Sugarcane tissue culture seedling cultivation system

Abstract

This invention provides a sugarcane tissue culture seedling cultivation system, belonging to the field of sugarcane cultivation technology. It includes a sterile environment cultivation chamber, an environmental factor synergistic regulation system, a machine vision-based intelligent management system for culture media and cultures, culture bottles, and culture racks. The culture bottles are placed on the culture racks, which are located within the sterile environment cultivation chamber. This invention establishes a light-temperature-humidity-air linkage model to simulate the optimal growth microenvironment, significantly promoting the robust growth of sugarcane tissue culture seedlings, shortening the cultivation cycle, and increasing the proliferation coefficient and rooting rate. It utilizes machine vision technology to automatically monitor growth parameters such as seedling height, bud number, and root system, accurately determining the optimal time for transfer and termination of rooting culture, eliminating weak, deformed, and contaminated seedlings, and greatly improving the uniformity and quality of seedlings. By optimizing the design of the lighting system and the standardized placement of culture bottles, it ensures uniform light distribution and reduces energy consumption; through precise control, it reduces resource waste.

Description

Technical Field

[0001] This invention relates to the field of sugarcane cultivation technology, and more particularly to a sugarcane tissue culture seedling cultivation system. Background Technology

[0002] Sugarcane is my country's main sugar crop, accounting for approximately 90% of the total sugar crop planting area and producing over 90% of my country's sugar production. Traditionally, sugarcane seedling production relies on asexual reproduction, resulting in a low propagation coefficient, which hinders the rapid promotion of new varieties. Furthermore, sugarcane is repeatedly infected by various pathogens during production, leading to the accumulation of pathogens on the seed stalks after many years of planting. Some of these pathogens cannot be eliminated with chemical agents, inhibiting sugarcane growth and causing a significant decline in yield and quality. Healthy sugarcane seedling technology, obtained through stem tip meristem cultivation combined with molecular detection techniques, can remove accumulated pathogens on the seed stalks, restore the superior characteristics of improved varieties, and increase sugarcane yield and sugar content. Applying healthy seedling technology to breed superior new varieties can increase the propagation coefficient by more than 1000 times compared to traditional methods, accelerating their widespread application. Existing cultivation methods mostly rely on simple manual fixation, resulting in inconsistent quality of healthy plants. The cultivation environment cannot be precisely controlled, and contamination control depends on fixed disinfection cycles and manual experience, leading to untimely responses and high risks. The cultivation process is poorly managed, relying on manual observation and experience to determine the timing of transplantation. Standardization is low, resulting in inconsistent seedling quality. Insufficient light uniformity on the cultivation racks causes lower-layer seedlings to become leggy or weak due to decreased light intensity. Therefore, a sugarcane tissue culture seedling cultivation system needs to be designed. Summary of the Invention

[0003] The purpose of this invention is to provide a sugarcane tissue culture seedling cultivation system that solves the technical problems of inaccurate environmental control, reliance on manual observation and experience to determine the timing of transplantation, and inconsistent seedling quality in existing sugarcane tissue culture seedling cultivation systems. It upgrades key elements such as aseptic environment control, light, temperature, humidity, gas composition, and culture medium status from a static, isolated set of parameters into a dynamically correlated, intelligently feedback-based, and synergistic organic control system. Furthermore, it introduces real-time monitoring of growth status based on image recognition to achieve standardization, precision, and efficiency in the cultivation process.

[0004] To achieve the above objectives, the technical solution adopted by the present invention is as follows: The sugarcane tissue culture seedling cultivation system includes a sterile environment culture chamber, an environmental factor synergistic regulation system, a machine vision-based intelligent management system for culture media and cultures, culture media, and culture racks. The culture media is placed on the culture racks, which are located within the sterile environment culture chamber. The machine vision-based intelligent management system for culture media and cultures, located on the culture racks, performs visual recognition and management of the culture media and cultures in the culture bottles. The environmental factor synergistic regulation system, located within the sterile environment culture chamber, is used to regulate the cultivation environment within the sterile environment culture chamber.

[0005] Furthermore, the sterile environment incubation room is an independent and enclosed clean space, adjacent to and zoned with the inoculation room and sterilization room, and connected by a physically separated material transfer window. It is regularly sterilized by fumigation with formaldehyde and potassium permanganate. The sterile environment incubation room is equipped with an ozone generator and an ultraviolet circulating air system. Based on real-time data detected by particulate and microbial sensors installed in the room, dynamic disinfection is automatically initiated during non-operation periods, and the frequency and results of sedimentation bacteria testing are incorporated into the disinfection control strategy.

[0006] Furthermore, culture bottles and culture media in the sterile environment culture room are sterilized by high-pressure sterilization at a set temperature for 20-30 minutes. Personnel entering the room must change into sterile clothing and disinfect their hands. Non-sterile items are strictly prohibited from being brought in to prevent fungal and bacterial contamination of the cultures. Any culture bottles found to be contaminated in the sterile environment culture room are immediately removed from the culture room and disposed of separately to avoid cross-contamination. At the same time, the cause of contamination is investigated, and the cause of contamination is determined to be culture media, explants, or operation.

[0007] Furthermore, the environmental factor coordinated regulation system includes a light parameter setting device, a temperature precision control device, and a humidity and gas environment regulation device. The light parameter setting device is used to adjust the light data in the sterile environment culture chamber, the temperature precision control device is used to control the temperature in the sterile environment culture chamber, and the humidity and gas environment regulation device is used to regulate the humidity and airflow in the sterile environment culture chamber. Furthermore, a precise temperature control device is used to maintain a constant temperature of 25±2℃ throughout the sterile culture chamber, ensuring the optimal temperature for sugarcane tissue culture seedling cell division, differentiation, and growth. The temperature difference between different areas within the sterile culture chamber does not exceed 2℃, preventing localized high or low temperatures and avoiding uneven seedling growth, aging, or cessation of growth. The device is equipped with a temperature controller, heating and cooling equipment, which monitors and adjusts the temperature in real time, eliminating temperature fluctuations.

[0008] Furthermore, the humidity and gas environment regulation device is used to control the relative humidity of the sterile culture room and inoculation room to below 45%. If the humidity is higher than the set value, it will easily cause condensation on the inner wall of the culture bottle, which will cause the culture medium to become contaminated. If the humidity is lower than the set value, the moisture in the culture medium will evaporate, which will affect the growth of seedlings. It can be adjusted by humidifiers and dehumidifiers to avoid water accumulation on the ground. Slight ventilation is required in the sterile culture room to ensure fresh air and remove harmful gases such as ethylene produced by the respiration of tissue culture seedlings. If the accumulation of harmful gases exceeds the response value, it will inhibit growth and cause seedlings to turn yellow. However, when ventilating, it is necessary to avoid airflow blowing directly onto the culture bottles and to avoid damaging the sterile environment.

[0009] Furthermore, the light parameter setting device is used to set the light intensity to 2000~3000 lx during the initial culture and subculture propagation stages to meet the needs of bud differentiation and seedling propagation. During the rooting culture stage, the light intensity is increased to 3000~4000 lx to promote chlorophyll synthesis in leaves and cultivate robust seedlings. A light cycle of 12~16 h / d is adopted, and complete darkness is ensured during the dark period. This follows the photoperiodic law of plants and avoids insufficient light leading to seedling etiolation and yellowing, or excessive light affecting the growth rhythm. White fluorescent lamps and LED plant growth lamps are used as light sources. The light sources are evenly suspended to ensure that each layer of culture rack and each culture bottle receives consistent light. Aged light sources are replaced regularly.

[0010] Furthermore, the machine vision-based intelligent management system for culture media and cultivars is used to automatically identify phenomena such as drying, mold, and acidification of the culture medium through regular image scanning, and to issue warnings. It automatically collects images of each culture bottle every week and uses image recognition algorithms to non-contactly measure seedling height, number of tillers, root length and density. When the seedling density reaches a set threshold or the rate of new biomass begins to decline, it prompts for transfer. When the average root length reaches 2-3 cm, the seedling height is 5-8 cm and the morphology is robust, it prompts that the seedlings can be removed from the bottle for hardening. It identifies and marks weak seedlings, etiolated seedlings, and deformed seedlings, and guides manual removal.

[0011] Furthermore, regularly check the culture medium in the culture bottles. If cracking, mold, or turbidity occurs, deal with it immediately to ensure that the culture medium is nutrient-rich and the pH value is stable at 5.8-6.0 to meet the growth requirements of sugarcane tissue culture seedlings. Subculture and proliferate every 15-20 days to prevent seedling aging and decreased differentiation ability. When the rooting culture reaches 2-3cm in root length and 5-8cm in seedling height, stop the culture in time and prepare for hardening.

[0012] Furthermore, the culture racks are made of several layers of steel, with a layer spacing of 25-30cm, to ensure sufficient light for the lower layers. The racks are stable and easy to clean and disinfect. The culture bottles are placed vertically with uniform spacing to avoid blocking light from each other. They are arranged in zones according to the culture stage, which facilitates management and transfer operations and improves culture efficiency. The present invention, by adopting the above-described technical solution, has the following beneficial effects: This invention establishes a light-temperature-humidity-air linkage model to simulate the optimal growth microenvironment, significantly promoting the robust growth of sugarcane tissue culture seedlings, shortening the culture cycle, and increasing the proliferation coefficient and rooting rate. It utilizes machine vision technology to automatically monitor growth parameters such as seedling height, bud number, and root system, accurately determining the optimal timing for transfer and termination of rooting culture, eliminating weak, deformed, and contaminated seedlings, and greatly improving the uniformity and quality of seedlings. By optimizing the design of the lighting system and the standardized placement of culture bottles, it ensures uniform light distribution and reduces energy consumption; precise control further reduces water and fertilizer waste. Detailed Implementation

[0013] To make the objectives, technical solutions, and advantages of this invention clearer, preferred embodiments are described below to further illustrate the invention in detail. However, it should be noted that many details listed in the specification are merely to provide the reader with a thorough understanding of one or more aspects of the invention, and these aspects can be achieved even without these specific details.

[0014] The sugarcane tissue culture seedling cultivation system includes a sterile environment culture chamber, an environmental factor synergistic regulation system, a machine vision-based intelligent management system for culture media and cultures, culture media, and culture racks. The culture media is placed on the culture racks, which are located within the sterile environment culture chamber. The machine vision-based intelligent management system for culture media and cultures, located on the culture racks, performs visual recognition and management of the culture media and cultures in the culture bottles. The environmental factor synergistic regulation system, located within the sterile environment culture chamber, is used to regulate the cultivation environment within the sterile environment culture chamber.

[0015] The sterile culture room is an independent, enclosed Class 10,000 cleanroom, physically separated from the inoculation and sterilization rooms by a material transfer window. It not only undergoes regular formaldehyde and potassium permanganate fumigation for thorough sterilization, but also integrates an ozone generator and an ultraviolet circulating air system. Based on real-time data from particulate and microbial sensors installed indoors, it automatically initiates dynamic disinfection during non-operational periods, incorporating the frequency and results of settling bacteria testing into an automatic disinfection strategy adjustment algorithm. The system automatically calculates and analyzes disinfection strategies using existing AI algorithms and notifies users accordingly.

[0016] The sterile environment culture room includes the following: (1) Space sterility: The culture room must be independent and closed, adjacent to and zoned with the inoculation room and sterilization room; ozone generator disinfection is performed regularly, and formaldehyde + potassium permanganate fumigation is used for thorough sterilization every month, and sedimentation bacteria are tested regularly. (2) Operational sterility: Culture bottles and culture media are sterilized by high temperature and high pressure (121℃, 20~30min); personnel entering the room must change into sterile clothes and disinfect their hands, and it is strictly forbidden to bring in non-sterile items to prevent fungal and bacterial contamination of the culture. (3) Contamination treatment: Culture bottles found to be contaminated are immediately removed from the culture room and disposed of separately to avoid cross-contamination, and the cause of contamination (culture media, explants, operation, etc.) is investigated.

[0017] Once the high-definition camera system or sensor detects suspected contamination characteristics in a culture bottle, such as abnormal turbidity or bacterial colonies, the system immediately alarms and instructs a robotic arm or staff to move the bottle into a dedicated sealed contamination bottle handling box. At the same time, it automatically records the location and batch number, initiates the contamination source tracing and analysis process, and investigates the culture medium, explants, and operation time period.

[0018] The environmental factor synergistic regulation system includes the following control contents: (1) Programmable light regulation: Multi-layer suspended full-spectrum tunable LED plant growth lamps are used. The system automatically adjusts according to the cultivation stage. Primary and subgeneration proliferation stage: The light intensity is set to 2000-3000 lx, and the photoperiod is 12-16h / d. Rooting stage: The light intensity is increased to 3000-4000 lx. According to the temperature sensor data, if the temperature of a certain area is close to the upper limit (27℃), the light intensity of that area can be automatically fine-tuned to prevent photothermal stress.

[0019] Light intensity: 2000-3000 lx for initial culture and subculture propagation to meet the needs of bud differentiation and seedling proliferation; for rooting culture, it can be appropriately increased to 3000-4000 lx to promote chlorophyll synthesis in leaves and cultivate robust seedlings. Photoperiod: A photoperiod of 12-16 h / d is used, ensuring complete darkness during the dark period, following the plant's photoperiodic pattern to avoid insufficient light leading to etiolation and yellowing, or excessive light affecting the growth rhythm. Light source selection: Commonly used are white fluorescent lamps and LED plant growth lights. The light source should be evenly suspended to ensure consistent light exposure on each culture rack and in each culture bottle. Aged light sources should be replaced regularly.

[0020] (2) A central air conditioning system combined with a culture rack shelf auxiliary temperature control module is used to ensure that the temperature of the entire culture room is stable at 25±1℃ (better than ±2℃ in the document), with a temperature difference of ≤1℃ between different areas. Temperature data is linked with the lighting and humidification systems. A constant temperature of 25±2℃ is maintained throughout the process, which is the optimal temperature for cell division, differentiation and growth of sugarcane tissue culture seedlings. Temperature difference control: The temperature difference between different areas in the culture room does not exceed 2℃ to avoid local high or low temperatures, and to prevent uneven growth, aging or cessation of growth of seedlings; a temperature controller and heating / cooling equipment are provided to monitor and adjust the temperature in real time, with no temperature fluctuations.

[0021] (3) By using a dehumidifier and humidifier, the air humidity is precisely controlled within the optimized range of 40%-50%, which prevents condensation and excessive water loss of the culture medium. A micro gas circulation purification device is added to monitor and remove excess ethylene, CO2 and other substances produced by the respiration of tissue culture seedlings in real time, keeping the air fresh. The gas exchange rate is automatically adjusted by the system according to the culture density and growth stage.

[0022] The relative humidity in the culture and inoculation rooms should be controlled below 45%. Excessive humidity can cause condensation on the inner walls of the culture bottles, leading to culture medium contamination; insufficient humidity will cause the culture medium to evaporate too quickly, affecting seedling growth. Humidity can be adjusted using humidifiers or dehumidifiers, avoiding water accumulation on the floor. The culture room requires slight ventilation to ensure fresh air and remove harmful gases such as ethylene (produced by the respiration of tissue culture seedlings; excessive accumulation can inhibit growth and cause yellowing of seedlings). However, ventilation should avoid direct airflow onto the culture bottles and should not disrupt the sterile environment.

[0023] The intelligent management system for culture media and cultured organisms based on machine vision includes automatic monitoring of culture media status and automatic measurement and decision-making of growth parameters. Through regular image scanning, existing AI algorithms automatically identify phenomena such as drying, mold growth, and acidification of the culture media and issue warnings.

[0024] The system automatically collects images of each culture bottle weekly, using image recognition algorithms to non-contactly measure seedling height, number of tillers, root length, and density. Based on the following models, the system automatically prompts optimal timing for subculturing / transfer decisions: When seedling density reaches a set threshold or the rate of new biomass begins to decline (usually after 15-20 days of culture), a transfer prompt is given. For rooting culture termination, when the average root length reaches 2-3 cm, seedling height is 5-8 cm, and the seedlings are robust, a prompt is given indicating they can be removed from the bottle for hardening. Automatic sorting identifies and marks weak, etiolated, and deformed seedlings, guiding manual removal.

[0025] For the cultivation process using culture media, check the condition of the media regularly. If the media in the culture bottles becomes dry, cracked, moldy, or turbid, deal with it immediately. Ensure the media has sufficient nutrients and a stable pH value of 5.8-6.0 to meet the growth requirements of sugarcane tissue culture seedlings. Strictly control the transfer cycle, transferring every 15-20 days during subculture to prevent seedling aging and decreased differentiation ability. When the rooting culture reaches 2-3 cm in root length and 5-8 cm in seedling height, stop the culture in time to prepare for hardening. Observe the growth of seedlings regularly every week, record seedling height, number of buds, and rooting status, and promptly select strong seedlings and remove weak, overgrown, and deformed seedlings.

[0026] Culture rack design: A multi-layer steel culture rack is used, with moderate spacing between layers (25-30cm) to ensure sufficient light exposure for the lower layers; the rack is sturdy and easy to clean and disinfect. Placement guidelines: Culture bottles are placed vertically with uniform spacing to avoid mutual shading of light; they are arranged in sections according to culture stages (primary generation, subculture, rooting) for easy management and transfer operations, improving culture efficiency. Increased light transmittance through the bottle caps ensures sufficient light exposure throughout the bottle, resulting in robust and uniform seedling growth.

[0027] Matters not covered in this invention are common knowledge.

[0028] 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 principle 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 sugarcane tissue culture seedling cultivation system, characterized in that: It includes a sterile culture chamber, an environmental factor synergistic regulation system, a machine vision-based intelligent management system for culture media and cultures, culture media and culture racks. The culture media is placed on the culture racks, which are located in the sterile culture chamber. The machine vision-based intelligent management system for culture media and cultures is located on the culture racks and performs visual recognition and management of the culture media and cultures in the culture bottles. The environmental factor synergistic regulation system is located in the sterile culture chamber and is used to regulate the culture environment within the sterile culture chamber.

2. The sugarcane tissue culture seedling cultivation system according to claim 1, characterized in that: The sterile environment incubation room is an independent and enclosed clean space, adjacent to and zoned with the inoculation room and sterilization room. It is physically separated but connected by a material transfer window. It is regularly sterilized by fumigation with formaldehyde and potassium permanganate. The sterile environment incubation room is equipped with an ozone generator and an ultraviolet circulating air system. Based on real-time data detected by particulate and microbial sensors installed in the room, dynamic disinfection is automatically initiated during non-operation periods, and the frequency and results of sedimentation bacteria testing are incorporated into the disinfection control strategy.

3. The sugarcane tissue culture seedling cultivation system according to claim 1, characterized in that: Culture bottles and culture media in the sterile environment culture room are sterilized by high temperature and high pressure for 20-30 minutes. Personnel entering the room must change into sterile clothing and disinfect their hands. Non-sterile items are strictly prohibited from being brought in to prevent fungal and bacterial contamination of the culture. If any contaminated culture bottle is found in the sterile environment culture room, it should be immediately removed from the culture room and disposed of separately to avoid cross-contamination. At the same time, the cause of contamination should be investigated. The cause of contamination may be the culture medium, explants, or the operator.

4. The sugarcane tissue culture seedling cultivation system according to claim 1, characterized in that: The environmental factor coordinated regulation system includes a light parameter setting device, a temperature precision control device, and a humidity and gas environment regulation device. The light parameter setting device is used to adjust the light data in the sterile environment culture chamber, the temperature precision control device is used to control the temperature in the sterile environment culture chamber, and the humidity and gas environment regulation device is used to regulate the humidity and airflow in the sterile environment culture chamber.

5. The sugarcane tissue culture seedling cultivation system according to claim 1, characterized in that: A precise temperature control device maintains a constant temperature of 25±2℃ throughout the sterile culture chamber, ensuring optimal temperature for sugarcane tissue culture seedling cell division, differentiation, and growth. Temperature differences within the sterile culture chamber are limited to no more than 2℃, preventing localized high or low temperatures and thus avoiding uneven seedling growth, aging, or cessation of growth. The device is equipped with a temperature controller and heating and cooling equipment to monitor and adjust the temperature in real time, eliminating temperature fluctuations.

6. The sugarcane tissue culture seedling cultivation system according to claim 1, characterized in that: The humidity and gas environment control device is used to control the relative humidity of the sterile culture room and inoculation room to below 45%. If the humidity is higher than the set value, condensation will easily form on the inner wall of the culture bottle, causing culture medium contamination. If the humidity is lower than the set value, the moisture in the culture medium will evaporate, affecting the growth of seedlings. It can be adjusted by humidifiers and dehumidifiers to avoid water accumulation on the ground. Slight ventilation is required in the sterile culture room to ensure fresh air and remove harmful ethylene gas produced by the respiration of tissue culture seedlings. Accumulation of harmful gas exceeding the response value will inhibit growth and cause seedlings to turn yellow. However, ventilation should avoid direct airflow onto the culture bottles and should not disrupt the sterile environment.

7. The sugarcane tissue culture seedling cultivation system according to claim 1, characterized in that: The light parameter setting device is used to set the light intensity to 2000~3000 lx during the initial culture and subculture propagation stages to meet the needs of bud differentiation and seedling propagation. During the rooting culture stage, the light intensity is increased to 3000~4000 lx to promote chlorophyll synthesis in leaves and cultivate robust seedlings. A light cycle of 12~16 h / d is adopted, and complete darkness is ensured during the dark period. This follows the photoperiodic law of plants and avoids insufficient light leading to seedling etiolation and yellowing, or excessive light affecting the growth rhythm. White fluorescent lamps and LED plant growth lamps are used as light sources. The light sources are evenly suspended to ensure that each layer of culture rack and each culture bottle receives consistent light. Aged light sources are replaced regularly.

8. The sugarcane tissue culture seedling cultivation system according to claim 1, characterized in that: The machine vision-based intelligent management system for culture media and cultivars is used to automatically identify phenomena such as drying, cracking, mold, and acidification of culture media through regular image scanning and to issue warnings. It automatically collects images of each culture bottle every week and uses image recognition algorithms to non-contactly measure seedling height, number of tillers, root length and density. When the seedling density reaches a set threshold or the rate of new biomass begins to decline, it prompts for transfer. When the average root length reaches 2-3 cm, the seedling height is 5-8 cm and the morphology is robust, it prompts that the seedlings can be removed from the bottle for hardening. It identifies and marks weak seedlings, etiolated seedlings, and deformed seedlings and guides manual removal.

9. The sugarcane tissue culture seedling cultivation system according to claim 8, characterized in that: Regularly check the culture medium in the culture bottles. If it becomes dry, cracked, moldy, or turbid, deal with it immediately. Ensure that the culture medium is nutrient-rich and that the pH value is stable at 5.8-6.0 to meet the growth requirements of sugarcane tissue culture seedlings. Subculture and proliferate the culture every 15-20 days to prevent seedling aging and decreased differentiation ability. Stop the culture when the root system is 2-3 cm long and the seedling height is 5-8 cm, and prepare for hardening off.

10. The sugarcane tissue culture seedling cultivation system according to claim 1, characterized in that: The culture rack consists of several layers of steel culture racks with a layer spacing of 25-30cm to ensure sufficient light for the lower layers. The rack is stable and easy to clean and disinfect. Culture bottles are placed vertically with uniform spacing to avoid mutual shading of light. They are arranged in zones according to the culture stage for easy management and transfer operations, thus improving culture efficiency.