A method for inducing rapid germination of sugarcane mutant buds by heavy ion irradiation

Abstract

The application discloses a method for fast germination of sugarcane mutant sprouts induced by heavy ion irradiation, and relates to the technical field of crop mutation breeding and seedling cultivation, and specifically comprises the following steps: first, homogenizing and regularizing sugarcane seed stems, then dynamically irradiating the sugarcane sprouts according to the size of the sugarcane sprouts by using a low-dose heavy ion beam, then placing the sugarcane sprouts in a constant-temperature light-proof environment at 25-30 DEG C for static curing, then gradually adjusting beam parameters to complete gradient irradiation mutation, and finally integrating temperature and humidity control and ventilation in a steady environment at 25-30 DEG C. The method constructs a full-process integrated processing system through homogenization and regularization, beam irradiation, closed constant-temperature curing, gradient irradiation mutation and low-temperature steady cultivation, eliminates the beam action blind area, integrates multi-stage data by combining algorithms, accurately optimizes process parameters, effectively improves the germination efficiency and uniformity of the sugarcane sprouts, enhances the targeting and stability of the mutant induction, avoids exogenous pollution, and improves the success rate of the sugarcane mutant cultivation and the large-scale application level.

Description

Technical Field

[0001] This invention relates to the field of crop mutation breeding and seedling cultivation technology, specifically a method for inducing rapid germination of sugarcane mutant buds using heavy ion beam irradiation. Background Technology

[0002] Sugarcane is a globally important sugar and economic crop. Germplasm resource innovation and variety improvement are core links supporting the high-quality development of the sugarcane industry. Heavy ion beam mutagenesis, with its rich mutation types and outstanding mutagenesis efficiency, has become a highly valuable technology in modern crop breeding. The germination efficiency and quality of sugarcane shoots directly affect the survival, trait expression, and breeding screening progress of sugarcane mutants after heavy ion beam mutagenesis. It is a key link in the sugarcane mutagenesis breeding process. Currently, research on heavy ion beam-induced sugarcane mutants in the industry mainly focuses on mutagenesis dose screening. In the selection and identification of variant traits, a complete processing system adapted to the physiological characteristics of sugarcane buds has not yet been formed. The pretreatment of seed stalks, the regulation of heavy ion beam action, and subsequent maintenance and cultivation are fragmented, lacking integrated process connection and data coordination. At the same time, the environmental control of sugarcane bud germination and the matching degree of heavy ion beam action parameters are insufficient, making it difficult to achieve precise control based on quantitative logic. This limits the depth and scope of application of heavy ion beam mutagenesis technology in the field of sugarcane breeding. The industry urgently needs to improve the specialized sugarcane bud germination regulation technology to meet the needs of large-scale and standardized sugarcane mutation breeding.

[0003] Traditional sugarcane bud germination treatment methods, lacking specific optimization tailored to the characteristics of heavy ion beam mutagenesis, suffer from numerous application limitations. The seed stalk treatment process lacks standardized procedures, leading to inconsistent physiological states and cleanliness levels, resulting in varying basic bud germination conditions and impacting the consistency of subsequent mutagenesis treatments. The heavy ion beam treatment employs a single parameter application mode, failing to dynamically adjust the treatment method based on the bud's developmental state, easily causing uneven tissue action and physiological damage imbalances. This reduces mutagenesis effectiveness and delays bud germination. Furthermore, the environmental management during bud care and cultivation is rudimentary, lacking refined control measures such as airtight disinfection and constant temperature / light avoidance. External microbial contamination and environmental parameter fluctuations easily interfere with the bud's physiological repair and germination process. Each process step is executed independently without data collection and integrated analysis, lacking algorithmic control support. The beam effect and environmental adaptability cannot be synergistically optimized, ultimately resulting in long bud germination cycles, low germination uniformity, and poor mutant phenotypic stability, failing to meet the requirements of efficient and precise sugarcane mutagenesis breeding. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a method for inducing rapid germination of sugarcane mutant buds using heavy ion beam irradiation. This method first homogenizes and standardizes the sugarcane seed stalks to unify their physiological basis; then, it uses a low-dose heavy ion beam to directionally stimulate the sugarcane buds, dynamically matching the beam density according to the bud size; next, it is statically nurtured in a closed, constant-temperature, and light-protected environment to promote physiological repair; then, a second medium-dose mutagenesis is performed using gradient irradiation, and an irradiation repair coupling algorithm is introduced to integrate process parameters; finally, temperature, humidity, and ventilation are comprehensively controlled in a low-temperature, stable-state cultivation space to achieve integrated parameter matching.

[0005] To solve the above-mentioned technical problems, the present invention provides the following technical solution: a method for inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation, the specific steps of which are as follows:

[0006] S100, Homogeneous and Regular Stems: Select the main stems of mature sugarcane plants with uniform growth characteristics, and perform standardized cutting and purification treatment to unify the basic physiological state of the stem segments.

[0007] S200, Low-dose beam excitation: The homogenized and regularized sugarcane stalks are arranged in the heavy ion beam operation area. The beam current is dynamically matched according to the size of the sugarcane buds using a beam current targeted adaptation and adjustment mechanism. Low-dose heavy ion beams are used to act on the sugarcane bud tissue in the whole area to regulate the internal physiological state of the buds. Various process parameters generated during the operation are collected simultaneously to form a primary processing data reserve.

[0008] S300, Closed Constant Temperature Curing: For sugarcane stalks after low-volume beam intensification, place them in a closed space after compound disinfection, and constantly regulate the internal temperature and humidity index of the space. Maintain a light-proof and static storage mode throughout the process, continuously collect information on the duration of the static curing period, and accumulate basic data on environmental control.

[0009] S400, gradient irradiation mutagenesis: Combined with sugarcane stalks after closed constant temperature curing, the heavy ion beam action parameters are progressively adjusted, the beam current action dose is changed to complete the secondary beam current treatment, and the irradiation repair coupling algorithm is used to integrate and collect the multi-stage process data from the previous stage.

[0010] S500, Low-Temperature Steady-State Cultivation: Sugarcane stem segments after mutagenesis treatment are placed in a closed cultivation space. The temperature, humidity and ventilation of the cultivation area are controlled in a coordinated manner. Environmental data and periodic data are continuously collected during the cultivation stage. The habitat germination adaptation algorithm is used to match environmental parameters and complete the integrated parameter control of the entire process.

[0011] Furthermore, the standardized cutting process employs sterile specialized cutting tools for fixed-length segmentation, with the length of the cut sugarcane stalk segments controlled within the range of 10cm to 15cm. The cutting operation must avoid buds and surrounding thin-walled tissues, maintaining a flat and neat cut surface, and preserving the complete sugarcane bud structure in each segment. The purification process relies on a multi-stage progressive operation. First, surface solid impurities are removed by rinsing with flowing clean water. Then, surface soluble substances are dissolved by static soaking in sterile clean water. Finally, a second rinsing purification is completed using sterile soft water. After purification, the material is placed in a dark and ventilated area to naturally dissipate surface free moisture, thus unifying the cutting specifications and standardizing the purification process, and constraining the overall shape and surface cleanliness of the material.

[0012] Furthermore, the beam-targeting adaptation and adjustment mechanism divides the beam density control into three levels based on the outline size and development volume of the sugarcane buds: the nascent level, the conventional level, and the mature level. The nascent level corresponds to small, newly formed sugarcane buds, with a beam density range of 12–16 pA / cm². The conventional level corresponds to medium-sized sugarcane buds with standard morphology, with a beam density range of 18–22 pA / cm². The mature level corresponds to fully developed large sugarcane buds, with a beam density range of 24–28 pA / cm². Each control level has an independent and fixed beam density range. The corresponding level is matched based on the real-time identified sugarcane bud morphology parameters. The output density per unit area of ​​the heavy ion beam is limited according to the level setting standard. The beam action boundary is locked according to the actual coverage area of ​​the sugarcane buds. This abandons the uniform beam output mode across the entire area and simultaneously saves the original information of the matching and parameter settings, enriching the categories of basic parameters collected during the operation phase.

[0013] Furthermore, the homogenized and regularized sugarcane stalks are laid out in a single layer in the heavy ion beam operation area, with a uniform interval of 2cm to 3cm between individual stalks. All sugarcane buds are placed facing upwards in a uniform orientation, and the horizontal angle of the irradiation platform is adjusted simultaneously to maintain the vertical incidence of the heavy ion beam. During the low-dose heavy ion beam operation phase, the irradiation dose range is set to 0.2Gy to 0.5Gy, and the beam output energy parameters are stabilized. The equipment's full-area scanning mode is used to fully cover the meristematic tissue and surrounding cortical tissue of the sugarcane buds, eliminating local blind spots and irradiation deviations. The ambient temperature in the operation area is kept constant at 20℃ to 25℃ to maintain a continuous and stable beam output intensity. The directional action is completed in a uniform full-area coverage operation form, and the actual operation parameters such as beam dose and layout spacing are recorded simultaneously.

[0014] Furthermore, the sugarcane stalks after low-volume beam intensification are uniformly collected into a sealed curing chamber after composite disinfection treatment. The chamber employs a dual-mode full-area disinfection system combining ultraviolet irradiation and ozone diffusion to complete the full-area purification of the inner walls and supporting equipment, removing residual bacteria and external pollutants. The sealed space continuously maintains environmental control indicators, keeping the temperature constant at 25-30℃ and the relative humidity at 75%-85%, while maintaining a completely light-proof and enclosed structure. The stalks remain stationary within the space, without material movement or human intervention during the curing period, isolating them from external airflow, light, and environmental fluctuations. The entire cycle of static curing is continuously recorded, and the original environmental control data for the curing stage is collected to maintain the curing environment parameters continuously constant.

[0015] Furthermore, the duration information is collected in a full-time segmented recording mode, dividing the time scale nodes of the closed constant temperature curing stage, marking the start and end nodes of each interval in sequence, continuously capturing the original information of the entire time dimension, and unifying the collection standards and scale specifications of time recording; the complete time trajectory of static storage is retained without interruption throughout the entire process, anchoring the time nodes of the entire curing process, establishing the corresponding relationship between time data and the closed static storage stage, and completely collecting the original records of the entire cycle duration.

[0016] Furthermore, the sugarcane stalks after the sealed constant temperature curing stage retain the same arrangement and spacing as in the low-volume beam intensification treatment stage, maintaining a uniform beam action reference height. The heavy ion beam operating parameters are progressively corrected based on a graded gradient control logic, and a dedicated medium-gradient irradiation dose threshold is set. Unlike the low-dose range of the initial base beam, the secondary beam treatment uses a stable medium-dose range, keeping the secondary beam treatment dose stably controlled within the 55Gy–70Gy limit. Simultaneously, the beam output power and particle beam density are coordinated and adjusted to maintain a constant beam vertical incident angle and sugarcane bud target coverage area, achieving precise secondary beam application from the same source and location. Through progressive switching of differentiated beam parameters between the two stages, a hierarchical gradient irradiation control architecture is constructed, forming a segmented precise beam action mode, and orderly completing the step-by-step superposition of multiple gradient beams.

[0017] Furthermore, the mathematical expression for the irradiation repair coupling algorithm is:

[0018] ;

[0019] In the formula, The gradient irradiation effect value is a comprehensive quantitative indicator that integrates multiple preceding processes. The low-dose pre-irradiation activation coefficient is used to calibrate the physiological response correction ratio of low-dose beam effects, with a fixed value range of 0.3 to 0.5. The actual dose of low-dose pre-irradiation corresponds to the actual effect value of the heavy ion beam in the substrate beam activation process. This is the medium-dose mutagenic synergistic coefficient, used to define the proportion of the effect correction of medium-dose beam flow, with a fixed value range of 0.6 to 0.8; is the actual dose for medium-dose mutagenesis, corresponding to the actual effect value of the heavy ion beam in the secondary beam treatment process; e is the natural constant, used as the basic constant for exponential operations to characterize the temporal decay change law of physiological repair in sugarcane buds. The actual duration of intermittent recovery maintenance refers to the timing parameters for the entire closed, constant-temperature, and quiet curing process. The duration of physiological repair characteristics of sugarcane buds is a fixed reference parameter, with a constant value of 24h.

[0020] Furthermore, the mathematical expression for the habitat germination adaptation algorithm is:

[0021] ;

[0022] In the formula, G is the synergistic germination index, which is used to quantify the comprehensive germination potential of sugarcane shoots after irradiation treatment and steady-state cultivation, and its value range is limited to 0 to 1; This represents the gradient irradiation effect value. The temperature regulation weighting coefficient represents the proportion of the influence of the cultivation temperature on the germination process, and is fixed at 0.4; T is the actual ambient temperature obtained by monitoring inside the mutation cultivation space. The optimal reference temperature for sugarcane bud germination is 27℃, which is a fixed process parameter. The humidity control weighting coefficient represents the proportion of the influence of environmental humidity on the germination state, and is fixed at 0.3; H is the actual relative humidity of the environment detected in real time in the cultivation space. The optimal baseline humidity for sugarcane bud germination is set at a constant value of 85% for a fixed process parameter. The cultivation duration weighting coefficient represents the proportion of the cultivation cycle's influence on germination effectiveness, and is fixed at 0.3; t represents the actual number of days of mutagenesis-induced low-temperature cultivation. The maximum control period for the cultivation stage is preset, and it is a fixed process parameter with a constant value of 10 days.

[0023] Compared with existing technologies, this method for inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation has the following advantages:

[0024] I. This invention unifies the physiological basis of sugarcane buds through homogenization and standardization of seed stalks, achieves precise action of heavy ion beams through targeted adaptation and adjustment mechanisms, stabilizes the internal physiological state of sugarcane buds through closed constant temperature cultivation, completes segmented beam treatment with gradient irradiation, and finally matches the germination needs of sugarcane buds with low-temperature steady-state cultivation, constructing a fully integrated sugarcane bud treatment and cultivation system. This method abandons the uniform beam action mode across the entire area, dynamically adjusts the beam action form according to the actual state of sugarcane buds, eliminates treatment blind spots and irradiation deviations, ensures uniform effect on sugarcane bud tissue, collects process and environmental data at each stage throughout the process, and optimizes treatment parameters by integrating multi-dimensional information through algorithms, effectively regulating the internal physiological activity of sugarcane buds, accelerating the germination process, improving the targeting and stability of heavy ion beam-induced mutants, reducing ineffective treatment steps, shortening the sugarcane bud germination cycle, and improving the efficiency and success rate of sugarcane mutant cultivation.

[0025] II. This invention isolates external interference through a closed-loop environment controlled by comprehensive disinfection, and provides stable cultivation conditions for sugarcane buds by constant temperature, humidity, and ventilation. Combined with irradiation repair coupling and habitat germination adaptation algorithms, it achieves synergistic integration of process data and environmental parameters, accurately quantifies the germination potential of sugarcane buds, and optimizes control strategies. The entire process adopts standardized operating procedures for material handling and environmental control, avoiding the impact of exogenous pollution and environmental fluctuations on sugarcane bud growth. The graded gradient beam treatment takes into account both the physiological tolerance of sugarcane buds and the mutagenic effect, improving the accuracy of mutant induction. Continuous collection of full-cycle data forms a closed-loop control mechanism, continuously optimizing the cultivation habitat and treatment process, significantly improving the uniformity and consistency of sugarcane bud germination, ensuring stable expression of mutant traits, reducing material loss during cultivation, and making the germination of sugarcane mutant buds more efficient and the control more precise, adapting to the application needs of large-scale cultivation.

[0026] Other advantages, objectives and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination or study, or may be learned from the practice of the invention. Attached Figure Description

[0027] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without any creative effort.

[0028] Figure 1 A flowchart of a method for inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation;

[0029] Figure 2This is a schematic diagram of data transmission for a method of inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation.

[0030] Figure 3 This is a schematic diagram of data transmission in the low-volume beam excitation step of the present invention. Detailed Implementation

[0031] To further illustrate the technical means and effects of the present invention in achieving its intended purpose, the following detailed description of the specific implementation methods, structures, features, and effects of the present invention, in conjunction with the accompanying drawings and preferred embodiments, is provided below.

[0032] Example 1:

[0033] This embodiment is specifically applied to small-scale sugarcane germplasm mutation breeding in the laboratory, focusing on the precise creation of sugarcane mutant materials and the targeted improvement of sugarcane shoot germination efficiency. Based on the complete technical process of this invention, it provides a standardized and reproducible treatment scheme for laboratory sugarcane mutation breeding experiments, ensuring accurate and reliable experimental data. Simultaneously, it lays a solid technical foundation for subsequent large-scale breeding applications. The entire process strictly follows the operating specifications and parameter requirements of this invention, without adding any additional external parameters. Each step revolves around optimizing the physiological state of sugarcane shoots, stabilizing treatment effects, and ensuring complete data collection. Figure 1 As shown, the specific implementation steps are as follows:

[0034] S100, Homogeneous and Uniform Sugarcane Stems: Mature sugarcane stems with uniform growth characteristics are selected as raw materials. Standardized cutting and length segmentation are carried out using sterile cutting tools, precisely controlling the length of sugarcane stem segments to within the range of 10cm-15cm. The cutting process avoids buds and surrounding thin-walled tissues throughout, preserving the complete sugarcane bud structure of each segment while ensuring a smooth and uniform cut surface. This eliminates processing deviations caused by differences in raw material morphology at the source, ensuring all sugarcane buds have a consistent physical morphology. Purification is achieved through a multi-stage progressive process: first, the surface solid impurities of the sugarcane stem segments are thoroughly rinsed with running water, then they are statically soaked in sterile water to fully dissolve... Soluble substances adhering to the surface are removed, and a second rinsing purification is performed using sterile soft water to thoroughly remove exogenous pollutants and residual impurities from the stem segments. After purification, the material is placed in a dark and ventilated area to allow the surface free moisture to dissipate slowly and evenly, bringing the surface humidity of the stem segments to a state suitable for subsequent processing. Through uniform cutting specifications and standardized purification processes, the overall shape and surface cleanliness of the material are comprehensively constrained, ensuring that all sugarcane stem segments have a uniform physiological basis and physical state. This provides stable and consistent raw material conditions for subsequent heavy ion beam treatment, avoiding uneven treatment effects due to differences in raw materials and ensuring the accuracy and repeatability of small-scale laboratory experiments.

[0035] S200, Low-volume Beam Intensification: Homogenized and well-formed sugarcane stalks are neatly arranged in a single layer in the heavy ion beam operation area, with a uniform 2cm-3cm interval between individual stalks. All sugarcane buds are placed facing upwards in a uniform orientation. The horizontal angle of the irradiation platform is simultaneously and precisely adjusted to ensure that the heavy ion beam acts on the sugarcane buds in a vertical incidence manner, resulting in more uniform and deviation-free beam coverage. A beam-targeted adaptation adjustment mechanism is adopted, dividing the beam density into three fixed levels based on the outline size and development volume of the sugarcane buds. By identifying the morphological parameters of the sugarcane buds in real time, the corresponding level is matched, and the output density per unit area of ​​the heavy ion beam is limited according to the level setting standard. The beam action boundary is locked according to the actual coverage area of ​​the sugarcane buds, abandoning the uniform beam output mode across the entire area, and allowing the beam output to be highly adapted to the development state of the sugarcane buds themselves. Low-dose heavy ion beams of 0.2 Gy to 0.5 Gy are used to treat the entire sugarcane bud tissue. The ambient temperature in the operating area is kept constant at 20℃ to 25℃. Utilizing the equipment's full-area scanning mode, the meristematic tissue and surrounding cortex of the sugarcane bud are fully covered, completely eliminating blind spots and irradiation deviations. This stabilizes the beam output intensity and energy parameters, gently regulating the internal physiological state of the sugarcane bud, activating the physiological activity of the meristematic tissue, and bringing the sugarcane bud into a physiological state suitable for subsequent mutagenesis treatment. Simultaneously, various process parameters such as beam dose, beam spacing, beam level matching, and parameter settings are collected during the operation. Complete operational information and raw data are preserved, forming a primary treatment data reserve. This enriches the categories of basic parameters collected during the operation stage, providing detailed data support for subsequent process integration and parameter optimization. Figure 3 As shown.

[0036] S300, Sealed Constant Temperature Curing: Sugarcane stalks that have undergone low-volume beam intensification are uniformly stored in a sealed curing chamber that has undergone dual disinfection using ultraviolet irradiation combined with ozone diffusion. This dual disinfection mode thoroughly purifies the entire space's interior walls and supporting equipment, completely eliminating residual bacteria and external pollutants, providing a sterile and clean environment for the sugarcane buds to repair. The sealed space continuously maintains controlled environmental parameters, keeping the temperature constant at 25–30℃ and the relative humidity at 75%–85%. The entire space maintains a completely light-proof, enclosed structure, with the stalks placed statically within the space. No material movement or human intervention is performed during the curing period, completely isolating the sugarcane buds from external airflow, light, and environmental fluctuations, allowing them to grow in a stable and gentle environment. The system completes internal physiological repair within a controlled environment, smoothly transitions to the physiological changes brought about by beam treatment, stabilizes its own metabolic rhythm, and avoids interference from external factors in the repair process. A full-time segmented recording mode is used to collect complete duration information, dividing the time scale nodes of the closed constant-temperature curing stage, sequentially marking the start and end nodes of each interval, continuously capturing raw information in the entire time dimension, unifying the collection standards and scale specifications for time recording, and continuously retaining the complete time-series trajectory of static storage throughout the entire process. Time-series nodes are anchored throughout the entire curing process, establishing a corresponding relationship between time-series data and the closed static storage stage, completely collecting raw records of the entire cycle duration, and accumulating basic data for environmental control. This provides crucial temporal and environmental basis for the subsequent data integration of the irradiation repair coupling algorithm.

[0037] S400 Gradient Irradiation Mutagenesis: Sugarcane stalks that have completed closed-loop constant-temperature curing strictly adhere to the same placement and spacing as those in the low-volume beam intensification treatment stage, maintaining a consistent beam action reference height. This ensures complete consistency in position and orientation between the two beam treatments. The heavy ion beam operating parameters are progressively corrected based on a graded gradient control logic, with a dedicated medium-gradient irradiation dose threshold set to stably control the dose of the second beam treatment within the defined range of 55 Gy to 70 Gy. Simultaneously, the beam output power and particle beam density are coordinated and adjusted to maintain a constant beam vertical incident angle and sugarcane bud target coverage area. This process achieves precise secondary beam application from the same source and location. By progressively switching between differentiated beam parameters in the two stages, a hierarchical gradient irradiation control architecture is constructed, forming a segmented precise beam action mode. This orderly completes the step-by-step superposition of multiple gradient beams, significantly improving the targeting and effectiveness of mutagenesis while ensuring that the activity of sugarcane buds is not damaged, allowing stable and controllable mutations to occur in the genetic material inside the sugarcane buds. Simultaneously, an irradiation repair coupling algorithm is used to integrate and collect process data from multiple stages, including early seed stalk regularization, low-volume beam intensification, and closed constant temperature curing. The mathematical expression of the irradiation repair coupling algorithm is:

[0038] ;

[0039] In the formula, This represents the gradient irradiation effect value. The activation coefficient for low-dose pre-irradiation; This is the actual dose of the low-dose pre-irradiation. The synergistic effect coefficient of medium-dose mutagenesis; The actual dose for medium-dose mutagenesis; e is the natural constant; The actual duration of intermittent recovery maintenance; To determine the duration of physiological repair characteristics of sugarcane buds, process parameters, environmental data, and time-series data from each stage are organically integrated. This allows the gradient irradiation parameters to be set more closely to the actual physiological state of sugarcane buds, optimizes the precision of the mutagenesis treatment, and ensures stable and consistent mutagenesis effects, laying a solid foundation for the rapid germination of sugarcane buds in the future.

[0040] S500, Low-Temperature Steady-State Cultivation: Sugarcane stem segments after gradient irradiation mutagenesis treatment are centrally placed in a closed cultivation space. Temperature, humidity, and ventilation are controlled to create a stable and suitable growth environment for bud germination. Environmental and periodic data are continuously collected during the cultivation process to comprehensively understand environmental changes and growth cycle information during bud germination. A habitat germination adaptation algorithm is used to match environmental parameters. The mathematical expression of the habitat germination adaptation algorithm is as follows:

[0041] ;

[0042] In the formula, G is the co-germination index; This represents the gradient irradiation effect value. The temperature control weighting coefficient is T; T is the actual ambient temperature obtained from monitoring inside the mutation cultivation space. The optimal reference temperature for sugarcane bud germination; H represents the humidity control weighting coefficient; H is the actual relative humidity of the cultivation space as detected in real time. The optimal baseline humidity for sugarcane bud germination; The cultivation duration is the weighting coefficient; t is the actual number of days of mutagenesis and low-temperature cultivation. To ensure the maximum control period during the cultivation stage, key conditions such as temperature, humidity, and ventilation in the cultivation space are dynamically optimized by combining all previous process data and real-time cultivation data. This ensures that the cultivation environment always meets the physiological needs of sugarcane bud germination, promoting the rapid breakthrough of dormancy and initiation of the germination process, while maintaining a uniform germination rhythm. Through integrated parameter control, process parameters and environmental parameters throughout the entire process are synergistically optimized, allowing sugarcane buds to complete germination in the optimal habitat. This significantly shortens the germination cycle, improves the germination rate and uniformity, and provides high-quality, uniform germination material for small-scale screening of sugarcane mutants in the laboratory. It fully leverages the technical advantages of heavy ion beam mutagenesis, facilitating the efficient advancement of sugarcane germplasm innovation and variety improvement in the laboratory.

[0043] This embodiment, in a small-scale sugarcane mutation breeding scenario in the laboratory, utilizes a complete process including homogenization and regularization of seed stalks, low-volume beam intensification, closed constant-temperature cultivation, gradient irradiation mutation, and low-temperature steady-state cultivation. This process continuously optimizes the physiological state of sugarcane buds. By relying on irradiation repair coupling algorithms and habitat germination adaptation algorithms, data integration and parameter control are achieved, completely eliminating the impact of differences in raw materials and environment. The germination cycle of sugarcane buds is significantly shortened, and the uniformity of germination is significantly improved. The heavy ion beam mutation effect is stable and controllable, providing a standardized and reproducible technical solution for sugarcane germplasm innovation in the laboratory, and ensuring the accuracy and reliability of small-scale breeding experiments.

[0044] Example 2:

[0045] This embodiment is adapted to the mass mutant cultivation and production scenario of large-scale sugarcane breeding bases. Addressing the actual needs of large-scale sugarcane seed stalk processing, standardized mutagenesis, and uniform germination at these bases, it utilizes the technical system of this invention to conduct full-process batch operations. This enables large-scale, standardized, and efficient mutagenesis and germination of sugarcane buds, significantly improving the production efficiency of breeding bases, reducing material loss during batch processing, and ensuring the uniformity of batch bud germination and the stability of the mutagenesis effect. The entire process strictly follows the parameter specifications and operating procedures of this invention, without introducing any external parameters. Each step of the operation balances the uniformity and efficiency of batch processing with the protection of the physiological state of the sugarcane buds. Figure 2 As shown, the specific implementation steps are as follows:

[0046] S100. Homogeneous and Uniform Sugarcane Stems: Mature sugarcane stems with uniform growth characteristics are selected in batches as raw materials for batch processing. This ensures that the growth cycle and physiological state of all raw materials remain highly consistent. Standardized cutting and length segmentation are carried out using sterile cutting tools, uniformly controlling the length of all sugarcane stem segments to within the range of 10cm to 15cm. The cutting operation precisely avoids buds and surrounding thin-walled tissues, preserving the complete sugarcane bud structure of each stem segment. This ensures that all cut surfaces are flat and uniform, making the physical morphology of the batch materials completely uniform and eliminating the impact of individual morphological differences during batch processing. The batch materials are purified using a multi-stage progressive purification process. First, flowing clean water is used to centrally rinse away the solid impurities on the surface of the sugarcane stem segments. The sugarcane stalks are first subjected to static soaking in sterile water to dissolve soluble substances adhering to their surface. Finally, sterile soft water is used for a second rinsing purification process, achieving comprehensive cleaning of the entire batch of sugarcane stalks and thoroughly removing surface bacteria and contaminants. The purified batch of materials is then placed in a dark and ventilated area to allow the surface free moisture to dissipate evenly, ensuring that the surface humidity of all stalks remains consistent. Through uniform cutting specifications and standardized purification processes, the overall shape and surface cleanliness of the batch of materials are comprehensively constrained, ensuring that the large batch of sugarcane stalks have a uniform physiological basis and physical state. This provides stable and uniform raw material conditions for subsequent batch heavy ion beam treatment, ensuring the consistency of batch treatment results and meeting the batch production needs of large-scale breeding bases.

[0047] S200, Low-Quantity Beam Intensification: Homogenized and well-formed sugarcane stalks are neatly arranged in a single layer within the heavy ion beam treatment area, with a uniform 2-3 cm interval between individual stalks. All sugarcane buds are placed facing upwards in a uniform orientation. Simultaneously, the horizontal angle of the irradiation platform is precisely adjusted to ensure vertical incidence of the heavy ion beam, providing uniform beam conditions for the batch of sugarcane buds. A beam-targeted adaptation adjustment mechanism is employed, rapidly dividing the batch of sugarcane buds into three fixed beam density control levels based on their outline size and developmental size. By real-time identification of the morphological parameters of the batch of sugarcane buds, the corresponding level is matched, and the heavy ion beam output density per unit area is uniformly limited according to the level setting standards. The beam action boundary is locked according to the actual coverage area of ​​the batch of sugarcane buds, abandoning the uniform beam output mode across the entire area and allowing the beam output to precisely adapt to the developmental state of the batch of sugarcane buds. Low-dose heavy ion beams of 0.2 Gy to 0.5 Gy were used to treat the entire area of ​​sugarcane bud tissue in batches. The ambient temperature in the operating area was kept constant at 20℃ to 25℃. The equipment's full-area scanning mode was used to achieve full coverage treatment of the meristematic tissue and surrounding cortex of the batch sugarcane buds, eliminating local blind spots and irradiation deviations in batch treatment, stabilizing beam output intensity and energy parameters, gently regulating the internal physiological state of the batch sugarcane buds, activating the activity of the meristematic tissue, and allowing the large batch of sugarcane buds to simultaneously enter a physiological state suitable for subsequent mutagenesis treatment, ensuring the consistency of the physiological response of the batch sugarcane buds. Simultaneously, various process parameters such as beam dose, beam spacing, beam level matching, and parameter settings were collected during the batch operation, and the original information of the batch treatment was completely preserved to form a primary treatment data reserve, providing detailed basic data for subsequent batch process integration and parameter optimization.

[0048] S300, Sealed Constant Temperature Curing: Sugarcane stalks that have undergone low-volume beam intensification are uniformly stored in a large, sealed curing chamber that undergoes dual disinfection using ultraviolet irradiation and ozone diffusion. This dual disinfection mode thoroughly purifies the internal space and supporting equipment of the chamber, completely eliminating bacteria and external pollutants from the batch processing environment, providing a sterile and clean centralized repair environment for large batches of sugarcane buds. The sealed curing chamber continuously and constantly regulates temperature and humidity, maintaining a stable temperature of 25–30℃ and a constant relative humidity of 75%–85%. The chamber maintains a completely light-proof, sealed structure, and the sugarcane stalks are kept in a uniform, static position inside the chamber. No material movement or human intervention is performed during the curing period, completely isolating external airflow and light. The study aims to mitigate the impact of environmental fluctuations on large-scale sugarcane buds, ensuring they undergo internal physiological repair synchronously in a stable and uniform environment. This allows them to smoothly adapt to the physiological changes brought about by the beam treatment and maintain a consistent metabolic rhythm. A full-time, segmented recording mode is employed to collect complete batch curing duration information, defining time scale nodes for the closed-loop constant-temperature curing stage. This process continuously captures raw information across the entire time dimension, standardizes the time recording criteria and scale specifications for batch curing, and continuously retains the complete time-series trajectory of batch static storage. A corresponding relationship is established between time-series data and the closed-loop static storage stage, comprehensively collecting raw records of the entire batch curing cycle and accumulating basic data for batch environmental control. This provides crucial support for the subsequent batch data integration of the irradiation repair coupling algorithm.

[0049] S400 Gradient Irradiation Mutagenesis: Sugarcane stalks that have undergone batch-wise closed-loop constant-temperature curing strictly adhere to the layout and spacing from the low-volume beam intensification treatment stage, maintaining a uniform beam application reference height. This ensures complete consistency in the position and orientation of the two beam treatments for the batch of sugarcane buds. Utilizing a graded gradient control logic, the heavy ion beam operating parameters are progressively corrected, stabilizing the secondary beam treatment dose within the defined range of 55 Gy to 70 Gy. Simultaneously, the beam output power and particle beam density are coordinated and adjusted to maintain a constant beam vertical incident angle and sugarcane bud target coverage area. This achieves precise secondary beam application to the batch of sugarcane buds, ensuring homologous and in-situ targeting. Through progressive switching of differentiated beam parameters between the two stages, a hierarchical structure is constructed for the batch of sugarcane buds. A gradient irradiation control architecture is adopted to form a segmented, precise beam action mode, which orderly completes the step-by-step superposition of multiple gradient beams. While protecting the activity of batch sugarcane buds, it significantly improves the targeting and efficiency of batch mutagenesis, enabling large batches of sugarcane buds to produce stable and controllable genetic variations. Simultaneously, an irradiation repair coupling algorithm is used to integrate and collect process data from multiple stages of batch treatment, such as seed stalk regularization, low-volume beam intensification, and closed constant temperature curing. Various parameters, environmental data, and time-series data of batch treatment are organically integrated, allowing gradient irradiation parameters to be precisely adapted to the physiological state of batch sugarcane buds, optimizing the accuracy of batch mutagenesis treatment, ensuring the stability and consistency of batch sugarcane bud mutagenesis effects, and meeting the batch mutagenesis needs of large-scale breeding bases.

[0050] S500 Low-Temperature Steady-State Cultivation: Sugarcane stalks that have undergone gradient irradiation mutation in batches are centrally placed in a large, enclosed cultivation space. The temperature, humidity, and ventilation of the cultivation area are controlled in a coordinated manner to create a uniform and stable germination environment for a large number of sugarcane buds. Environmental and periodic data are continuously collected during the batch cultivation stage to comprehensively understand the environmental changes and growth cycle information during the germination process of the batch sugarcane buds. A habitat germination adaptation algorithm is used to match the environmental parameters of the batch cultivation. Combined with the previous batch process data and real-time cultivation data, the key conditions such as temperature, humidity, and ventilation in the cultivation space are dynamically optimized to ensure that the cultivation environment always conforms to the needs of the large batch. To meet the germination needs of large quantities of sugarcane buds, this technology promotes the synchronous and rapid initiation of the germination process for batches of buds, ensuring a highly uniform germination rhythm. Through integrated parameter control of the overall process, the entire process parameters and environmental parameters of batch processing are synergistically optimized, allowing large quantities of sugarcane buds to complete germination in an optimal and uniform habitat. This significantly shortens the batch germination cycle, improves the batch germination rate and uniformity, reduces material loss in large-scale processing, and provides a large amount of uniform and high-quality mutant germination material for large-scale sugarcane breeding bases. This fully releases the large-scale application value of heavy ion beam mutagenesis technology and promotes the efficient and stable development of the sugarcane mutation breeding industry.

[0051] This embodiment is adapted to the mass production needs of large-scale sugarcane breeding bases. It standardizes the entire process for processing large quantities of sugarcane stalks, unifies beam action and environmental control conditions, integrates batch process data with the help of irradiation repair coupling algorithm, and optimizes the cultivation environment through habitat germination adaptation algorithm. This achieves synchronous, rapid, and uniform germination of batch sugarcane buds, with stable mutagenesis effect and low material loss. It perfectly matches the efficiency and quality requirements of large-scale breeding, greatly improves the breeding capacity of the base, and provides solid support for the widespread application of heavy ion beam mutagenesis technology in sugarcane industrial breeding.

[0052] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-disclosed technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A method for inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation, characterized in that, The specific steps of this method are as follows: S100, Homogeneous and Regular Stems: Select the main stems of mature sugarcane plants with uniform growth characteristics, and perform standardized cutting and purification treatment to unify the basic physiological state of the stem segments. S200, Low-dose beam excitation: The homogenized and regularized sugarcane stalks are arranged in the heavy ion beam operation area. The beam current is dynamically matched according to the size of the sugarcane buds using a beam current targeted adaptation and adjustment mechanism. Low-dose heavy ion beams are used to act on the sugarcane bud tissue in the whole area to regulate the internal physiological state of the buds. Various process parameters generated during the operation are collected simultaneously to form a primary processing data reserve. S300, Closed Constant Temperature Curing: For sugarcane stalks after low-volume beam intensification, place them in a closed space after compound disinfection, and constantly regulate the internal temperature and humidity index of the space. Maintain a light-proof and static storage mode throughout the process, continuously collect information on the duration of the static curing period, and accumulate basic data on environmental control. S400, gradient irradiation mutagenesis: Combined with sugarcane stalks after closed constant temperature curing, the heavy ion beam action parameters are progressively adjusted, the beam current action dose is changed to complete the secondary beam current treatment, and the irradiation repair coupling algorithm is used to integrate and collect the multi-stage process data from the previous stage. S500, Low-Temperature Steady-State Cultivation: Sugarcane stem segments after mutagenesis treatment are placed in a closed cultivation space. The temperature, humidity and ventilation of the cultivation area are controlled in a coordinated manner. Environmental data and periodic data are continuously collected during the cultivation stage. The habitat germination adaptation algorithm is used to match environmental parameters and complete the integrated parameter control of the entire process.

2. The method for inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation according to claim 1, characterized in that, In step S100, the standardized cutting is carried out using sterile special cutting tools to perform fixed-length segmentation. The length of the cut sugarcane stalk segments is controlled within the range of 10cm to 15cm. The cutting operation must avoid the buds and surrounding thin-walled tissues to keep the cut surface flat and neat, and each segment of raw material retains the complete sugarcane bud structure. The purification treatment is completed by multi-stage progressive operation. First, the surface solid impurities are removed by rinsing with running clean water. Then, the surface soluble substances are dissolved by static soaking in sterile clean water. Finally, a second rinsing purification is completed using sterile soft water. After the material is purified, it is placed in a light-proof and ventilated area to naturally dissipate the surface free moisture, so as to unify the cutting specifications and standardize the purification process, and constrain the overall shape and surface cleanliness of the material.

3. The method for inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation according to claim 1, characterized in that, In step S200, the beam targeting and adaptation adjustment mechanism divides the beam density control into three levels based on the outline size and development volume of the sugarcane buds: the nascent level, the conventional level, and the mature level. The nascent level corresponds to small-sized newly formed sugarcane buds, with a beam density range of 12–16 pA / cm². The conventional level corresponds to medium-sized sugarcane buds with standard morphology, with a beam density range of 18–22 pA / cm². The mature level corresponds to fully developed large sugarcane buds, with a beam density range of 24–28 pA / cm². Each control level has an independent and fixed beam density range. The corresponding level is matched based on the real-time identified sugarcane bud morphology parameters. The output density per unit area of ​​the heavy ion beam is limited according to the level setting standard, and the beam action boundary is locked according to the actual coverage area of ​​the sugarcane buds.

4. The method for inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation according to claim 1, characterized in that, In step S200, the homogenized and regularized sugarcane stalks are laid out in a single layer in the heavy ion beam operation area, with a uniform interval of 2cm to 3cm between individual stalks. All sugarcane buds are placed facing upwards in a uniform orientation. The horizontal angle of the irradiation platform is adjusted simultaneously to maintain the vertical incidence of the heavy ion beam. In the low-dose heavy ion beam operation stage, the irradiation dose range is set to 0.2Gy to 0.5Gy, and the beam output energy parameters are stabilized. The equipment's full-area scanning mode is used to fully cover the meristematic tissue and surrounding cortical tissue of the sugarcane buds, eliminating local blind spots and irradiation deviations. The ambient temperature in the operation area is kept constant at 20℃ to 25℃ to maintain a continuous and stable beam output intensity. The directional operation is completed in a uniform full-area coverage manner, and the actual operation parameters are recorded simultaneously.

5. The method for inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation according to claim 1, characterized in that, In step S300, the sugarcane stalks after low-volume beam excitation are uniformly collected into a sealed curing chamber after composite disinfection treatment. The chamber uses a dual full-area disinfection mode of ultraviolet irradiation combined with ozone diffusion to complete the full-area purification treatment of the inner wall of the space and the supporting equipment, and remove residual bacteria and external pollutants in the space. The environmental control indicators inside the sealed space are continuously locked to maintain constant range of temperature and humidity values, and the space as a whole maintains a completely light-proof closed structure. The stalks are kept in a static state inside the space, and no material movement or human intervention is performed during the curing period to isolate external airflow, light and environmental fluctuations. The time nodes of the entire static curing cycle are continuously recorded, and the original environmental control data of the curing stage are collected.

6. The method for inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation according to claim 1, characterized in that, In step S300, the duration information is collected in a full-time segmented recording mode, dividing the time scale nodes of the closed constant temperature curing stage, marking the start and end nodes of each interval in sequence, continuously capturing the original information of the entire time dimension, and unifying the collection standards and scale specifications of time recording; the complete time trajectory of static storage is retained without interruption throughout the entire process, the time nodes of the entire curing process are anchored, the corresponding relationship between time data and closed static storage stage is established, and the original records of the entire cycle duration are completely collected.

7. The method for inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation according to claim 1, characterized in that, In step S400, the sugarcane stalks after the sealed constant temperature curing adopt the same layout posture and arrangement spacing as in the low-volume beam intensification treatment stage. The heavy ion beam operation parameters are progressively corrected based on the graded gradient control logic, and a dedicated medium gradient irradiation dose threshold is set. Unlike the low dose range of the initial base beam, the secondary beam treatment adopts a medium dose stable range, and the dose of the secondary beam treatment is stably controlled within the limited range of 55Gy to 70Gy. Synchronous matching of beam output power and particle beam density is used to maintain a constant beam vertical incident angle and sugarcane bud target coverage area. By progressively switching between two segments of differentiated beam parameters, a hierarchical gradient irradiation control architecture is constructed to form a segmented precise beam action mode, and to orderly complete the step-by-step superposition of multi-gradient beams.

8. The method for inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation according to claim 1, characterized in that, In step S400, the mathematical expression of the irradiation repair coupling algorithm is: ； In the formula, This represents the gradient irradiation effect value. The activation coefficient for low-dose pre-irradiation; This is the actual dose of the low-dose pre-irradiation. The synergistic effect coefficient of medium-dose mutagenesis; The actual dose for medium-dose mutagenesis; e is the natural constant; This refers to the actual duration of intermittent recovery maintenance. The duration of physiological repair characteristics of sugarcane buds.

9. The method for inducing rapid germination of sugarcane mutant shoots using heavy ion beam irradiation according to claim 1, characterized in that, In step S500, the mathematical expression of the habitat germination adaptation algorithm is: ； In the formula, G is the co-germination index; This represents the gradient irradiation effect value. The temperature control weighting coefficient is T; T is the actual ambient temperature obtained from monitoring inside the mutation cultivation space. The optimal reference temperature for sugarcane bud germination; H represents the humidity control weighting coefficient; H is the actual relative humidity of the cultivation space as detected in real time. The optimal baseline humidity for sugarcane bud germination; The cultivation duration is the weighting coefficient; t is the actual number of days of mutagenesis and low-temperature cultivation. The maximum control period is preset for the cultivation phase.

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