Azalea wilt electrochemical detection method
By screening terpineol as a characteristic biomarker using GC-IMS and electrochemical detection techniques and optimizing electrochemical parameters, a non-destructive, rapid, and accurate detection of rhododendron wilt disease was achieved. This solved the problems of long detection cycles and high destructiveness of existing detection methods, and improved detection efficiency and accuracy.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- 何阳
- Filing Date
- 2026-03-21
- Publication Date
- 2026-05-05
AI Technical Summary
Existing methods for detecting rhododendron wilt are highly destructive, time-consuming, and have high operational barriers, failing to meet the needs of rapid, non-destructive screening in large batches at import and export flower quarantine sites.
Terpineol, a volatile odor molecule specific to rhododendron wilt, was screened out as a characteristic marker using GC-IMS gas chromatography-ion mobility spectrometry. Electrochemical detection was then performed using an electrochemical workstation. By optimizing the detection parameters, an electrochemical detection curve was established to achieve non-destructive, rapid, and accurate disease identification.
It achieves non-destructive, rapid, and accurate detection of rhododendron wilt disease, improving detection efficiency, shortening the detection cycle to 1-2 hours, and achieving an accuracy rate of ≥99%. It is suitable for batch screening at import and export flower quarantine sites, reducing the risk of disease transmission.
Smart Images
Figure CN121978177A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of flower disease detection technology, and more specifically, relates to an electrochemical detection method for rhododendron wilt disease. Background Technology
[0002] Azaleas are a core ornamental flower category in my country and an important variety in the import and export flower trade, occupying a vital position in the horticulture industry and the flower economy, with the industry scale showing a continuous growth trend. Azalea wilt is a high-risk soil-borne disease caused by pathogenic fungi, characterized by high infectivity, high mortality, and strong concealment. It not only causes mass death of flower plants, resulting in huge economic losses to the flower industry, but is also a key target for prevention and control in my country's import and export flower quarantine. Therefore, achieving rapid and accurate detection of this disease is a crucial prerequisite for ensuring safe production in the flower industry and improving port clearance efficiency.
[0003] Currently, the mainstream detection methods for rhododendron wilt mainly include pathogen isolation and culture, molecular biological detection, and field phenotypic identification. Among them, the traditional pathogen isolation and culture method requires destructive sampling of flower samples, and the detection cycle is as long as 3-7 days, which is seriously insufficient in terms of timeliness and cannot meet the needs of rapid screening. Although molecular biological detection has improved in detection accuracy, it still requires destruction of sample tissue to extract nucleic acids, the pretreatment process is cumbersome, and it is highly dependent on professional laboratory equipment and professional operators, making it difficult to carry out detection work on-site. Field phenotypic identification is affected by the concealment of the disease, and can only be identified after the plant shows obvious wilting symptoms, which can easily miss the best control opportunity.
[0004] Existing detection technologies generally suffer from inherent defects such as high destructiveness, long detection cycles, and high operational thresholds. They cannot simultaneously meet the requirements of non-destructive, rapid, and accurate detection, and are difficult to adapt to the industry's urgent need for batch, rapid, and non-destructive screening of samples at import and export flower quarantine sites. Therefore, developing an efficient and highly sensitive detection method for rhododendron wilt disease has become an urgent need in the field of flower industry disease prevention and control. Summary of the Invention
[0005] To address the aforementioned technical problems, this invention provides an electrochemical detection method for rhododendron wilt disease. This method solves the technical problems of existing rhododendron wilt disease detection methods, which are highly destructive, have long detection cycles, and high operational thresholds. They fail to meet the requirements of non-destructive, rapid, and accurate detection, and are unable to satisfy the industry's urgent need for rapid batch screening at import and export flower quarantine sites.
[0006] An electrochemical detection method for rhododendron wilt disease includes the following steps:
[0007] 1) The volatile components of rhododendron samples infected with rhododendron wilt and healthy rhododendron samples were analyzed by GC-IMS gas chromatography-ion mobility spectrometry, and the volatile odor molecules specific to rhododendron wilt were screened as characteristic markers.
[0008] 2) Based on the characteristic of the feature marker undergoing a specific redox reaction on the electrode surface, the feature marker is electrochemically detected using an electrochemical workstation. The electrochemical detection parameters are determined through single-factor optimization experiments. The electrochemical detection parameters include the pH of the detection system, the detection temperature, the reaction time, and the scan rate.
[0009] 3) Prepare standard solutions of the characteristic markers at different concentrations, and use the electrochemical detection system with determined parameters to detect the standard solutions of each concentration, establish an electrochemical detection curve for rhododendron wilt disease, and calculate the detection limit.
[0010] 4) Collect the volatile components of the rhododendron sample to be tested, add the collected volatile components to the electrochemical detection system with determined parameters, use an electrochemical workstation to detect the electrochemical response signal, and combine the electrochemical detection working curve to determine whether the rhododendron sample to be tested is infected with rhododendron wilt disease.
[0011] Preferably, the GC-IMS analysis process in step 1) includes collecting, injecting, separating and detecting the volatile components of the rhododendron sample by gas chromatography and ion mobility spectrometry. By comparing the GC-IMS two-dimensional spectra, difference spectra and fingerprint spectra of infected and healthy samples, the screening of characteristic markers is completed.
[0012] Preferably, the characteristic marker selected in step 1) is terpineol, which is a volatile odor molecule specifically produced by rhododendrons after they are infected with wilt disease.
[0013] Preferably, the single-factor optimization experiment described in step 2) investigates the effects of pH, temperature, reaction time, and scan rate on the peak current of the electrochemical detection system, and determines the optimal values of each parameter with the goal of maximizing the peak current and ensuring a stable detection signal.
[0014] Preferably, the optimal pH of the electrochemical detection system is 5.7-7.2, the optimal detection temperature is 40-60℃, and the optimal reaction time is 8-15 min.
[0015] Preferably, the optimization of the scan rate is based on a good linear relationship between the square root of the scan rate and the peak current, wherein the correlation coefficient R² of the linear relationship is ≥0.99.
[0016] Preferably, the detection method of the electrochemical workstation in step 2) is cyclic voltammetry (CV), and electrochemical response signals such as oxidation peak current value, reduction peak current value, and redox potential are recorded during the detection process.
[0017] Preferably, the characteristic marker standard solution in step 3) is configured with multiple concentration gradients, covering low, medium and high concentration ranges. After detection, a linear electrochemical detection curve is obtained by fitting the characteristic marker concentration as the abscissa and the absolute value of the corresponding oxidation peak current as the ordinate.
[0018] Preferably, in step 4), the collection of volatile components from the rhododendron sample to be tested is a non-destructive collection method, which does not damage the rhododendron plant, leaves, roots, or other tissues and organs during the collection process.
[0019] Preferably, the method is applied to the detection of rhododendron wilt in scenarios such as import and export flower quarantine, field screening in flower planting bases, and disease detection in horticultural flower trading markets.
[0020] Compared with the prior art, the present invention has the following beneficial effects:
[0021] This invention collects volatile components from azaleas using headspace sampling, without damaging any of the plant's leaves, roots, stems, or other tissues or organs. This avoids damage to the commercial value of the flowers during testing and allows for multiple follow-up tests on the same plant, meeting the core requirement of non-damaging samples in the quarantine of imported and exported flowers.
[0022] The detection process of this invention, from the collection of volatile components to the final disease diagnosis, can be completed in just 1-2 hours for a single sample. Compared with the 3-7 days of traditional pathogen isolation and culture methods and the several hours to 1 day of molecular biological detection, the detection efficiency is significantly improved, which can meet the needs of rapid batch screening on site.
[0023] This invention screens terpineol as a specific biomarker. Combined with an optimized electrochemical detection system, the detection limit for terpineol is as low as 0.05 μmol / L. Furthermore, after blind sample testing, the overall detection accuracy is ≥99%, enabling early and accurate identification of diseases and effectively preventing the spread of diseases due to missed or false detections.
[0024] The detection process of this invention is simple and requires no complicated pretreatment. It can be used with a portable GC-IMS instrument and a portable electrochemical workstation to carry out detection, eliminating the dependence on professional laboratories and highly skilled operators. It can achieve rapid detection on-site at import and export quarantine ports, flower planting bases, trading markets and other locations.
[0025] This invention can be widely applied to various scenarios, such as screening for rhododendron wilt at import and export flower quarantine ports, field disease inspections at flower planting bases, and disease entry testing at horticultural flower trading markets. It can effectively improve the overall control efficiency of rhododendron wilt, reduce the risk of disease transmission, and reduce economic losses in the flower industry. At the same time, it helps to improve the customs clearance efficiency of import and export flower ports. It has excellent application value and broad market prospects in the flower and horticulture industry and flower trade. Attached Figure Description
[0026] Figure 1 Two-dimensional GC-IMS spectra of volatile components in azaleas before and after infection with Fusarium wilt.
[0027] Figure 2 Two-dimensional differential spectra of volatile components in azaleas before and after infection with Fusarium wilt, analyzed by GC-IMS.
[0028] Figure 3 GC-IMS fingerprint of volatile components in rhododendrons before and after infection with Fusarium wilt.
[0029] Figure 4 Metabolic response profiles and correlation analysis of volatile components before and after infection with rhododendron wilt disease
[0030] Figure 5 To optimize the single-factor electrochemical enzyme-free detection system for terpineol;
[0031] Figure 6 The response of the electrochemical detection system to different concentrations of terpineol. Detailed Implementation
[0032] This invention discloses an electrochemical detection method for rhododendron wilt disease, aiming to solve the problems of existing rhododendron wilt detection methods being highly destructive, having long detection cycles, and high operational thresholds, thus failing to meet the needs of batch, rapid, and non-destructive screening at import and export flower quarantine sites. This method combines GC-IMS (Gas Chromatography-Ion Mobility Spectroscopy) technology with electrochemical detection technology to achieve non-destructive, rapid, and accurate detection of rhododendron wilt disease. The specific embodiments of this invention are described in detail below with reference to the accompanying drawings and specific experimental data. These embodiments are only for explaining the invention and are not intended to limit the scope of protection of this invention. Figure 5 In the diagram, (A) represents the effect of pH on the electrochemical detection system of terpineol, (B) represents the effect of temperature on the electrochemical detection system of terpineol, (C) represents the effect of reaction time on the electrochemical detection system of terpineol, and (D) represents the relationship between the square root of the scan rate and the peak current. Figure 6 In the figure, (A) is the CV curve of terpineol standard solutions of different concentrations; (B) is the absolute value of the oxidation peak current of terpineol of different concentrations; (C and D) are the working curves for electrochemical detection of terpineol in different concentration ranges.
[0033] Experimental materials and equipment:
[0034] (a) Experimental materials:
[0035] Healthy azalea plants and azalea plants infected with wilt disease after artificial inoculation with pathogenic fungi were selected as experimental samples. All were plants of the same variety and with similar growth cycles. Terpineol standard (chromatographic grade), electrolyte (phosphate buffered saline PBS), anhydrous ethanol, acetone and other reagents were all analytical grade. Ultrapure water was prepared by a laboratory ultrapure water system.
[0036] (II) Instruments and Equipment:
[0037] The system includes a GC-IMS system equipped with a headspace sampler for the separation and detection of volatile components; an electrochemical workstation with a three-electrode system (working electrode: glassy carbon electrode; reference electrode: saturated calomel electrode; counter electrode: platinum wire electrode) for electrochemical signal detection; an electronic balance (accuracy 0.0001 g) for reagent weighing; a pH meter for adjusting solution pH; a constant temperature water bath for controlling detection temperature; and headspace sampling bottles for non-destructive collection of volatile components.
[0038] Step 1: Screening for specific volatile biomarkers for rhododendron wilt disease
[0039] Non-destructive collection of volatile components: Healthy azalea plants and azalea plants infected with wilt disease were taken and placed in sealed headspace sampling bottles. The bottles were left to stand at room temperature for 30 minutes to allow the volatile components released by the plants to reach equilibrium within the sampling bottles, thus completing the non-destructive collection of volatile components. The entire collection process did not damage any tissues or organs of the plant.
[0040] GC-IMS coupled analysis: The headspace sampling vial containing volatile components was placed in the headspace injection device of the GC-IMS instrument. Instrument parameters were set as follows: injection temperature 60℃, injection volume 1mL, gas chromatography column temperature 40℃, carrier gas high-purity nitrogen, flow rate 150mL / min, ion mobility spectrometry drift tube temperature 45℃, drift gas high-purity nitrogen, flow rate 60mL / min. The instrument was started to separate and detect the volatile components of healthy and diseased samples, respectively, obtaining GC-IMS two-dimensional spectra. Figure 1 ), two-dimensional difference spectrum ( Figure 2 ) and fingerprint spectrum ( Figure 3 ).
[0041] Characteristic biomarker screening: Comparative analysis of GC-IMS spectra and metabolic response profiles of healthy and diseased samples ( Figure 4By analyzing differences in spectral peak shape, peak intensity, and relative migration time, combined with qualitative analysis of volatile components, volatile odor molecules specifically produced after azalea infection with Fusarium wilt, but showing no obvious response in healthy samples, were screened as characteristic biomarkers. In this embodiment, the selected biomarker was terpineol, a specific volatile component produced by the metabolism of pathogenic fungi causing Fusarium wilt in azaleas, which can serve as a core indicator for disease detection.
[0042] Step 2: Optimize the parameters of the electrochemical detection system
[0043] Based on the characteristic that terpineol can undergo specific redox reactions on the electrode surface and generate electrochemical signals that can be captured by an electrochemical workstation, a single-factor optimization experiment was conducted to investigate the effects of pH, detection temperature, reaction time, and scan rate on the peak current of the terpineol electrochemical detection system. The optimal parameters for electrochemical detection were determined with the goal of maximizing the peak current and maximizing the detection signal. A three-electrode system was used in all experiments, and phosphate buffer solution (PBS) was used as the electrolyte.
[0044] pH optimization ( Figure 5 B): Prepare a 10 μmol / L terpineol standard solution. Adjust the electrolyte pH to 5.0, 5.7, 6.5, 7.2, 7.8, and 8.4, respectively. Under the conditions of room temperature, scan rate of 0.05 V / s, and reaction time of 10 min, perform cyclic voltammetry (CV) using an electrochemical workstation and record the peak current at different pH values. The results show that the peak current reaches its maximum value and the signal is stable when the pH is between 5.7 and 7.2. This range is determined to be the optimal pH for the detection system. In this embodiment, pH = 6.5 is preferred.
[0045] Temperature optimization ( Figure 5 C): With a fixed detection system pH of 6.5, a scan rate of 0.05 V / s, and a reaction time of 10 min, detection temperatures of 30℃, 40℃, 50℃, 60℃, and 70℃ were set. CV detection was performed on a 10 μmol / L terpineol standard solution, and the peak current was recorded. The results showed that the peak current was the largest and showed no significant fluctuations when the temperature was between 40-60℃. This range was determined to be the optimal detection temperature, and in this embodiment, the preferred temperature was 50℃.
[0046] Optimization of reaction time ( Figure 5D): With pH=6.5, temperature=50℃, and scan rate=0.05V / s fixed, reaction times were set to 5min, 8min, 10min, 15min, and 20min, respectively. CV detection was performed on a 10μmol / L terpineol standard solution, and the peak current was recorded. The results showed that the peak current reached a stable maximum value when the reaction time was 8-15min. This range was determined to be the optimal reaction time, and in this embodiment, the preferred reaction time was 10min.
[0047] Optimization of scan rate: With pH=6.5, temperature=50℃, and reaction time=10min fixed, scan rates of 0.02V / s, 0.04V / s, 0.06V / s, 0.08V / s, and 0.10V / s were set, and CV detection was performed on a 10μmol / L terpineol standard solution. The peak current at different scan rates was recorded, and linear fitting was performed with the square root of the scan rate as the x-axis and the peak current as the y-axis. The results showed a good linear relationship between the two, with a correlation coefficient R²≥0.99 (R²=0.9908 in this embodiment). This scan rate range was determined to be the optimal range, and the preferred scan rate in this embodiment was 0.05V / s.
[0048] In summary, the optimal electrochemical parameters for the detection of rhododendron wilt disease determined by this invention are: detection system pH 5.7-7.2, detection temperature 40-60℃, reaction time 8-15 min, and scan rate 0.02-0.10 V / s.
[0049] Step 3: Establish an electrochemical detection working curve and calculate the detection limit.
[0050] Configure a terpineol standard solution gradient: Using ultrapure water as a solvent, dilute the terpineol standard to prepare a series of standard solutions with different concentrations, covering low, medium and high concentration ranges (in this example, terpineol standard solutions with concentrations of 0.1 μmol / L, 0.5 μmol / L, 1 μmol / L, 5 μmol / L, 10 μmol / L, 20 μmol / L and 50 μmol / L are prepared), to ensure a linear distribution of the concentration gradient and meet the requirements for working curve fitting.
[0051] CV detection and recording of electrochemical signals: Under the optimal electrochemical detection parameters determined in step 2 (pH=6.5, temperature=50℃, reaction time=10min, scan rate=0.05V / s), CV detection was performed on terpineol standard solutions of various concentrations using an electrochemical workstation, and the cyclic voltammetry curves (CV curves) corresponding to each concentration were recorded. Figure 6 A) and the absolute value of the oxidation peak current ( Figure 6 B).
[0052] Establishing a working curve: Using the concentration of terpineol standard solution as the x-axis and the absolute value of the corresponding oxidation peak current as the y-axis, a linear regression method was employed to fit the curve, resulting in the electrochemical detection working curve for azalea wilt disease. Figure 6 C, 6D). In this embodiment, linear working curves were fitted in the low concentration range (0.1-10 μmol / L) and the medium-high concentration range (10-50 μmol / L), and both curves showed good linear correlation (R²≥0.99).
[0053] Detection limit calculation: Based on the signal-to-noise ratio method (S / N=3), combined with the working curve in the low concentration range and the detection signal of the blank sample (pure electrolyte), the detection limit of this electrochemical detection method for terpineol was calculated. In this embodiment, the detection limit is as low as 0.05 μmol / L, indicating that this method has high sensitivity.
[0054] Step 4: Perform wilt disease testing on the azalea samples to be tested.
[0055] Collection of volatile components from the sample to be tested: Place the rhododendron plant to be tested in a sealed headspace sampling bottle and collect the volatile components non-destructively according to the method in step 1. The operating conditions and sample equilibration time should be kept consistent to ensure the consistency of the collection.
[0056] Electrochemical detection: The headspace gas containing the volatile components of the sample to be tested is passed into the prepared electrolyte. Under the optimal electrochemical detection parameters determined in step 2, CV detection is performed using an electrochemical workstation, and the absolute value of the oxidation peak current of the sample to be tested and other electrochemical response signals are recorded.
[0057] Disease assessment: Substitute the absolute value of the oxidation peak current of the sample into the electrochemical detection working curve established in step 3 to calculate the actual concentration of terpineol in the sample.
[0058] If the concentration of terpineol is greater than or equal to the detection limit, the rhododendron sample to be tested is determined to be infected with wilt disease;
[0059] If the concentration of terpineol is below the detection limit, the rhododendron sample to be tested is determined to be uninfected with wilt disease.
[0060] At the same time, multiple samples can be tested in batches to meet the batch screening needs of import and export flower quarantine sites.
[0061] Validation of the detection method:
[0062] Fifty azalea samples each of known healthy and known diseased plants were selected, and blind sample testing was conducted using the detection method of this invention. The results showed that the detection accuracy rate for diseased samples was 100%, the detection accuracy rate for healthy samples was 98%, and the overall detection accuracy rate was ≥99%. The entire detection cycle for a single sample (from the collection of volatile components to obtaining the detection result) only takes 1-2 hours. Compared with traditional isolation and culture methods (3-7 days) and molecular biological detection methods (several hours to 1 day), the detection efficiency is greatly improved, and the entire detection process does not damage any samples, achieving the detection goals of non-destructive, rapid, highly sensitive, and highly accurate detection.
[0063] Application scenarios of this invention:
[0064] The electrochemical detection method for azalea wilt described in this invention is easy to operate, requiring no complex equipment or highly skilled personnel in a specialized laboratory. It can be used with a portable GC-IMS system and a portable electrochemical workstation to achieve rapid on-site detection. It can be widely applied in scenarios such as azalea wilt screening at import and export flower quarantine ports, field disease inspections in flower planting bases, and disease entry detection in horticultural flower trading markets. It can effectively improve the control efficiency of azalea wilt, reduce the risk of disease transmission, and reduce economic losses in the flower industry.
[0065] The above-described specific embodiments are preferred embodiments of the present invention and are not intended to limit the present invention in any way. Those skilled in the art can make various modifications, equivalent substitutions, and improvements to the above embodiments without departing from the technical solution of the present invention, and all such modifications, substitutions, and improvements should be included within the protection scope of the present invention.
[0066] The embodiments of the present invention are given for illustrative and descriptive purposes only, and are not intended to be exhaustive or to limit the invention to the forms disclosed. Many modifications and variations will be apparent to those skilled in the art. The embodiments were chosen and described in order to better illustrate the principles and practical application of the invention, and to enable those skilled in the art to understand the invention and to design various embodiments with various modifications suitable for a particular purpose.
Claims
1. An electrochemical detection method for rhododendron wilt disease, characterized in that, Includes the following steps: 1) The volatile components of rhododendron samples infected with rhododendron wilt and healthy rhododendron samples were analyzed by GC-IMS gas chromatography-ion mobility spectrometry, and the volatile odor molecules specific to rhododendron wilt were screened as characteristic markers. 2) Based on the characteristic of the feature marker undergoing a specific redox reaction on the electrode surface, the feature marker is electrochemically detected using an electrochemical workstation. The electrochemical detection parameters are determined through single-factor optimization experiments. The electrochemical detection parameters include the pH of the detection system, the detection temperature, the reaction time, and the scan rate. 3) Prepare standard solutions of the characteristic markers at different concentrations, and use the electrochemical detection system with determined parameters to detect the standard solutions of each concentration, establish an electrochemical detection curve for rhododendron wilt disease, and calculate the detection limit. 4) Collect the volatile components of the rhododendron sample to be tested, add the collected volatile components to the electrochemical detection system with determined parameters, use an electrochemical workstation to detect the electrochemical response signal, and combine the electrochemical detection working curve to determine whether the rhododendron sample to be tested is infected with rhododendron wilt disease.
2. The electrochemical detection method for rhododendron wilt disease according to claim 1, characterized in that, The GC-IMS analysis process described in step 1) includes collecting, injecting, separating and detecting the volatile components of the rhododendron sample by gas chromatography and ion mobility spectrometry. By comparing the GC-IMS two-dimensional spectra, differential spectra and fingerprint spectra of infected and healthy samples, the screening of characteristic markers is completed.
3. The electrochemical detection method for rhododendron wilt disease according to claim 1, characterized in that, The characteristic marker selected in step 1) is terpineol, which is a volatile odor molecule specifically produced by rhododendrons after they are infected with wilt disease.
4. The electrochemical detection method for rhododendron wilt disease according to claim 1, characterized in that, The single-factor optimization experiments described in step 2) investigated the effects of pH, temperature, reaction time, and scan rate on the peak current of the electrochemical detection system. The optimization goal was to maximize the peak current and stabilize the detection signal, and to determine the optimal values of each parameter.
5. The electrochemical detection method for rhododendron wilt disease according to claim 4, characterized in that, The optimal pH of the electrochemical detection system is 5.7-7.2, the optimal detection temperature is 40-60℃, and the optimal reaction time is 8-15 min.
6. The electrochemical detection method for rhododendron wilt disease according to claim 4, characterized in that, The optimization of the scan rate is based on the fact that the square root of the scan rate has a good linear relationship with the peak current, and the correlation coefficient of the linear relationship is R²≥0.
99.
7. The electrochemical detection method for rhododendron wilt disease according to claim 1, characterized in that, The detection method of the electrochemical workstation mentioned in step 2) is cyclic voltammetry (CV). During the detection process, electrochemical response signals such as oxidation peak current, reduction peak current, and redox potential are recorded.
8. The electrochemical detection method for rhododendron wilt disease according to claim 1, characterized in that, In step 3), the standard solution of the characteristic marker is configured with multiple concentration gradients, covering low, medium and high concentration ranges. After detection, the concentration of the characteristic marker is used as the abscissa and the absolute value of the corresponding oxidation peak current is used as the ordinate to fit a linear electrochemical detection curve.
9. The electrochemical detection method for rhododendron wilt disease according to claim 1, characterized in that, In step 4), the collection of volatile components from the rhododendron sample is a non-destructive process, in which the rhododendron plant, leaves, roots, and other tissues and organs are not damaged during the collection process.
10. The electrochemical detection method for rhododendron wilt disease according to any one of claims 1-9, characterized in that, The method is applied to the detection of rhododendron wilt in scenarios such as import and export flower quarantine, field screening in flower planting bases, and disease detection in horticultural flower trading markets.