 1. Experimental Methods

[0032] (1) Preparation of cinnamyl alcohol preparations

[0033] Using dimethyl sulfoxide as a solvent, cinnamyl alcohol was prepared to a concentration of 0.1 g·mL in a fume hood. -1 The mother liquor should be prepared and used immediately.

[0034] (2) Activation of microbial strains

[0035] The soft rot pathogen was inoculated onto PDA medium and cultured in a constant temperature incubator at 28°C for 3 days. Once the mycelium had covered the surface of the culture dish, it was used as a starter culture.

[0036] (3) Preparation of plates with different concentration gradients

[0037] Plates of different concentrations were prepared using a serial dilution method with a concentration gradient of 8:4:2:1. A specific volume of cinnamyl alcohol or stock solution was accurately transferred to a 50 mL centrifuge tube using a pipette. PDA medium, heated to 70°C, was then added to the centrifuge tube, bringing the volume to 40 mL. The centrifuge tube cap was tightened, and the tube was inverted to mix thoroughly. 20 mL of the mixed PDA medium was poured into a 9 cm diameter transparent plastic petri dish. The dish was gently shaken to level the surface, and then the dish was lifted to a suitable height and dropped to break air bubbles, forming a uniform, transparent plate free of air bubbles. The medium was then added to the centrifuge tube to bring the volume to 40 mL, the cap was tightened, and the plate was mixed thoroughly before being inverted again. This process was repeated three times to obtain four plates with a concentration gradient of 8:4:2:1. After the plates cooled and solidified, the petri dish caps were closed, and the plates were inverted for later use.

[0038] (4) Result measurement and data analysis

[0039] The soft rot pathogen was inoculated onto plates containing different concentration gradients of cinnamyl alcohol using the inoculation disc method. The growth status of each colony was recorded by photographing at 0, 12, 16, 20 and 24 h after inoculation. Finally, the results were compiled into a chart to observe the growth rate of the soft rot pathogen after the addition of antibacterial substances.

[0040] The 24-hour inhibition rate of this substance was determined based on the colony growth status at 24 hours. The formula for calculating the inhibition rate is as follows:

[0041]

[0042] ImageJ software (v1.53) is used to measure the diameter of bacterial colonies.

[0043] R version 4.3.2 was used to perform one-way ANOVA (Analysis of variance), with the significance threshold set to P < 0.05. The graphs were plotted using the ggplot2 package.

[0044] 2. Results and Analysis

[0045] Based on the exponential growth characteristics of microorganisms, the experiment was conducted by photographing the culture dishes at 0, 12, 16, 20, and 24 hours after inoculation. The treatment groups with different concentrations of cinnamyl alcohol were then arranged and combined into a graph to visually represent its antibacterial effect. The results are as follows: Figures 1-2 As shown. The results indicate that at the highest concentration treatment (400 μg·ml) -1 At this concentration, no mycelia could grow on the culture medium within 24 hours, indicating that the growth of the soft rot pathogen was completely inhibited. At 200 μg / ml... -1 At the same concentration, cinnamyl alcohol also showed good antibacterial effects, with noticeable mycelia appearing only 20 hours after inoculation. Therefore, it can be preliminarily and roughly considered that the minimum inhibitory concentration of cinnamyl alcohol against soft rot pathogens is between 200 μg / ml. -1 Up to 400 μg·ml -1 Between. And 50 μg·ml -1 The cinnamyl alcohol treatment did not show significant differences in mycelial growth throughout the entire period, and the colony size was not significantly reduced compared to the control group. However, this indicates that the antibacterial effect of cinnamyl alcohol at this concentration was not ideal. (100 μg·ml) -1 The mycelial growth of cinnamyl alcohol-treated colonies was observed to be smaller compared to the control group, but the antibacterial effect was not as high as that of the 200 μg / ml treatment. -1 Or 400 μg·ml -1 The effect of cinnamyl alcohol treatment is obvious.

[0046] The antibacterial rates of cinnamyl alcohol at different concentrations after 24 hours are shown in Table 1. Figure 3 As shown.

[0047] Table 1. Analysis of the antibacterial rate of cinnamyl alcohol at different concentration gradients over 24 hours.

[0048]

[0049] Note: For cinnamyl alcohol, no colonies were observed to grow in repeated experiments when treated at the maximum concentration. Therefore, it is assumed that the antibacterial rate of these substances reaches 100% and the variance is zero at this concentration.

[0050] As can be seen from the data in Table 1, 200 μg·mL -1 and 400 μg·mL -1 Cinnamyl alcohol treatment of Rhizopus stolonifera for 24 h showed a significant antibacterial effect, resulting in slowed mycelial growth, abnormal colony morphology, and inhibited fungal growth. Furthermore, the inhibition rate increased significantly with increasing concentration, reaching a maximum of 400 μg / mL. -1 The results showed that low concentrations of cinnamyl alcohol (200, 400) maintained good antibacterial effects, which is a significant advantage for sweet potato storage conditions. This provides a scientific basis for developing it as a green bactericide and preservative for sweet potato preservation. It also indicates that even if the concentration of the active ingredient decreases due to oxidative decomposition or other reasons, the residual low-dose bactericide and preservative can still exert its antibacterial and preservative effects. Therefore, cinnamyl alcohol is a suitable choice for long-term storage of sweet potatoes.

[0051] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. Application of cinnamyl alcohol in prevention and treatment of soft rot of sweet potato.

2. Application of cinnamyl alcohol in prevention and treatment of Rhizopus stolonifer.

3. Application of cinnamyl alcohol in preparation of a postharvest storage preservative for sweet potato.

4. Application of cinnamyl alcohol in preparation of a biological preparation for prevention and treatment of soft rot or Rhizopus stolonifer of sweet potato.

5. A sweet potato postharvest storage preservative, characterized by comprising: consists of 200-400 μg·ml -1 of cinnamyl alcohol and a solvent for dissolving the cinnamyl alcohol.

6. The sweet potato postharvest storage preservative according to claim 5, wherein The solvent comprises dimethyl sulfoxide.

7. A biological agent for controlling soft rot of sweet potato or Rhizopus stolonifer. characterized by, consists of 200-400 μg·ml -1 of cinnamyl alcohol and a solvent for dissolving the cinnamyl alcohol.

8. The biological preparation of claim 7, wherein, The solvent comprises dimethyl sulfoxide.

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