Efficient screening method of bamboo shoot postharvest preservative
Through big data analysis and molecular simulation technology, the inhibitor of the ethylene signal receptor PeETR1 was screened, which solved the problem of difficulty and high cost in the screening of fresh preservatives after harvest of bamboo shoots, and achieved efficient and safe fresh preservative screening and evaluation.
Patent Information
- Application Number
- CN202510089305.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-20
- Publication Date
- 2025-05-16
AI Technical Summary
The existing bamboo shoots' post-harvest preservative screening process has problems such as difficult to uniform operation, poor repeatability, and high screening cost, and lacks efficient screening and evaluation methods.
Through big data analysis and molecular simulation technology, the inhibitors of the ethylene signal receptor PeETR1 were simulated and screened, combined with 1-MCP to regulate the quality of fresh bamboo shoots, and the extraction of experimental data, meta-analysis and storage experiment evaluation, so as to achieve efficient screening of fresh bamboo shoots preservatives.
It realizes efficient screening of bamboo shoots after harvest fresh preservatives, reduces R&D costs, obtains safe and effective fresh preservatives, and has the advantages of low cost and efficient screening.
Smart Images
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Abstract
Description
Technical Field
[0001] The invention relates to the field of plant physiology, and in particular to a method for efficiently screening a post-harvest preservative for moso bamboo shoots. Background Art
[0002] China is a big country in the bamboo industry, and its bamboo shoot cultivation area and output rank first in the world. On the one hand, most bamboo shoots are planted in suburbs or mountainous areas far away from cities, and the maturity period is relatively concentrated; on the other hand, aging and decay are the main factors causing the quality deterioration of bamboo shoots. Therefore, efficient and reasonable storage and transportation technology is needed to regulate the contradiction between the supply and demand of the fresh bamboo shoot market of "centralized listing" and "long-term supply". Post-harvest preservation technology of bamboo shoots is mainly divided into preservatives / chemical treatment, physical treatment and biological treatment according to the characteristics of the treated material. Chemical treatment refers to the preservation technology of post-harvest treatment of fruits and vegetables through treatment methods such as soaking, coating, fumigation, etc. of chemicals; physical treatment refers to the preservation technology of post-harvest treatment of fruits and vegetables through physical means such as high temperature, high pressure, gas conditioning, etc.; biological treatment generally refers to the preservation technology of inhibiting the disease of fruits and vegetables through the antagonism between exogenous bacteria and pathogens. Although physical treatment and biological treatment are safe and reduce the amount of residual agents in fruits, they have the characteristics of high facility construction cost and complex operation. Therefore, chemical preservative treatment has the advantages of low cost, easy operation and easy promotion, and is the most widely used preservation technology today. As people pay more and more attention to healthy diet, screening and developing more non-toxic, pollution-free, pollution-free and low-cost chemical preservative treatment agents is the main research direction at present.
[0003] In the past 20 years, with the support of multiple research projects such as the National Natural Science Foundation, the Ministry of Science and Technology, and the Chinese Academy of Sciences, my country has developed a number of preservatives with potential for application in the market, such as 1-methylcyclopropene (1-MCP) (Jiang et al., 1999; Huang et al., 2019), which can be used to preserve bananas, kiwis, cantaloupes, broccoli, etc., and extend the post-harvest storage period. However, the screening process of these preservatives has problems such as difficult to unify operations, poor repeatability, and high screening costs. The horticultural product materials required to screen one preservative are tens or even hundreds of kilograms. Because there are many types of bamboo shoots, the preservatives suitable for them are also different. How to improve the screening speed of preservatives and reduce R&D costs has become an urgent problem to be solved. Moreover, there have been no reports or patents published on methods for efficiently screening and evaluating the effects of bamboo shoot preservatives through big data analysis and standardized batch experiments. Summary of the invention
[0004] The object of the present invention is to provide a method for efficiently screening a post-harvest preservative for bamboo shoots, so as to overcome the defects in the prior art.
[0005] To achieve the above object, the technical solution of the present invention is: a method for efficiently screening a preservative for post-harvest bamboo shoots, taking the screening of a preservative for moso bamboo shoots as an example, and implementing the method according to the following steps:
[0006] Step S1: 1- Literature search strategy and exclusion criteria for MCP regulation of postharvest quality of fresh bamboo shoots;
[0007] Step S2: Extraction and correction of experimental data on 1-MCP regulating postharvest quality of fresh bamboo shoots;
[0008] Step S3: 1-Meta-analysis of MCP regulation of postharvest quality of fresh bamboo shoots;
[0009] Step S4: 3D structure simulation and molecular docking of bamboo ethylene receptor PeETR1 protein;
[0010] Step S5: exploring preservatives by virtual screening of inhibitors of the bamboo ethylene signaling receptor PeETR1;
[0011] Step S6: Storage experiment to evaluate the effect of PeETR1 inhibitor on the preservation of bamboo shoots.
[0012] In step S1, we conducted in-depth studies to evaluate the different effects of 1-MCP on the biochemical and physiological activities of postharvest bamboo shoots. This search included three databases: PubMed, ISI Web of Knowledge, and China National Knowledge Infrastructure (CNKI). In order to obtain as many relevant studies as possible, we used the keywords "1-MCP" and "1-methylcyclopropene" as well as "bamboo" in the queries of these databases. Our selection process followed the PRISMA guidelines and adopted the following exclusion criteria: 1. Literature lacking original data, such as reviews, meta-analyses, book chapters, and protocols. 2. Publications that did not clearly define the experimental and control groups or were conducted under inconsistent storage conditions. 3. Studies using cereals and cooked foods as experimental materials.
[0013] In step S2, we selected the data reported during different storage periods and selected the final data available during the storage process. When the study did not include numerical data, we extracted data from bar charts and line graphs using the webplot digitization tool. The extracted information included references, bamboo species, 1-MCP treatment concentration, storage temperature, and selected physiological parameters at the end of storage with their means and standard deviations (SDs). The data were extracted and reported according to the PRISMA guidelines (http: / / prisma-statement.org / ). The extracted data tables were validated to ensure precision and accuracy before further analysis.
[0014] The meta-analysis of 1-MCP regulating the postharvest quality of fresh bamboo shoots described in step S3 was specifically to use the restricted maximum likelihood method to perform a Meta-analysis on the extracted data, including the mean, standard deviation and sample size; heterogeneity was analyzed using the chi-square test, Tau 2 and 95% confidence intervals, with high heterogeneity defined as Tau 2 and I 2 >75%; 1-MCP treatment was considered significant if the 95% confidence interval did not intersect the zero line, and mixed effects were represented by dotted lines; potential publication bias was tested using funnel plots, and sensitivity analysis was performed to determine the robustness of the summary results.
[0015] The post-harvest quality of fresh bamboo shoots in step S3 includes aging index, antioxidant capacity, and post-harvest secondary wall biosynthesis capacity. The aging index includes ethylene production rate, decay rate, and respiration rate. The antioxidant capacity determination includes determining the enzyme activities of ascorbate peroxidase, peroxidase, and superoxide dismutase. The post-harvest secondary wall biosynthesis capacity includes determining the hardness, cellulose, lignin, and PAL and CAD activities of the post-harvest bamboo shoots.
[0016] Step S4 specifically includes the establishment of the three-dimensional structure of the bamboo ethylene receptor ETR1 protein and the analysis of the active binding site of PeETR1.
[0017] The PeETR1 active binding site analysis specifically includes the following steps: using Autodock4.2 and Vina4 software to predict the binding mode of PeETR1 with ethylene or 1-MCP; then using ChimeraX and LigPlot+ to visualize the final results and analyze the active binding site residues of PeETR1 with ethylene or 1-MCP.
[0018] Step S5 specifically includes: using the EasyVS platform to perform virtual screening of bamboo shoot preservatives; the screening criteria are: less than 30 atoms, a maximum of three ring structures, selecting protein targets with known structures, and performing docking simulation and ADMET analysis.
[0019] Step S6: Through virtual screening and comprehensive analysis of existing research, it is confirmed that kynurenine may serve as an inhibitor of the ethylene signaling receptor PeETR1, and its preservation effect is further verified through storage experiments on winter bamboo shoots at room temperature.
[0020] Step S6 specifically includes the following steps: directly placing the bamboo shoots into a sealed can, adding 1 μL / L 1-MCP for sealing and fumigation for 16 hours; or immersing the bamboo shoots into a 100 μM / L kynurenine solution, taking them out after immersion for 5 minutes, and standing them to dry for 1 hour; or soaking the bamboo shoots in distilled water for 5 minutes, taking them out, and standing them to dry for 1 hour, as a control group; and then storing them at room temperature for 10 days to observe the changing trend of the appearance quality of the bamboo shoots.
[0021] The beneficial effects of the present invention are as follows: the present invention proposes an efficient screening method for post-harvest preservatives for bamboo shoots. Literature collection and experimental data extraction on 1-MCP regulating the post-harvest quality of fresh bamboo shoots; Meta-analysis of 1-MCP regulating the post-harvest quality of fresh bamboo shoots; 3D structural simulation and molecular docking comparison of binding affinity of the bamboo ethylene receptor PeETR1 protein; mining of bamboo shoot preservatives based on the competitive inhibition mechanism. The present invention takes bamboo shoots as the research object, and through big data analysis and molecular simulation technology, simulates and screens ethylene signal receptor inhibitors to achieve the preservation of bamboo shoots, which has the advantages of low cost and efficient screening to obtain safe preservatives. BRIEF DESCRIPTION OF THE DRAWINGS
[0022] Figure 1 It is a flow chart of the present invention.
[0023] Figure 2 Forest plot of random effect analysis of 1-MCP treatment on ethylene production rate, decay index and respiration rate of Group 1.
[0024] Figure 3 Forestplot of random effect analysis of 1-MCP treatment on APX activity, POD activity, and SOD activity in group 1.
[0025] Figure 4 Forest plot of random effect analysis of 1-MCP treatment on cellulose, hardness, lignin, PAL activity and CAD activity in Group 3.
[0026] Figure 5 The mechanism of 1-MCP in delaying postharvest senescence and maintaining quality of bamboo shoots was analyzed based on meta-analysis.
[0027] Figure 6 This is the interaction mode between PeETR1 protein and ethylene and 1-MCP, and the mechanism of competitive inhibition mediating post-harvest senescence of bamboo shoots.
[0028] Figure 7 To investigate the interaction pattern of kynurenine with ethylene receptor PeETR1 protein, its safety evaluation and its effect on the appearance quality of winter bamboo shoots. DETAILED DESCRIPTION
[0029] The technical solution of the present invention is described in detail below in conjunction with the accompanying drawings.
[0030] The present invention proposes an efficient screening method for post-harvest preservatives for bamboo shoots. Literature collection and experimental data extraction on 1-MCP regulating the post-harvest quality of fresh bamboo shoots; Meta-analysis of 1-MCP regulating the post-harvest quality of fresh bamboo shoots; 3D structural simulation and molecular docking comparison of binding affinity of the bamboo ethylene receptor PeETR1 protein; mining of bamboo shoot preservatives based on the competitive inhibition mechanism. Taking bamboo shoots as an example, the present invention simulates and screens ethylene signal receptor inhibitors through big data analysis and molecular simulation technology to achieve the preservation of bamboo shoots, which has the advantages of low cost and efficient screening of safe preservatives.
[0031] Example 1 A method for efficiently screening postharvest preservatives for bamboo shoots
[0032] The steps include:
[0033] Step S1: 1- Literature search strategy and exclusion criteria for MCP regulation of postharvest quality of fresh bamboo shoots;
[0034] Step S2: Extraction and correction of experimental data on 1-MCP regulating postharvest quality of fresh bamboo shoots;
[0035] Step S3: 1-Meta-analysis of MCP regulation of postharvest quality of fresh bamboo shoots;
[0036] Step S4: 3D structure simulation and molecular docking of bamboo ethylene receptor PeETR1 protein;
[0037] Step S5: exploring preservatives by virtual screening of inhibitors of the bamboo ethylene signaling receptor PeETR1;
[0038] Step S6: Storage experiment to evaluate the effect of PeETR1 inhibitor on the preservation of bamboo shoots.
[0039] In order to allow those skilled in the art to further understand the method proposed in the present invention, the specific implementation method of the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. In this embodiment, the method for investigating bamboo shoot experimental data, preservative screening process and evaluating the safety of preservatives is universal. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.
[0040] In this embodiment, if Figure 1 The flowchart of the method is shown in FIG.
[0041] In step S1, we conducted in-depth studies to evaluate the different effects of 1-MCP on the biochemical and physiological activities of postharvest bamboo shoots. This search included three databases: PubMed, ISI Web of Knowledge, and China National Knowledge Infrastructure (CNKI). In order to obtain as many relevant studies as possible, we used the keywords "1-MCP" and "1-methylcyclopropene" as well as "bamboo" in the queries of these databases. Our selection process followed the PRISMA guidelines and adopted the following exclusion criteria: 1. Literature lacking original data, such as reviews, meta-analyses, book chapters, and protocols. 2. Publications that did not clearly define the experimental and control groups or were conducted under inconsistent storage conditions. 3. Studies using cereals and cooked foods as experimental materials.
[0042] In step S2, we selected the data reported during different storage periods and selected the final data available during the storage process. When the study did not include numerical data, we extracted data from bar charts and line graphs using the webplot digitization tool. The extracted information included references, bamboo species, 1-MCP treatment concentration, storage temperature, and selected physiological indicators at the end of storage and their means and standard deviations (SDs). The data were extracted and reported according to the PRISMA guidelines (http: / / prisma-statement.org / ). The extracted data tables were validated before further analysis to ensure precision and accuracy. The extracted results are shown in Table 1.
[0043] Table 1.1 - Contribution of MCP treatments to the main characteristics of selected reports in the meta-analysis.
[0044]
[0045] Note: APX, ascorbate peroxidase activity; BI, browning index; CAD, cinnamyl alcohol dehydrogenase activity; CI, chilling injury index; EP, ethylene yield; FAA, free amino acid; malondialdehyde content; O2-·, superoxide anion; PAL, phenylalanine ammonia lyase activity; POD, peroxidase; PPO, polyphenol oxidase activity; RR, respiration rate; RS, reducing sugar content; SOD, superoxide dismutase; SP, soluble protein content; TFC, total flavonoids content; TS, total soluble sugar content; TPC, total phenolic compounds; WL, weight loss rate.
[0046] In step S3, we used a random effects model (restricted maximum likelihood method) to conduct a detailed meta-analysis of the extracted data, including mean, standard deviation and sample size. In the Forest plot, the length of each point represents the sample size. In order to evaluate the source of heterogeneity, subgroup analysis based on the physiological and biological functions of the subjects was performed, and the results were reported as standard mean difference (SMD). Heterogeneity was analyzed using chi-square (I 2 ) test, Tau 2 and 95% confidence intervals (CI), and high heterogeneity was defined as Tau 2 and I 2 >75%. If the 95% confidence interval did not intersect the zero line, the effect of 1-MCP treatment was considered significant, and mixed effects were represented by dotted lines. Funnel plots were used to test for potential publication bias, and sensitivity analysis was performed to determine the stability of the summary results. All statistical analyses and visualizations were performed using the “meta” package in R software version 4.1.1, and statistical significance was indicated when the p value was <0.05. As shown in Table 2, we summarize the overall heterogeneity, inconsistency, and confidence intervals associated with the effect of 1-MCP treatment on the preservation of fresh bamboo shoots.
[0047] Table 2 Summary of subgroup analysis results
[0048]
[0049] Note: APX, ascorbate peroxidase; CAD, cinnamyl alcohol dehydrogenase; PAL, phenylalanine ammonia lyase; POD, peroxidase; SOD, superoxide dismutase.
[0050] In one embodiment of the present invention, step S3 further includes the following steps:
[0051] Step S31: Analysis of the effect of 1-MCP treatment on postharvest senescence of bamboo shoots
[0052] like Figure 2 , the overall effect of 1-MCP treatment on the postharvest senescence index of fresh bamboo shoots, including ethylene production rate, decay rate, and respiration rate. Subgroup analysis showed that the ethylene production rate was significantly inhibited in the 1-MCP treatment group (SMD-2.33, 95%CI[-2.72,-1.94]; sample size=870; studies=4; I 2 =81.7%; p<0.01). Regarding the incidence of postharvest diseases, 1-MCP treatment significantly reduced the decay rate (SMD 0.05, 95% CI [-4.79, 4.90]; sample size = 720; studies = 2; I 2=99.9%; p<0.01). The respiration rate of the postharvest bamboo shoots in the 1-MCP treatment group also showed a significant decrease (SMD -1.69, 95% CI [-2.26, -1.13]; sample size = 780; studies = 3; I 2 =96.2%; p<0.01). Overall, 1-MCP treatment effectively delayed postharvest senescence of bamboo shoots, as reflected in the reduction of ethylene production rate, respiration rate, and decay rate (SMD 2.63, 95% CI [-2.67, -0.54]; sample size = 2370; studies = 5; I 2 =100%; p=0).
[0053] Step S32: Analysis of the effect of 1-MCP on the antioxidant capacity system of post-harvest bamboo shoots
[0054] like Figure 3 As shown in the results, 1-MCP treatment affected the antioxidant capacity of bamboo shoots, with a particular focus on the enzyme activities of ascorbate peroxidase (APX), peroxidase (POD), and superoxide dismutase (SOD). All studies observed an increase in APX activity induced by 1-MCP (SMD 6.23, 95% CI [-5.68, 6.78]; sample size = 150; studies = 2; I 2 =0%; p=0.81). Similarly, 1-MCP treatment significantly stimulated SOD activity (SMD 7.27, 95% CI [-0.53, 15.07]; sample size = 150; studies = 2; I 2 =100%; p<0.01), while the peroxidase POD activity was significantly inhibited (SMD-3.39; 95%CI [-6.06, -0.73]; sample size = 840; studies = 4; I 2 =99.3%; p<0.01). In summary, 1-MCP can effectively increase the APX and SOD activities, inhibit POD activity, and reduce the risk of oxidation during storage of postharvest bamboo shoots. Subgroup analysis of Forestplot supported the antioxidant effect of 1-MCP on postharvest bamboo shoots (SMD 1.66, 95% CI [-2.57, 5.89]; sample size = 1140; studies = 5; I 2 =100%; p=0). Therefore, 1-MCP can regulate the antioxidant system and thus affect the postharvest quality and shelf life of bamboo shoots during storage.
[0055] Step S33: Analysis of the effect of 1-MCP on secondary wall biosynthesis of postharvest bamboo shoots
[0056] Figure 4 The effects of exogenous 1-MCP on the hardness, cellulose, lignin, and the activities of PAL and CAD in postharvest bamboo shoots were studied. Figure 4 Results showed that 1-MCP treatment significantly reduced cellulose content in all studies (SMD -3.26, 95% CI [-7.24, -0.73]; sample size = 780; studies = 3; I 2 =99%; p<0.01). As an indicator of lignification, lignin content showed a slower increase in bamboo shoots treated with 1-MCP postharvest, as reported by 4 studies (SMD -3.94, 95% CI [-8.31, -0.43]; sample size = 840; studies = 4; I 2 =99%; p<0.01). In addition, 1-MCP significantly reduced the firmness of postharvest bamboo shoots (SMD -3.41, 95% CI [-5.62, -1.19]; sample size = 930; studies = 5; I 2 =99%; p<0.01). Four studies with a total sample size of 4170 showed that the activities of PAL and CAD were significantly reduced in bamboo shoots treated with 1-MCP. Therefore, the biosynthesis of secondary wall in postharvest bamboo shoots was significantly inhibited (SMD -3.37, 95% CI [-4.76, -1.99]; sample size = 4170; studies = 5; I 2 =98%; p<0.01). In conclusion, 1-MCP treatment effectively inhibited the lignification of postharvest bamboo shoots during storage.
[0057] Step S34: 1-Overall analysis of MCP on postharvest physiological regulation of bamboo shoots
[0058] As attached Figure 5 As shown in the meta-analysis, 1-MCP treatment has a preservation effect on bamboo shoots, including reduced decay rate, respiration rate, and ethylene production, which together delay postharvest senescence of bamboo shoots. In addition, the results also showed that the preservation effect of 1-MCP is related to enhanced antioxidant capacity and reduced secondary wall biosynthesis.
[0059] In the step S4, structural simulation and molecular docking of the bamboo ethylene receptor ETR1 are performed.
[0060] In one embodiment of the present invention, the step S4 further includes the following steps:
[0061] Step S41: Establishment of the three-dimensional structure of the bamboo ethylene receptor ETR1 protein.
[0062] The genome of bamboo was downloaded from Bamboobase (https: / / bamboo.genobank.org / ). In addition, the Arabidopsis genome was obtained from the TAIR database (https: / / www.arabidopsis.org / ). The AtETR1 gene sequence from Arabidopsis (AT1G66340) was aligned with the bamboo genome sequence to identify PeETR1 (PH01000744G0730) as a homologous protein. The trRosetta server (https: / / yanglab.qd.sdu.edu.cn / trRosetta / ) was then used to perform 3D structural modeling of the PeETR1 protein.
[0063] Step S42: To compare the binding affinity, the 3D structures of ethylene and 1-MCP molecules were obtained from the NCBI website. After hydrogenation, the binding modes of PeETR1 with ethylene or 1-MCP were predicted using Autodock4.2 and Vina4 software; ChimeraX and LigPlot+ were then used to visualize the final results. Figure 6 As shown, the interaction analysis of PeETR1 with ethylene revealed that the active binding site of PeETR1 includes four residues: Phe49, Val54, Tyr51 and Leu55, with an affinity index of -1.6 kcal / mol. In addition, 1-MCP effectively binds to the same active site, including five residues: Phe49, Tyr51, Val54, Leu55 and Ile108, with a higher affinity-index of 3.0 kcal / mol ( Figure 6 C&D). This suggests a more stable interaction between PeETR1 and 1-MCP than between ethylene.
[0064] Step S5: Discover bamboo shoot preservatives through virtual screening of PeETR1 inhibitors.
[0065] The EasyVS platform (https: / / biosig.lab.uq.edu.au / easyvs / ) was used to simplify the selection of molecular libraries and facilitate the virtual screening of bamboo shoot preservatives. The screening criteria included less than 30 atoms, a maximum of three ring structures, etc. Protein targets with known structures were selected for docking simulation and ADMET (absorption, distribution, metabolism, excretion and toxicity of drugs) analysis. Some of the results of molecular docking and ADMET evaluation are shown in Table 3. Then, further storage and preservation experiments were performed to verify that these compounds had a specific inhibitory effect on PeETR1 and a certain preservation effect on bamboo shoots.
[0066]
[0067] Step S6: Through virtual screening and comprehensive analysis of existing studies, we confirmed that kynurenine may be an inhibitor of the ethylene signaling receptor PeETR1; through molecular docking analysis, we determined that kynurenine interacts with the active site of the PeETR1 protein, showing a higher binding free energy of 6.1 kcal / mol than that of 1-MCP and ethylene, which includes hydrogen bonds and various non-covalent interactions, involving 7 residues: Arg356, Ser397, Asn352, Mse355, Mse359, Ala393 and Thr394 ( Figure 7 A&B); ADMET analysis showed that kynurenine has relatively stable compound properties, and toxicity tests on humans showed its safety ( Figure 7 C&D); and further verified its preservation effect by conducting storage experiments on winter bamboo shoots at room temperature ( Figure 7 E). The specific steps are as follows:
[0068] Winter bamboo shoots were collected from bamboo forests in Renhua County, Shaoguan City, China, and sent to the laboratory for processing. The selected bamboo shoots were ensured to have uniform shape and color, and any samples with defects or lesions were removed and divided into three groups, each containing 10 bamboo shoots: (1) Treatment group 1 (1-MCP): The bamboo shoots were directly placed in a sealed jar, and 1 μL / L 1-MCP was added for sealed fumigation for 16 hours. (2) Treatment group (kynurenine): The bamboo shoots were immersed in a 100 μM / L kynurenine (CAS: 2922-83-0) solution, soaked for 5 minutes, taken out, and left to dry for 1 hour. (3) Control group: Sterile water was used instead of solution, soaked for 5 minutes, taken out, and left to dry for 1 hour.
[0069] After all treatments were completed, the bamboo shoots were packed in 0.03 mm polyethylene bags and stored at room temperature for 10 days with relative humidity maintained at 85-90%. The appearance changes were observed at 0, 1, 4, 7 and 10 days. Figure 7 The experimental results showed that the senescence and browning degree of bamboo shoots and fruits in the 1-MCP and kynurenine treatment groups were significantly lower than those in the control group, and the appearance freshness of bamboo shoots and fruits in the kynurenine treatment group was slightly higher than that in the 1-MCP treatment group, indicating that kynurenine has a better preservation effect on bamboo shoots and fruits, further verifying the reliability of this research method.
[0070] The above description is only a preferred embodiment of the present invention. All equivalent changes and modifications made according to the scope of the patent application of the present invention should be covered by the present invention.
Claims
1. A method for efficiently screening a post-harvest preservative for bamboo shoots, characterized in that: The following steps are involved: Step S1: Determination of literature search strategy and exclusion criteria for 1-MCP regulation of postharvest quality of fresh bamboo shoots; Step S2: Extraction and correction of experimental data on 1-MCP regulating postharvest quality of fresh bamboo shoots; Step S3: 1-Meta-analysis of MCP regulation of postharvest quality of fresh bamboo shoots; Step S4: 3D structure simulation and molecular docking of bamboo ethylene receptor PeETR1 protein; Step S5: exploring preservatives by virtual screening of inhibitors of the bamboo ethylene signaling receptor PeETR1; Step S6: Experimental verification of the inhibitor and evaluation of the bamboo shoot preservation effect.
2. The efficient screening method for post-harvest bamboo shoots preservative according to claim 1, characterized in that: The meta-analysis of 1-MCP regulating the postharvest quality of fresh bamboo shoots described in step S3 was specifically to use the restricted maximum likelihood method to perform a Meta-analysis on the extracted data, including the mean, standard deviation and sample size; heterogeneity was analyzed using the chi-square test, Tau 2 and 95% confidence intervals, with high heterogeneity defined as Tau 2 and I 2 >75%; 1-MCP treatment was considered significant if the 95% confidence interval did not intersect the zero line, and mixed effects were represented by dotted lines; potential publication bias was tested using funnel plots, and sensitivity analysis was performed to determine the robustness of the summary results.
3. The efficient screening method for post-harvest bamboo shoots preservative according to claim 1, characterized in that: The post-harvest quality of fresh bamboo shoots in step S3 includes aging index, antioxidant capacity, and post-harvest secondary wall biosynthesis capacity.
4. The efficient screening method for post-harvest bamboo shoots preservative according to claim 3, characterized in that: The aging index includes ethylene production rate, decay rate and respiration rate.
5. The efficient screening method for post-harvest bamboo shoots preservative according to claim 3, characterized in that: The antioxidant capacity assay includes assaying the enzymatic activities of ascorbate peroxidase, peroxidase and superoxide dismutase.
6. The efficient screening method for post-harvest bamboo shoots preservative according to claim 3, characterized in that: The post-harvest secondary wall biosynthesis capacity includes measuring the hardness, cellulose, lignin, and PAL and CAD activities of the post-harvest bamboo shoots.
7. The efficient screening method for post-harvest bamboo shoots preservative according to claim 1, characterized in that: Step S4 specifically includes the establishment of the three-dimensional structure of the bamboo ethylene receptor ETR1 protein and the analysis of the active binding site of PeETR1.
8. The efficient screening method for post-harvest bamboo shoots preservative according to claim 7, characterized in that: The PeETR1 active binding site analysis specifically includes the following steps: using Autodock4.2 and Vina4 software to predict the binding mode of PeETR1 with ethylene or 1-MCP; then using ChimeraX and LigPlot+ to visualize the final results and analyze the active binding site residues of PeETR1 with ethylene or 1-MCP.
9. The efficient screening method for post-harvest bamboo shoots preservative according to claim 1, characterized in that: Step S5 specifically includes: using the EasyVS platform to perform virtual screening of bamboo shoot preservatives; the screening criteria are: less than 30 atoms, a maximum of three ring structures, selecting protein targets with known structures, and performing docking simulation and ADMET analysis.
10. The efficient screening method for post-harvest bamboo shoots preservative according to claim 1, characterized in that: The evaluation of the bamboo shoot preservation effect in S6 is specifically to verify the preservation effect of the screened compound through a room temperature storage and preservation experiment; specifically comprising the following steps: directly placing the bamboo shoots into a sealed can, adding 1 μL / L 1-MCP for sealed fumigation for 16 hours; or immersing the bamboo shoots in a 100 μM / L kynurenine solution, taking out after soaking for 5 minutes, and letting it stand and dry for 1 hour; or soaking the bamboo shoots in distilled water for 5 minutes and then taking out, letting it stand and dry for 1 hour, which is the control group; then storing at room temperature for 10 days, and observing the changing trend of the appearance quality of the bamboo shoots.