Optimization method and system for regulation and control of ratooning rice narrow-row hormone

Abstract

The invention discloses a ratooning rice narrow-row hormone regulation and control optimization method and system, particularly relates to the technical field of rice cultivation, and aims to solve the problem of yield loss caused by asynchronous germination of wide-row and narrow-row regenerated buds in the prior art. The method comprises the following steps: measuring an anaerobic index of narrow-row rhizosphere soil, and screening microbial active substances containing ACC deaminase when the anaerobic index exceeds a threshold value; secondly, analyzing absorption characteristics of regenerated buds through near infrared spectroscopy to realize dormancy depth grading; synchronously tracking a transmission time sequence of ethylene concentration from a root system to a bud to determine an inhibition window period; dynamically adjusting the compounding proportion of gibberellin GA3, cytokinin 6-BA and microbial active substances based on dormancy grading and a time window; and finally, directionally spraying the compound solution to the narrow-row area in the time window. By eliminating rhizosphere ethylene accumulation and accurately matching the physiological state and action time sequence of buds, the narrow-row regeneration bud germination efficiency and wide-row synchronization are realized.

Description

Technical Field

[0001] The present invention relates to the technical field of rice cultivation, and more particularly to a method and system for optimizing hormone regulation of narrow rows of regenerated rice. Background Art

[0002] The wide-narrow row cultivation model for regeneration rice has become a key agronomic measure for increasing yields in the Yangtze River Basin by improving light energy utilization through altering the field microenvironment. After the first rice harvest, wide rows experience early bud germination and rapid growth due to their superior ventilation and light conditions, while narrow rows experience a significant delay in bud germination due to the shaded, high-humidity environment. Existing technologies typically employ exogenous spraying of gibberellins (GA3) and cytokinins (6-BA) to promote bud germination in narrow rows, attempting to narrow the growth gap between wide and narrow rows.

[0003] In the existing technology, the difference in rhizosphere microenvironment between wide and narrow rows leads to an imbalance in the response efficiency of exogenous hormones, making it difficult to achieve synchronous growth of regenerated buds. The narrow rows form a special rhizosphere environment due to long-term shading, which triggers the accumulation of endogenous inhibitory substances and interferes with the physiological effects of exogenous GA3 and 6-BA, resulting in the sensitivity and metabolic response of the regenerated buds in narrow rows to hormones being significantly weaker than those in wide rows. This environmental dependence effect causes the germination efficiency of the same dose of hormones in narrow rows to decrease, and the difference in growth rate of regenerated buds in wide and narrow rows to widen, which ultimately restricts the improvement of tillering 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 of narrow-row regenerated rice to solve the problems raised in the above-mentioned background technology.

[0005] To achieve the above object, the present invention provides the following technical solutions:

[0006] A method for optimizing hormone regulation of narrow-row regenerated rice comprises the following steps:

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

[0008] S2. When the anaerobic index exceeds a preset threshold, screening for active substances of rhizosphere microorganisms containing ACC deaminase;

[0009] S3. Analyze the spectral absorption characteristics of the narrow row regenerated buds by near-infrared spectroscopy, and classify the dormancy depth into light dormancy and deep dormancy according to the absorption threshold;

[0010] S4. Track the timing of the transfer of the peak ethylene concentration from the root system to the shoot in the narrow row area to determine the time window of the ethylene inhibitory effect;

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

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

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

[0014] Rhizosphere soil samples were collected at the location where regeneration shoots were delayed in the narrow row area;

[0015] Determine real-time oxygen partial pressure in rhizosphere soil samples from narrow row areas;

[0016] Obtain the oxygen partial pressure of rhizosphere soil samples from wide-row areas at the same growth period as the wide-row baseline value;

[0017] The anaerobic index was calculated based on the deviation ratio of the real-time oxygen partial pressure of rhizosphere soil samples in the narrow row area to the wide row baseline value.

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

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

[0020] When the anaerobic index exceeds a preset threshold, the Pseudomonas strain that secretes ACC deaminase is selected;

[0021] The ACC deaminase activity of Pseudomonas strains was detected by enzyme-linked immunosorbent assay;

[0022] The strains whose ACC deaminase activity reached the standard value were screened to prepare rhizosphere microbial active substances.

[0023] Furthermore, the spectral absorption characteristics of the narrow row regeneration buds were analyzed by near-infrared spectroscopy, and the dormancy depth was divided into light dormancy and deep dormancy according to the absorption threshold, including:

[0024] The leaf sheath tissue at the base of the narrow row regenerated buds was collected as the test sample;

[0025] The sample was scanned and tested using a near-infrared spectrometer to obtain the absorbance at a wavelength of 1450 nm;

[0026] The absorbance is compared with the preset absorption threshold to divide the sleep depth into light sleep level and deep sleep level.

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

[0028] Furthermore, the timing of the transfer of ethylene concentration peaks from roots to shoots in narrow row areas was tracked to determine the time window of ethylene inhibition effects, including:

[0029] Arrange ethylene gas collection devices at the root level in narrow row areas;

[0030] An ethylene gas collection device is synchronously arranged at the base of the narrow row of regenerated shoots;

[0031] The ethylene concentrations at the root and base were measured at regular time intervals;

[0032] The time difference of the peak ethylene concentration from the root position to the base position was recorded;

[0033] The time window of ethylene inhibitory effect was determined based on the time difference.

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

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

[0036] Adjust the concentration ratio of gibberellin GA3 and cytokinin 6-BA according to the results of light dormancy or deep dormancy in the dormancy depth classification;

[0037] Adjust the addition ratio of rhizosphere microbial active substances according to the time window of ethylene inhibition effect;

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

[0039] Furthermore, within the time window, the compound solution is sprayed in a targeted manner on the narrow row area, including:

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

[0041] Position the spray target of the spray device to the location of the regenerating buds above the ground surface in the narrow row area;

[0042] Spray the compound solution in a mist form to the location of regenerating buds above the ground surface in the narrow row area;

[0043] Control the duration of the spraying time window.

[0044] In another aspect, the present invention provides a narrow-row hormone regulation optimization system for regenerated rice, comprising the following modules:

[0045] The rhizosphere anaerobic testing module is used to obtain rhizosphere soil samples from the narrow row area of ​​replanted rice and measure the anaerobic index of the rhizosphere soil samples;

[0046] ACC bacteria screening module, used to screen rhizosphere microbial active substances containing ACC deaminase when the anaerobic index exceeds the preset threshold;

[0047] The dormancy classification module is used to analyze the spectral absorption characteristics of narrow-row regeneration buds through near-infrared spectroscopy and classify the dormancy depth into light dormancy and deep dormancy according to the absorption threshold;

[0048] The ethylene time window module is used to track the timing of the transfer of ethylene concentration peaks from roots to shoots in narrow row areas and determine the time window of ethylene inhibition effects;

[0049] The compound solution adjustment 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 directional spraying module is used to spray the compound solution in a targeted manner on the narrow row area within the time window.

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

[0052] 1. The present invention solves the problem of asynchronous germination of regenerated buds in wide and narrow rows by establishing a collaborative diagnostic mechanism between the narrow row microenvironment and the dormancy state of regenerated buds. Aiming at the ethylene inhibitory effect caused by the anaerobic environment in the rhizosphere of narrow rows, ACC deaminase active substances are introduced to selectively 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 ethylene transmission timing, the window period of exogenous hormone action can be accurately captured. This three-level regulatory system of "microenvironment repair-dormancy state analysis-time-targeted intervention" can improve the physiological efficacy of gibberellins and cytokinins in narrow rows to the same level as wide rows, ensuring that the uniformity of regenerated bud germination meets the requirements of synchronous growth.

[0053] 2. Through the collaborative decision-making mechanism of dormancy depth classification and time window, the inherent limitations of traditional uniform spraying are broken through. Based on the dynamic adjustment of hormone ratios in light dormancy / deep dormancy levels, the precise matching of exogenous substances and the physiological state of the buds is achieved; and the directional spraying controlled by the time window enables the active substances of rhizosphere microorganisms to effectively block the inhibition chain transmission before the peak of ethylene concentration. This compound scheme with the dormancy state of the bud as the core of regulation and the timing of ethylene transmission as the axis of action not only significantly improves the response sensitivity of narrow row regeneration buds to hormones, but also fundamentally eliminates the metabolic response differences between wide and narrow rows, providing a core guarantee for the uniform tillering of regenerated rice. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 This is a flow chart of a method for optimizing hormone regulation of narrow rows of regenerated rice according to the present invention;

[0055] Figure 2 This is a schematic structural diagram of a narrow-row hormone regulation optimization system for regenerated rice according to the present invention. DETAILED DESCRIPTION

[0056] The following will provide a clear and complete description of the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0057] Example 1: Figure 1 The present invention provides a method for optimizing hormone regulation of narrow-row regenerated rice, which comprises the following steps:

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

[0059] S2. When the anaerobic index exceeds a preset threshold, screening for active substances of rhizosphere microorganisms containing ACC deaminase;

[0060] S3. Analyze the spectral absorption characteristics of the narrow row regenerated buds by near-infrared spectroscopy, and classify the dormancy depth into light dormancy and deep dormancy according to the absorption threshold;

[0061] S4. Track the timing of the transfer of the peak ethylene concentration from the root system to the shoot in the narrow row area to determine the time window of the ethylene inhibitory effect;

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

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

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

[0065] The location of delayed bud germination is determined by the following criteria: the measured length of the narrow row bud is less than or equal to 50% of the length of the wide row bud measured during the same period. Specifically, at 9:00 AM on the third day after rice harvest, the length of the buds in the narrow row is measured using a digital caliper. A location where the length of the bud at a particular point does not exceed 50% of the average length of the buds of five randomly selected plants in the adjacent wide row is considered delayed. At this location, a 2-cm inner diameter oxygen-free stainless steel soil drill is used to vertically drill a soil sample within 5 cm of the base of the rice stem. The sampling depth is from the surface to a depth of 20 cm. Soil particles attached to the root surface are retained as the rhizosphere soil sample. The single-point sampling volume should be no less than 200 grams. Immediately after sampling, the sample is sealed in a nitrogen-filled bag and stored at 4 degrees Celsius.

[0066] A portable soil gas analyzer was used to measure the real-time oxygen partial pressure in rhizosphere soil samples from narrow row areas. Prior to testing, the instrument underwent a two-point calibration: first, zero calibration in a 99.99% high-purity nitrogen atmosphere, and second, full-scale calibration in standard air with an oxygen concentration of 20.9%. The soil sample was transferred to a 25°C constant-temperature testing chamber. After temperature equilibration, a zirconium oxide solid electrolyte oxygen electrode sensor was inserted 2 cm below the center of the sample. The data collection interval was set to 10 seconds. After excluding the first 30 seconds of data, the arithmetic mean of the stable readings over the subsequent 90 seconds was taken as the real-time oxygen partial pressure value, expressed in kilopascals. The measurement was repeated three times for each sample. Retesting was required if the range of the three measurements exceeded 1 kilopascal.

[0067] When obtaining the oxygen partial pressure of rhizosphere soil samples in the wide row area of ​​the same growth period as the wide row benchmark value, the same growth period refers to the same number of days after rice harvest and the same light period. Sampling points are arranged at the center line of the wide row adjacent to the target narrow row area, with the principle that the straight-line distance does not exceed 5 meters. The oxygen partial pressure data is obtained using the same sampling tools and detection procedures as the narrow row area. Five sampling points are selected for each field. When the coefficient of variation of the oxygen partial pressure detection values ​​at the five points exceeds 15%, additional sampling is performed at eight points and recalculated. Finally, the arithmetic mean of all valid detection values ​​is taken as the wide row benchmark value.

[0068] The anaerobic index is calculated based on the deviation between the real-time oxygen partial pressure of rhizosphere soil samples in narrow-row areas and the wide-row baseline value. The following steps are followed: First, the difference between the wide-row baseline value and the real-time oxygen partial pressure in the narrow row is calculated; second, this difference is divided by the wide-row baseline value to obtain the proportionality factor; and finally, the proportionality factor is multiplied by 100% to convert the value into a percentage. The specific calculation formula is: Anaerobic Index = [(Wide-row Baseline Value - Narrow-row Real-time Oxygen Partial Pressure) ÷ Wide-row Baseline Value] × 100%. A soil texture correction factor, K, is incorporated into the calculation: 0.95 for sandy soil, 1.05 for clay soil, and 1.00 for loam soil. The corrected formula is: Anaerobic Index = [(Wide-row Baseline Value - Narrow-row Real-time Oxygen Partial Pressure) × K ÷ Wide-row Baseline Value] × 100%. The preset thresholds are dynamically adjusted. For example, the threshold is 30% when the average daily temperature is below 20°C, 40% when the average daily temperature is above 28°C, and 35% under other temperature conditions. When the wide row baseline value is lower than 8 kPa, the whole field is judged to be in an oxygen-deficient state and proceeds directly to the next step.

[0069] S2. When the anaerobic index exceeds a preset threshold, screening for active substances of rhizosphere microorganisms containing ACC deaminase is performed as follows:

[0070] When determining whether the anaerobic index exceeds a preset threshold, the threshold is dynamically adjusted based on the ambient temperature. When the average daily ambient temperature is below 20°C, the preset threshold is set to 30%; when the average daily ambient temperature is above 28°C, the preset threshold is set to 40%; and under other temperature conditions, the preset threshold is set to 35%. This determination is made by comparing the anaerobic index value with the preset threshold for the corresponding temperature range. When the anaerobic index exceeds the preset threshold, it is considered to have exceeded the preset threshold. The anaerobic index is derived from the calculation results of step S1. Each determination requires recording the timestamp, temperature value, and corresponding preset threshold parameters. The determination process for whether the anaerobic index exceeds the preset threshold includes a data verification mechanism: When the anaerobic index exceeds the preset threshold, the raw oxygen partial pressure data from step S1 must be reviewed. This verification includes ensuring that the range of the three replicates of the narrow-row real-time oxygen partial pressure test is less than 1 kPa and that the coefficient of variation of the wide-row baseline value is less than 15%. If any of these verification conditions are not met, resamples are collected and tested. The temperature relevance of the preset thresholds was determined through field trials. For example, in a three-year trial, it was found that when the anaerobic index exceeded 30% under low temperature conditions, the inhibition rate of regenerated buds reached 92%, and when it exceeded 40% under high temperature conditions, the inhibition rate reached 89%.

[0071] When the anaerobic index exceeds a preset threshold, Pseudomonas strains that secrete ACC deaminase are selected. Pseudomonas strains are isolated from rhizosphere soil samples from the target field. The specific procedure is as follows: 10 grams of the rhizosphere soil sample collected in step S1 is added to 90 milliliters of sterile saline and suspended with shaking for 30 minutes. The supernatant is then diluted to a gradient concentration of 10 to the power of -3, 10 to the power of -4, and 10 to the power of -5. 0.1 milliliter of each dilution is spread on an ACC screening plate containing 10 grams of tryptone, 5 grams of yeast extract, 10 grams of sodium chloride, and 15 grams of agar per liter, with 3 millimolar ACC added as the sole nitrogen source. After 48 hours of incubation in a 28-degree Celsius incubator, single colonies with a growth diameter greater than 2 mm are selected and verified as Pseudomonas strains by Gram staining and oxidase testing for storage. A strain traceability mechanism is established for the selection of Pseudomonas strains: each isolated strain must record its source field, sampling location coordinates, and isolation time. The ACC concentration in the ACC screening plate was optimized through preliminary experiments. For example, a concentration gradient experiment demonstrated that 3 millimolar ACC effectively inhibited the growth of non-target bacteria while ensuring normal proliferation of Pseudomonas. Colony morphology was observed daily during the plate culture period. Pseudomonas is typically characterized by round colonies with neat edges and a moist surface. Gram staining was performed using crystal violet solution; a positive result was a red bacilli. The oxidase test was performed using tetramethyl-p-phenylenediamine test paper; a positive result was a dark blue color within 30 seconds.

[0072] To detect ACC deaminase activity in Pseudomonas strains using an enzyme-linked immunosorbent assay (ELISA), a bacterial lysate is prepared: a preserved Pseudomonas strain is inoculated into LB liquid medium and cultured at 28°C with shaking at 180 rpm for 24 hours. 10 mL of the culture is then centrifuged at 10,000 rpm for 10 minutes at 4°C to collect the cells. The cells are washed twice with 0.1 M Tris-HCl buffer and resuspended in 1 mL of the same buffer containing 0.5 mM dithiothreitol. Ultrasonication is performed at 300 W for 10 seconds with 20 seconds intervals for a total of 5 minutes. The lysate is then centrifuged at 12,000 rpm for 20 minutes at 4°C, and the supernatant is used as the enzyme extract. The enzyme-linked immunosorbent assay (ELISA) uses a 96-well microplate. To each well, 50 μL of enzyme extract and 150 μL of reaction mixture containing 0.2 M Tris-HCl buffer, 10 mM ACC substrate, and 0.2 mM pyridoxal phosphate are added. After incubation at 30°C for 30 minutes, the reaction is terminated by adding 100 μL of a chromogenic stop solution consisting of 0.5 M citric acid buffer containing 1 mg / mL o-phenylenediamine and 0.3% hydrogen peroxide. The absorbance is measured at 490 nm on a microplate reader, and ACC deaminase activity is calculated based on a standard curve. One unit of activity is defined as the amount of enzyme that catalyzes the production of 1 μmole of α-ketobutyrate per minute. The standardized ELISA protocol includes the following: a standard control well is set up for each assay. The standard consists of purified ACC deaminase solution at concentrations of 0, 5, 10, 15, and 20 activity units per mg of protein. A correlation coefficient greater than 0.99 is required for the standard curve to be valid. Samples were tested in triplicate. Absorbance differences exceeding 15% between replicates required retesting. The reaction temperature was controlled to an accuracy of ±0.5°C and maintained in a constant-temperature water bath. Color development was strictly limited to 10 minutes; any time exceeding this time would result in elevated background values.

[0073] When screening strains for ACC deaminase activity that meets the standard value for preparing rhizosphere microbial active substances, the standard value is set at 15 activity units per milligram of protein. The screening process includes calculating the specific activity of ACC deaminase for each strain, which is equal to the total activity units divided by the protein concentration. Protein concentration is determined using the Coomassie Brilliant Blue method, using bovine serum albumin as the standard protein to generate a standard curve. Strains with an ACC deaminase specific activity of 15 activity units per milligram of protein or greater are considered to meet the standard. To prepare rhizosphere microbial active substances, strains that meet the standard are inoculated into a fermentation medium containing 10 grams of glucose, 5 grams of peptone, 1 gram of potassium dihydrogen phosphate, and 0.5 grams of magnesium sulfate per liter. Fermentation is carried out at 28 degrees Celsius with an aeration rate of 1 volume per minute for 48 hours. After fermentation, the cells are harvested by centrifugation at 8,000 rpm for 15 minutes at 4 degrees Celsius. After washing the cells twice with sterile saline, they are resuspended in a protective solution containing 10% glycerol. The concentration is adjusted to 10^9 viable cells per milliliter. The cells are then aliquoted and frozen for storage. This is the active rhizosphere microbial substance. Quality control is implemented during the preparation of the active rhizosphere microbial substance: the cell concentration at the end of fermentation must be at least 10^9 viable cells per milliliter. The viability rate is determined by plate count, with a requirement of >95%. The protective solution contains 100g sucrose, 10g sodium glutamate, 5g ascorbic acid, and 10ml glycerol per liter. Cryopreservation utilizes a programmed cooling process: first, cooling at 1°C / min to 4°C for 2 hours; then cooling at 1°C / min to -20°C for 4 hours; and finally, transferring to -80°C for long-term storage. Each batch of product is tested for residual ACC deaminase activity, which is required to remain at least 90% of the initial activity after three months of frozen storage.

[0074] S3. Analyze the spectral absorption characteristics of the narrow row regenerated buds by near-infrared spectroscopy, and classify the dormancy depth into light dormancy and deep dormancy according to the absorption threshold. The specific implementation is as follows:

[0075] When collecting basal leaf sheath tissue from regenerating shoots in narrow rows for testing, this is defined as the sheath segment no higher than 3 cm from the ground. The procedure is as follows: Between 10:00 AM and 2:00 PM on the fifth day after rice harvest, select plants within the narrow row whose regenerating shoot length is less than or equal to 60% of the corresponding regenerating shoot length in the wide row. Use sterile surgical scissors to remove the basal leaf sheath tissue. Sampling is performed between 0.5 cm and 1.5 cm from the basal node of the regenerating shoot, using a 1 cm long by 0.5 cm wide block. Immediately after sampling, the block is placed in a pre-cooled vacuum desiccator, stored at -20°C, and transported to the laboratory. The interval between collection and testing should not exceed 24 hours. The collection of basal leaf sheath tissue is spatially controlled: sampling points are arranged at a density of one point per square meter within the narrow row, with three representative plants selected at each sampling point. When harvesting basal leaf sheath tissue, avoid mechanically damaged areas. After every five samples, disinfect the sampling tool with 75% ethanol. Store samples in brown, light-proof bottles filled with silica gel desiccant to maintain relative humidity below 10%. Maintain a temperature of -20°C ± 2°C during transportation. Samples will be discarded if the temperature fluctuates by more than 3°C.

[0076] When scanning samples using a near-infrared spectrometer to measure absorbance at 1450 nm, pre-treat the sample: thaw frozen tissue blocks at 4°C for 2 hours, then remove surface moisture with filter paper. Spread the pre-treated tissue blocks flat on a quartz slide, ensuring a uniform thickness of 2 to 3 mm. After the near-infrared spectrometer has warmed up for 30 minutes, perform a baseline calibration using a standard white plate as the reference for 100% reflectance and a light shield as the reference for 0% reflectance. Scanning parameters are set to: wavelength range 1300 to 1600 nm, 4 nm resolution, 32 scans, and average. During testing, maintain a laboratory temperature of 25°C ± 1°C and a relative humidity of 40% ± 5%. Place the slide in the fixed position of the sample cell, initiate the scanning program, and record the characteristic absorbance value at 1450 nm. Absorbance values ​​are rounded to three decimal places. Each sample is scanned three times. Retesting is required if the range of the three measurements exceeds 0.01. Spectral detection introduces a live in-situ measurement mode: a non-destructive testing scheme is adopted for some samples. After the plant to be tested is placed in a dark room to adapt for 30 minutes, the fiber optic probe is directly contacted with the surface of the basal sheath tissue. The probe applies a constant pressure of 50 grams to ensure close contact. Before measurement, a refractive index matching liquid is applied to the contact surface of the probe to eliminate interface reflection. The number of scans is increased to 64 times during in-situ measurement, and the spectral data is processed by five-point cubic smoothing filtering. The absorbance value at 1450 nm is adjusted according to the temperature correction formula: the corrected absorbance is equal to the difference between the measured value multiplied by 1 plus 0.003 multiplied by 25 minus the detection environment temperature T, where T is the temperature value in Celsius.

[0077] When comparing absorbance to a preset absorbance threshold, the threshold is dynamically set. The baseline threshold is set at 0.85, based on a three-year field trial: statistics show that samples with an absorbance of 0.85 or less have a germination rate of over 95%, while samples with an absorbance greater than 0.85 have a germination rate of less than 30%. The dynamic adjustment rule is: when the ambient daily average temperature is below 18°C, the threshold is lowered by 0.05; when the ambient daily average temperature is above 30°C, the threshold is raised by 0.05. The comparison is performed using data processing software: the sample absorbance measurement and the current ambient temperature are input, and the system automatically matches the preset absorbance threshold for the corresponding temperature range for comparison. The temperature compensation mechanism for the preset absorbance threshold is mathematically modeled: the temperature compensation coefficient Wp is equal to 0.002 multiplied by the difference between the ambient daily average temperature T and 24, where T is the temperature in degrees Celsius. The adjusted threshold is equal to the baseline threshold plus Wp. For example, when the average daily ambient temperature is 15 degrees Celsius, Wp equals 0.002 times the difference between 15 and 24, or negative 0.018. The corrected 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 times the difference between 32 and 24, or 0.016. The corrected threshold is 0.85 plus 0.016, which equals 0.866. The basic threshold of 0.85 was verified by collecting 500 samples over three consecutive years, measuring absorbance, and tracking the germination of regenerated buds. Statistics show that the germination success rate of samples with an absorbance of 0.85 or less is, for example, 96.2%, while the germination success rate of samples with an absorbance greater than 0.85 is, for example, 28.7%. Variety-specific correction of the preset absorbance threshold: A threshold library is established for different rice varieties. For example, the basic threshold of variety A is 0.82, and the temperature coefficient is 0.0018; the basic threshold of variety B is 0.88, and the temperature coefficient is 0.0022.

[0078] When the absorbance is less than or equal to the preset absorption threshold, it is judged as light dormancy. The criteria for light dormancy include: the measured absorbance value is not greater than the preset absorption threshold after correction for the current temperature. The judgment process records three parameters: ambient temperature, measured absorbance, and correction threshold, and generates a judgment report for archiving. The physiological characteristics of the regenerated buds corresponding to light dormancy samples are: the bud scales are light green, the cell division activity index of the basal internode is higher than 0.7, and the endogenous gibberellin concentration is higher than 1.2 nanograms per milligram fresh weight. Additional verification indicators for light dormancy judgment: 20% of the samples judged to be light dormancy are randomly sampled for microscopic observation. Paraffin sections are made from the basal leaf sheath tissue, and the vascular bundle structure is observed under a 400x microscope after staining. Light dormancy samples should have the following characteristics: sieve tube cell wall thickness is less than 2 microns, companion cell mitochondrial density is higher than 15 per square micron, and the degree of vacuolation of phloem parenchyma cells is less than 30%. If the microscopic characteristics do not meet the requirements, the judgment conclusion is invalid and resampling is required. A quick screening method for light-dormant samples: Samples identified as light-dormant are immediately tested for germination potential. Base leaf sheath tissue is immersed in a 10 mg / L gibberellin solution and incubated in a dark incubator at 28°C for 48 hours. After incubation, the elongation of the shoots is measured. Any elongation exceeding 2 mm is considered true light-dormant; any elongation less than 0.5 mm is revised to deep-dormant.

[0079] Deep dormancy is determined when the absorbance exceeds the preset threshold. Deep dormancy is determined when the absorbance value exceeds the preset threshold after correction for the current temperature. The determination is correlated with the sample's physiological status: The bud scales of deep dormancy samples are yellow-brown, the mitotic activity index of the basal internodes is less than 0.3, and the endogenous abscisic acid concentration is greater than 5.6 ng / mg fresh weight. All determinations are subject to data review. If any of the three scan values ​​disagree with the determination, resampling is required. Emergency procedures for deep dormancy determination: When the proportion of deep dormancy samples exceeds 70% of the total sample volume, a field-wide alert is activated. Within 24 hours of the alert, three backup sample points must be retested, with the retest points located at least 5 meters from the initial test points. If the retest results still indicate an excessive deep dormancy ratio, subsequent control steps are automatically initiated. Storage and retesting of deep dormancy samples: Samples determined to be deep dormancy are split into two aliquots: one is immediately tested for endogenous hormones, and the other is stored in a liquid nitrogen tank. The absorbance of the preserved samples was retested on the 5th day after the control measures were implemented. When the retested value dropped by more than 15% compared with the initial value, the dormancy depth classification of the plant was updated to light dormancy.

[0080] Optimizing the sample collection window: During periods of continuous rain, the sampling time is adjusted to 4 to 8 hours after the rain stops. Sampling personnel wear static-free protective clothing and use ceramic scissors to prevent interference from metal ions. Sample bottles are filled with high-purity nitrogen to displace the air, and the bottles are sealed with screw caps containing polytetrafluoroethylene gaskets. Sample transport boxes are equipped with temperature recorders. Upon arrival at the laboratory, the temperature profile is checked. If the sample remains below -20°C for less than 90% of the total transport time, the sample is discarded. Variety identification is assisted by molecular markers: leaf DNA is collected simultaneously with sampling, and the OsS1 gene fragment is amplified by PCR. Sequencing results are then matched to a variety database. When testing new varieties, a default threshold of 0.85 is used, and a learning mode is initiated: the relationship between absorbance and germination rate of 50 samples is continuously tracked. The optimal threshold is automatically calculated and entered into the database. All judgment data is uploaded to a central database, generating a spatial distribution heat map to guide precision operations. All judgment changes are recorded with timestamps and environmental parameters, forming a dormancy state migration map for model optimization. Test results are fed back to the threshold optimization system to dynamically calibrate the judgment model.

[0081] S4. Track the transmission sequence of the peak ethylene concentration in the narrow row area from the root system to the shoot body to determine the time window of the ethylene inhibition effect. The specific implementation is as follows:

[0082] When laying out ethylene gas collection devices at the root position in narrow row areas, the root position is defined as a soil layer area 10 cm horizontally and 15 to 25 cm vertically from the base of the main stem of rice. The specific laying operation is: use a 5 mm diameter stainless steel probe to vertically insert the soil to a depth of 20 cm, and connect the end of the probe to a gas collection chamber. The gas collection chamber is made of polytetrafluoroethylene material, has a volume of 5 ml, and contains activated alumina particles with a particle size of 1 to 2 mm. When laying out, ensure that the collection chamber is completely buried in the soil, the pipe mouth is 2 cm above the ground and a dust cap is installed. Three gas collection devices are laid out in each narrow row area, distributed in a triangle, with a distance of 50 cm between the devices. After the laying is completed, let it stand for 24 hours to allow the device to balance with the environment, and maintain the normal water level in the field during this period.

[0083] When the ethylene gas collection device is synchronously laid out at the base position of the narrow row regeneration buds, the base position is defined as the internode area at the base of the regeneration bud at a height of 2 cm to 3 cm from the ground surface. The specific operation is: use a special fixing clamp to attach the micro collection cover to the surface of the base internode. The collection cover is made of a breathable silicone membrane with an effective adsorption area of ​​0.5 square centimeters. The rear end of the collection cover is connected to a polyetheretherketone catheter with an inner diameter of 1 mm, and the catheter extends to the ground to connect to the vacuum pump. Two collection devices are laid out at each monitoring point, located on both sides of the internode, with a distance of 1 cm between the devices. Keep the time synchronized with the layout of the root position device, and the time difference should not exceed 5 minutes. Check the sealing after layout: apply a positive pressure test of 0.1 MPa, and the pressure drop within 1 minute does not exceed 5% to be qualified.

[0084] Ethylene concentrations at the root and basal levels were measured at fixed intervals of 30 minutes. The measurement process involved first drawing gas from the root gas collection chamber. Using a 50-ml gastight syringe, 10 ml of gas was drawn at a flow rate of 5 ml / min and immediately injected into the gas chromatograph inlet. Gas chromatograph testing conditions included: a 30-meter capillary column with an inner diameter of 0.32 mm; high-purity nitrogen carrier gas at a flow rate of 1.5 ml / min; a flame ionization detector at 200°C; and a column oven temperature program from 40°C for 2 minutes, then increasing at 10°C / min to 100°C. The same method was used to measure basal gas. Before each measurement, the instrument was calibrated with standard ethylene gas at concentration gradients of 0.1, 0.5, and 1.0 μL / L. The calibration curve correlation coefficient was required to be greater than 0.995. Each measurement was repeated three times, and the arithmetic mean was taken as the ethylene concentration at that point in time, expressed in μL / L.

[0085] When recording the time difference for the peak ethylene concentration to be transferred from the root position to the base position, the peak ethylene concentration is defined as the highest ethylene concentration value among three consecutive measurement time points. The identification method is as follows: draw a curve of the ethylene concentration change over time at the root position, find the maximum concentration value and the corresponding time point T1; use the same method to find the maximum concentration value at the base position and the corresponding time point T2. The time difference Δt is calculated as the absolute value of T2 minus T1, measured in minutes. When the peak value at the root position is not obvious (the difference between the highest and second highest values ​​is less than 10%), take the midpoint of the highest concentration period of three consecutive time points as T1; the same method is used to determine T2 at the base position. The time difference is recorded to an accuracy of 0.5 minutes, and the weather conditions are marked: the actual sunshine intensity is recorded on sunny days, and the start and end time of rainfall is recorded on rainy days.

[0086] When the time window for ethylene inhibition is determined based on the time difference as the period before the peak reaches the base position, the time window is defined as the period of Δt calculated from T2 forward. For example, when Δt is 120 minutes, the time window is 120 minutes before T2 to T2. Rules for handling special situations: when Δt is less than 60 minutes, the time window is extended 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 time of the time window is 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, marked to minute accuracy, and the average soil temperature, humidity and light intensity parameters during this period are recorded.

[0087] Maintenance procedures for the ethylene gas collection device: Replace the adsorbent every 24 hours, and purge the pipeline with nitrogen for 3 minutes before replacement. Replace the micro-collection cover with a new one every 8 hours. The old cover should be ultrasonically cleaned with methanol and then dried for use. The gas chromatograph should be verified by injecting standard gas after measuring every 10 samples, and recalibrated if the drift exceeds 5%. Time difference verification method: During the time window period, the ethylene concentration at the base position is supplemented every 15 minutes to verify whether the concentration shows a monotonically increasing trend. If a downward inflection point occurs, the end point of the time window is corrected to the inflection point. All data recording tables contain fields such as device number, installation time, measurement time, concentration value, peak time, time difference, window period, etc., forming an electronic tracking log.

[0088] Selection criteria for root location points: Prioritize the layout in areas where the anaerobic index exceeds the standard as determined in step S1. When the length of the narrow row area exceeds 5 meters, add a monitoring unit for every additional 2 meters. Adsorbent pretreatment method: Activated alumina particles are calcined in a muffle furnace at 550 degrees Celsius for 4 hours, immersed in a 0.1 mol per liter hydrochloric acid solution for 12 hours after cooling, rinsed with deionized water until neutral, and dried at 120 degrees Celsius for use. The insertion angle of the probe tube remains vertical, and the tilt error does not exceed 3 degrees. Avoid areas with dense plant roots when laying out the device, and select areas with fewer lateral roots.

[0089] Microenvironmental control at the base: Apply silicone grease around the collection hood to prevent gas leakage. Clamp pressure is controlled between 50 and 80 grams to avoid compression and tissue deformation. The vacuum pump flow rate is set to 0.2 ml per minute, and the vacuum duration is 10 minutes. Each vacuum is allowed to equilibrate for 20 minutes before the next collection. A built-in temperature sensor in the synchronous monitoring device records contact surface temperature changes in real time, and this temperature data is used for subsequent concentration correction.

[0090] Quality control for ethylene concentration determination: Gas samples are treated to remove water in a cold trap before injection, maintained at -20°C. Chromatographic peak identification criteria: peak height greater than three times the baseline noise, and peak shape symmetry factors between 0.8 and 1.2. Concentration is calculated using an external standard method: sample peak areas are compared with a standard curve, rounded to three significant figures. Replicate samples are resampled and measured if relative deviation exceeds 8%. Each batch includes a blank control: ethylene-free air is drawn to measure background values. If background values ​​exceed 0.05 μL / L, the system is shut down and the source of contamination is investigated.

[0091] Statistical algorithm for peak transit time difference: When multiple monitoring units are present, the median of the Δt values ​​for each unit is used as the final value. Outliers are removed before median calculation: if a unit's Δt deviates from the median by more than 30%, and the peak concentration for that unit is less than 70% of the mean, the data is considered invalid. The time difference Δt is correlated with soil physical properties: a correction factor is added based on soil type: clay soil is multiplied by 1.1, sand soil by 0.9, and loam soil remains unchanged. The corrected time difference is used in the final time window calculation.

[0092] Verification of the time window's suitability: Within the time window, samples were taken every 30 minutes to measure ethylene receptor protein expression in basal internode tissue. Western blotting was performed using an anti-ethylene receptor (ETR1) antibody. When the expression level increased by more than 50%, the time window was considered valid. If the increase was less than 20%, the peak transition process was re-tracked. The time window output format was: start time (hour:minute) to end time (hour:minute), with an accompanying confidence rating: A (validated), B (partially validated), C (unvalidated, requiring review). All time window data were linked to the field number, monitoring date, and weather station code, and uploaded to a central database to generate a spatiotemporal distribution map.

[0093] Recovery and disinfection of gas collection devices: After monitoring, the stainless steel probe is soaked in 0.5% sodium hypochlorite solution for 30 minutes, rinsed with deionized water, and dried. The silica gel collection cover is ultrasonically cleaned with ultrapure water for 15 minutes, dried at 60 degrees Celsius, and stored. The connecting pipes are purged with high-purity nitrogen for 10 minutes and then sealed for storage. The data storage medium is packaged in a waterproof and anti-magnetic package with a shelf life of not less than three years. Within 12 hours after the time window is determined, fluorescent markers are placed in the field. The sign displays the start and end times of the window period, with a font height of 5 cm, ensuring a visible distance of more than 10 meters.

[0094] 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, which is specifically implemented as follows:

[0095] When adjusting the concentration ratio of gibberellin GA3 and cytokinin 6-BA according to the results of the light dormancy level or deep dormancy level in the dormancy depth classification, the dormancy depth classification result is derived from the judgment conclusion of step S3. When the dormancy depth classification is the light dormancy level, the concentration ratio of gibberellin GA3 and cytokinin 6-BA is set to 3:1. The specific operation is: the concentration of gibberellin GA3 mother liquor is 100 mg per liter, the concentration of cytokinin 6-BA mother liquor is 50 mg per liter, 75 ml of gibberellin GA3 mother liquor is mixed with 25 ml of cytokinin 6-BA mother liquor to form a mixed hormone solution with a total concentration of 85 mg per liter. When the dormancy depth classification is the deep dormancy level, the concentration ratio of gibberellin GA3 and cytokinin 6-BA is adjusted to 5:1. The specific operation is: 83.3 ml of gibberellin GA3 mother liquor is mixed with 16.7 ml of cytokinin 6-BA mother liquor to form a mixed hormone solution with a total concentration of 90 mg per liter. The ratio adjustment is based on potted plant experiments, which have shown that light-dormant shoots are more sensitive to cytokinins, so the 6-BA ratio is increased. Deep-dormant shoots require higher concentrations of gibberellins to break dormancy. Ratio setting verification is performed by sampling and testing the cytokinesis index of shoots 48 hours after adjustment. The light-dormant combination must reach a value of at least 0.8, while the deep-dormant combination must reach a value of at least 0.6.

[0096] The addition ratio of rhizosphere microbial active substances was adjusted based on the time window of ethylene inhibition. The time window was defined as the start time Ts to the end time Te, determined in step S4. The time window was divided into three equal periods: early (Ts to Ts + 1 / 3 (Te-Ts)), mid (Ts + 1 / 3 (Te-Ts) to Ts + 2 / 3 (Te-Ts)), and late (Ts + 2 / 3 (Te-Ts) to Te). The addition ratio in the early period was 5% of the volume of the mixed hormone solution; 10% in the mid-period; and 15% in the late period. The rhizosphere microbial active substances were derived from the bacterial suspension prepared in step S2, which was brought to room temperature and vortexed before use. The addition ratio was determined based on the following: low ethylene concentrations in the early period of the time window allowed a small amount of microorganisms to degrade ACC; high ethylene accumulation in the late period necessitated an increased microbial dosage. The ratio was verified by sampling the rhizosphere ACC content at each time point during the time window, requiring a degradation rate of at least 70% within 24 hours of addition.

[0097] When the adjusted gibberellin GA3, cytokinin 6-BA, and rhizosphere microbial active substances are mixed in proportion to form a composite 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 mixing tank and start the magnetic stirrer at 300 rpm. Then, slowly add the rhizosphere microbial active substances at a controlled addition rate of 50 ml / min. Finally, add deionized water to the total volume. Maintain the mixing temperature at 20°C ± 2°C for 30 minutes. The final composition of the composite solution is controlled to have a gibberellin GA3 concentration range of 50 to 70 mg / L, a cytokinin 6-BA concentration range of 10 to 15 mg / L, and a rhizosphere microbial active substance viable count of 10⁷ to 10⁸ bacteria per milliliter. Immediately after mixing, check the solution pH and adjust it to a range of 6.0 to 6.5 with 0.1 mol / L hydrochloric acid or sodium hydroxide. Shelf life of the reconstituted solution: Store at 20 degrees Celsius in a dark environment for no more than 6 hours. Reconstitute if the shelf life is exceeded.

[0098] Additional rules for concentration ratio adjustment: When the proportion of light-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 deep-dormant proportion exceeds 70%, the ratio is changed to 6:1. Ratio adjustment range limit: The ratio change within each regulation cycle shall not exceed ±0.5. Correction of the addition ratio of rhizosphere microbial active substances under special conditions: When the soil pH value is lower than 5.5, the addition ratio in each period is increased by 2%; when the soil temperature is higher than 30 degrees Celsius, the addition ratio is reduced by 3%. Environmental control of mixing operations: It is carried out in a clean workbench with an air cleanliness level of 10,000. The mixing container is made of brown glass or stainless steel, and plastic containers are prohibited to avoid adsorption.

[0099] Quality inspection indicators of the compound solution: Take samples immediately after mixing for biological activity testing. Take tissue sections from the base of the regenerated buds, immerse them in the compound solution, and place them in a 25-degree Celsius incubator for 24 hours. The qualification criteria are: the cell length growth rate of the light-dormant sample treatment group exceeds 50% of the control group; the cell division index of the deep-dormant sample treatment group reaches above 0.5. Physical indicator detection: the turbidity of the solution does not exceed 10NTU, and the viscosity is in the range of 1.2 to 1.5 centipoise. Microbial activity detection: Take 0.1 ml of the compound solution and spread it on the ACC plate. After 48 hours of incubation, the colony-forming unit is not less than 106 bacteria per ml. The labeling information of each batch of compound solution includes: preparation time, hormone concentration ratio, microbial addition ratio, applicable dormancy depth classification type, and corresponding time window period.

[0100] Preparation standards for the mother solutions of gibberellin GA3 and cytokinin 6-BA: Dissolve the gibberellin GA3 mother solution in 70% ethanol and then add distilled water to make up the volume; dissolve the cytokinin 6-BA mother solution in 0.1 mol / L sodium hydroxide and then add distilled water to make up the volume. The shelf life of the mother solution shall not exceed 15 days, and the storage temperature shall be 4 degrees Celsius. Filter and sterilize before use. Activation treatment of active substances of rhizosphere microorganisms: Thaw the frozen bacterial suspension in a 28-degree Celsius water bath for 5 minutes, add an equal volume of nutrient activation solution (containing 0.5% glucose and 0.1% yeast extract), and let it stand at 30 degrees Celsius for 30 minutes before use. Traceability mechanism for ratio adjustment: Record the reason for each adjustment, the original ratio, the adjusted ratio and the verification result to form a ratio adjustment history database.

[0101] Packaging and labeling of compound solutions: After mixing, the solution is packaged into 1-liter brown reagent bottles. The bottle label should indicate: solution type (light sleep type / deep sleep type), time window applicable period (early / middle / late period), preparation batch number. The packaging process is carried out under nitrogen protection, the liquid level is 1 cm away from the bottle mouth, and the bottle mouth is sealed with a polytetrafluoroethylene gasket. Transportation conditions: temperature 15 to 25 degrees Celsius, light-proof and shock-proof, transportation time not exceeding 2 hours. Shake well and check before use: when stratification or precipitation occurs, centrifuge at 2000 rpm for 5 minutes to take the supernatant. Treatment of residual 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 contain fields such as raw material batch number, dosage, operator, ambient temperature and humidity, test data, etc., and the storage period is not less than three years.

[0102] Field verification method for ratio setting: set up experimental plots of different ratios in the same dormancy depth graded area, and measure the elongation of regenerated buds 72 hours after regulation. Criteria for determining the optimal ratio: the ratio of the treatment group with the largest elongation and the smallest coefficient of variation. Biological basis for time window division: sampling and detection of ethylene receptor expression in the early stage is lower 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 compound solution: storage at 40 degrees Celsius for 24 hours is equivalent to storage at 20 degrees Celsius for 6 hours. During this period, the decrease in the number of viable bacteria does not exceed 10%, and the hormone degradation rate does not exceed 5% is considered qualified. All compound solutions need to be tested for compatibility before use: take 5 ml of solution and add it to a test tube. Observe for 2 hours without precipitation, flocculation, or phase separation before use.

[0103] S6. Within the time window, spray the compound solution in a targeted manner in the narrow row area. The specific implementation is as follows:

[0104] When the spray system is activated within the time window for ethylene inhibition, the time window is derived from the start time Ts to the end time Te determined in step S4. The startup procedure is as follows: 10 minutes before the start time Ts of the time window, the spray system is activated. A system self-check is performed, including pressure sensor calibration, nozzle clogging detection, and solution level monitoring. After passing the self-check, the spray program is initiated promptly at time Ts. The time window conversion mechanism is as follows: the time window data output in step S4 (formatted as "start hour: minute - end hour: minute") is input into the spray control terminal, which automatically converts it to local time and sets a countdown. Environmental constraints: When wind speed exceeds 3 meters per second or rainfall exceeds 2 mm per hour, the start is delayed and the time window is recalculated. After the delay, the time window is compressed to 50% of its original length and the system starts no later than 120 minutes after Ts.

[0105] When positioning the spray target area of ​​the spray device to the position of the regenerated buds above the surface of the narrow row area, the positioning operation is achieved through the following steps: first, obtain the coordinates of the field geographic information system and import the narrow row area boundary data. The sprayer is equipped with a differential global positioning system with a positioning accuracy of centimeters. The spray target area is set as a three-dimensional space layer with a height of 15 cm to 25 cm from the ground surface, and the width is the same as the narrow row area. Positioning calibration method: preset reference points in the field, use a laser rangefinder to measure the height and horizontal inclination of the spray boom, and ensure that the error of the parallelism of the spray boom to the ground is less than 1 degree. Regenerated bud position identification: based on the dormancy depth classification map of step S3, the deep dormancy level dense area is set as the priority target area. Positioning accuracy verification: after spraying the fluorescent tracer, randomly select points outside the target area to detect the deposition amount, and the deposition amount in the non-target area is required to not exceed 5% of the target area.

[0106] When spraying the compound solution above the ground surface at the location of regenerating buds in narrow rows, atomization parameters are controlled as follows: droplet median diameter 150 to 200 μm, atomization pressure 0.3 MPa to 0.5 MPa, and flow rate 0.8 to 1.2 L / min. Spraying method: Use fan-shaped atomizing nozzles with a nozzle spacing of 30 cm and a height of 20 cm above the ground. The compound solution is derived from the solution prepared in step S5. Shake well and preheat to 20°C ± 2°C before use. Environmental monitoring during spraying: Temperature and humidity sensors are placed in the target area, providing 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 losses. When the relative humidity falls below 50%, anti-drift mode is triggered, increasing the droplet median diameter to 250 μm. Spray coverage requirement: A continuous film of liquid forms on the surface of the regenerating buds without dripping, with a deposition rate of 20 to 30 μl per square centimeter of leaf surface.

[0107] When controlling the spray duration to cover the duration of the time window, the spray duration is set to 1.2 to 1.5 times the time window length (Te-Ts). Specific implementation: When the time window length is 60 minutes, the spray duration is 72 to 90 minutes. The spraying pattern is intermittent: 30 seconds of operation followed by 10 seconds of rest to ensure sufficient absorption of the solution. Time window coverage mechanism: Spraying starts 5 minutes earlier and ends 10 minutes later. Dynamic adjustment rule: When real-time monitoring detects that the ethylene concentration peak arrives early, the spray duration is automatically shortened to 80% of the original plan; if the peak arrives late, the spray duration is extended to 120%. The formula for calculating spray volume is: The total spray volume is equal to the narrow row area multiplied by the design dosage per hectare, then multiplied by the time window correction factor. For example, if the narrow row area is 0.5 hectares, the design dosage is 50 liters per hectare, and the time window correction factor is 1.3, the total spray volume is 0.5 times 50 times 1.3, which equals 32.5 liters.

[0108] Maintenance procedures for the spray device: Clean the pipeline after daily operation, first circulate it with 5% citric acid solution for 10 minutes, and then rinse it with deionized water 3 times. The nozzle should be disassembled and inspected weekly, and replaced when the wear exceeds 10% of the nominal aperture value. Daily calibration of the positioning system: Check the positioning deviation in the standard test field, and recalibrate when the deviation exceeds 3 cm. Verification of the spraying effect: Take samples 2 hours after spraying to detect the residual amount of solution on the surface of the regenerated buds, and use high-performance liquid chromatography to determine the amount of gibberellin GA3 deposition, which is required to be no less than 85% of the designed dose. Abnormal handling: When the coefficient of variation of the droplet deposition uniformity is monitored to exceed 25%, it will automatically pause and prompt to check for nozzle blockage.

[0109] Dynamic tracking of the spray target area: A machine vision system is installed on the spray boom to identify the location of the regenerated buds in real time and adjust the nozzle direction. Visual system recognition rate calibration: 1,000 images of the regenerated buds are taken under typical lighting conditions to establish a recognition model with an accuracy requirement of greater than 95%. Positioning compensation algorithm: The nozzle advance is calculated based on the locomotive's travel speed (10 to 15 meters per minute). 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 times 0.2, which equals 0.04 meters.

[0110] Atomization quality assurance measures: Droplet distribution is checked every 30 minutes, using a laser particle size analyzer for online monitoring. Acceptable droplet distribution criteria: Volume median diameter between 150 and 200 microns, and span factor (Dv0.9 minus Dv0.1) divided by Dv0.5 less than 1.5. If the test fails, the pressure is automatically adjusted: if the droplets are too large, the pressure is increased by 0.05 MPa; if the droplets are too small, the pressure is reduced by 0.03 MPa. Filtration of the compound solution: Install a 5-micron filter at the liquid inlet and replace the filter every 500 liters of solution processed.

[0111] Precise control of spraying duration: The control system has a built-in multi-level timing module. The first-level timer controls the total duration, the second-level timer divides the intervals, and the third-level timer manages the nozzle opening and closing timing. Time window exception handling: If the spraying is interrupted by heavy rain, the spraying time is recorded, and 150% of the remaining amount is re-sprayed within 24 hours after the time window closes. Before re-spraying, the residual amount on the surface of the buds must be tested. If the residual amount is higher than 50% of the designed dose, the re-spraying is cancelled. Data recording system: Completely record the start and end time of spraying, actual duration, environmental parameters, interruption events and handling measures to form a traceable spraying log.

[0112] Post-spraying effect evaluation: 24 hours after spraying, 20 randomly selected regenerated shoots are tested for changes in endogenous hormones. Light-dormant samples require a 40% or greater increase in gibberellin GA3 content, while deep-dormant samples require a 30% or greater increase in cytokinin 6-BA receptor expression. Areas that fail the evaluation are marked as abnormal and an additional 30% dose is applied during the next spraying window. Daily maintenance reports are generated for all spraying equipment, including nozzle status, positioning accuracy, solution consumption, and other data, and are kept for at least three years.

[0113] Example 2: Figure 2 The present invention provides a schematic structural diagram of a regeneration rice narrow row hormone regulation optimization system, which includes the following modules:

[0114] The rhizosphere anaerobic testing module is used to obtain rhizosphere soil samples from the narrow row area of ​​replanted rice and measure the anaerobic index of the rhizosphere soil samples;

[0115] ACC bacteria screening module, used to screen rhizosphere microbial active substances containing ACC deaminase when the anaerobic index exceeds the preset threshold;

[0116] The dormancy classification module is used to analyze the spectral absorption characteristics of narrow-row regeneration buds through near-infrared spectroscopy and classify the dormancy depth into light dormancy and deep dormancy according to the absorption threshold;

[0117] The ethylene time window module is used to track the timing of the transfer of ethylene concentration peaks from roots to shoots in narrow row areas and determine the time window of ethylene inhibition effects;

[0118] The compound solution adjustment 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 directional spraying module is used to spray the compound solution in a targeted manner on the narrow row area within the time window.

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

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

[0122] Those skilled in the art will appreciate that the modules and algorithm steps of each example 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 performed in hardware or software depends on the specific application of the technical solution and the invention constraints. Professional and technical personnel 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, each functional module in each embodiment of the present application may be integrated into one processing module, or each module may exist physically separately, or two or more modules may be integrated into one module.

[0124] In the several embodiments provided in this application, it should be understood that the disclosed systems, devices and methods can be implemented in other ways. For example, the device embodiments described above are merely schematic. For example, the division of the modules is only a logical function division. In actual implementation, there may be other division methods, such as multiple modules or components can be combined or integrated into another system, or some features can be ignored or not executed. Another point is that the mutual coupling or direct coupling or communication connection shown or discussed can be through some interfaces, indirect coupling or communication connection of devices or modules, which can be electrical, mechanical or other forms.

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

[0126] Finally: 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 in the scope of protection of the present invention.

Claims

1. A method for optimizing hormone regulation of narrow-row regeneration rice, characterized in that: The steps include: S1. Obtain rhizosphere soil samples from the narrow row area of ​​ratoon rice and measure the anaerobic index of the rhizosphere soil samples; S2. When the anaerobic index exceeds a preset threshold, screening for active substances of rhizosphere microorganisms containing ACC deaminase; S3. Analyze the spectral absorption characteristics of the narrow row regenerated buds by near-infrared spectroscopy, and classify the dormancy depth into light dormancy and deep dormancy according to the absorption threshold; S4. Track the timing of the transfer of the peak ethylene concentration from the root system to the shoot in the narrow row area to determine the time window of the ethylene inhibitory effect; S5. Based on the dormancy depth classification and time window, determine the ratio of gibberellin GA3, cytokinin 6-BA and rhizospheric microbial active substances to form a compound solution; S6. Within the time window, spray the compound solution in a targeted manner in the narrow row area.

2. The method for optimizing hormone regulation of narrow row regeneration rice according to claim 1, characterized in that: Obtain rhizosphere soil samples from the narrow row area of ​​ratoon rice and measure the anaerobic index of the rhizosphere soil samples, including: Rhizosphere soil samples were collected at the location where regeneration shoots were delayed in the narrow row area; Determine real-time oxygen partial pressure in rhizosphere soil samples from narrow row areas; Obtain the oxygen partial pressure of rhizosphere soil samples from wide-row areas at the same growth period as the wide-row baseline value; The anaerobic index was calculated based on the deviation ratio of the real-time oxygen partial pressure of rhizosphere soil samples in the narrow row area to the wide row baseline value.

3. The method for optimizing hormone regulation of narrow row regeneration rice according to claim 2, characterized in that: When the anaerobic index exceeds the preset threshold, the active substances of rhizosphere microorganisms containing ACC deaminase are screened, including: Determine whether the anaerobic index exceeds a preset threshold; When the anaerobic index exceeds a preset threshold, the Pseudomonas strain that secretes ACC deaminase is selected; The ACC deaminase activity of Pseudomonas strains was detected by enzyme-linked immunosorbent assay; The strains whose ACC deaminase activity reached the standard value were screened to prepare rhizosphere microbial active substances.

4. The method for optimizing hormone regulation of narrow row regeneration rice according to claim 3, characterized in that: The spectral absorption characteristics of narrow row regeneration buds were analyzed by near-infrared spectroscopy, and the dormancy depth was divided into light dormancy and deep dormancy according to the absorption threshold, including: The leaf sheath tissue at the base of the narrow row regenerated buds was collected as the test sample; The sample was scanned and tested using a near-infrared spectrometer to obtain the absorbance at a wavelength of 1450 nm; The absorbance is compared with the preset absorption threshold to divide the sleep depth into light sleep level and deep sleep level.

5. The method for optimizing hormone regulation of narrow row regeneration 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 a light sleep level; when the absorbance is greater than the preset absorption threshold, it is determined to be a deep sleep level.

6. The method for optimizing hormone regulation of narrow row regeneration rice according to claim 4, characterized in that: Track the timing of the transfer of ethylene concentration peaks from roots to shoots in narrow row areas and determine the time window of ethylene inhibition effects, including: Arrange ethylene gas collection devices at the root level in narrow row areas; An ethylene gas collection device is synchronously arranged at the base of the narrow row of regenerated shoots; The ethylene concentrations at the root and base were measured at regular time intervals; The time difference of the peak ethylene concentration from the root position to the base position was recorded; The time window of ethylene inhibitory effect was determined based on the time difference.

7. The method for optimizing hormone regulation of narrow row regeneration rice according to claim 6, characterized in that: The time window is the period before the peak reaches the base position.

8. The method for optimizing hormone regulation of narrow row regeneration rice according to claim 6, characterized in that: Based on the dormancy depth classification and time window, the ratio of gibberellin GA3, cytokinin 6-BA and rhizospheric microbial active substances is determined to form a compound solution, including: Adjust the concentration ratio of gibberellin GA3 and cytokinin 6-BA according to the results of light dormancy or deep dormancy in the dormancy depth classification; Adjust the addition ratio of rhizosphere microbial active substances according to the time window of ethylene inhibition effect; The adjusted gibberellin GA3, cytokinin 6-BA and rhizosphere microbial active substances are mixed in proportion to form a compound solution.

9. The method for optimizing hormone regulation of narrow-row regenerated rice according to claim 8, characterized in that: Within the time window, target narrow row areas with a targeted spray application of a formulated solution consisting of: Start the spray device within the time window of ethylene inhibition effect; Position the spray target of the spray device to the location of the regenerating buds above the ground surface in the narrow row area; Spray the compound solution in a mist form to the location of regenerating buds above the ground surface in the narrow row area; Control the duration of the spraying time window.

10. A system for optimizing hormone regulation of narrow rows of regenerated rice, used to implement the method for optimizing hormone regulation of narrow rows of regenerated rice according to any one of claims 1 to 9, characterized in that: Includes the following modules: The rhizosphere anaerobic testing module is used to obtain rhizosphere soil samples from the narrow row area of ​​replanted rice and measure the anaerobic index of the rhizosphere soil samples; ACC bacteria screening module, used to screen rhizosphere microbial active substances containing ACC deaminase when the anaerobic index exceeds the preset threshold; The dormancy classification module is used to analyze the spectral absorption characteristics of narrow-row regeneration buds through near-infrared spectroscopy and classify the dormancy depth into light dormancy and deep dormancy according to the absorption threshold; The ethylene time window module is used to track the timing of the transfer of ethylene concentration peaks from roots to shoots in narrow row areas and determine the time window of ethylene inhibition effects; The compound solution adjustment 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 directional spraying module is used to spray the compound solution in a targeted manner on the narrow row area within the time window.

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