A hormone-free culture growth monitoring method for Dendrobium protocorm stem cells

Abstract

The present invention relates to the technical field of plant growth monitoring, and in particular to a hormone-free culture growth monitoring method for Dendrobium protocorm stem cells, which not only improves culture efficiency and effect, but also better controls growth conditions to ensure the quality and yield of Dendrobium protocorm stem cells. The method comprises: collecting a Dendrobium protocorm stem cell culture feature set and a plant growth regulator content during a historical culture process of the Dendrobium protocorm stem cells, wherein the Dendrobium protocorm stem cell culture feature set and the plant growth regulator content have a one-to-one correspondence; performing a correlation analysis on the Dendrobium protocorm stem cell culture feature set and the plant growth regulator content to obtain a culture growth feature correlation matrix; in the culture growth feature correlation matrix, correlation feature values ​​exist between the plant growth regulator content corresponding to each set content range and each Dendrobium protocorm stem cell culture feature; and traversing the culture growth feature correlation matrix.

Description

Technical Field

[0001] The present invention relates to the technical field of plant growth monitoring, in particular to a hormone-free culture growth monitoring method for Dendrobium protocorm stem cells. Background Art

[0002] Dendrobium protocorms are embryonic tissues obtained through plant tissue culture using seeds or stems as explants. These cells possess the ability to both proliferate and differentiate into complete plants, sharing the characteristics of stem cells. If not properly regulated during culture, these cells will simultaneously proliferate and differentiate. This differentiation process not only slows down the proliferation rate and yield of these cells, but also affects their appearance, morphology, and quality.

[0003] Therefore, how to monitor the content of plant growth regulators and cell proliferation rate in Dendrobium protocorm stem cells in real time without the addition of exogenous hormones has become a major challenge facing current Dendrobium protocorm tissue culture technology. Existing monitoring methods, most of which rely on cumbersome chemical testing and manual observation, are not only time-consuming and labor-intensive, but also difficult to achieve real-time monitoring of the growth process of Dendrobium protocorm stem cells. Summary of the Invention

[0004] To solve the above technical problems, the present invention provides a hormone-free culture growth monitoring method for Dendrobium protocorm stem cells, which not only improves the culture efficiency and effect, but also can better control the growth conditions to ensure the quality and yield of Dendrobium protocorm stem cells.

[0005] In a first aspect, the present invention provides a method for monitoring the growth of protocorm stem cells in a hormone-free culture, the method comprising:

[0006] Collecting a set of culture characteristics of Dendrobium protocorm stem cells and a plant growth regulator content during the historical culture process of Dendrobium protocorm stem cells, wherein the set of culture characteristics of Dendrobium protocorm stem cells and the plant growth regulator content correspond one to one;

[0007] Correlation analysis was performed on the culture characteristic set of Dendrobium protocorm stem cells and the content of plant growth regulators to obtain a culture growth characteristic correlation matrix. In the culture growth characteristic correlation matrix, there were correlation characteristic values ​​between the plant growth regulator content corresponding to each set content range and each Dendrobium protocorm stem cell culture characteristic.

[0008] Traversing the culture growth feature correlation matrix, extracting the Dendrobium protocorm stem cell culture features whose correlation feature values ​​are greater than a preset correlation threshold, and summarizing them to obtain a set of key growth culture features of the Dendrobium protocorm stem cells;

[0009] Real-time collection of key culture characteristics of the growth of Dendrobium protocorm stem cells;

[0010] Inputting the key growth culture characteristic set into a pre-built plant growth hormone prediction model to obtain the real-time content of plant growth regulators in the Dendrobium protocorm stem cells;

[0011] Collect microscopic images of the protocorm tissue of Dendrobium officinale at both ends of a preset time period;

[0012] The proliferation trend of the two groups of collected microscopic images of Dendrobium protocorm tissues was analyzed to obtain the cell proliferation rate of Dendrobium protocorm stem cells;

[0013] The cell proliferation rate and real-time content of plant growth regulators of Dendrobium protocorm stem cells were weighted to obtain the culture growth index of Dendrobium protocorm stem cells.

[0014] The culture growth index is compared with the preset standard index to determine whether the hormone-free culture growth of the Dendrobium protocorm stem cells is normal.

[0015] Furthermore, the Dendrobium protocorm stem cell culture characteristic set includes culture medium component characteristics, culture environment characteristics and culture time characteristics; culture medium component characteristics include basal culture medium type, sugar type and concentration, vitamin type and concentration, trace element type and concentration, and initial addition amount of plant growth regulator; culture environment characteristics include temperature, light intensity, light cycle, humidity, CO2 concentration, pH value and oxygen supply; culture time characteristics include time points of different growth stages, overall culture time and differences between different batches.

[0016] Furthermore, the culture growth characteristic correlation matrix is:

[0017]

[0018] Where m represents the number of culture characteristics, n represents the number of samples, and r mn It represents the correlation coefficient between the mth cultural characteristic and the nth plant growth regulator content.

[0019] Furthermore, the formula for calculating the culture growth index of the Dendrobium protocorm stem cells is:

[0020] I=w V V+w C ·C

[0021] Wherein, I represents the culture growth index of the protocorm stem cells of Dendrobium officinale, V represents the cell proliferation rate, C represents the real-time content of the plant growth regulator, and w V represents the weight of cell proliferation rate, w CIndicates the weight of the real-time content of plant growth regulators.

[0022] Furthermore, factors affecting the setting of the preset correlation threshold include application background, data quality, sample size, computational complexity, economic cost and environmental factors.

[0023] Furthermore, the method for constructing the plant growth hormone prediction model includes:

[0024] Collect historical data, including the culture characteristics of Dendrobium protocorm stem cells under different culture conditions and the corresponding plant growth regulator content;

[0025] Preprocess the collected data, including data cleaning and data transformation;

[0026] Divide the dataset into training and validation sets;

[0027] Selecting a deep learning model as the infrastructure of the plant growth hormone prediction model; the deep learning model includes linear regression, decision tree, random forest, support vector machine and neural network;

[0028] Input the training set into the selected model to perform model training;

[0029] After training is completed, the model is verified and evaluated using the validation set;

[0030] After training and validation, the model is deployed into the real-time monitoring system.

[0031] Furthermore, the method for collecting microscopic images of the protocorm tissue of Dendrobium officinale at two end points of a preset time period includes:

[0032] Select a high-resolution, high-magnification microscope for image acquisition;

[0033] Configure the lighting system to provide stable and uniform lighting conditions;

[0034] According to the size and morphology of the protocorm stem cells of Dendrobium officinale, the magnification was set for image acquisition;

[0035] Adjusting the exposure time can clearly show the details of the cells;

[0036] Set the microscope to focus correctly to obtain a clear image;

[0037] At the two end points of the predetermined time period, samples were taken from the protocorm culture of Dendrobium officinale;

[0038] Place the processed sample on the microscope stage, adjust the position and angle of the sample, start the microscope, and capture the microscopic image of the Dendrobium protocorm tissue according to the preset parameter settings;

[0039] Save the acquired images to the computer.

[0040] Furthermore, the method for obtaining the cell proliferation rate of Dendrobium protocorm stem cells comprises:

[0041] Enhance the collected microscopic images to improve the contrast and clarity of the images;

[0042] Remove noise from images to reduce interference with cell identification;

[0043] Perform color correction on images;

[0044] Using edge detection algorithms, the outlines of the protocorm stem cells of Dendrobium candidum were identified;

[0045] Segment cells from the image based on cell outlines;

[0046] Extract cell morphological features, including area, perimeter, circularity, and aspect ratio;

[0047] Extract cell texture features to distinguish cells in different proliferation states;

[0048] Count the segmented cells to obtain the number of cells at each time point;

[0049] Calculate the cell proliferation rate based on the number of cells at the two end points of the preset time segment;

[0050] The cell number and proliferation rate at two time points are plotted into a trend graph to visually display the cell proliferation trend;

[0051] The cell number and proliferation rate were statistically analyzed to obtain the cell proliferation rate of Dendrobium protocorm stem cells.

[0052] Furthermore, factors influencing the setting of the preset standard index include historical culture data analysis, culture conditions, cell type, cell growth stage, limitations of monitoring technology and technological updates.

[0053] Compared with the existing technology, the present invention has the following advantages: by collecting key growth and culture characteristics of Dendrobium protocorm stem cells in real time and using a pre-established plant growth hormone prediction model, the method can predict and monitor the content of plant growth regulators in Dendrobium protocorm stem cells in real time; it avoids the time-consuming and labor-intensive problems of tedious chemical testing and manual observation in traditional methods, and improves the efficiency and accuracy of monitoring;

[0054] By combining advanced technologies such as data analysis, model prediction, and image processing, the system has achieved intelligent and automated monitoring of the growth process of Dendrobium protocorm stem cells. This not only reduces the burden of manual operation but also improves the accuracy and reliability of monitoring.

[0055] By collecting microscopic images of Dendrobium protocorm tissue for proliferation trend analysis, this method enables non-destructive monitoring of cell proliferation rate, helping to protect sample integrity while providing accurate proliferation rate data.

[0056] This method not only considers the content of plant growth regulators but also incorporates multiple factors such as cell proliferation rate to calculate the culture growth index of Dendrobium protocorm stem cells through weighted calculation. This comprehensive evaluation method can more comprehensively and accurately reflect the growth status of Dendrobium protocorm stem cells.

[0057] This method is based on data analysis and technical means, and does not rely on specific hardware equipment or complex experimental conditions. Therefore, it has high operability and scalability. It is suitable for Dendrobium protocorm tissue culture laboratories of different sizes and conditions, and will help promote the further development of Dendrobium protocorm tissue culture technology.

[0058] In summary, this method has obvious advantages in improving the accuracy, efficiency and operability of hormone-free culture growth monitoring of Dendrobium protocorm stem cells. It not only improves the culture efficiency and effect, but also can better control the growth conditions and ensure the quality and yield of Dendrobium protocorm stem cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Figure 1 is a flow chart of the present invention;

[0060] Figure 2 It is a flow chart of the method for constructing a plant growth hormone prediction model. DETAILED DESCRIPTION

[0061] The present application is described below in conjunction with the accompanying drawings.

[0062] Example 1: Figures 1 to 2 As shown, the present invention provides a method for monitoring the growth of Dendrobium protocorm stem cells without hormone culture, which specifically comprises the following steps:

[0063] S1. Collecting a set of culture characteristics of Dendrobium protocorm stem cells and a plant growth regulator content during the historical culture process of Dendrobium protocorm stem cells, wherein the set of culture characteristics of Dendrobium protocorm stem cells and the plant growth regulator content correspond to each other; the set of culture characteristics of Dendrobium protocorm stem cells includes culture medium component characteristics, culture environment characteristics, and culture time characteristics;

[0064] A collection of culture characteristics of Dendrobium protocorm stem cells and a method for collecting plant growth regulator content, including:

[0065] From past Dendrobium protocorm stem cell culture experiments, we compiled the culture characteristic data and plant growth regulator content data recorded during each culture process;

[0066] Select representative samples for data analysis to ensure that the samples cover different culture conditions, growth stages, and plant growth regulator content ranges;

[0067] Preprocess the collected data, including removing outliers, filling missing values, and normalizing the data to ensure consistency and comparability;

[0068] Using database software or spreadsheet software to establish a data storage system for storing data on the culture characteristic set of Dendrobium protocorm stem cells and the content of plant growth regulators;

[0069] Add labels to each data point, including culture batch, timestamp, culture conditions, etc., to facilitate subsequent data analysis and query;

[0070] Back up your database or spreadsheet regularly to prevent data loss or corruption;

[0071] Verify the collected data to ensure its accuracy and reliability;

[0072] The characteristics of culture medium components include the type of basal culture medium, the type and concentration of sugars, the type and concentration of vitamins, the type and concentration of trace elements, and the initial addition amount of plant growth regulators;

[0073] Culture environment characteristics include temperature, light intensity, photoperiod, humidity, CO2 concentration, pH value, and oxygen supply;

[0074] Culture time characteristics include the time points of different growth stages, the overall culture duration, and the differences between different batches.

[0075] In this step, by collating data from past Dendrobium protocorm stem cell culture experiments and selecting representative samples for data analysis, it is ensured that the collected data can fully cover different culture conditions, growth stages and plant growth regulator content ranges; the collected data are preprocessed, including removing outliers, filling missing values ​​and data normalization, which effectively improves the quality and consistency of the data; this helps to reduce data noise and improve the accuracy and efficiency of data analysis; a data storage system is established using database software or spreadsheet software, and labels are added to each data point, making data storage and query more convenient; this helps subsequent researchers quickly access and analyze data and improve work efficiency; databases or spreadsheets are backed up regularly to prevent data loss or damage , ensuring the security and reliability of the data; at the same time, the collected data are verified to ensure the accuracy and reliability of the data, providing a solid foundation for subsequent data analysis and model establishment; the Dendrobium protocorm stem cell culture feature set is refined into culture medium component characteristics, culture environment characteristics and culture time characteristics, and the specific content of each feature is clarified; this helps to more deeply understand the growth laws and influencing factors of Dendrobium protocorm stem cells, and provides a more detailed and accurate basis for subsequent correlation analysis and model establishment; this step helps to achieve real-time monitoring and precise regulation of the growth process of Dendrobium protocorm stem cells by comprehensively and systematically collecting data on Dendrobium protocorm stem cell culture feature set and plant growth regulator content, and preprocessing, storing and verifying them.

[0076] S2. performing a correlation analysis on the Dendrobium protocorm stem cell culture characteristic set and the plant growth regulator content to obtain a culture growth characteristic correlation matrix; in the culture growth characteristic correlation matrix, there is a correlation characteristic value between the plant growth regulator content corresponding to each set content range and each Dendrobium protocorm stem cell culture characteristic;

[0077] The method for obtaining a culture growth characteristic correlation matrix includes:

[0078] The Pearson correlation coefficient is used for correlation analysis. It measures the degree of linear correlation between two variables, and its value range is between -1 and 1. When the correlation coefficient is 1, it means that the two variables are completely positively correlated; when the correlation coefficient is -1, it means that the two variables are completely negatively correlated; when the correlation coefficient is 0, it means that there is no linear correlation between the two variables.

[0079] According to the selected correlation analysis method, the correlation coefficient between each feature in the Dendrobium protocorm stem cell culture feature set and the content of plant growth regulators is calculated;

[0080] The calculated correlation coefficients were organized into a matrix, namely the culture growth characteristic correlation matrix; in this matrix, each row represents a culture characteristic of the Dendrobium protocorm stem cells, and each column represents a set content range of the plant growth regulator. The elements in the matrix are the correlation coefficients between the corresponding characteristics and the content ranges;

[0081] The correlation matrix of culture growth characteristics is:

[0082]

[0083] Where m represents the number of culture characteristics, n represents the number of samples, and r mn It represents the correlation coefficient between the mth cultural characteristic and the nth plant growth regulator content.

[0084] In this step, by performing correlation analysis on the culture characteristic set of Dendrobium protocorm stem cells and the content of plant growth regulators, a culture growth characteristic correlation matrix was successfully obtained. This step not only revealed the potential relationship between each culture characteristic and the content of plant growth regulators, but also provided an important data basis for the subsequent growth monitoring of Dendrobium protocorm stem cells. The use of Pearson correlation coefficient for correlation analysis can quantitatively measure the degree of linear correlation between two variables. This analysis method has a clear range of values, so that the strength of the correlation can be intuitively judged by the size of the value. When the correlation coefficient is 1 or -1, it means that there is a complete positive or negative correlation between the two variables, which provides clear guidance for researchers to help them identify which culture characteristics are most important for plants. The influence of growth regulator content is the most significant; the calculated correlation coefficients are organized into a matrix, namely the culture growth characteristic correlation matrix, to make the data more structured and easier to analyze; in this matrix, each row represents a culture characteristic of Dendrobium protocorm stem cells, and each column represents a set content range of plant growth regulator content. The elements in the matrix are the correlation coefficients between the corresponding characteristics and content ranges; this representation method is not only convenient for data storage and retrieval, but also for subsequent data mining and the application of machine learning algorithms; step S2 successfully constructed a culture growth characteristic correlation matrix through correlation analysis, providing strong data support for hormone-free culture growth monitoring of Dendrobium protocorm stem cells; it helps to reveal the intrinsic connection between culture characteristics and plant growth regulator content.

[0085] S3. Traversing the culture growth feature correlation matrix, extracting the culture features of the Dendrobium protocorm stem cells whose correlation feature values ​​are greater than a preset correlation threshold, and summarizing them to obtain a set of key growth culture features of the Dendrobium protocorm stem cells;

[0086] A method for obtaining a set of key growth culture characteristics of Dendrobium protocorm stem cells, including:

[0087] Set a preset correlation threshold; the preset correlation threshold is used to determine which cultural characteristics are significantly correlated with the content of plant growth regulators. The selection of the threshold is usually based on statistical significance tests and practical application requirements. A higher threshold means that we only select those cultural characteristics that are highly correlated with the content of plant growth regulators, while a lower threshold may include more characteristics that are somewhat correlated with the content of plant growth regulators but not significantly correlated.

[0088] During the matrix traversal process, the correlation eigenvalues ​​between each culture characteristic and the plant growth regulator content are checked one by one. If a certain eigenvalue is greater than a preset correlation threshold, it is considered that there is a significant correlation between the culture characteristic and the plant growth regulator content, and thus it is considered to be a key culture characteristic for the growth of Dendrobium protocorm stem cells. The key culture characteristics are extracted and summarized to form a set of key culture characteristics for the growth of Dendrobium protocorm stem cells.

[0089] After extracting key culture characteristics, further verification and adjustment are carried out; this includes checking whether the extracted characteristics are indeed closely related to the growth of Dendrobium protocorm stem cells, and whether other important characteristics may have been missed;

[0090] The key growth culture characteristics of the Dendrobium protocorm stem cells are output and applied to the subsequent growth monitoring and regulation process;

[0091] The factors affecting the setting of the preset relevance threshold include:

[0092] Application Background: Different application scenarios require different levels of accuracy. In a production environment, a higher threshold is preferred to ensure that the selected features have stronger relevance, thereby improving the reliability and stability of the prediction model.

[0093] Data quality: High-quality data provides a clearer correlation signal, so a higher threshold is used in this case; if the data is noisy or has missing values, a lower threshold may be more appropriate to capture as many potential correlations as possible;

[0094] Sample size: A larger sample size can increase the power of the statistical test, allowing even weaker correlations to be detected; in the case of small samples, the threshold needs to be lowered to avoid missing important features;

[0095] Computational complexity: Choosing a higher threshold reduces the number of features that need to be processed, thereby reducing computational cost; however, if the threshold is too high, important information will be lost;

[0096] Economic cost: Considering the cost of experiments and analysis, the economic feasibility of data collection and processing needs to be weighed;

[0097] Environmental factors: External factors such as environmental changes and seasonal differences may also affect the results of correlation analysis and need to be considered when setting thresholds.

[0098] In this step, by setting a preset correlation threshold and traversing the culture growth feature correlation matrix, the culture features of Dendrobium protocorm stem cells that are significantly correlated with the content of plant growth regulators can be accurately extracted; the extracted key culture feature set can be used as input parameters of the prediction model to predict the real-time content of plant growth regulators and cell proliferation rate of Dendrobium protocorm stem cells; since these features are based on historical data and correlation analysis, they can more accurately reflect the growth status of Dendrobium protocorm stem cells, thereby improving the accuracy and reliability of the prediction model; by setting a reasonable preset correlation threshold, this step can balance the relationship between the number of features and prediction performance; a higher threshold can reduce the number of features that need to be processed, reducing the computational cost and time cost; while a lower threshold can capture more potential associations, but may increase computational complexity and data processing difficulty; therefore, when setting the threshold, it is necessary to consider multiple factors such as application background, data quality, sample size, etc. to optimize resource utilization; the extracted key culture feature set provides a scientific basis for hormone-free culture growth monitoring of Dendrobium protocorm stem cells; by real-time monitoring and analysis of these features, problems in the growth process can be discovered and solved in a timely manner, culture conditions can be optimized, and yield and quality can be improved; at the same time, these features can also serve as important indicators for evaluating the effectiveness of different culture strategies, providing a scientific basis for decision-making; this step considers multiple influencing factors when setting the preset correlation threshold, including application background, data quality, sample size, computational complexity, and economic cost; this makes this step have good adaptability and flexibility in different application scenarios; by adjusting the threshold and related parameters, it can adapt to different data characteristics and experimental requirements, and achieve more accurate and efficient feature extraction and monitoring.

[0099] S4, real-time collection of key growth and culture characteristics of Dendrobium protocorm stem cells;

[0100] The method for collecting a set of key growth culture characteristics of Dendrobium protocorm stem cells comprises:

[0101] Determine the reasonable sampling frequency based on experimental requirements and the dynamic characteristics of the growth of Dendrobium protocorm stem cells;

[0102] For rapidly changing parameters, sensors are used for continuous monitoring to obtain more detailed data;

[0103] Select sensors and image acquisition equipment, wherein the sensors include temperature sensors, humidity sensors, light intensity sensors, CO2 sensors, pH sensors, dissolved oxygen sensors and conductivity sensors;

[0104] Using a high-definition camera and microscope, real-time image information of Dendrobium protocorm stem cells is captured; the acquired images are used for subsequent image analysis to monitor cell growth status and morphological changes;

[0105] Regularly extract samples of Dendrobium protocorm stem cells from the culture container for subsequent chemical analysis to determine the content of plant growth regulators;

[0106] Transmit data acquired by sensors and image acquisition devices to the central data processing system in real time;

[0107] Preprocess the collected raw data, including data cleaning, denoising, and calibration, to improve data reliability and accuracy, providing strong support for subsequent analysis and prediction;

[0108] The processed data is stored in a dedicated database for subsequent data analysis and mining; at the same time, an effective data management mechanism is established to ensure the security and integrity of the data.

[0109] In this step, by setting a reasonable sampling frequency and using sensors for continuous monitoring, key data in the process of culturing Dendrobium protocorm stem cells can be obtained in real time; at the same time, the use of high-definition cameras and microscopes makes the acquisition of cell image information more precise, which helps to monitor the growth status and morphological changes of cells; this step selects a variety of sensors and image acquisition equipment, including temperature sensors, humidity sensors, light intensity sensors, CO2 sensors, pH sensors, dissolved oxygen sensors and conductivity sensors, etc., as well as high-definition cameras and microscopes; they can comprehensively cover various environmental parameters and cell characteristics in the process of culturing Dendrobium protocorm stem cells, providing rich data support for subsequent analysis and prediction; the original data After the preprocessing step, the reliability and accuracy of the data are improved; it helps to eliminate outliers and noise in the data, providing a more reliable basis for subsequent analysis and prediction; the processed data is stored in a dedicated database, and an effective data management mechanism is established; this ensures the security and integrity of the data, facilitating subsequent data analysis and mining; at the same time, the use of the database also improves the accessibility and reusability of the data; this step improves the real-time, accuracy, comprehensiveness and reliability of the data by collecting a set of key culture characteristics of the growth of Dendrobium protocorm stem cells in real time, providing strong support for subsequent analysis and prediction, and helping to achieve accurate monitoring and regulation of the hormone-free culture growth of Dendrobium protocorm stem cells.

[0110] S5. Inputting the key growth culture characteristic set into a pre-established plant growth hormone prediction model to obtain the real-time content of plant growth regulators in the Dendrobium protocorm stem cells;

[0111] The method for constructing the plant growth hormone prediction model comprises:

[0112] Collect a large amount of historical data, including the culture characteristics of Dendrobium protocorm stem cells under different culture conditions and the corresponding plant growth regulator content;

[0113] Preprocess the collected data, including data cleaning and data transformation; the preprocessed data will be used for model training and verification;

[0114] Divide the dataset into training and validation sets;

[0115] Selecting a deep learning model as the infrastructure of the plant growth hormone prediction model; the deep learning model includes linear regression, decision tree, random forest, support vector machine and neural network;

[0116] Input the training set into the selected model for model training. During the training process, the model parameters and structure need to be continuously adjusted to minimize the prediction error and improve the generalization ability of the model. At the same time, methods such as cross-validation can be used to evaluate the performance of the model and prevent overfitting.

[0117] After training is completed, the validation set is used to verify and evaluate the model; this is achieved by calculating indicators such as the error between the predicted value and the true value, the accuracy rate, and the recall rate;

[0118] After training and validation, the model is deployed into the real-time monitoring system.

[0119] In this step, the plant growth hormone prediction model constructed can receive the key growth culture feature set of Dendrobium protocorm stem cells in real time and quickly predict the real-time content of plant growth regulators; in the process of model construction, a large amount of historical data is collected and preprocessed to ensure the accuracy and completeness of the data; at the same time, by selecting a deep learning model as the basic architecture of the prediction model and using the training set for model training, the parameters and structure are continuously adjusted to minimize the prediction error, thereby improving the prediction accuracy of the model; in the process of model training, cross-validation and other methods are used to evaluate the performance of the model and prevent overfitting, thereby enhancing the generalization ability of the model; the model can not only accurately predict the content of plant growth regulators in historical data, but also effectively predict new data; the model parts are used to train the model. The method is deployed in a real-time monitoring system to realize the automated monitoring and intelligent prediction of the culture process of Dendrobium protocorm stem cells; this reduces manual intervention and errors, and improves monitoring efficiency and accuracy; by real-time monitoring of the content of plant growth regulators and the rate of cell proliferation, abnormal conditions in the culture process can be discovered in time, and the culture conditions can be adjusted accordingly to optimize the culture process of Dendrobium protocorm stem cells; the implementation of this method helps scientific researchers to obtain experimental data more quickly, reduce the number of experiments and time costs, and thus improve research efficiency; this step and the plant growth hormone prediction model construction method described therein provide strong technical support for hormone-free culture growth monitoring of Dendrobium protocorm stem cells, and realize multiple beneficial effects such as real-time monitoring, accurate prediction, automation and intelligence, and optimization of culture conditions.

[0120] S6. collecting microscopic images of the protocorm tissue of Dendrobium officinale at two end points of a preset time segment;

[0121] The method for collecting microscopic images of Dendrobium protocorm tissue at two end points of a preset time period comprises:

[0122] Select a microscope with high resolution and high magnification to clearly observe the microstructure and morphology of the protocorm stem cells of Dendrobium officinale;

[0123] Ensure that the microscope is equipped with a highly sensitive image sensor to capture high-quality image data;

[0124] Configure an appropriate lighting system to provide stable and uniform lighting conditions to ensure consistent and accurate image acquisition;

[0125] According to the size and morphology of the protocorm stem cells of Dendrobium officinale, select the appropriate magnification for image acquisition;

[0126] Adjust the exposure time to ensure that the image is neither overexposed nor too dark, and that the cell details are clearly visible.

[0127] Set a high resolution to capture more details, but also consider image processing efficiency and storage space limitations;

[0128] Ensure that the microscope system is correctly focused to obtain a clear image;

[0129] At the two end points of the preset time period, appropriate samples are taken from the protocorm culture of Dendrobium officinale and processed as necessary to highlight the cell structure;

[0130] Place the processed sample on the microscope stage and adjust the position and angle of the sample to ensure the best image acquisition effect;

[0131] Start the image acquisition function of the microscope system and capture the microscopic image of the Dendrobium protocorm tissue according to the preset parameter settings;

[0132] The collected images were saved in a computer for subsequent proliferation trend analysis;

[0133] After the image acquisition is completed, the image quality is checked to ensure that the image is clear, without blur, noise and other defects.

[0134] In this step, by selecting a high-resolution, high-magnification microscope and a high-sensitivity image sensor, the microstructure and morphology of the protocorm stem cells of Dendrobium can be clearly captured, providing high-quality image data for subsequent proliferation trend analysis; configuring a suitable lighting system to provide stable and uniform lighting conditions ensures the consistency and accuracy of image acquisition and avoids image quality differences caused by uneven lighting; according to the size and morphology of the protocorm stem cells of Dendrobium, the appropriate magnification, exposure time and resolution are selected to capture sufficient detailed information while taking into account the efficiency and storage of image processing. The spatial limitation is eliminated to achieve the optimization of image acquisition; after the image acquisition is completed, a quality check is carried out to ensure that the image is clear, without defects such as blur and noise, which provides a reliable basis for subsequent image analysis; by collecting microscopic images of Dendrobium protocorm tissue at the two end points of a preset time segment, it provides important data support for the subsequent proliferation trend analysis, which helps to understand the growth and proliferation rate of Dendrobium protocorm stem cells; this method can realize real-time monitoring of the growth process of Dendrobium protocorm stem cells, which helps to timely discover growth abnormalities or problems, and carry out corresponding regulation and optimization to improve culture efficiency and cell quality.

[0135] S7. Analyze the proliferation trend of the two groups of collected microscopic images of Dendrobium protocorm tissues to obtain the cell proliferation rate of Dendrobium protocorm stem cells;

[0136] The method for obtaining the cell proliferation rate of Dendrobium protocorm stem cells comprises:

[0137] Enhance the collected microscopic images to improve the contrast and clarity of the images, facilitating subsequent cell identification and segmentation;

[0138] A filtering algorithm is used to remove noise from the image to reduce interference with cell identification;

[0139] Perform color correction on the image to ensure the accuracy of the image's color information, which is helpful for subsequent cell feature extraction;

[0140] Using edge detection algorithms, the outlines of the protocorm stem cells of Dendrobium candidum were identified;

[0141] Based on the cell outline, an image segmentation algorithm is used to segment the cells from the image;

[0142] Extracting cell morphological characteristics, including area, perimeter, circularity, and aspect ratio, can reflect the proliferation status of cells;

[0143] Extracting cell texture features helps distinguish cells in different proliferation states;

[0144] Count the segmented cells to obtain the number of cells at each time point;

[0145] Calculate the cell proliferation rate based on the number of cells at the two end points of the preset time segment;

[0146] The cell number and proliferation rate at two time points are plotted into a trend graph to visually display the cell proliferation trend;

[0147] Statistical analysis was performed on the cell number and proliferation rate to obtain the cell proliferation rate of Dendrobium protocorm stem cells in order to evaluate the cell proliferation stability and consistency.

[0148] In this step, by performing enhancement processing, filtering denoising and color correction on the microscopic image, the contrast and clarity of the image are significantly improved, noise interference is reduced, the accuracy of the image color information is ensured, and a high-quality image foundation is provided for subsequent cell identification and segmentation; the edge detection algorithm and image segmentation algorithm can accurately identify the outline of the Dendrobium protocorm stem cells and segment them from the image; by extracting the morphological and texture characteristics of the cells, the proliferation state of the cells can be fully reflected, which helps to distinguish cells in different proliferation states; reliable data support is provided for subsequent cell proliferation rate calculation and proliferation trend analysis; based on the cell number at the two end points of the preset time segment, the cell proliferation rate is accurately calculated, which can intuitively display the cell proliferation trend; plotting the cell number and proliferation rate at the two time points into a trend graph can intuitively display the cell proliferation trend, which helps researchers better understand the growth status of the Dendrobium protocorm stem cells; statistical analysis of the cell number and proliferation rate can obtain more in-depth cell proliferation information, providing strong support for evaluating cell proliferation performance and optimizing culture conditions.

[0149] S8. performing weighted calculation on the cell proliferation rate of the Dendrobium protocorm stem cells and the real-time content of the plant growth regulator to obtain the culture growth index of the Dendrobium protocorm stem cells;

[0150] The method for obtaining the culture growth index of Dendrobium protocorm stem cells comprises:

[0151] According to the importance of cell proliferation rate and plant growth regulator content in the growth process of Dendrobium protocorm stem cells, corresponding weights were assigned; cell proliferation rate reflects the growth vitality and proliferation ability of cells, while the plant growth regulator content directly affects the growth direction and speed of cells;

[0152] Using historical data, we analyzed the correlation between cell proliferation rate and plant growth regulator content, as well as their relationship with the growth status of Dendrobium protocorm stem cells, to determine a more reasonable weight distribution;

[0153] The cell proliferation rate and the plant growth regulator content are multiplied by their corresponding weights, and then the results are added together to obtain the culture growth index of the Dendrobium protocorm stem cells;

[0154] Before calculation, the cell proliferation rate and plant growth regulator content were standardized to eliminate the influence of dimension difference and numerical range difference on the results;

[0155] The calculated culture growth index can be used to determine the hormone-free growth status of Dendrobium protocorm stem cells. The higher the index, the better the cell growth status and the more ideal the proliferation capacity and growth direction regulation.

[0156] The formula for calculating the culture growth index of Dendrobium protocorm stem cells is:

[0157] I=w V V+w C ·C

[0158] Wherein, I represents the culture growth index of the protocorm stem cells of Dendrobium officinale, V represents the cell proliferation rate, C represents the real-time content of the plant growth regulator, and w V represents the weight of cell proliferation rate, w C Indicates the weight of the real-time content of plant growth regulators.

[0159] In this step, the culture growth index comprehensively considers two key indicators, cell proliferation rate and plant growth regulator content, and can comprehensively and objectively reflect the growth status of Dendrobium protocorm stem cells; it not only takes into account the growth vitality and proliferation ability of the cells, but also takes into account the regulation of the growth direction, providing scientific researchers with a more accurate growth status assessment tool; by utilizing historical data and analyzing correlations, a more reasonable weight distribution is determined; this data-based weight distribution method can reduce the influence of subjective factors and improve the accuracy and reliability of the evaluation results; at the same time, the adjustability of weight distribution also enables this method to adapt to the needs of different culture conditions and growth stages; before calculating the culture growth index, the cell proliferation rate and plant growth regulator content are standardized to eliminate the influence of dimensional differences and numerical range differences on the results; the accuracy and comparability of the calculation results are improved, so that data from different batches and different time points can be more accurately compared. Reflect the growth status of cells; according to the calculated culture growth index, the hormone-free culture growth status of Dendrobium protocorm stem cells can be judged, and growth regulation strategies can be formulated accordingly; when the index is low, it indicates that the cell growth status is poor or the proliferation ability is limited, and the culture conditions need to be adjusted to improve the growth status; when the index is high, it indicates that the cell growth status is good, and the culture conditions can be maintained or further optimized; by real-time monitoring and evaluation of the growth status of Dendrobium protocorm stem cells, problems in the growth process can be discovered and solved in a timely manner, and the continuous development and improvement of Dendrobium protocorm stem cell culture technology can be promoted; it is helpful to improve the yield and quality of Dendrobium protocorm stem cells, and provide strong support for the sustainable utilization of Dendrobium medicinal materials; step S8 comprehensively considers the two key indicators of cell proliferation rate and plant growth regulator content, and based on data analysis and standardization processing, obtains a culture growth index that can comprehensively and objectively reflect the growth status of Dendrobium protocorm stem cells.

[0160] S9. comparing the culture growth index with a preset standard index to determine whether the hormone-free culture growth of the Dendrobium protocorm stem cells is normal;

[0161] The factors affecting the setting of the preset standard index include:

[0162] Historical culture data analysis: Statistical analysis of historical data was performed to determine the range of the culture growth index of Dendrobium protocorm stem cells under normal growth conditions; this helps to set a reasonable standard index to reflect normal growth conditions;

[0163] Culture conditions: The composition of the culture medium significantly affects the growth of Dendrobium protocorm stem cells. Therefore, when setting the standard index, the ratio and concentration of various nutrients in the culture medium need to be considered. The temperature, light, humidity and other conditions of the culture environment also affect cell growth. These environmental factors should be used as a reference for setting the standard index.

[0164] Cell type: Different types of Dendrobium protocorm stem cells may have different growth characteristics and requirements; therefore, the differences in cell types need to be considered when setting the standard index;

[0165] Cell growth stage: The growth rate and regulator requirements of cells in different growth stages will also vary; therefore, the setting of the standard index should take into account the growth stage of the cells;

[0166] Limitations of monitoring technology: Different monitoring technologies have different accuracy and limitations. When setting the standard index, the accuracy and reliability of the monitoring technology used need to be considered to ensure the accuracy and practicality of the standard index;

[0167] Technological updates: With the continuous development of monitoring technology, new methods and technologies may improve the accuracy and efficiency of monitoring; therefore, when setting standard indices, it is necessary to pay attention to technological updates and development trends.

[0168] In this step, by comparing the culture growth index calculated in real time with the preset standard index, it is possible to accurately determine whether the growth state of the Dendrobium protocorm stem cells under hormone-free culture conditions is normal; this helps to promptly detect growth abnormalities and take corresponding intervention measures to ensure the healthy growth of cells; this step uses a pre-constructed plant growth hormone prediction model and proliferation trend analysis technology to achieve real-time monitoring of the growth process of Dendrobium protocorm stem cells; compared with traditional chemical detection and manual observation methods, it can greatly shorten the monitoring cycle and improve monitoring efficiency; by setting a reasonable preset standard index and combining it with real-time monitoring data, the culture conditions of Dendrobium protocorm stem cells can be finely regulated; it helps to optimize the culture medium composition, culture environment and other conditions, increase the growth rate and yield of cells, and at the same time ensure the cell quality and appearance; when setting the preset standard index, factors such as cell type and growth stage are fully considered, so that the standard index is more in line with the growth characteristics of Dendrobium protocorm stem cells; it helps to ensure the accuracy and reliability of the monitoring results, and provide strong support for subsequent cell culture and research; with the continuous development of monitoring technology, this step also pays attention to the update and development trend of technology; by continuously introducing new technologies and methods, the accuracy and efficiency of monitoring can be further improved, and the continuous innovation and development of Dendrobium protocorm stem cell culture technology can be promoted; this step S9 realizes the accurate judgment and optimal regulation of the hormone-free culture growth state of Dendrobium protocorm stem cells by setting a reasonable preset standard index and combining it with real-time monitoring technology, which provides strong support for cell culture and research, and also promotes the innovation and development of related technologies.

[0169] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the technical principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for monitoring the growth of Dendrobium protocorm stem cells without hormone culture, characterized in that: The method comprises: Collecting a set of culture characteristics of Dendrobium protocorm stem cells and a plant growth regulator content during the historical culture process of Dendrobium protocorm stem cells, wherein the set of culture characteristics of Dendrobium protocorm stem cells and the plant growth regulator content correspond one to one; Correlation analysis was performed on the culture characteristic set of Dendrobium protocorm stem cells and the content of plant growth regulators to obtain a culture growth characteristic correlation matrix. In the culture growth characteristic correlation matrix, there were correlation characteristic values ​​between the plant growth regulator content corresponding to each set content range and each Dendrobium protocorm stem cell culture characteristic. Traversing the culture growth feature correlation matrix, extracting the Dendrobium protocorm stem cell culture features whose correlation feature values ​​are greater than a preset correlation threshold, and summarizing them to obtain a set of key growth culture features of the Dendrobium protocorm stem cells; Real-time collection of key culture characteristics of the growth of protocorm stem cells of Dendrobium officinale cultured without hormones; Inputting the key growth culture characteristic set into a pre-built plant growth hormone prediction model to obtain the real-time content of plant growth regulators in the Dendrobium protocorm stem cells; Collect microscopic images of the protocorm tissue of Dendrobium officinale at both ends of a preset time period; The proliferation trend of the two groups of collected microscopic images of Dendrobium protocorm tissues was analyzed to obtain the cell proliferation rate of Dendrobium protocorm stem cells; The cell proliferation rate and real-time content of plant growth regulators of Dendrobium protocorm stem cells were weighted to obtain the culture growth index of Dendrobium protocorm stem cells. Compare the culture growth index with the preset standard index to determine whether the hormone-free culture growth of the Dendrobium protocorm stem cells is normal; The formula for calculating the culture growth index of Dendrobium protocorm stem cells is: I=w V ·V+w C ·C Wherein, I represents the culture growth index of the protocorm stem cells of Dendrobium officinale, V represents the cell proliferation rate, C represents the real-time content of the plant growth regulator, and w V represents the weight of cell proliferation rate, w C Indicates the weight of the real-time content of plant growth regulators.

2. The method for monitoring the growth of Dendrobium protocorm stem cells without hormone culture according to claim 1, wherein: The Dendrobium protocorm stem cell culture characteristic set includes culture medium component characteristics, culture environment characteristics and culture time characteristics; culture medium component characteristics include basal culture medium type, sugar type and concentration, vitamin type and concentration, trace element type and concentration, and initial addition amount of plant growth regulator; culture environment characteristics include temperature, light intensity, light cycle, humidity, CO2 concentration, pH value and oxygen supply; culture time characteristics include time points of different growth stages, overall culture time and differences between different batches.

3. The method for monitoring the growth of Dendrobium protocorm stem cells without hormone culture according to claim 1, wherein: The correlation matrix of culture growth characteristics is: Where m represents the number of culture characteristics, n represents the number of samples, and r mn represents the correlation coefficient between the mth cultural characteristic and the nth plant growth regulator content.

4. The method for monitoring the growth of Dendrobium protocorm stem cells without hormone culture according to claim 1, wherein: The factors affecting the setting of the preset correlation threshold include application background, data quality, sample size, computational complexity, economic cost and environmental factors.

5. The method for monitoring the growth of Dendrobium protocorm stem cells without hormone culture according to claim 1, wherein: The method for constructing the plant growth hormone prediction model comprises: Collect historical data, including the culture characteristics of Dendrobium protocorm stem cells under different culture conditions and the corresponding plant growth regulator content; Preprocess the collected data, including data cleaning and data transformation; Divide the dataset into training and validation sets; Selecting a deep learning model as the infrastructure of the plant growth hormone prediction model; the deep learning model includes linear regression, decision tree, random forest, support vector machine and neural network; Input the training set into the selected model to perform model training; After training is completed, the model is verified and evaluated using the validation set; After training and validation, the model is deployed into the real-time monitoring system.

6. The method for monitoring the growth of Dendrobium protocorm stem cells without hormone culture according to claim 1, wherein: The method for collecting microscopic images of Dendrobium protocorm tissue at two end points of a preset time period comprises: Select a high-resolution, high-magnification microscope for image acquisition; Configure the lighting system to provide stable and uniform lighting conditions; According to the size and morphology of the protocorm stem cells of Dendrobium officinale, the magnification was set for image acquisition; Adjusting the exposure time can clearly show the details of the cells; Set the microscope to focus correctly to obtain a clear image; At the two end points of the predetermined time period, samples were taken from the protocorm culture of Dendrobium officinale; Place the processed sample on the microscope stage, adjust the position and angle of the sample, start the microscope, and capture the microscopic image of the Dendrobium protocorm tissue according to the preset parameter settings; Save the acquired images to the computer.

7. The method for monitoring the growth of Dendrobium protocorm stem cells without hormone culture according to claim 1, wherein: The method for obtaining the cell proliferation rate of Dendrobium protocorm stem cells comprises: Enhance the collected microscopic images to improve the contrast and clarity of the images; Remove noise from images to reduce interference with cell identification; Perform color correction on images; Using edge detection algorithms, the outlines of the protocorm stem cells of Dendrobium candidum were identified; Segment cells from the image based on cell outlines; Extract cell morphological features, including area, perimeter, circularity, and aspect ratio; Extract cell texture features to distinguish cells in different proliferation states; Count the segmented cells to obtain the number of cells at each time point; Calculate the cell proliferation rate based on the number of cells at the two end points of the preset time segment; The cell number and proliferation rate at two time points are plotted into a trend graph to visually display the cell proliferation trend; The cell number and proliferation rate were statistically analyzed to obtain the cell proliferation rate of Dendrobium protocorm stem cells.

8. The method for monitoring the growth of Dendrobium protocorm stem cells without hormone culture according to claim 1, wherein: The factors affecting the setting of the preset standard index include historical culture data analysis, culture conditions, cell type and cell growth stage.

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