A method for controlling the clonal proliferation of Huoshan Dendrobium stem cells

Abstract

This invention relates to the technical field of plant cultivation, and in particular to a method for controlling the clonal proliferation of Dendrobium huoshanense stem cells, which improves proliferation efficiency and quality and enhances resource conservation. The method includes: based on the design requirements for the clonal proliferation of Dendrobium huoshanense stem cells, obtaining a target proliferation feature quantification set for each preset culture stage; at the end of each preset culture stage, collecting culture environment data of Dendrobium huoshanense stem cells to obtain a culture environment data set and a real-time proliferation feature quantification set; calculating the deviation distance between the real-time proliferation feature quantification set and the corresponding target proliferation feature quantification set to obtain a real-time proliferation feature deviation vector; and inputting the real-time proliferation feature deviation vector and the corresponding culture environment data set into a pre-constructed culture environment regulation and analysis model to obtain a culture environment regulation strategy for the next preset culture stage.

Description

Technical Field

[0001] This invention relates to the technical field of plant cultivation, and in particular to a method for controlling the clonal proliferation of Dendrobium huoshanense stem cells. Background Technology

[0002] Dendrobium huoshanense, a rare medicinal plant, possesses extremely high medicinal and economic value. Its stem cell cloning and proliferation technology is of great significance for the large-scale propagation and resource conservation of Dendrobium huoshanense.

[0003] In existing clonal proliferation processes of Dendrobium huoshanense stem cells, fixed culture modes and parameters are mostly employed. Typically, a predetermined culture protocol is followed, with uniform culture medium composition, culture temperature, and light conditions, which remain largely unchanged from the start to the end of the culture. However, the environmental requirements of Dendrobium huoshanense stem cells vary significantly at different culture stages. In the initial culture stage, stem cells may require a specific ratio of nutrients to initiate division and proliferation; while in subsequent differentiation and maturation stages, the requirements for light intensity, temperature, and other conditions change.

[0004] Existing fixed culture methods cannot precisely meet the unique needs of Dendrobium officinale stem cells at each culture stage. On the one hand, this can easily lead to insufficient or excessive nutrient supply at certain stages, affecting the normal proliferation and development of stem cells, resulting in low proliferation efficiency and difficulty in achieving the expected proliferation quantity. On the other hand, an unsuitable culture environment may also induce abnormal differentiation of stem cells, reducing their quality and affecting their subsequent medicinal value and application effects. Furthermore, due to the lack of real-time monitoring and dynamic adjustment mechanisms during the culture process, it is difficult to detect and correct mismatches between the culture environment and stem cell growth requirements in a timely manner, further increasing the risk of culture failure. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides a method for controlling the clonal proliferation of Dendrobium huoshanense stem cells, which improves proliferation efficiency and quality and enhances resource conservation.

[0006] In a first aspect, the present invention provides a method for controlling the clonal proliferation of Dendrobium huoshanense stem cells, the method comprising:

[0007] Based on the design requirements for the clonal proliferation of Dendrobium huoshanense stem cells, a quantitative set of target proliferation characteristics for each preset culture stage was obtained.

[0008] At the end of each preset culture stage, the culture environment data of Dendrobium huoshanense stem cells are collected to obtain a culture environment data set and a real-time proliferation characteristic quantification set.

[0009] Calculate the deviation distance between the real-time proliferation feature quantization set and the corresponding target proliferation feature quantization set to obtain the real-time proliferation feature deviation vector;

[0010] The real-time proliferation feature deviation vector and the corresponding culture environment data set are input into a pre-constructed culture environment regulation and analysis model to obtain a culture environment regulation strategy for the next preset culture stage.

[0011] The culture environment for the clonal proliferation of Dendrobium huoshanense stem cells was regulated based on the aforementioned culture environment regulation strategy.

[0012] Furthermore, the formula for calculating the deviation distance between the real-time proliferation feature quantization set and the corresponding target proliferation feature quantization set is as follows:

[0013]

[0014] Where d represents the deviation distance, x i y represents the value of the i-th feature in the real-time proliferation feature quantization set. i represents the value of the i-th feature in the target proliferation feature quantization set, and n represents the total number of dimensions of the features.

[0015] Furthermore, the target proliferation feature quantification set includes the target stem cell quantity, target stem cell quality, target proliferation rate, target differentiation inhibition rate, and target Dendrobium polysaccharide content.

[0016] Furthermore, the preset culture stages include an initiation period, a proliferation period, and a differentiation period.

[0017] Furthermore, the culture environment data set includes culture medium component concentration, light intensity, light exposure time, culture temperature, culture humidity, and aeration rate.

[0018] Furthermore, the real-time proliferation feature quantification set includes real-time stem cell quantity, real-time stem cell quality, real-time proliferation rate, real-time differentiation inhibition rate, and real-time Dendrobium polysaccharide content.

[0019] Furthermore, the method for constructing the culture environment regulation and analysis model includes:

[0020] Collect growth data for each pre-set culture stage;

[0021] The collected data is cleaned to remove records with too many errors, duplicates, and missing values; and different types of data are standardized.

[0022] The model is constructed using machine learning algorithms, including neural network algorithms, convolutional neural network algorithms, and decision tree algorithms.

[0023] The preprocessed data is divided into training set, validation set and test set. The selected model is trained using the training set data, and the training process is monitored using the validation set data to prevent the model from overfitting.

[0024] The trained model is evaluated using test set data to measure its performance; the model is then optimized based on the evaluation results.

[0025] Deploy the trained and optimized model to real-world application scenarios.

[0026] On the other hand, this application also provides a Dendrobium huoshanense stem cell clonal proliferation control system, the system comprising:

[0027] The target proliferation feature quantification set acquisition module, based on the design requirements for Dendrobium huoshanense stem cell clonal proliferation, acquires the target proliferation feature quantification set for Dendrobium huoshanense stem cell clonal proliferation for each preset culture stage;

[0028] The data acquisition module collects culture environment data of Dendrobium officinale stem cells at the end time of each preset culture stage, and obtains a culture environment data set and a real-time proliferation characteristic quantification set.

[0029] The deviation calculation module calculates the deviation distance between the real-time proliferation feature quantization set and the corresponding target proliferation feature quantization set to obtain the real-time proliferation feature deviation vector.

[0030] The culture environment regulation and analysis module inputs the real-time proliferation feature deviation vector and the corresponding culture environment data set into a pre-constructed culture environment regulation and analysis model to obtain a culture environment regulation strategy for the next preset culture stage.

[0031] The environmental regulation execution module regulates the culture environment for the clonal proliferation of Dendrobium huoshanense stem cells based on the aforementioned culture environment regulation strategy.

[0032] Thirdly, this application provides an electronic device including a bus, a transceiver, a memory, a processor, and a computer program stored in the memory and executable on the processor. The transceiver, the memory, and the processor are connected via the bus, and the computer program, when executed by the processor, implements the steps in the method described above.

[0033] Fourthly, this application also provides a computer-readable storage medium having a computer program stored thereon, which, when executed by a processor, implements the steps in the method described above.

[0034] Compared with existing technologies, the beneficial effects of this invention are as follows: This method sets specific target proliferation characteristic quantification sets for different culture stages of Dendrobium officinale stem cells; making the regulation more precise and enabling targeted adjustments according to the needs of different stages, thereby meeting the unique needs of stem cells at each growth stage; by collecting culture environment data and real-time proliferation characteristic data at the end time of each preset culture stage, this method can monitor the growth status of stem cells in real time; it helps to promptly detect deviations between the growth status and the target status, providing an accurate basis for subsequent regulation; using a pre-constructed culture environment regulation and analysis model, it can determine the growth status of stem cells based on the real-time proliferation characteristic deviation vector and the culture environment... This method uses environmental data sets to intelligently generate environmental control strategies for the next culture stage. This not only improves the efficiency and accuracy of control but also reduces the subjectivity and uncertainty of human intervention. Because this method can precisely meet the needs of stem cells at different growth stages, it helps improve the proliferation efficiency and quality of Dendrobium huoshanense stem cells. By optimizing the culture environment, it can promote normal stem cell proliferation and development, reduce abnormal differentiation, and thus improve the medicinal value and application effects of stem cells. This method features real-time data monitoring and intelligent control, enabling timely detection and correction of mismatches between the culture environment and stem cell growth needs. This helps reduce the risk of culture failure and improve the overall success rate of cultivation.

[0035] This method does not rely solely on fixed culture modes and parameters, but rather adjusts the culture environment in real time according to the specific needs of *Dendrobium huoshanense* stem cells at different culture stages. This ensures that stem cells receive the most suitable culture conditions at each growth stage, thus achieving precise regulation. By collecting culture environment data and stem cell proliferation characteristic data in real time and calculating the deviation distance, the method can quickly identify the difference between the current culture environment and the target state, and formulate regulatory strategies accordingly, greatly improving the accuracy and efficiency of regulation. Because this method can precisely meet the needs of stem cells at different growth stages, it significantly improves the proliferation efficiency of *Dendrobium huoshanense* stem cells. Under suitable culture conditions, stem cells can grow faster and more efficiently. The system achieves stable proliferation, reaching the desired proliferation quantity. By controlling key indicators such as differentiation inhibition rate, it can reduce abnormal differentiation of stem cells, maintaining their original characteristics and medicinal value. Through real-time monitoring and dynamic adjustment of the culture environment, this method significantly reduces the risk of culture failure. If the system detects a mismatch between the culture environment and the stem cell growth requirements, it immediately takes corrective measures, thus avoiding potential growth obstacles and losses. By optimizing the culture environment and improving proliferation efficiency, the system can produce more high-quality Dendrobium huoshanense stem cells. By improving the efficiency and stability of stem cell clonal proliferation, it can reduce dependence on wild resources, thereby protecting the ecological environment and biodiversity.

[0036] In summary, the above-mentioned methods for controlling the clonal proliferation of Dendrobium huoshanense stem cells can provide strong support for the large-scale breeding and resource protection of Dendrobium huoshanense; through precise regulation and intelligent management, the proliferation efficiency and quality can be improved, thereby enhancing the effectiveness of resource protection. Attached Figure Description

[0037] Figure 1 This is a flowchart of the present invention;

[0038] Figure 2 This is a flowchart illustrating the construction method of an environmental regulation and analysis model.

[0039] Figure 3 This is a structural diagram of the Dendrobium huoshanense stem cell cloning and proliferation control system. Detailed Implementation

[0040] As will be apparent to those skilled in the art from the description of this application, this application can be implemented as a method, apparatus, electronic device, and computer-readable storage medium. Therefore, this application can be specifically implemented in the following forms: entirely hardware, entirely software (including firmware, resident software, microcode, etc.), or a combination of hardware and software. Furthermore, in some embodiments, this application can also be implemented as a computer program product contained in one or more computer-readable storage media, which includes computer program code.

[0041] The aforementioned computer-readable storage medium may be any combination of one or more computer-readable storage media. Computer-readable storage media include: electrical, magnetic, optical, electromagnetic, infrared, or semiconductor systems, apparatuses, or devices, or any combination thereof. More specific examples of computer-readable storage media include: portable computer disks, hard disks, random access memory, read-only memory, erasable programmable read-only memory, flash memory, optical fiber, optical disc read-only memory, optical storage devices, magnetic storage devices, or any combination thereof. In this application, a computer-readable storage medium may be any tangible medium containing or storing a program that can be used by or in conjunction with an instruction execution system, apparatus, or device.

[0042] The acquisition, storage, use, and processing of data in this application all comply with relevant national laws and regulations.

[0043] This application describes the provided methods, apparatus, and electronic devices using flowcharts and / or block diagrams.

[0044] It should be understood that each block of a flowchart and / or block diagram, as well as combinations of blocks in a flowchart and / or block diagram, can be implemented by computer-readable program instructions. These computer-readable program instructions can be provided to a processor of a general-purpose computer, a special-purpose computer, or other programmable data processing apparatus to produce a machine that, when executed by a computer or other programmable data processing apparatus, creates means for implementing the functions / operations specified in the blocks of the flowchart and / or block diagram.

[0045] These computer-readable program instructions may also be stored in a computer-readable storage medium that enables a computer or other programmable data processing device to function in a particular manner. In this way, the instructions stored in the computer-readable storage medium produce an instruction apparatus product that includes the functions / operations specified in the blocks of a flowchart and / or block diagram.

[0046] Computer-readable program instructions may also be loaded onto a computer, other programmable data processing apparatus or other device to cause a series of operational steps to be performed on the computer, other programmable data processing apparatus or other device to produce a computer-implemented process, such that the instructions that execute on the computer or other programmable data processing apparatus provide a process for implementing the functions / operations specified in the blocks of the flowchart and / or block diagram.

[0047] This application will now be described with reference to the accompanying drawings.

[0048] Example 1: As Figures 1 to 2 As shown, the present invention provides a method for controlling the clonal proliferation of Dendrobium huoshanense stem cells, which specifically includes the following steps:

[0049] A method for controlling the clonal proliferation of Dendrobium huoshanense stem cells, characterized in that the method comprises:

[0050] S1. Based on the design requirements for the clonal proliferation of Dendrobium huoshanense stem cells, a quantitative set of target proliferation characteristics of Dendrobium huoshanense stem cell clonal proliferation is obtained for each preset culture stage;

[0051] The target proliferation feature quantification set includes the number of target stem cells, the quality of target stem cells, the target proliferation rate, the target differentiation inhibition rate, and the content of target Dendrobium polysaccharides;

[0052] The target number of stem cells is set based on the expected production scale and efficiency, indicating the ideal number of cells at the end of each stage.

[0053] The target stem cell quality is used to measure indicators such as stem cell health status and viability, ensuring a high-quality cell population.

[0054] The target proliferation rate is used to define the desired rate of cell division at each stage in order to optimize resource use and time management;

[0055] The target differentiation inhibition rate is during the proliferation phase, and it is necessary to control the occurrence of differentiation in order to focus on the increase in cell number.

[0056] As an important active ingredient of Dendrobium huoshanense, monitoring and optimizing the content of target Dendrobium polysaccharides is crucial to ensuring the efficacy of the final product.

[0057] The pre-defined culture stages include the initiation stage, the proliferation stage, and the differentiation stage;

[0058] During the initiation phase, based on the fundamental theory of plant cell division initiation, stem cells require a specific ratio of nutrients to stimulate their division activity. By studying the nutrient uptake patterns of Dendrobium huoshanense during its initial growth in natural environments, and combining this with commonly used nutrient addition methods in plant tissue culture, the target stem cell quantity, quality, and other indicators required during the initiation phase were determined. For example, by referring to the requirements of other orchid plants for macroelements such as nitrogen, phosphorus, and potassium, as well as microelements such as iron and zinc, during the initiation phase of tissue culture, and considering the characteristics of Dendrobium huoshanense itself, a suitable combination of nutrients for initiating stem cell division was determined. This allowed for setting a reasonable range for the target proliferation rate and the initial number of stem cells, providing a starting standard for subsequent culture.

[0059] Entering the proliferation phase, based on the principle of rapid plant cell proliferation, the cells' nutritional needs at this stage are more focused on meeting the energy supply and material synthesis required for massive cell division. Referring to research findings on the relationship between cell density, nutrient consumption, and proliferation rate in plant cell suspension culture, the target number of stem cells for Dendrobium huoshanense stem cells at this stage is determined, as well as the target stem cell quality requirements for maintaining normal cell physiological function and high-quality cell division. At the same time, based on the relationship between cell division cycle and accumulation of metabolites, a target differentiation inhibition rate is set to prevent premature differentiation of stem cells during the proliferation phase and ensure an efficient proliferation process.

[0060] During the differentiation period, based on the plant cell differentiation regulation mechanism, environmental factors such as light and temperature play a key role in the direction of cell differentiation. Combining research on plant photomorphogenesis and temperature-responsive gene expression, the appropriate polysaccharide content of the target Dendrobium during the differentiation period was determined, as polysaccharide content is closely related to the medicinal value of Dendrobium huoshanense. At the same time, based on the changes in cell morphology and function during differentiation, the reasonable range of the number and quality of target stem cells at this stage was determined to ensure that the differentiated cells have good physiological characteristics and medicinal potential.

[0061] In this step, by deeply studying the biological characteristics and growth requirements of Dendrobium huoshanense stem cells and combining them with the fundamental theories of plant cell culture, a scientific and precise set of target proliferation characteristics was established for different culture stages. These quantitative indicators can comprehensively and accurately reflect the growth status and physiological needs of stem cells at each stage, providing a reliable basis for subsequent culture environment control and real-time monitoring. By setting target proliferation rates and differentiation inhibition rates, this step helps optimize resource allocation and time management. During the proliferation phase, by controlling the supply of nutrients and the rate of cell division, it is possible to ensure that stem cells proliferate efficiently while avoiding resource waste and prolonged growth cycles. During the differentiation phase, by precisely controlling environmental factors such as light and temperature, stem cells can be guided to differentiate in a specific direction, improving differentiation efficiency and the accumulation of medicinal components. This step, by setting target stem cell quantity and quality, and... Targeting the polysaccharide content of Dendrobium officinale helps improve culture efficiency and product quality. Real-time monitoring and control of the culture environment ensures that stem cells reach predetermined growth targets and medicinal component content at each stage, thereby enhancing the efficacy and market competitiveness of the final product. This step facilitates the large-scale propagation of Dendrobium officinale stem cells. Precise control of culture conditions and growth parameters enables the mass production of high-quality stem cells to meet market demand. Simultaneously, large-scale propagation helps reduce dependence on wild resources, protecting the ecological environment and biodiversity. This step forms the basis for the clonal proliferation control process of Dendrobium officinale stem cells. Setting a scientific and precise target proliferation characteristic quantification set provides a clear direction and basis for subsequent culture environment control, real-time monitoring, and dynamic adjustment. This helps ensure the smooth progress of the entire culture process and the high-quality output of the final product.

[0062] S2. At the end of each preset culture stage, collect culture environment data of Dendrobium officinale stem cells to obtain a culture environment data set and a real-time proliferation characteristic quantification set.

[0063] The culture environment data set includes culture medium component concentration, light intensity, light duration, culture temperature, culture humidity, and aeration rate;

[0064] Culture medium component concentration: High-performance liquid chromatography (HPLC) and atomic absorption spectrometry (AAS) are used to detect the content of various nutrients in the culture medium. For example, HPLC can accurately determine the concentration of organic nutrients such as amino acids and vitamins in the culture medium; AAS is used to analyze the content of macro- and micro-elements. In plant tissue culture, the concentration of these nutrients directly affects cell growth and metabolism. Accurate data collection can help understand the consumption of nutrients in the culture medium in a timely manner, providing a basis for subsequent adjustments.

[0065] Light intensity and duration: A photometer was used to measure light intensity. The photometer can accurately sense the intensity of light at different wavelengths, ensuring that the acquired light intensity data accurately reflects the actual culture environment. At the same time, a timer was used to record the duration of light exposure. In the study of plant photomorphogenesis and photosynthesis, light intensity and duration have a significant impact on plant growth and development. For Dendrobium huoshanense stem cell culture, appropriate light conditions at different stages can promote normal cell division, differentiation, and accumulation of active ingredients.

[0066] Culture temperature and humidity: Using high-precision temperature and humidity sensors, temperature and humidity affect physiological processes such as the activity of intracellular enzymes and water metabolism during plant growth. A suitable temperature and humidity environment is the guarantee for the normal proliferation and differentiation of Dendrobium huoshanense stem cells. By collecting data in real time, abnormal fluctuations in temperature and humidity can be detected in time to avoid adverse effects on stem cell growth.

[0067] Aeration: A gas flow meter is used to measure the aeration rate in the culture container. In plant cell culture, adequate oxygen supply and removal of harmful gases are crucial for cell respiration and metabolism. Proper control of aeration rate can maintain a good growth environment for cells, and accurate collection of aeration rate data helps to adjust gas exchange conditions according to the growth stage of stem cells.

[0068] The real-time proliferation feature quantification set includes real-time stem cell quantity, real-time stem cell quality, real-time proliferation rate, real-time differentiation inhibition rate, and real-time Dendrobium polysaccharide content;

[0069] Real-time stem cell count: The number of cells per unit area or volume is counted by observing under a microscope and using image analysis software.

[0070] Real-time stem cell quality: Cell viability is assessed using fluorescent dye labeling technology, such as measuring cell health by detecting cell membrane integrity;

[0071] Real-time proliferation rate: Calculates the proliferation rate per unit time based on changes in the number of stem cells; helps to understand the growth rate of stem cells under current culture conditions.

[0072] Real-time differentiation inhibition rate: The degree of inhibition of stem cell differentiation is assessed by detecting the expression levels of differentiation-related genes in stem cells;

[0073] Real-time Dendrobium polysaccharide content: The content of Dendrobium polysaccharides in stem cells was detected using chemical analysis methods such as high performance liquid chromatography or gas chromatography. Dendrobium polysaccharides are one of the important medicinal components of Dendrobium huoshanense, and their content directly affects the medicinal value of stem cells.

[0074] In this step, from the perspective of culture environment control, the collection of data on culture environment factors such as culture medium concentration, light intensity, light duration, culture temperature, culture humidity, and aeration provides crucial information for precise control of the culture environment. Detecting the concentration of culture medium components using equipment such as high-performance liquid chromatography (HPLC) and atomic absorption spectrometry (AAS) allows for timely monitoring of nutrient consumption, enabling reasonable adjustments to the culture medium formula to meet the nutritional needs of stem cells at different stages. Using a photometer and timer to collect light data, and based on the principles of plant photomorphogenesis and photosynthesis, suitable light conditions can be set according to different culture stages to promote normal cell division, differentiation, and the accumulation of active components. High-precision temperature and humidity sensors monitor temperature and humidity in real time, ensuring normal intracellular enzyme activity and water metabolism, and avoiding the negative impact of abnormal temperature and humidity on stem cell growth. A gas flow meter measures aeration to ensure the gaseous environment required for cell respiration and metabolism, maintaining a favorable growth environment. Regarding stem cell growth monitoring, real-time monitoring of stem cell quantity, quality, proliferation rate, and differentiation inhibition... The acquisition of real-time proliferation characteristic quantification sets, such as the growth rate and Dendrobium polysaccharide content, enabled comprehensive monitoring of stem cell growth status. Microscopic observation combined with image analysis software to count cell numbers and fluorescent dye labeling technology to assess cell viability allowed for accurate understanding of the dynamic changes in stem cell quantity and quality. Calculating the proliferation rate based on stem cell quantity changes, detecting the expression levels of differentiation-related genes to assess differentiation inhibition rate, and using high-performance liquid chromatography or gas chromatography to detect Dendrobium polysaccharide content helped to gain a deeper understanding of stem cell growth rate, differentiation status, and medicinal value under current culture conditions, providing strong support for subsequent culture strategy adjustments. Overall, this step effectively solved the problems of traditional fixed culture modes failing to accurately meet the needs of each stage and lacking real-time monitoring and dynamic adjustment. Comprehensive and accurate data acquisition provided a detailed data foundation for subsequent deviation calculation, regulation strategy formulation, and culture environment control, enabling precise control of the Dendrobium huoshanense stem cell culture process and greatly improving the success rate and quality of stem cell clonal proliferation.

[0075] S3. Calculate the deviation distance between the real-time proliferation feature quantization set and the corresponding target proliferation feature quantization set to obtain the real-time proliferation feature deviation vector;

[0076] The real-time proliferation characteristic deviation vector includes stem cell quantity deviation, stem cell quality deviation, proliferation rate deviation, differentiation inhibition rate deviation, and Dendrobium polysaccharide content deviation.

[0077] The formula for calculating the deviation distance between the real-time proliferation feature quantization set and the corresponding target proliferation feature quantization set is as follows:

[0078]

[0079] Where d represents the deviation distance, xi y represents the value of the i-th feature in the real-time proliferation feature quantization set. i This represents the value of the i-th feature in the target proliferation feature quantization set, where n represents the total number of dimensions of the features;

[0080] Stem cell number deviation: This reflects the gap between the actual number of Dendrobium huoshanense stem cells and the target number. A positive deviation indicates that the actual number exceeds the target, while a negative deviation indicates that the number is insufficient. In plant cultivation, the number of stem cells directly affects the final propagation scale and yield. For example, during the proliferation period, if the stem cell number deviation is large and negative, it means that the expected proliferation effect may not be achieved, and the cause needs to be found in time.

[0081] Stem cell quality deviation: Stem cell quality is related to its differentiation ability, activity, and subsequent medicinal value; quality deviation reflects the difference between the current actual quality of stem cells and the target quality; if the quality deviation is too large, it indicates that there are factors affecting the quality of stem cells during the culture process, which will affect subsequent differentiation and development, and thus affect the quality of Dendrobium huoshanense.

[0082] Proliferation rate deviation: This deviation reflects the difference between the actual proliferation rate and the target proliferation rate. In the process of plant cultivation, if the proliferation rate deviation is negative, it indicates that the proliferation rate is too slow. This may be because the supply of nutrients cannot meet the needs of rapid cell division, or environmental factors such as light and temperature limit the metabolic activities of cells.

[0083] Differentiation inhibition rate deviation: The differentiation inhibition rate reflects the ability of stem cells to maintain an undifferentiated state. When the differentiation inhibition rate deviation is large, it indicates that the actual differentiation inhibition situation is inconsistent with the target. If the differentiation inhibition rate is too low, it may cause stem cells to differentiate prematurely, affecting the subsequent proliferation. If the differentiation inhibition rate is too high, it may hinder stem cells from differentiating into the required tissues or organs, affecting their application value.

[0084] Deviation in Dendrobium polysaccharide content: Dendrobium polysaccharides are important medicinal components of Dendrobium huoshanense. Deviation in Dendrobium polysaccharide content reflects the gap between the current actual content and the target content. In plant cultivation, environmental factors such as light intensity and culture medium composition can affect the synthesis of Dendrobium polysaccharides. If the content deviation is large, it is necessary to adjust the culture environment, such as optimizing light time and intensity and adjusting the proportion of nutrients such as carbon sources in the culture medium, in order to promote the synthesis of Dendrobium polysaccharides.

[0085] In this step, by calculating the deviation distance, the difference between real-time proliferation characteristics and target proliferation characteristics can be precisely quantified. This provides a scientific basis for subsequent regulation strategies, making regulation more precise and effective. The real-time proliferation characteristic deviation vector includes multiple dimensions such as stem cell quantity deviation, stem cell quality deviation, proliferation rate deviation, differentiation inhibition rate deviation, and Dendrobium polysaccharide content deviation. These deviation components help identify problems in the culture process, thereby guiding targeted adjustment measures. Based on the analysis results of the real-time proliferation characteristic deviation vector, the culture program can be adjusted in a timely manner to optimize the culture environment and improve the proliferation efficiency and medicinal value of Dendrobium huoshanense stem cells. This dynamic adjustment mechanism helps adapt to the unique needs of stem cells at different culture stages, achieving efficient and stable clonal proliferation. By precisely quantifying differences and making targeted adjustments, this step helps improve the reproduction efficiency and quality of Dendrobium huoshanense stem cells. This not only helps meet the needs of large-scale breeding but also ensures the medicinal value and application effects of the final product. The analysis of the real-time proliferation characteristic deviation vector helps to promptly identify and correct mismatches between the culture environment and stem cell growth needs, thereby reducing the risk of culture failure. This preventive regulation strategy helps improve the stability and reliability of the entire culture process.

[0086] S4. Input the real-time proliferation feature deviation vector and the corresponding culture environment data set into the pre-constructed culture environment regulation and analysis model to obtain the culture environment regulation strategy for the next preset culture stage;

[0087] The real-time proliferation feature deviation vector contains key information such as stem cell quantity deviation, stem cell quality deviation, proliferation rate deviation, differentiation inhibition rate deviation, and Dendrobium polysaccharide content deviation. These deviation data intuitively reflect the gap between the actual growth status of Dendrobium huoshanense stem cells and the expected target. For example, some cell quantity deviations are negative and have large absolute values, indicating that the current stem cell quantity is far from the expected level. This may be due to insufficient nutrient supply or factors in the culture environment that inhibit cell division. Inputting these deviation data into the model provides the model with a clear problem orientation, enabling it to focus on the key problems that occur in the current culture process.

[0088] The culture environment dataset encompasses various aspects such as culture medium component concentration, light intensity, light duration, culture temperature, culture humidity, and aeration. This data provides a detailed description of the current culture environment of *Dendrobium huoshanense* stem cells. For example, the culture medium component concentration data reflects the levels of various nutrients, while light intensity and duration data reflect the light environment conditions. By analyzing this environmental data and combining it with deviation vector information, the model can deeply uncover the root causes of environmental factors leading to current growth deviations. For instance, if a significant deviation in proliferation rate is found along with a low nitrogen concentration in the culture medium, the model may infer that nitrogen deficiency may be one of the key factors affecting the proliferation rate.

[0089] After receiving the real-time proliferation feature deviation vector and the culture environment data set, the model performs calculations and inferences based on its internal algorithm. The data is transmitted and processed in each neuron layer of the model. By adjusting the weights and thresholds, the model finally outputs a culture environment regulation strategy for the next preset culture stage. These strategies may include adjusting the concentration of certain nutrients in the culture medium, changing the light conditions, optimizing the culture temperature and humidity range, and adjusting the aeration rate. If the model analysis shows that the current large deviation in Dendrobium polysaccharide content is due to insufficient light intensity and lack of carbon source in the culture medium, the generated regulation strategy may be to appropriately increase the light intensity and increase the concentration of carbon source in the culture medium to promote the synthesis of Dendrobium polysaccharide and meet the needs of stem cell growth and medicinal value accumulation in the next stage.

[0090] The method for constructing the culture environment regulation and analysis model includes:

[0091] Clonal proliferation experiments of Dendrobium officinale stem cells were carried out under different culture conditions, and growth data of each preset culture stage were collected in a long-term and systematic manner. These data include proliferation characteristics such as stem cell quantity, quality, proliferation rate, differentiation inhibition rate and Dendrobium officinale polysaccharide content. At the same time, corresponding culture environment data, such as culture medium component concentration, light intensity, light time, culture temperature, culture humidity and aeration, were recorded.

[0092] The collected data is cleaned to remove data records with errors, duplicates, and excessive missing values. For data with a small number of missing values, methods such as mean imputation and regression prediction are used to fill in the missing values. For example, if the potassium concentration data in the culture medium of a certain culture sample is missing, it can be filled in based on the mean potassium concentration of samples under other similar culture conditions. Different types of data are standardized to make them have the same scale and distribution range.

[0093] Based on the complex nonlinear relationship between the culture environment and growth characteristics of Dendrobium huoshanense stem cells, suitable machine learning algorithms were selected to construct the model, including neural network algorithms, convolutional neural networks, and decision tree algorithms.

[0094] The preprocessed and feature-engineered data is divided into training, validation, and test sets, with proportions of 70%, 15%, and 15%, respectively. The selected model is trained using the training set data. By continuously adjusting the model's parameters, the model can accurately learn the relationship between the culture environment data and the propagation feature bias. During training, the backpropagation algorithm is used to calculate the loss function, and the model parameters are updated using gradient descent to minimize the loss function. Simultaneously, the validation set data is used to monitor the training process and prevent overfitting. For example, if the loss function on the validation set stops decreasing or even increases, it indicates that the model may be overfitting. In this case, measures such as early termination of training and adding regularization terms can be taken.

[0095] The trained model is evaluated using test set data, and multiple evaluation metrics are used to measure the model's performance. The model is then optimized based on the evaluation results. If the model performance is not ideal, the model structure can be adjusted, hyperparameters can be changed, or different model algorithms can be tried until the model achieves satisfactory performance metrics.

[0096] The trained and optimized model is deployed to real-world application scenarios; the model is integrated into the automated control system for the cloning and proliferation of Dendrobium officinale stem cells in Huoshan, so as to receive real-time proliferation characteristic deviation vectors and culture environment data, and output culture environment regulation strategies.

[0097] In this step, the real-time proliferation characteristic deviation vector accurately captures the gap between the current growth status of Dendrobium huoshanense stem cells and the expected target, providing a clear problem-oriented approach for subsequent regulation. Combined with the culture environment data set, the model can deeply explore the root causes of environmental factors leading to the current growth deviation. Through data analysis of various aspects such as culture medium component concentration, light intensity, light duration, culture temperature, culture humidity, and aeration, the model can accurately determine which environmental factors are key factors affecting stem cell growth. Based on real-time data and internal algorithms, the model can intelligently generate culture environment regulation strategies for the next preset culture stage. These strategies include adjusting the culture medium... By adjusting the composition, light conditions, temperature and humidity ranges, and ventilation, this step aims to optimize the culture environment, promote the healthy growth of stem cells, and accumulate medicinal value. The integrated model of the automated control system can receive data in real time and output regulatory strategies, significantly improving experimental efficiency and accuracy. Compared to traditional manual analysis and control methods, this approach is faster, more accurate, and reduces human error. In summary, this step, through the construction and application of a culture environment regulation and analysis model, achieves precise identification of the growth status of Dendrobium huoshanense stem cells, in-depth analysis of environmental factors, and generation of intelligent regulation strategies, thereby improving experimental efficiency and accuracy and promoting the sustainable development of stem cell culture.

[0098] S5. The culture environment for the clonal proliferation of Dendrobium huoshanense stem cells is regulated based on the aforementioned culture environment regulation strategy;

[0099] The culture environment control strategy includes regulating the composition of the culture medium, the intensity and duration of light exposure, the culture temperature, the culture humidity, and the aeration rate.

[0100] Culture medium composition control: Based on the control strategy, precisely change the concentration of various nutrients in the culture medium; if the strategy indicates that nitrogen source needs to be increased, select a suitable nitrogen-containing compound and add it to the culture medium according to the calculated amount; if the carbon source needs to be adjusted, commonly used carbon sources such as glucose and sucrose should be increased or decreased in proportion in the culture medium as needed; at the same time, pay attention to adjusting the concentration of various trace elements to ensure that the element ratio in the culture medium is appropriate to meet the growth needs of Dendrobium officinale stem cells in the next stage; in addition, plant growth regulators such as auxins and cytokinins can be added as needed to regulate cell division and differentiation;

[0101] Light intensity and duration control: Adjust light intensity and duration according to the control strategy; if it is necessary to increase light intensity, replace with a higher power light source, or adjust the distance between the light source and the culture container to make the light intensity reach a suitable range; if it is necessary to change the light duration, a timer can be used to precisely control the on and off time of the light source; in addition, a suitable type of light source can be selected, such as LED lights, whose spectrum can be customized according to the plant growth needs to provide more favorable light conditions for the growth of Dendrobium officinale stem cells;

[0102] Temperature control during cultivation: Use a constant temperature incubator or other temperature control equipment to adjust the temperature of the cultivation environment to the range required by the control strategy; the suitable temperature for Dendrobium officinale stem cells varies at different stages. During the proliferation phase, the temperature needs to be controlled at 25-28℃; during the differentiation phase, the temperature needs to be adjusted to 22-25℃; monitor the temperature of the cultivation environment in real time through a temperature sensor, and adjust the heating or cooling device in time if the temperature deviates from the set range to ensure temperature stability.

[0103] Culture humidity control: Use a humidifier or dehumidifier to regulate the humidity of the culture environment; when the control strategy requires an increase in humidity, turn on the humidifier to increase the water vapor content in the air; if it is necessary to reduce the humidity, turn on the dehumidifier; at the same time, monitor the humidity of the culture environment in real time through a humidity sensor to ensure that the humidity is maintained at a suitable level. The suitable humidity for Dendrobium officinale stem cell culture is between 60% and 80%.

[0104] Ventilation control: Use equipment such as air pumps to regulate the ventilation of the culture environment; if the control strategy requires increasing the ventilation, increase the power of the air pump appropriately to allow more fresh air to enter the culture environment; if it is necessary to reduce the ventilation, reduce the power of the air pump; by controlling the ventilation, ensure that the oxygen and carbon dioxide content in the culture environment is appropriate, and promote the respiration and photosynthesis of Dendrobium officinale stem cells.

[0105] In this step, by precisely adjusting the culture medium composition, light intensity and duration, culture temperature, culture humidity, and aeration, the most suitable growth environment for *Dendrobium huoshanense* stem cells was provided. This ensured that stem cells received optimal growth conditions at each culture stage, thereby promoting their healthy and rapid proliferation. Under the optimized culture environment, the proliferation efficiency of *Dendrobium huoshanense* stem cells was significantly improved. Precise nutrient ratios and suitable environmental conditions help accelerate cell division and increase cell number, thus improving overall culture efficiency. By regulating the culture environment, not only can stem cell proliferation be promoted, but their quality can also be improved. Suitable light, temperature, and humidity conditions help maintain the normal physiological state of stem cells, reduce the occurrence of abnormal differentiation, and thus improve stem cell quality. This step improves the quality and medicinal value of Dendrobium huoshanense stem cells; precise control strategies reduce unnecessary resource waste, lowering culture costs and improving resource utilization efficiency; real-time monitoring and adjustment of the culture environment allows for timely detection and correction of deviations, ensuring the stability of the culture environment; it helps reduce uncertainties in the culture process, increasing the success rate and reliability of culture; the implementation of this step not only enhances the technical level of Dendrobium huoshanense stem cell clonal proliferation but also provides valuable lessons and references for the cultivation of other medicinal plants or crops; this step, through precise control of the culture environment for Dendrobium huoshanense stem cell clonal proliferation, achieves multiple beneficial effects, including optimizing growth conditions, improving proliferation efficiency, enhancing stem cell quality, reducing culture costs, enhancing culture stability, and promoting technological progress.

[0106] Example 2: Figure 3 As shown, the present invention provides a Dendrobium huoshanense stem cell cloning and proliferation control system, which specifically includes the following modules;

[0107] The target proliferation feature quantification set acquisition module, based on the design requirements for Dendrobium huoshanense stem cell clonal proliferation, acquires the target proliferation feature quantification set for Dendrobium huoshanense stem cell clonal proliferation for each preset culture stage;

[0108] The data acquisition module collects culture environment data of Dendrobium officinale stem cells at the end time of each preset culture stage, and obtains a culture environment data set and a real-time proliferation characteristic quantification set.

[0109] The deviation calculation module calculates the deviation distance between the real-time proliferation feature quantization set and the corresponding target proliferation feature quantization set to obtain the real-time proliferation feature deviation vector.

[0110] The culture environment regulation and analysis module inputs the real-time proliferation feature deviation vector and the corresponding culture environment data set into a pre-constructed culture environment regulation and analysis model to obtain a culture environment regulation strategy for the next preset culture stage.

[0111] The environmental regulation execution module regulates the culture environment for the clonal proliferation of Dendrobium huoshanense stem cells based on the aforementioned culture environment regulation strategy.

[0112] The system sets corresponding target proliferation characteristic quantification sets for different culture stages of Dendrobium huoshanense stem cells. This staged management approach can more accurately reflect the specific needs of stem cells at different growth stages, thereby avoiding the problem of insufficient or excessive nutrient supply caused by the unchanging parameters in the traditional fixed culture mode.

[0113] Through the data acquisition module, the system can acquire real-time data on the culture environment and proliferation characteristics of stem cells, enabling the system to promptly detect and quantify the deviation between the growth status of stem cells and the target status; this helps the system to quickly respond to environmental changes and adjust the culture strategy in a timely manner.

[0114] The culture environment regulation and analysis module utilizes a pre-built analysis model to intelligently generate the environment regulation strategy for the next culture stage based on the real-time proliferation characteristic deviation vector and the culture environment data set. This intelligent regulation method not only improves the accuracy of regulation but also reduces the subjectivity and uncertainty of human intervention.

[0115] The system can dynamically adjust the culture environment parameters according to the actual needs of stem cell growth. This strong dynamic adaptability allows the system to better adapt to the changing needs of Dendrobium officinale stem cells at different growth stages, thereby improving proliferation efficiency and stem cell quality. Because the system has the characteristics of real-time data monitoring, intelligent regulation and strong dynamic adaptability, it can promptly detect and correct situations where the culture environment does not match the needs of stem cell growth, which helps to reduce the risk of culture failure and improve the overall success rate of culture.

[0116] In summary, this Dendrobium huoshanense stem cell cloning and proliferation control system effectively solves the problems existing in the cloning and proliferation process of Dendrobium huoshanense stem cells through precise control, real-time monitoring, and dynamic adjustment, thereby improving proliferation efficiency and quality and enhancing the effectiveness of resource protection.

[0117] The various variations and specific embodiments of the Dendrobium huoshanense stem cell clonal proliferation control method in the aforementioned Example 1 are also applicable to the Dendrobium huoshanense stem cell clonal proliferation control system of this embodiment. Through the foregoing detailed description of the Dendrobium huoshanense stem cell clonal proliferation control method, those skilled in the art can clearly understand the implementation method of the Dendrobium huoshanense stem cell clonal proliferation control system of this embodiment. Therefore, for the sake of brevity, it will not be described in detail here.

[0118] In addition, this application also provides an electronic device, including a bus, a transceiver, a memory, a processor, and a computer program stored in the memory and executable on the processor. The transceiver, the memory, and the processor are respectively connected via the bus. When the computer program is executed by the processor, it implements the various processes of the above-described method embodiment for controlling output data and achieves the same technical effect. To avoid repetition, it will not be described again here.

[0119] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for controlling the clonal proliferation of Dendrobium huoshanense stem cells, characterized in that, The method includes: Based on the design requirements for the clonal proliferation of Dendrobium huoshanense stem cells, a quantitative set of target proliferation characteristics of Dendrobium huoshanense stem cell clonal proliferation was obtained for each preset culture stage; At the end of each preset culture stage, culture environment data of Dendrobium huoshanense stem cells are collected to obtain a culture environment data set and a real-time proliferation characteristic quantification set; the culture environment data set includes culture medium component concentration, light intensity, light duration, culture temperature, culture humidity, and aeration rate; Calculate the deviation distance between the real-time proliferation feature quantization set and the corresponding target proliferation feature quantization set to obtain the real-time proliferation feature deviation vector; The real-time proliferation feature deviation vector and the corresponding culture environment data set are input into a pre-constructed culture environment regulation and analysis model to obtain a culture environment regulation strategy for the next preset culture stage. The culture environment for the clonal proliferation of Dendrobium huoshanense stem cells was regulated based on the aforementioned culture environment regulation strategy.

2. The method for controlling the clonal proliferation of Dendrobium huoshanense stem cells as described in claim 1, characterized in that, The formula for calculating the deviation distance between the real-time proliferation feature quantization set and the corresponding target proliferation feature quantization set is as follows: Where d represents the deviation distance, x i y represents the value of the i-th feature in the real-time proliferation feature quantization set. i represents the value of the i-th feature in the target proliferation feature quantization set, and n represents the total number of dimensions of the features.

3. The method for controlling the clonal proliferation of Dendrobium huoshanense stem cells as described in claim 1, characterized in that, The target proliferation feature quantification set includes the number of target stem cells, the quality of target stem cells, the target proliferation rate, the target differentiation inhibition rate, and the content of target Dendrobium polysaccharides.

4. The method for controlling the clonal proliferation of Dendrobium huoshanense stem cells as described in claim 1, characterized in that, The pre-set culture stages include the initiation period, the proliferation period, and the differentiation period.

5. The method for controlling the clonal proliferation of Dendrobium huoshanense stem cells as described in claim 1, characterized in that, The real-time proliferation feature quantification set includes real-time stem cell quantity, real-time stem cell quality, real-time proliferation rate, real-time differentiation inhibition rate, and real-time Dendrobium polysaccharide content.

6. The method for controlling the clonal proliferation of Dendrobium huoshanense stem cells as described in claim 1, characterized in that, The method for constructing the culture environment regulation and analysis model includes: Collect growth data for each pre-set culture stage; The collected data is cleaned to remove records with too many errors, duplicates, and missing values; and different types of data are standardized. The model is constructed by selecting machine learning algorithms, including neural network algorithms and decision tree algorithms. The preprocessed data is divided into training set, validation set and test set. The selected model is trained using the training set data, and the training process is monitored using the validation set data to prevent the model from overfitting. The trained model is evaluated using test set data to measure its performance; the model is then optimized based on the evaluation results. Deploy the trained and optimized model to real-world application scenarios.

7. A control system for the cloning and proliferation of Dendrobium huoshanense stem cells, characterized in that, The system is applied to the method for controlling the clonal proliferation of Dendrobium officinale stem cells as described in claim 1, and the system comprises: The target proliferation feature quantification set acquisition module, based on the design requirements for Dendrobium huoshanense stem cell clonal proliferation, acquires the target proliferation feature quantification set for Dendrobium huoshanense stem cell clonal proliferation for each preset culture stage; The data acquisition module collects culture environment data of Dendrobium officinale stem cells at the end time of each preset culture stage, and obtains a culture environment data set and a real-time proliferation characteristic quantification set. The deviation calculation module calculates the deviation distance between the real-time proliferation feature quantization set and the corresponding target proliferation feature quantization set to obtain the real-time proliferation feature deviation vector. The culture environment regulation and analysis module inputs the real-time proliferation feature deviation vector and the corresponding culture environment data set into a pre-constructed culture environment regulation and analysis model to obtain a culture environment regulation strategy for the next preset culture stage. The environmental regulation execution module regulates the culture environment for the clonal proliferation of Dendrobium huoshanense stem cells based on the aforementioned culture environment regulation strategy.

8. An electronic device comprising a bus, a transceiver, a memory, a processor, and a computer program stored in the memory and executable on the processor, wherein the transceiver, the memory, and the processor are connected via the bus, characterized in that, When the computer program is executed by the processor, it implements the steps of the method as described in claim 6.

9. A computer-readable storage medium having a computer program stored thereon, characterized in that, When the computer program is executed by a processor, it implements the steps of the method as described in claim 6.

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