A method and system for optimizing hormone regulation of narrow row in rice regeneration

Abstract

The application discloses a kind of narrow row hormone regulation optimization methods and systems of regeneration rice, specifically related to rice cultivation technical field, for solving the yield loss problem caused by the different step of wide and narrow row regeneration bud germination in prior art;By determining the anaerobic index of narrow row rhizosphere soil, when exceeding threshold value, screening ACC deaminase-containing microbial active substance;Secondly, by near infrared spectroscopy analysis regeneration bud absorption characteristics, realize dormancy depth grading;Synchronous tracking ethylene concentration from root system to bud body transmission timing determines inhibition window period;Based on dormancy grading and time window, dynamically allocate gibberellin GA3, cell division factor 6-BA and the compounding ratio of microbial active substance;Finally, in time window, directional spray compound solution to narrow row area;By eliminating rhizosphere ethylene accumulation, accurate matching bud body physiological state and action timing, realize the synchronization of narrow row regeneration bud germination efficiency and wide row.

Description

Technical Field

[0001] This invention relates to the field of rice cultivation technology, and more specifically, to a method and system for optimizing hormone regulation in narrow-row regenerated rice. Background Technology

[0002] The wide-narrow row cultivation model for ratooning rice, by altering the field microenvironment to improve light energy utilization, has become an important agronomic measure for increasing yields in the Yangtze River Basin. After the first rice harvest, wide rows, due to their superior ventilation and light penetration, result in earlier and faster regenerated shoot germination and growth; narrow rows, constrained by a shady and humid environment, experience significantly delayed regenerated shoot germination. Current techniques typically employ exogenous spraying of a combination of gibberellin (GA3) and cytokinin (6-BA) to promote regenerated shoot germination in narrow rows, attempting to reduce the growth difference between wide and narrow rows.

[0003] In existing technologies, the difference in rhizosphere microenvironment between wide and narrow rows leads to an imbalance in the efficiency of exogenous hormone response, making it difficult to achieve synchronous growth of regenerated shoots. Narrow rows, due to long-term shading, form a special rhizosphere environment, which causes the accumulation of endogenous inhibitory substances and interferes with the physiological effects of exogenous GA3 and 6-BA. As a result, the sensitivity and metabolic response of regenerated shoots in narrow rows to hormones are significantly weaker than those in wide rows. This environment-dependent effect causes a decrease in the germination-promoting efficiency of the same dose of hormones in narrow rows, widens the difference in the growth rate of regenerated shoots between wide and narrow rows, and ultimately restricts the improvement of tiller uniformity. Summary of the Invention

[0004] In order to overcome the above-mentioned defects of the prior art, the present invention provides a method and system for optimizing hormone regulation in narrow-row regenerated rice to solve the problems mentioned in the background art.

[0005] To achieve the above objectives, the present invention provides the following technical solution:

[0006] A method for optimizing hormone regulation in narrow-row rice regeneration includes the following steps:

[0007] S1. Obtain rhizosphere soil samples from the narrow row area of ​​the regenerated rice and determine the anaerobic index of the rhizosphere soil samples;

[0008] S2. When the anaerobic index exceeds the preset threshold, screen for rhizosphere microbial active substances containing ACC deaminase.

[0009] S3. The spectral absorption characteristics of narrow-row regenerated buds were analyzed by near-infrared spectroscopy, and the dormancy depth was divided into shallow dormancy and deep dormancy based on the absorption threshold.

[0010] S4. Track the transmission time of peak ethylene concentration from roots to shoots in narrow-row areas to determine the time window of ethylene inhibition effect.

[0011] S5. Based on the dormancy depth classification and time window, determine the ratio of gibberellin GA3, cytokinin 6-BA and rhizosphere microbial active substances to form a compound solution.

[0012] S6. Within the time window, apply the compound solution in a targeted manner to the narrow row area.

[0013] Furthermore, rhizosphere soil samples were obtained from the narrow-row area of ​​the ratooning rice, and the anaerobic index of the rhizosphere soil samples was measured, including:

[0014] Rhizosphere soil samples were collected from locations where regenerated shoots were delayed in the narrow-row area.

[0015] Real-time oxygen partial pressure was measured in rhizosphere soil samples from narrow-row areas;

[0016] Oxygen partial pressure was obtained from rhizosphere soil samples from wide-row areas with the same growth period as the baseline value for wide-row planting.

[0017] The anaerobic index is calculated based on the deviation ratio between the real-time oxygen partial pressure of rhizosphere soil samples from narrow-row areas and the baseline value for wide-row areas.

[0018] Furthermore, when the anaerobic index exceeds a preset threshold, rhizosphere microbial bioactive substances containing ACC deaminase are screened, including:

[0019] Determine whether the anaerobic index exceeds a preset threshold;

[0020] When the anaerobic index exceeds the preset threshold, select a Pseudomonas strain that secretes ACC deaminase.

[0021] The ACC deaminase activity of Pseudomonas strains was detected by enzyme-linked immunosorbent assay (ELISA).

[0022] Rhizosphere microbial bioactive substances were prepared by screening strains with ACC deaminase activity reaching the standard value.

[0023] Furthermore, by analyzing the spectral absorption characteristics of narrow-row regenerated buds using near-infrared spectroscopy, dormancy depth was classified into shallow dormancy and deep dormancy levels based on absorption thresholds, including:

[0024] Leaf sheath tissue from the base of narrow-row regenerated buds was collected as a test sample;

[0025] The absorbance at a wavelength of 1450 nm was obtained by scanning the sample using a near-infrared spectrometer.

[0026] The sleep depth is divided into light sleep level and deep sleep level by comparing the absorbance with the preset absorption threshold.

[0027] Furthermore, when the absorbance is less than or equal to the preset absorption threshold, it is determined to be in a light sleep state; when the absorbance is greater than the preset absorption threshold, it is determined to be in a deep sleep state.

[0028] Furthermore, by tracing the timing of the peak ethylene concentration in the narrow-row region from the roots to the shoots, the time window of the ethylene inhibitory effect was determined, including:

[0029] Ethylene gas collection devices are installed at the root system location in the narrow row area;

[0030] Ethylene gas collection devices are simultaneously installed at the base of the narrow row of regenerated buds;

[0031] The ethylene concentration at the root and basal positions was measured at fixed time intervals.

[0032] Record the time difference in which the peak ethylene concentration is transmitted from the root region to the basal region;

[0033] The time window for determining the ethylene inhibition effect is based on the time difference.

[0034] Furthermore, the time window is the period before the peak reaches the base position.

[0035] Furthermore, based on dormancy depth classification and time windows, the ratio of gibberellin GA3, cytokinin 6-BA, and rhizosphere microbial active substances was determined to form a compound solution, including:

[0036] The concentration ratio of gibberellin GA3 to cytokinin 6-BA was adjusted according to the results of the shallow or deep dormancy level in the dormancy depth classification.

[0037] The proportion of rhizosphere microbial active substances added should be adjusted according to the time window stage of the ethylene inhibition effect.

[0038] The adjusted gibberellin GA3, cytokinin 6-BA, and rhizosphere microbial active substances were mixed in a certain proportion to form a compound solution.

[0039] Furthermore, within the time window, the compound solution is sprayed directionally onto the narrow-row area, including:

[0040] Start the spraying device within the time window of the ethylene inhibition effect;

[0041] Position the spraying target area of ​​the spraying device at the location of the regenerated buds above the surface of the narrow row area;

[0042] The compound solution was sprayed in a mist form onto the regenerated buds above the surface of the narrow row area;

[0043] Control the duration of the spraying time window to cover the area.

[0044] On the other hand, the present invention provides a narrow-row hormone regulation and optimization system for regenerated rice, comprising the following modules:

[0045] The rhizosphere anaerobic index measurement module is used to acquire rhizosphere soil samples from narrow-row areas of regenerated rice and to measure the anaerobic index of the rhizosphere soil samples.

[0046] The ACC microbial screening module is used to screen for active substances of rhizosphere microorganisms containing ACC deaminase when the anaerobic index exceeds a preset threshold.

[0047] The dormancy grading module is used to analyze the spectral absorption characteristics of narrow-row regenerated buds through near-infrared spectroscopy, and to classify the dormancy depth into shallow dormancy and deep dormancy based on the absorption threshold.

[0048] The ethylene time window module is used to track the transmission time of the peak ethylene concentration from the root to the shoot in narrow-row areas and determine the time window of the ethylene inhibitory effect.

[0049] The compound solution preparation module is used to determine the ratio of gibberellin GA3, cytokinin 6-BA and rhizosphere microbial active substances based on dormancy depth classification and time window to form a compound solution;

[0050] The targeted spraying module is used to apply the compound solution to a narrow row area within a time window.

[0051] Compared with the prior art, the present invention has the following beneficial effects:

[0052] 1. This invention solves the problem of asynchronous germination of regenerated buds in narrow and wide rows by establishing a synergistic diagnostic mechanism between the narrow-row microenvironment and the dormancy state of regenerated buds. Targeting the ethylene inhibition effect caused by the anaerobic rhizosphere environment in narrow rows, it introduces ACC deaminase active substances to directionally degrade ethylene synthesis precursors, significantly weakening the interference of endogenous inhibitory substances on hormone signals. Combined with near-infrared spectroscopy to quantify the dynamic coupling relationship between dormancy depth and the timing of ethylene transfer, it achieves precise capture of the window of action of exogenous hormones. This three-level regulatory system of "microenvironment repair - dormancy state analysis - timing-targeted intervention" enhances the physiological efficacy of gibberellin and cytokinin in narrow rows to a level comparable to that in wide rows, ensuring that the uniformity of regenerated bud germination meets the requirements of synchronous growth.

[0053] 2. By using a collaborative decision-making mechanism based on dormancy depth grading and time window, the inherent limitations of traditional uniform pesticide application are overcome. Based on the dynamic adjustment of hormone ratios between light and deep dormancy levels, precise matching between exogenous substances and the physiological state of the buds is achieved. Furthermore, the targeted spraying controlled by the time window enables rhizosphere microbial active substances to efficiently block the transmission of inhibitory chains before the peak ethylene concentration. This compounding scheme, which uses the dormancy state of the buds as the core of regulation and the ethylene transmission sequence as the axis of action, not only significantly improves the sensitivity of narrow-row regenerated buds to hormone responses but also fundamentally eliminates the differences in metabolic responses between wide and narrow rows, providing a core guarantee for uniform tillering in regenerated rice. Attached Figure Description

[0054] Figure 1 This is a flowchart of a method for optimizing hormone regulation in narrow-row regenerated rice according to the present invention;

[0055] Figure 2 This is a schematic diagram of the structure of a narrow-row hormone regulation and optimization system for regenerated rice according to the present invention. Detailed Implementation

[0056] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0057] Example 1: Figure 1 This invention provides a method for optimizing hormone regulation in narrow-row regenerated rice, comprising the following steps:

[0058] S1. Obtain rhizosphere soil samples from the narrow row area of ​​the regenerated rice and determine the anaerobic index of the rhizosphere soil samples;

[0059] S2. When the anaerobic index exceeds the preset threshold, screen for rhizosphere microbial active substances containing ACC deaminase.

[0060] S3. The spectral absorption characteristics of narrow-row regenerated buds were analyzed by near-infrared spectroscopy, and the dormancy depth was divided into shallow dormancy and deep dormancy based on the absorption threshold.

[0061] S4. Track the transmission time of peak ethylene concentration from roots to shoots in narrow-row areas to determine the time window of ethylene inhibition effect.

[0062] S5. Based on the dormancy depth classification and time window, determine the ratio of gibberellin GA3, cytokinin 6-BA and rhizosphere microbial active substances to form a compound solution.

[0063] S6. Within the time window, apply the compound solution in a targeted manner to the narrow row area.

[0064] S1. Obtain rhizosphere soil samples from the narrow-row area of ​​the ratooning rice and determine the anaerobic index of the rhizosphere soil samples. The specific implementation is as follows:

[0065] The location of delayed germination of regenerated shoots was determined by the following criteria: the measured length of regenerated shoots in the narrow row was less than or equal to 50% of the measured length of regenerated shoots in the wide row area at the same time. Specifically, at 9:00 AM on the third day after rice harvest, the length of regenerated shoots in the narrow row area was measured using a digital caliper. If the length of a regenerated shoot at a certain point did not exceed 50% of the average length of five randomly selected regenerated shoots from the adjacent wide row area, it was determined to be a location of delayed germination. At this location, a 2 cm inner diameter oxygen-free stainless steel soil drill was used to vertically drill a soil sample within 5 cm of the base of the rice main stem. The sampling depth was from the surface to a soil layer of 20 cm, retaining soil particles attached to the root surface as rhizosphere soil samples. The sample weight at a single point was no less than 200 grams. Immediately after sampling, the samples were placed in a pre-filled nitrogen-sealed bag and stored at 4 degrees Celsius.

[0066] To determine the real-time oxygen partial pressure of rhizosphere soil samples from narrow-row areas, a portable soil gas analyzer was used. Before testing, the instrument underwent two-point calibration: first, zero-point calibration in a 99.99% high-purity nitrogen environment, and second, full-scale calibration in standard air with an oxygen concentration of 20.9%. The soil samples were transferred to a 25°C constant-temperature testing chamber. After temperature equilibrium, the zirconia solid electrolyte oxygen electrode sensor was inserted 2 cm below the center of the sample. Data acquisition intervals were set to 10 seconds. After discarding the data from the first 30 seconds, the arithmetic mean of the stable readings over the subsequent 90 seconds was taken as the real-time oxygen partial pressure value, expressed in kPa. Each sample was measured three times; if the range of the three measurements exceeded 1 kPa, a retest was required.

[0067] When obtaining the oxygen partial pressure (OPP) of rhizosphere soil samples from wide-row areas with the same growth period as the baseline value for wide-row areas, "same growth period" refers to the same number of days after rice harvest and the same period of sunlight. Sampling points are set up along the centerline of adjacent wide-row areas, with a straight-line distance not exceeding 5 meters. Oxygen POP data are obtained using the exact same sampling tools and testing procedures as in the narrow-row areas. Five sampling points are selected for each field. If the coefficient of variation of the oxygen POP values ​​at the five points exceeds 15%, additional sampling is conducted to bring the total to eight points, and the results are recalculated. The arithmetic mean of all valid values ​​is then taken as the baseline value for wide-row areas.

[0068] When calculating the anaerobic index based on the deviation ratio between the real-time oxygen partial pressure of rhizosphere soil samples from narrow-row areas and the baseline value for wide-row areas, the following steps are performed: First, calculate the difference between the baseline value for wide-row areas and the real-time oxygen partial pressure for narrow-row areas; second, divide this difference by the baseline value for wide-row areas to obtain a proportionality coefficient; finally, multiply the proportionality coefficient by 100% to convert it into a percentage value. The specific calculation formula is: Anaerobic Index = [(Width-row baseline value - Narrow-row real-time oxygen partial pressure) ÷ Width-row baseline value] × 100%. A soil texture correction coefficient K is introduced in the calculation process, where 0.95 is used for sandy soil, 1.05 for clay soil, and 1.00 for loam. The corrected formula is: Anaerobic Index = [(Width-row baseline value - Narrow-row real-time oxygen partial pressure) × K ÷ Width-row baseline value] × 100%. A dynamic adjustment mechanism is used for the preset threshold. For example, the threshold is 30% when the average daily temperature is below 20 degrees Celsius, 40% when the average daily temperature is above 28 degrees Celsius, and 35% under other temperature conditions. When the baseline value for wide-row conditions is below 8 kPa, the field is determined to be in a state of hypoxia and proceeds directly to the next step.

[0069] S2. When the anaerobic index exceeds a preset threshold, rhizosphere microbial active substances containing ACC deaminase are screened. The specific implementation is as follows:

[0070] When determining whether the anaerobic index exceeds a preset threshold, the preset threshold is dynamically adjusted based on the ambient temperature. When the average daily ambient temperature is below 20 degrees Celsius, the preset threshold is set to 30%; when the average daily ambient temperature is above 28 degrees Celsius, the preset threshold is set to 40%; and under other temperature conditions, the preset threshold is set to 35%. The determination process is achieved by comparing the anaerobic index value with the preset threshold for the corresponding temperature range. When the anaerobic index is greater than the preset threshold, it is determined that the preset threshold has been exceeded. The anaerobic index is derived from the calculation results of step S1. Each determination requires recording the timestamp, temperature value, and corresponding preset threshold parameter. The determination process for the anaerobic index exceeding the preset threshold includes a data verification mechanism: when the anaerobic index exceeds the preset threshold, the original oxygen partial pressure data from step S1 needs to be reviewed. The review includes whether the range of the three replicate values ​​of the narrow-row real-time oxygen partial pressure detection is less than 1 kPa, and whether the coefficient of variation of the wide-row baseline value is less than 15%. If any review condition is not met, the sample is re-collected for testing. The temperature correlation of the preset threshold was determined through field trials. For example, in a three-year trial, it was found that the anaerobic index was more than 30% under low temperature conditions, and the inhibition rate of regenerated shoots reached 92% under high temperature conditions, and more than 40% under high temperature conditions, the inhibition rate reached 89%.

[0071] When the anaerobic index exceeds a preset threshold, *Pseudomonas* strains secreting ACC deaminase are selected. *Pseudomonas* strains are isolated from rhizosphere soil samples from the target field. The specific procedure is as follows: 10 g of rhizosphere soil sample collected in step S1 is added to 90 mL of sterile physiological saline and suspended by shaking for 30 minutes. The supernatant is then serially diluted to concentrations of 10^-3, 10^-4, and 10^-5. 0.1 mL of each dilution is plated onto ACC screening plates. The plate composition is: 10 g tryptone, 5 g yeast extract, 10 g sodium chloride, and 15 g agar per liter, with 3 mmol of ACC added as the sole nitrogen source. After incubation at 28°C for 48 hours, single colonies with a diameter greater than 2 mm are selected and verified as *Pseudomonas* strains by Gram staining and oxidase assays for preservation. A strain traceability mechanism is established for the selection of *Pseudomonas* strains: the source field, sampling location coordinates, and isolation time of each isolated strain are recorded. The ACC concentration in the ACC screening plates was optimized through preliminary experiments. For example, concentration gradient experiments showed that 3 mmol / L ACC effectively inhibited the growth of non-target bacteria while ensuring normal proliferation of Pseudomonas. Colony morphology was observed daily during plate culture; typical characteristics of Pseudomonas include round colonies with neat edges and moist surfaces. Gram staining verification was performed using crystal violet staining solution; a positive result indicated red bacilli. The oxidase test used tetramethyl-p-phenylenediamine (TMP) test paper; a positive result was indicated by a deep blue color change within 30 seconds.

[0072] When detecting the ACC deaminase activity of *Pseudomonas* strains using enzyme-linked immunosorbent assay (ELISA), a cell lysis buffer was first prepared: The preserved *Pseudomonas* strain was inoculated into LB liquid medium and cultured at 28°C with shaking at 180 rpm for 24 hours. 10 mL of the culture was collected by centrifugation at 4°C for 10 minutes at 10,000 rpm. The cells were washed twice with 0.1 mol / L Tris-HCl buffer and resuspended in 1 mL of the same buffer containing 0.5 mmol / L dithiothreitol. The cells were then sonicated at 300 W for 10 seconds with a 20-second interval, for a total of 5 minutes. The lysate was centrifuged at 4°C for 20 minutes at 12,000 rpm, and the supernatant was used as the enzyme extraction buffer. Enzyme-linked immunosorbent assay (ELISA) was performed using a 96-well microplate. Each well contained 50 μL of enzyme extraction buffer and 150 μL of reaction mixture, which included 0.2 mol / L Tris-HCl buffer, 10 mM ACC substrate, and 0.2 mM pyridoxal phosphate. After reacting at 30°C for 30 minutes, 100 μL of stop solution was added to terminate the reaction. The stop solution was 0.5 mol / L citrate buffer containing 1 mg / mL o-phenylenediamine and 0.3% hydrogen peroxide. The absorbance was measured at 490 nm using a microplate reader. The ACC deaminase activity units were calculated based on the standard curve. One activity unit was defined as the amount of enzyme that catalyzes the production of 1 μmol of α-ketobutyrate per minute. The standardized procedure for ELISA included setting up control wells for each assay. The standard was purified ACC deaminase solution with concentration gradients of 0, 5, 10, 15, and 20 activity units per milligram of protein. A correlation coefficient greater than 0.99 was required for the standard curve to be valid. Sample testing is performed in triplicate; if the absorbance difference between duplicates exceeds 15%, the test must be repeated. The reaction temperature is controlled with an accuracy of ±0.5 degrees Celsius, and a constant-temperature water bath is used to maintain temperature stability. The color development time is strictly controlled to 10 minutes; exceeding this time will result in increased background values.

[0073] When preparing rhizosphere microbial bioactive substances, strains with ACC deaminase activity meeting the standard value were selected, with the standard value set at 15 activity units per milligram of protein. The selection process included: calculating the specific activity of ACC deaminase for each strain, which was equal to the total activity units divided by the protein concentration; the protein concentration was determined using the Coomassie brilliant blue method, with bovine serum albumin as the standard protein to create a standard curve. Strains with an ACC deaminase specific activity greater than or equal to 15 activity units per milligram of protein were considered to have met the standard. For the preparation of rhizosphere microbial bioactive substances, the qualified strains were inoculated into a fermentation medium containing 10 g glucose, 5 g peptone, 1 g dipotassium hydrogen phosphate, and 0.5 g magnesium sulfate per liter. Fermentation was carried out at 28°C and an aeration rate of 1 v / v for 48 hours. After fermentation, the cells were collected by centrifugation at 8000 rpm for 15 minutes at 4°C. After washing the bacterial cells twice with sterile physiological saline, they were resuspended in a protective solution containing 10% glycerol. The cell concentration was adjusted to 10⁹ viable cells per milliliter, aliquoted, and frozen to obtain the rhizosphere microbial active substance. Quality control was implemented throughout the preparation process of the rhizosphere microbial active substance: the cell concentration at the end of fermentation must reach at least 10⁹ viable cells per milliliter, and the viability rate was tested using the plate count method, requiring a viability rate greater than 95%. The protective solution consisted of 100g sucrose, 10g monosodium glutamate, 5g ascorbic acid, and 10ml glycerol per liter. Cryopreservation employed a programmed cooling method: first, the temperature was lowered to 4°C at a rate of 1°C per minute and maintained for 2 hours; then, it was lowered to -20°C at a rate of 1°C per minute and maintained for 4 hours; finally, it was transferred to -80°C for long-term storage. The residual ACC deaminase activity of each batch of product was tested, requiring the activity to remain above 90% of the initial activity after three months of frozen storage.

[0074] S3. Near-infrared spectroscopy analysis of the spectral absorption characteristics of narrow-row regenerated buds is used to classify dormancy depth into shallow dormancy and deep dormancy levels based on absorption thresholds. The specific implementation is as follows:

[0075] When collecting leaf sheath tissue from the base of regenerated buds in narrow rows as test samples, the basal leaf sheath tissue is defined as a segment of leaf sheath no more than 3 cm above the ground. The specific procedure is as follows: On the 5th day after rice harvest, between 10:00 AM and 2:00 PM, select plants within the narrow row area whose regenerated bud length is less than or equal to 60% of the corresponding regenerated bud length in the wide row area, and use sterile surgical scissors to cut the basal leaf sheath tissue. The sampling location is a segment 0.5 cm to 1.5 cm from the base of the regenerated bud, and the sample size is a tissue block 1 cm long and 0.5 cm wide. Immediately after sampling, place the tissue block in a pre-cooled vacuum drying bottle, store it at -20 degrees Celsius, and transport it to the laboratory. The time interval from collection to testing should not exceed 24 hours. Spatial positioning control is implemented for the collection of basal leaf sheath tissue: sampling points are arranged at a density of 1 point per square meter within the narrow row area, and 3 representative plants are selected at each sampling point. When harvesting basal leaf sheath tissue, avoid areas of mechanical damage. Disinfect sampling tools by wiping with 75% ethanol after every five samples. Preserve samples in brown, light-proof bottles filled with silica gel desiccant to maintain a relative humidity below 10%. Maintain a temperature of -20°C ± 2°C during transport; samples must be discarded if temperature fluctuations exceed 3°C.

[0076] When using a near-infrared spectrometer to scan and obtain absorbance at 1450 nm, the samples are pretreated before detection: frozen tissue blocks are thawed at 4°C for 2 hours, and then surface moisture is absorbed with filter paper. The treated tissue blocks are then laid flat on a quartz sample dish, ensuring uniform tissue thickness within the range of 2 mm to 3 mm. After the near-infrared spectrometer has been powered on and warmed up for 30 minutes, baseline calibration is performed: a standard white plate is used as the 100% reflectance reference, and a light-shielding plate is used as the 0% reflectance reference. Scanning parameters are set as follows: wavelength range 1300 nm to 1600 nm, resolution 4 nm, and 32 scans with the average value taken. During detection, the laboratory temperature is maintained at 25°C ± 1°C, and the relative humidity is 40% ± 5%. The sample dish is placed in the fixed position in the sample cell, and the scanning program is started to record the characteristic absorbance value at 1450 nm wavelength. The absorbance value is retained to three decimal places. Each sample is scanned three times. If the range of the three measurements exceeds 0.01, the test must be repeated. Introducing in-situ in vivo measurement mode for spectral detection: a non-destructive testing method was adopted for some samples. After the plant to be tested was placed in a dark room for 30 minutes to acclimatize, a fiber optic probe was used to directly contact the surface of the basal leaf sheath tissue. A constant pressure of 50 grams was applied to the probe to ensure tight contact. Before measurement, a refractive index matching solution was applied to the probe contact surface to eliminate interface reflection. The number of scans was increased to 64 during in-situ measurement, and the spectral data were processed by a five-point, three-stage smoothing filter. The absorbance value at 1450 nm was adjusted according to the temperature correction formula: the corrected absorbance equals the difference between the measured value multiplied by 1 and 0.003 multiplied by 25 minus the ambient temperature T, where T is the temperature value in Celsius.

[0077] When comparing absorbance with a preset absorption threshold, the preset absorption threshold employs a dynamic setting mechanism. The base threshold is set at 0.85, a value derived from a three-year field trial: statistics show that samples with absorbance less than or equal to 0.85 achieved a germination rate of over 95%, while samples with absorbance greater than 0.85 had a germination rate of less than 30%. The dynamic adjustment rule is as follows: when the average daily ambient temperature is below 18 degrees Celsius, the threshold is lowered by 0.05; when the average daily ambient temperature is above 30 degrees Celsius, the threshold is raised by 0.05. The comparison operation is performed using data processing software: inputting the sample absorbance measurement value and the current ambient temperature value, the system automatically matches the preset absorption threshold for the corresponding temperature range for numerical comparison. The temperature compensation mechanism for the preset absorption threshold is established using a mathematical model: the temperature compensation coefficient Wp equals 0.002 multiplied by the difference between the average daily ambient temperature T and 24, where T is the temperature value in Celsius. The corrected threshold equals the base threshold plus Wp. For example, when the average daily ambient temperature is 15 degrees Celsius, Wp equals 0.002 multiplied by 15 minus 24, which is -0.018. The correction threshold is 0.85 minus 0.018, which equals 0.832. When the average daily ambient temperature is 32 degrees Celsius, Wp equals 0.002 multiplied by 32 minus 24, which is 0.016. The correction threshold is 0.85 plus 0.016, which equals 0.866. The verification method for the basic threshold of 0.85 is as follows: 500 samples are collected for three consecutive years, and the absorbance is measured and the germination of regenerated shoots is tracked. Statistical analysis shows that the germination success rate of samples with absorbance less than or equal to 0.85 is, for example, 96.2%, and the germination success rate of samples with absorbance greater than 0.85 is, for example, 28.7%. Variety-specific correction of the preset absorption threshold: A threshold library is established for different rice varieties. For example, the basic threshold for variety A is 0.82, and the temperature coefficient is 0.0018; the basic threshold for variety B is 0.88, and the temperature coefficient is 0.0022.

[0078] When the absorbance is less than or equal to a preset absorption threshold, the sample is classified as being in a state of light dormancy. The criteria for light dormancy include: the measured absorbance value not exceeding the preset absorption threshold corrected for the current temperature. The determination process records three parameters: ambient temperature, measured absorbance, and the correction threshold, generating a determination report for archiving. The physiological characteristics of regenerated buds corresponding to light dormant samples are: pale green bud scales, a basal internode cell division activity index higher than 0.7, and an endogenous gibberellin concentration greater than 1.2 ng / mg fresh weight. Additional verification indicators for light dormancy determination: 20% of the samples determined to be in light dormancy are randomly selected for microscopic observation. Paraffin sections are prepared from basal leaf sheath tissue, stained, and the vascular bundle structure is observed under a 400x microscope. Light dormant samples should possess the following characteristics: sieve tube cell wall thickness less than 2 μm, companion cell mitochondrial density greater than 15 cells / μm², and vacuole degree of phloem parenchyma cells less than 30%. If the microscopic characteristics do not meet the requirements, the determination is invalid, and resampling is required. A rapid screening method for shallow-dormant samples: Samples identified as shallow-dormant are immediately subjected to germination potential testing. Basal leaf sheath tissue is immersed in a 10 mg / L gibberellin solution and incubated in the dark at 28°C for 48 hours. After incubation, bud elongation is measured. Elongation exceeding 2 mm confirms true shallow dormancy; elongation less than 0.5 mm necessitates a correction to deep dormancy.

[0079] When the absorbance exceeds a preset absorption threshold, the sample is classified as being in deep dormancy. The criteria for deep dormancy are: the absorbance measurement value exceeds the preset absorption threshold corrected for the current temperature. The determination result is correlated with the physiological state of the sample: bud scales of deep dormant samples are yellowish-brown, the basal internode cell division activity index is below 0.3, and the endogenous abscisic acid concentration is above 5.6 ng / mg fresh weight. All determination results require verification of measurement data. If any value in three scans does not conform to the determination conclusion, resampling and testing are required. Emergency handling procedure for deep dormancy determination: When the proportion of deep dormant samples exceeds 70% of the total sample size, the whole-field early warning mechanism is activated. After the early warning is triggered, three backup sample points must be retested within 24 hours, with the retest points located at least 5 meters apart from the initial test points. If the retest results still show that the proportion of deep dormant samples exceeds the standard, subsequent control steps will be automatically executed ahead of schedule. Preservation and retesting of deep dormant samples: Samples determined to be in deep dormancy are divided into two portions; one portion is immediately tested for endogenous hormones, and the other portion is stored in a liquid nitrogen container. The absorbance of the preserved sample was retested on the 5th day after the control measures were implemented. When the retest value decreased by more than 15% compared with the initial value, the dormancy depth classification of the plant was updated to the shallow dormancy level.

[0080] Optimization of the time window for sample collection: During continuous rainy weather, the sampling time is adjusted to 4 to 8 hours after the rain stops. Sampling personnel wear anti-static protective clothing and use ceramic scissors to avoid metal ion interference. The sample bottle is filled with high-purity nitrogen to displace air, and the bottle mouth is sealed with a screw cap with a polytetrafluoroethylene gasket. The sample transport box is equipped with a temperature recorder. After arriving at the laboratory, check the temperature curve. If the duration of maintaining -20 degrees Celsius is less than 90% of the total transport duration, the sample is discarded. Variety identification is assisted by molecular markers: Leaf DNA is collected synchronously during sampling, the OsS1 gene fragment is amplified by PCR, and the variety database is matched according to the sequencing results. When detecting new varieties, the default threshold of 0.85 is adopted and the learning mode is started: Continuously track the relationship between the absorbance and germination rate of 50 samples, automatically calculate the optimal threshold and enter it into the database. All judgment data is uploaded to the central database to generate a spatial distribution heat map to guide precise operations. All judgment change records timestamp and environmental parameters to form a dormancy state migration map for model optimization. The test results are fed back to the threshold optimization system to dynamically calibrate the judgment model.

[0081] S4. Trace the transfer timing of the ethylene concentration peak from the root system to the bud in the narrow row area to determine the time window of the ethylene inhibitory effect. The specific implementation is as follows:

[0082] When arranging the ethylene gas collection device at the root position in the narrow row area, the root position is defined as the soil layer area 10 cm horizontally and 15 - 25 cm vertically deep from the base of the main rice stem. The specific arrangement operation is as follows: Use a stainless steel probe tube with a diameter of 5 mm to vertically insert into the soil to a depth of 20 cm, and connect the end of the probe tube to the gas collection chamber. The gas collection chamber is made of polytetrafluoroethylene material with a volume of 5 ml, and the adsorbent inside is activated alumina particles with a particle size of 1 - 2 mm. Ensure that the collection chamber is completely buried in the soil during arrangement, with the pipe mouth 2 cm above the ground and equipped with a dust cap. Three gas collection devices are arranged in each narrow row area in a triangular distribution, with a device spacing of 50 cm. After the arrangement, let it stand for 24 hours to balance the device with the environment, and keep the normal water level in the field during this period.

[0083] When arranging the ethylene gas collection device synchronously at the base position of the narrow row regenerated bud, the base position is defined as the internode area at the base of the regenerated bud 2 - 3 cm above the ground surface. The specific operation is as follows: Use a special fixing clip to fit the micro - collection cover on the surface of the base internode. The material of the collection cover is a breathable silicone membrane with an effective adsorption area of 0.5 square centimeters. The back end of the collection cover is connected to a polyether - ether - ketone catheter with an inner diameter of 1 mm, and the catheter extends to the ground and is connected to a vacuum pump. Two collection devices are arranged at each monitoring point, located on both sides of the internode respectively, with a device spacing of 1 cm. Keep the arrangement time synchronized with the device at the root position, with a time difference not exceeding 5 minutes. Check the airtightness after arrangement: Apply a positive pressure test of 0.1 MPa, and if the pressure drop does not exceed 5% within 1 minute, it is qualified.

[0084] When determining the ethylene concentration at the root and basal locations at fixed time intervals, the fixed time interval is set to 30 minutes. The measurement procedure includes: first, extracting gas from the gas collection chamber at the root location; then, using a 50 mL gas-tight syringe, extracting 10 mL of gas at a flow rate of 5 mL / min, and immediately injecting it into the gas chromatograph injection port. The gas chromatograph detection conditions are: a capillary column of 30 m in length and 0.32 mm in inner diameter; high-purity nitrogen as the carrier gas at a flow rate of 1.5 mL / min; a flame ionization detector at a temperature of 200°C; and a column oven temperature program that starts at 40°C and holds for 2 minutes, then increases to 100°C at a rate of 10°C / min. The same method is used to measure the gas at the basal location. Before each measurement, the instrument is calibrated with standard ethylene gas: the standard gas concentration gradient is 0.1, 0.5, and 1.0 μL / L, and the correlation coefficient of the standard curve is required to be greater than 0.995. Each measurement is repeated three times, and the arithmetic mean is taken as the ethylene concentration value at that time point, expressed in μL / L.

[0085] When recording the time difference of ethylene concentration peak transmission from the root system to the base, the ethylene concentration peak is defined as the highest ethylene concentration among three consecutive measurement time points. The identification method is as follows: plot the ethylene concentration change curve at the root system location over time, find the maximum concentration and the corresponding time point T1; similarly, find the maximum concentration at the base and the corresponding time point T2. The time difference Δt is calculated as the absolute value of T2 minus T1, in minutes. When the peak at the root system location is not obvious (the difference between the highest and second-highest values ​​is less than 10%), the midpoint of the period with the highest average concentration among three consecutive time points is taken as T1; T2 is determined similarly at the base location. The time difference is recorded accurate to 0.5 minutes, and weather conditions are noted: for sunny days, record the actual solar radiation intensity; for rainy days, record the start and end times of rainfall.

[0086] When determining the time window for the ethylene inhibition effect based on time difference as the period before the peak reaches the basal position, the time window is defined as the period of time Δt calculated backward from T2. For example, when Δt is 120 minutes, the time window is from 120 minutes before T2 to T2. Special case handling rules: when Δt is less than 60 minutes, the time window is expanded to 90 minutes before T2 to 30 minutes after T2; when Δt is greater than 180 minutes, the time window is compressed to 150 minutes before T2 to 30 minutes before T2. The boundary times of the time window are converted to local time and associated with GPS positioning coordinates. The final output is the start time Ts and end time Te of the time window, labeled to the minute accuracy, and the average soil temperature, humidity, and light intensity parameters within this period are recorded.

[0087] Maintenance procedures for the ethylene gas collection device: Replace the adsorbent every 24 hours, purging the pipeline with nitrogen for 3 minutes before replacement. Replace the miniature collection hood every 8 hours; clean the old hood ultrasonically with methanol and dry it for later use. Inject standard gas to verify the gas chromatograph after every 10 sample measurements; recalibrate if drift exceeds 5%. Verification method for time difference: Within the time window, measure the ethylene concentration at the base position every 15 minutes to verify if the concentration shows a monotonically increasing trend. If a decreasing inflection point is observed, adjust the time window termination point to the inflection point time. All data recording tables include: device number, deployment time, measurement time, concentration value, peak time, time difference, and window period, forming an electronic tracking log.

[0088] Selection criteria for root placement points: Prioritize placement in areas where the anaerobic index exceeds the limit as determined in step S1. When the length of the narrow row exceeds 5 meters, add one monitoring unit for every additional 2 meters. Adsorbent pretreatment method: Calcine activated alumina particles in a muffle furnace at 550°C for 4 hours, cool, then soak in a 0.1 mol / L hydrochloric acid solution for 12 hours, rinse with deionized water until neutral, and dry at 120°C for later use. The probe insertion angle should be kept vertical, with an inclination error not exceeding 3 degrees. The device should be placed away from areas with dense plant roots, selecting areas with fewer lateral roots.

[0089] Microenvironment control at the base: Apply silicone grease sealing rings around the collection hood to prevent gas leakage. Maintain clamp pressure between 50 and 80 grams to avoid tissue deformation. Set the vacuum pump flow rate to 0.2 ml / min for 10 minutes. Allow 20 minutes for equilibration after each vacuuming before the next collection. A built-in temperature sensor in the synchronous monitoring device records real-time temperature changes at the contact surface; temperature data is used for subsequent concentration correction.

[0090] Quality control for ethylene concentration determination: Gas samples are dehydrated in a cold trap before injection at a temperature of -20°C. Chromatographic peak identification criteria: Peak height greater than three times the baseline noise, peak shape symmetry factor between 0.8 and 1.2. Concentration calculation uses the external standard method: Sample peak area is compared with the standard curve, retaining three significant figures. Resampling and determination are performed when the relative deviation of parallel samples exceeds 8%. Each batch of determination includes a blank control: Background values ​​are measured using ethylene-free air; if the background value exceeds 0.05 μL / L, the instrument is stopped to investigate the source of contamination.

[0091] Statistical algorithm for peak propagation time difference: When multiple monitoring units exist, the median of Δt for each unit is taken as the final value. Outliers are removed before median calculation: If the Δt of a unit deviates from the median by more than 30%, and the peak concentration of that unit is less than 70% of the average, it is considered invalid data. Correlation between time difference Δt and soil physical properties: A correction factor is added according to soil type: clay is multiplied by 1.1, sandy soil by 0.9, and loam remains unchanged. The corrected time difference is used for the final time window calculation.

[0092] Validation of the time window's applicability: Within the time window period, samples were taken every 30 minutes to measure the expression level of ethylene receptor protein in the basal internodes. Western blotting was used, employing an anti-ethylene receptor ETR1 antibody for detection. The time window was considered valid when the expression level increased by more than 50%; if the increase was less than 20%, the peak transfer process was re-tracked. The time window output format was: start time (hour:minute) to end time (hour:minute), accompanied by a confidence rating: A (validated), B (partially validated), C (unvalidated and requiring review). All time window data were associated with the field number, monitoring date, and meteorological station code, and uploaded to the central database to generate a spatiotemporal distribution map.

[0093] Gas collection device recovery and disinfection: After monitoring, the stainless steel probe is soaked in 0.5% sodium hypochlorite solution for 30 minutes, rinsed with deionized water and dried. The silicone collection cover is ultrasonically cleaned with ultrapure water for 15 minutes, dried at 60 degrees Celsius and stored. The connecting pipeline is purged with high-purity nitrogen for 10 minutes and then sealed and stored. The data storage medium is waterproof and antimagnetic sealed and has a shelf life of no less than three years. Within 12 hours after the time window is determined, fluorescent markers are placed in the field, displaying the start and end times of the window period with a font height of 5 cm, ensuring a visibility distance of more than 10 meters.

[0094] S5. Based on dormancy depth classification and time window, determine the ratio of gibberellin GA3, cytokinin 6-BA, and rhizosphere microbial active substances to form a compound solution. The specific implementation is as follows:

[0095] When adjusting the concentration ratio of gibberellin GA3 to cytokinin 6-BA based on the dormancy depth classification (light or deep dormancy), the dormancy depth classification result is derived from the judgment conclusion of step S3. When the dormancy depth classification is light dormancy, the concentration ratio of gibberellin GA3 to cytokinin 6-BA is set at 3:1. Specifically, the gibberellin GA3 stock solution concentration is 100 mg / L, and the cytokinin 6-BA stock solution concentration is 50 mg / L. 75 ml of the gibberellin GA3 stock solution and 25 ml of the cytokinin 6-BA stock solution are mixed to form a mixed hormone solution with a total concentration of 85 mg / L. When the dormancy depth classification is deep dormancy, the concentration ratio of gibberellin GA3 to cytokinin 6-BA is adjusted to 5:1. Specifically, 83.3 ml of the gibberellin GA3 stock solution and 16.7 ml of the cytokinin 6-BA stock solution are mixed to form a mixed hormone solution with a total concentration of 90 mg / L. The ratio adjustment was based on the following: Pot experiments showed that lightly dormant buds are more sensitive to cytokinin, hence the increased 6-BA ratio; deeply dormant buds required a higher gibberellin concentration to break dormancy. Verification of the ratio setting: 48 hours after adjustment, samples were taken to measure the bud cell division index; the lightly dormant combination needed an index above 0.8, and the deeply dormant combination needed an index above 0.6.

[0096] When adjusting the addition ratio of rhizosphere microbial active substances according to the time window stage of the ethylene inhibition effect, the time window stage is derived from the start time Ts to the end time Te determined in step S4. The time window is divided into three periods: early stage (Ts to Ts+1 / 3(Te-Ts)), middle stage (Ts+1 / 3(Te-Ts) to Ts+2 / 3(Te-Ts)), and late stage (Ts+2 / 3(Te-Ts) to Te). The addition ratio in the early stage is 5% of the mixed hormone solution volume; the addition ratio in the middle stage is 10%; and the addition ratio in the late stage is 15%. The rhizosphere microbial active substances are derived from the bacterial suspension prepared in step S2, which is brought to room temperature and shaken to mix before use. The addition ratio is set based on the following: in the early stage of the time window, the ethylene concentration is low, and a small amount of microorganisms can degrade ACC; in the late stage, the ethylene accumulation is large, and the microbial dosage needs to be increased. The ratio verification method is to sample and measure the rhizosphere ACC content at each time point of the time window, requiring an ACC degradation rate of more than 70% within 24 hours after addition.

[0097] When mixing the adjusted gibberellin GA3, cytokinin 6-BA, and rhizosphere microbial active substances to form a compound solution, the mixing operation should be carried out in a light-proof container. The mixing sequence is as follows: first, place the mixed hormone solution in a 5-liter stirred tank and start the magnetic stirrer at 300 rpm; then, slowly add the rhizosphere microbial active substances at a rate of 50 mL per minute; finally, add deionized water to bring the total volume to a final volume. Maintain the mixing temperature at 20°C ± 2°C and mix for 30 minutes. The final composition of the compound solution should be controlled as follows: gibberellin GA3 concentration range of 50 to 70 mg / L, cytokinin 6-BA concentration range of 10 to 15 mg / L, and rhizosphere microbial active substances containing 10^7 to 10^8 viable bacteria per mL. Immediately after mixing, test the pH of the solution and adjust it to 6.0 to 6.5 using 0.1 mol / L hydrochloric acid or sodium hydroxide. Shelf life control of compound solutions: Store at 20 degrees Celsius in the dark for no more than 6 hours. If the shelf life is exceeded, the solution must be prepared again.

[0098] Additional rules for concentration ratio adjustment: When the proportion of lightly dormant samples exceeds 80% of the total number of monitoring points, the ratio of gibberellin GA3 to cytokinin 6-BA is changed to 2.5:1; when the proportion of deeply dormant samples exceeds 70%, the ratio is changed to 6:1. Limitation on ratio adjustment range: The ratio change within each control cycle shall not exceed ±0.5. Correction of the addition ratio of rhizosphere microbial active substances under special conditions: When the soil pH is below 5.5, the addition ratio is increased by 2% at each time period; when the soil temperature is above 30 degrees Celsius, the addition ratio is decreased by 3%. Environmental control for mixing operations: The mixing operation shall be carried out in a clean bench with an air cleanliness level of 10,000. The mixing container material shall be brown glass or stainless steel; plastic containers are prohibited to avoid adsorption.

[0099] Quality control indicators for the compound solution: Samples were taken immediately after mixing for bioactivity testing. Tissue sections from the base of regenerated buds were immersed in the compound solution and incubated at 25°C for 24 hours. The acceptable standards were: the cell length growth rate in the light dormancy group exceeded that of the control group by more than 50%; the cell division index in the deep dormancy group reached 0.5 or higher. Physical indicators: solution turbidity not exceeding 10 NTU, viscosity between 1.2 and 1.5 centipoise. Microbial activity testing: 0.1 ml of the compound solution was spread onto an ACC plate and incubated for 48 hours. The colony-forming units (CFU) were not less than 10⁶ CFU / ml. Information labeled for each batch of compound solution included: preparation time, hormone concentration ratio, microbial addition ratio, applicable dormancy depth classification type, and corresponding time window.

[0100] Preparation standards for stock solutions of gibberellin GA3 and cytokinin 6-BA: The stock solution of gibberellin GA3 is dissolved in 70% ethanol and then made up to volume with distilled water. The stock solution of cytokinin 6-BA is dissolved in 0.1 mol / L sodium hydroxide and then made up to volume with distilled water. The storage period of the stock solution shall not exceed 15 days, and the storage temperature is 4 °C. Filter and sterilize before use. Activation treatment of rhizosphere microbial active substances: Thaw the frozen bacterial suspension in a water bath at 28 °C for 5 minutes, add an equal volume of nutrient activation solution (containing 0.5% glucose and 0.1% yeast extract), and let it stand and activate at 30 °C for 30 minutes before use. Traceability mechanism for proportion adjustment: Record the reason for adjustment, original proportion, adjusted proportion and verification results each time an adjustment is made to form a historical database of proportion adjustment.

[0101] Sub-packaging and labeling of the compound solution: The mixed solution is sub-packaged into 1-L brown reagent bottles. The labels on the bottle body shall indicate: solution type (shallow dormancy type / deep dormancy type), applicable time window period (early stage / middle stage / late stage), and preparation batch number. The sub-packaging process is carried out under nitrogen protection, with the liquid level 1 cm from the bottle mouth, and the bottle mouth is sealed with a polytetrafluoroethylene gasket. Transportation conditions: Temperature is 15 to 25 °C, avoid light and shock, and the transportation duration shall not exceed 2 hours. Check by shaking well before use: When stratification or precipitation occurs, centrifuge at 2000 rpm for 5 minutes and take the supernatant. Treatment of the remaining solution: After the expiration date, add an equal volume of 10% sodium hypochlorite solution for disinfection for 30 minutes and then discharge. All preparation records include fields such as raw material batch number, dosage, operator, environmental temperature and humidity, and test data, and the storage period shall be no less than three years.

[0102] Field verification method for proportion setting: Set test plots with different proportions in areas with the same dormancy depth classification, and measure the elongation of regenerated buds 72 hours after regulation. Optimal proportion determination criteria: The proportion of the treatment group with the largest elongation and the smallest coefficient of variation. Biological basis for time window staging: The detected ethylene receptor expression level in the early stage is less than 10 ng / mg protein, 10 to 20 ng / mg protein in the middle stage, and higher than 20 ng / mg protein in the late stage. Accelerated stability test of the compound solution: Storing at 40 °C for 24 hours is equivalent to storing at 20 °C for 6 hours. If the viable bacteria count decreases by no more than 10% and the hormone degradation rate does not exceed 5% during this period, it is considered qualified. Compatibility test shall be carried out for all compound solutions before use: Take 5 mL of the solution and add it to a test tube, and observe for 2 hours. If there is no precipitation, flocculation or phase separation, it can be used.

[0103] S6. During the time window, spray the compound solution directionally on the narrow row area. The specific implementation is as follows:

[0104] When starting the spray system within the time window of the ethylene inhibition effect, the time window is derived from the start time Ts to the end time Te determined in step S4. The specific startup procedure is as follows: Activate the spray system 10 minutes before the start time Ts of the time window. The system performs self-checks, including pressure sensor calibration, nozzle blockage detection, and solution level monitoring. After passing the self-check, the spray program starts precisely at Ts. The time window conversion mechanism is as follows: Input the time window data output from step S4 (formatted as "start time: minute - end time: minute") into the spray control terminal. The terminal automatically converts it to local time and sets a countdown. Environmental constraints: When the wind speed exceeds 3 meters per second or the rainfall exceeds 2 millimeters per hour, startup is delayed and the time window is recalculated. After the delay, the time window is compressed to 50% of its original length, and startup must not be later than 120 minutes after Ts.

[0105] When positioning the spraying target area of ​​the spraying device to the location of regenerated buds above the surface of the narrow row area, the positioning operation is achieved through the following steps: First, obtain the field's geographic information system coordinates and import the boundary data of the narrow row area. The sprayer is equipped with a differential global positioning system, with positioning accuracy down to the centimeter level. The spraying target area is set as a three-dimensional spatial layer 15 to 25 centimeters above the ground surface, with a width equal to that of the narrow row area. Positioning calibration method: At a pre-set benchmark point in the field, use a laser rangefinder to measure the height and horizontal tilt angle of the spray boom, ensuring that the parallelism error between the spray boom and the ground is less than 1 degree. Regenerated bud location identification: Based on the dormancy depth classification map of step S3, the densely dormant area is set as the priority target area. Positioning accuracy verification: After spraying the fluorescent tracer, the deposition amount is measured at randomly selected points outside the target area, requiring that the deposition amount outside the target area does not exceed 5% of that in the target area.

[0106] When spraying the compound solution in atomized form onto the regenerated buds above the ground in a narrow row area, the atomization parameters are controlled as follows: droplet volume median diameter 150-200 μm, atomization pressure 0.3-0.5 MPa, and flow rate 0.8-1.2 L / min. Spraying method: Use fan-shaped atomizing nozzles with a nozzle spacing of 30 cm and a ground clearance of 20 cm. The compound solution is derived from the solution prepared in step S5, and should be shaken well and preheated to 20°C ± 2°C before use. Environmental monitoring during spraying: Temperature and humidity sensors are placed in the target area to provide real-time data feedback to the control system. When the temperature exceeds 30°C, the spray volume is automatically increased by 15% to compensate for evaporation loss; when the relative humidity is below 50%, an anti-drift mode is triggered, increasing the droplet volume median diameter to 250 μm. Spray coverage requirements: A continuous liquid film should be formed on the surface of the regenerated buds without dripping, with a deposition rate of 20-30 μL per square centimeter of leaf surface.

[0107] When controlling the spraying duration to cover the duration of the time window, the spraying duration is set to 1.2 to 1.5 times the time window length (Te-Ts). Specifically, when the time window length is 60 minutes, the spraying duration is 72 to 90 minutes. The spraying mode is intermittent: 30 seconds of operation followed by a 10-second pause to ensure sufficient solution absorption. The time window coverage mechanism is as follows: spraying starts 5 minutes earlier and ends 10 minutes later. Dynamic adjustment rules are in place: if the peak ethylene concentration is detected early, the spraying duration is automatically shortened to 80% of the original plan; if the peak is delayed, the spraying duration is extended to 120%. The formula for calculating the spraying volume is: the total spraying volume equals the narrow row area multiplied by the designed application rate per hectare, then multiplied by the time window correction factor. For example, if the narrow row area is 0.5 hectares, the designed application rate is 50 liters per hectare, and the time window factor is 1.3, then the total spraying volume equals 0.5 multiplied by 50 multiplied by 1.3, which equals 32.5 liters.

[0108] Maintenance procedures for the spraying device: Clean the pipeline after daily operation, first circulating a 5% citric acid solution for 10 minutes, then rinsing three times with deionized water. Disassemble and inspect the nozzles weekly; replace them when wear exceeds 10% of the nominal orifice diameter. Daily calibration of the positioning system: Verify positioning deviation at a standard test field; recalibrate when deviation exceeds 3 cm. Verification of spraying effect: Sample and test the residual solution on the surface of regenerated buds 2 hours after spraying; determine the gibberellin GA3 deposition amount using high-performance liquid chromatography (HPLC), requiring it to be no less than 85% of the designed dosage. Abnormal handling: When the coefficient of variation of droplet deposition uniformity exceeds 25%, the system will automatically pause and prompt for nozzle clogging inspection.

[0109] Dynamic tracking of the spray target area: A machine vision system is installed on the spray boom to identify the position of the regenerating buds in real time and adjust the nozzle direction. Vision system recognition rate calibration: 1000 images of the regenerating buds are captured under typical lighting conditions to establish a recognition model, with an accuracy requirement greater than 95%. Positioning compensation algorithm: The nozzle advance is calculated based on the locomotive's travel speed (10 to 15 meters per minute), and the compensation value is equal to the speed multiplied by 0.2 seconds. For example, at a speed of 12 meters per minute (0.2 meters per second), the compensation value is 0.2 multiplied by 0.2 equals 0.04 meters.

[0110] Measures to ensure atomization quality: Droplet distribution is monitored every 30 minutes using a laser particle size analyzer for online monitoring. Droplet distribution qualification standards: The median diameter is between 150 and 200 micrometers, and the span coefficient (Dv0.9 - Dv0.1) divided by Dv0.5 is less than 1.5. Automatic pressure adjustment is implemented when the test results are unsatisfactory: 0.05 MPa is increased for larger droplets, and 0.03 MPa is decreased for smaller droplets. Filtration of the compound solution: A 5-micron filter cartridge is installed at the inlet, and the cartridge is replaced every 500 liters of solution processed.

[0111] Precise control of spraying duration: The control system incorporates multi-level timing modules. A primary timer controls the total duration, a secondary timer manages zone intervals, and a tertiary timer manages the nozzle switching sequence. Handling of time window anomalies: If spraying is interrupted by heavy rain, the spraying time is recorded, and 150% of the remaining amount is sprayed within 24 hours after the time window closes. Before re-spraying, the residual amount on the bud surface must be checked; if the residual amount exceeds 50% of the designed dosage, re-spraying is cancelled. Data recording system: Completely records the start and end times of spraying, actual duration, environmental parameters, interruption events, and handling measures, forming a traceable spraying log.

[0112] Post-spray effect evaluation: 24 hours after spraying, 20 regenerated buds were randomly selected to test changes in endogenous hormones. For lightly dormant samples, a gibberellin GA3 content increase of over 40% was required; for deeply dormant samples, a cytokinin 6-BA receptor expression increase of over 30% was required. Areas failing the evaluation were marked as abnormal areas, and an additional 30% dose was sprayed in the next time window. Daily maintenance reports were generated for all spraying equipment, including nozzle status, positioning accuracy, and solution consumption data, and these reports were retained for at least three years.

[0113] Example 2: Figure 2 A schematic diagram of a hormone regulation and optimization system for narrow-row ratooning rice according to the present invention is provided. The system includes the following modules:

[0114] The rhizosphere anaerobic index measurement module is used to acquire rhizosphere soil samples from narrow-row areas of regenerated rice and to measure the anaerobic index of the rhizosphere soil samples.

[0115] The ACC microbial screening module is used to screen for active substances of rhizosphere microorganisms containing ACC deaminase when the anaerobic index exceeds a preset threshold.

[0116] The dormancy grading module is used to analyze the spectral absorption characteristics of narrow-row regenerated buds through near-infrared spectroscopy, and to classify the dormancy depth into shallow dormancy and deep dormancy based on the absorption threshold.

[0117] The ethylene time window module is used to track the transmission time of the peak ethylene concentration from the root to the shoot in narrow-row areas and determine the time window of the ethylene inhibitory effect.

[0118] The compound solution preparation module is used to determine the ratio of gibberellin GA3, cytokinin 6-BA and rhizosphere microbial active substances based on dormancy depth classification and time window to form a compound solution;

[0119] The targeted spraying module is used to apply the compound solution to a narrow row area within a time window.

[0120] All calculations involved in the embodiments are dimensionless numerical calculations, and the preset parameters and thresholds in the calculations are set by those skilled in the art according to the actual situation.

[0121] The above embodiments can be implemented, in whole or in part, by software, hardware, firmware, or any other combination thereof. When implemented using software, the above embodiments can be implemented, in whole or in part, in the form of a computer program product.

[0122] Those skilled in the art will recognize that the modules and algorithm steps of the various examples described in conjunction with the embodiments disclosed herein can be implemented in electronic hardware, or a combination of computer software and electronic hardware. Whether these functions are implemented in hardware or software depends on the specific application and inventive constraints of the technical solution. Those skilled in the art can use different methods to implement the described functions for each specific application, but such implementation should not be considered beyond the scope of this application.

[0123] In addition, the functional modules in the various embodiments of this application can be integrated into one processing module, or each module can exist physically separately, or two or more modules can be integrated into one module.

[0124] In the several embodiments provided in this application, it should be understood that the disclosed systems, apparatuses, and methods can be implemented in other ways. For example, the apparatus embodiments described above are merely illustrative; for instance, the division of modules is only a logical functional division, and in actual implementation, there may be other division methods. For example, multiple modules or components may be combined or integrated into another system, or some features may be ignored or not executed. Furthermore, the coupling or direct coupling or communication connection shown or discussed may be through some interfaces; the indirect coupling or communication connection between apparatuses or modules may be electrical, mechanical, or other forms.

[0125] The above description is merely a specific embodiment of this application, but the scope of protection of this application is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in this application should be included within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.

[0126] In conclusion, the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for optimizing hormone regulation in narrow-row regenerated rice, characterized in that, Includes the following steps: S1. Obtain rhizosphere soil samples from the narrow row area of ​​the regenerated rice and determine the anaerobic index of the rhizosphere soil samples; S2. When the anaerobic index exceeds the preset threshold, screen for rhizosphere microbial active substances containing ACC deaminase. S3. The spectral absorption characteristics of narrow-row regenerated buds were analyzed by near-infrared spectroscopy, and the dormancy depth was divided into shallow dormancy and deep dormancy based on the absorption threshold. S4. Track the transmission time of peak ethylene concentration from roots to shoots in narrow-row areas to determine the time window of ethylene inhibitory effect, including: Ethylene gas collection devices are installed at the root system location in the narrow row area; Ethylene gas collection devices are simultaneously installed at the base of the narrow row of regenerated buds; The ethylene concentration at the root and basal positions was measured at fixed time intervals. Record the time difference in which the peak ethylene concentration is transmitted from the root region to the basal region; Determining the time window for the ethylene inhibition effect based on time difference; The time window is the period before the peak reaches the base position; S5. Based on dormancy depth classification and time window, determine the ratio of gibberellin GA3, cytokinin 6-BA, and rhizosphere microbial active substances to form a compound solution, including: The concentration ratio of gibberellin GA3 to cytokinin 6-BA was adjusted according to the results of the shallow or deep dormancy level in the dormancy depth classification. When the dormancy depth is classified as light dormancy, the concentration ratio of gibberellin GA3 to cytokinin 6-BA is set to 3:1; when the dormancy depth is classified as deep dormancy, the concentration ratio of gibberellin GA3 to cytokinin 6-BA is adjusted to 5:

1. The proportion of rhizosphere microbial active substances added should be adjusted according to the time window stage of the ethylene inhibition effect. The time window is determined from a start time Ts to an end time Te; the time window is divided into three periods: early stage (Ts to Ts+1 / 3(Te-Ts)), middle stage (Ts+1 / 3(Te-Ts) to Ts+2 / 3(Te-Ts)), and late stage (Ts+2 / 3(Te-Ts) to Te); the addition ratio in the early stage is 5% of the mixed hormone solution volume; the addition ratio in the middle stage is 10%; and the addition ratio in the late stage is 15%. The adjusted gibberellin GA3, cytokinin 6-BA and rhizosphere microbial active substances were mixed in a certain proportion to form a compound solution; S6. Within the time window, apply the compound solution in a targeted manner to the narrow row area.

2. The method for optimizing hormone regulation in narrow-row regenerated rice according to claim 1, characterized in that, Obtain rhizosphere soil samples from the narrow-row area of ​​the ratooning rice, and determine the anaerobic index of the rhizosphere soil samples, including: Rhizosphere soil samples were collected from locations where regenerated shoots were delayed in the narrow-row area. Real-time oxygen partial pressure was measured in rhizosphere soil samples from narrow-row areas; Oxygen partial pressure was obtained from rhizosphere soil samples from wide-row areas with the same growth period as the baseline value for wide-row planting. The anaerobic index is calculated based on the deviation ratio between the real-time oxygen partial pressure of rhizosphere soil samples from narrow-row areas and the baseline value for wide-row areas.

3. The method for optimizing hormone regulation in narrow-row regenerated rice according to claim 2, characterized in that, When the anaerobic index exceeds a preset threshold, rhizosphere microbial bioactive substances containing ACC deaminase are screened, including: Determine whether the anaerobic index exceeds a preset threshold; When the anaerobic index exceeds the preset threshold, select a Pseudomonas strain that secretes ACC deaminase. The ACC deaminase activity of Pseudomonas strains was detected by enzyme-linked immunosorbent assay (ELISA). Rhizosphere microbial bioactive substances were prepared by screening strains with ACC deaminase activity reaching the standard value.

4. The method for optimizing hormone regulation in narrow-row regenerated rice according to claim 3, characterized in that, Near-infrared spectroscopy analysis was used to analyze the spectral absorption characteristics of narrow-row regenerated buds. Based on the absorption threshold, dormancy depth was divided into shallow dormancy and deep dormancy levels, including: Leaf sheath tissue from the base of narrow-row regenerated buds was collected as a test sample; The absorbance at a wavelength of 1450 nm was obtained by scanning the sample using a near-infrared spectrometer. The sleep depth is divided into light sleep level and deep sleep level by comparing the absorbance with the preset absorption threshold.

5. The method for optimizing hormone regulation in narrow-row regenerated rice according to claim 4, characterized in that, When the absorbance is less than or equal to the preset absorption threshold, it is determined to be in a light sleep state; when the absorbance is greater than the preset absorption threshold, it is determined to be in a deep sleep state.

6. The method for optimizing hormone regulation in narrow-row regenerated rice according to claim 4, characterized in that, Within the time window, the compound solution is sprayed directionally onto the narrow-row area, including: Start the spraying device within the time window of the ethylene inhibition effect; Position the spraying target area of ​​the spraying device at the location of the regenerated buds above the surface of the narrow row area; The compound solution was sprayed in a mist form onto the regenerated buds above the surface of the narrow row area; Control the duration of the spraying time window to cover the area.

7. A narrow-row hormone regulation and optimization system for ratooning rice, used to implement the narrow-row hormone regulation and optimization method for ratooning rice as described in any one of claims 1-6, characterized in that, Includes the following modules: The rhizosphere anaerobic index measurement module is used to acquire rhizosphere soil samples from narrow-row areas of regenerated rice and to measure the anaerobic index of the rhizosphere soil samples. The ACC microbial screening module is used to screen for active substances of rhizosphere microorganisms containing ACC deaminase when the anaerobic index exceeds a preset threshold. The dormancy grading module is used to analyze the spectral absorption characteristics of narrow-row regenerated buds through near-infrared spectroscopy, and to classify the dormancy depth into shallow dormancy and deep dormancy based on the absorption threshold. The ethylene time window module is used to track the transmission time of the peak ethylene concentration from the root to the shoot in narrow-row areas and determine the time window of the ethylene inhibitory effect. The compound solution preparation module is used to determine the ratio of gibberellin GA3, cytokinin 6-BA and rhizosphere microbial active substances based on dormancy depth classification and time window to form a compound solution; The targeted spraying module is used to apply the compound solution to a narrow row area within a time window.

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