A chickpea seed seedling cultivation method

Abstract

This invention discloses a method for cultivating robust seedlings from broad bean seeds, belonging to the field of agricultural planting and seed treatment technology. The method includes: rate-limited imbibition using a hypertonic solution at 12℃~15℃; initiating a synchronous induction program when the seed water absorption reaches 60%~70% of saturation; uniformly replacing the first solution with a second solution using a peristaltic pump within 20~30 minutes to achieve a smooth transition of water potential, while simultaneously applying red / far-red light pulsed cycles with an R / FR of 2.5~4.0 for induction; adjusting the temperature to 18℃~22℃ to maintain germination; and providing supplemental lighting after the radicle breaks through. This invention, by precisely capturing the physiological window and implementing synchronous photo-hyperbibition coupling, effectively overcomes the problems of severe imbibition damage and asynchronous germination in large broad bean seeds, significantly reduces solute leakage rate, shortens seedling uniformity deviation, accelerates endogenous GA3 synthesis, achieves synergistic effects of physical repair and biological induction, and significantly improves seedling vigor.

Description

Technical Field

[0001] This invention belongs to the field of agricultural planting and seed treatment technology, specifically relating to a method for osmotic regulation and photoperiod coupling treatment of large-grained legume seeds, and particularly to a method for cultivating robust seedlings of broad bean seeds. Background Technology

[0002] broad bean( Vicia faba L. As a typical large-grained legume, broad bean seed germination is extremely sensitive to the water environment. In production practice, soaking seeds is a common method to improve germination rate. However, due to the biological characteristics of broad bean seeds, such as thick seed coats, high cotyledon starch content, and deep dehydration, the following deep technical challenges exist in the seed soaking process.

[0003] Imbibitional damage is a core physical obstacle restricting the viability of large seeds. In the initial stages of dry seed contact with water, the cell membrane structure is in the gel phase. As water rapidly enters, the cell membrane must quickly transition to the liquid-crystalline phase. Due to the large size of broad bean seeds and the highly uneven osmotic gradient of water between tissues, if the imbibition rate is too fast, the cell membrane, which has not yet completed its remodeling, will physically rupture under immense turgor pressure. This damage not only leads to the massive leakage of intracellular electrolytes, sugars, and amino acids, providing a breeding ground for pathogens, but also directly disrupts the physiological integrity of the embryo, resulting in a significant decrease in germination rate. While existing technologies attempt to slow the imbibition rate using osmotic regulators (such as salts or sugars), they often employ a constant hyperosmotic environment. Although this protects the membrane structure, the excessive imbibition resistance in the later stages inhibits normal seed germination, making it difficult to simultaneously address both "repair" and "initiation."

[0004] Poor synchronicity in seed germination is a key logical flaw hindering precision sowing and robust seedling cultivation. Modern mechanized sowing requires "full and uniform seedling emergence," meaning a highly concentrated germination time distribution. However, individual seeds exhibit natural differences in maturity, vigor, and water absorption rate. Conventional soaking methods cannot provide real-time feedback control of the seed's internal physiological state, leading to uneven repair progress. Furthermore, the synthesis of endogenous hormones (such as gibberellin GA3) within the seed depends on specific water content thresholds and the activation state of phytochromes. If the timing of external induction signals (such as light or osmotic pressure changes) does not match the seed's internal physiological window (membrane fluidity recovery node), it will disrupt the metabolic kinetics of endogenous hormones, manifesting as large deviations in seedling emergence uniformity and uneven seedling growth.

[0005] The disconnect between changes in the physical environment and biological metabolic signals limits induction efficiency. Existing physical induction methods (such as specific wavelength illumination) are often independent of the water regulation process. In fact, the sensitivity of seeds to light signals is dynamically affected by their tissue water content and membrane permeability. In the early stages of imbibition, the phytochrome system is not yet fully activated, and early illumination is ineffective; while in the later stages of imbibition, the metabolic pathway is established, and the induction window closes. Currently, there is a lack of a technical solution that can precisely couple the dynamic switching of the osmotic pressure gradient with the light pulse signal on the time axis according to the real-time water absorption status of the seed.

[0006] In summary, accurately capturing the key physiological window for broad bean seed membrane repair and simultaneously activating endogenous hormone metabolism through controlled physical field changes, thereby effectively avoiding imbibition damage while significantly improving the synchronicity of population germination, is a technical bottleneck that urgently needs to be overcome in the current field of broad bean seedling cultivation. Summary of the Invention

[0007] To address the problems of severe imbibition damage, poor germination synchronicity, and insufficient seedling vigor during the soaking process of broad bean seeds, this invention provides a method for cultivating robust broad bean seedlings. The method aims to achieve synergistic membrane repair and metabolic activation by accurately capturing physiological windows and implementing synchronous photoperfusion induction.

[0008] To achieve the above-mentioned objectives, the present invention adopts the following technical solution:

[0009] A method for cultivating robust seedlings from broad bean seeds includes the following steps:

[0010] (1) Initial repair seed soaking: Place broad bean seeds in the first osmosis solution and carry out imbibition repair at 12℃~15℃;

[0011] (2) Induction node triggering: Real-time monitoring of seed water absorption. When the seed water absorption reaches 60% to 70% of its saturation water absorption, the synchronous induction program is started.

[0012] (3) Photoosmosis synchronous induction: The first osmosis solution is replaced with the second osmosis solution within 20 to 30 minutes, and red / far-red light pulse induction is applied at the same time as the solution replacement;

[0013] (4) Germination management: After replacement, adjust the ambient temperature to 18℃~22℃ and maintain it until germination, and provide supplemental light treatment after the radicle breaks through the seed coat.

[0014] Further, in step (1), the first permeation solution is a sorbitol solution containing calcium chloride, wherein the mass concentration of the sorbitol is 10% to 15% and the mass concentration of the calcium chloride is 0.1% to 0.2%.

[0015] Furthermore, the water potential of the first permeation solution is -1.5 MPa to -0.8 MPa.

[0016] Further, in step (3), the second permeation solution is a mannitol solution with a mass concentration of 3% to 5%.

[0017] Furthermore, the water potential of the second permeation solution is -0.3 MPa to -0.1 MPa.

[0018] Furthermore, in step (3), the red light / far-red light pulse induction adopts a narrow-band LED light source, with a red light wavelength of 660nm and a far-red light wavelength of 730nm.

[0019] Furthermore, the energy ratio R / FR of the red light to far-red light induced by the red light / far-red light pulse is 2.5 to 4.0.

[0020] Furthermore, the operation of the red light / far-red light pulse induction is as follows: red light irradiation for 3-5 minutes, followed by far-red light irradiation for 1-2 minutes, and the induction is repeated 3-5 times.

[0021] Furthermore, in step (3), the first permeation solution is replaced with the second permeation solution by a peristaltic pump, and the pumping flow rate of the second permeation solution is controlled by the peristaltic pump to be 0.5 to 2.0 mL / min, so as to achieve a smooth transition of the water potential of the soaking environment from the water potential of the first permeation solution to the water potential of the second permeation solution.

[0022] Furthermore, the supplementary lighting treatment in step (4) has a supplementary lighting intensity of 50-80. The duration is 1 hour.

[0023] Beneficial effects:

[0024] (1) This invention provides sufficient buffer time for the smooth reorganization of seed cell membranes from the gel phase to the liquid crystal phase by limiting the rate of imbibition under an initial low water potential environment (-1.5 MPa to -0.8 MPa). Combined with the peristaltic pump-guided smooth water potential transition mechanism, it effectively avoids the osmotic shock caused by drastic changes in water potential during traditional seed soaking, and significantly reduces the extravasation of intracellular solutes (the solute extravasation rate in Example 3 was only 5.4% ± 0.3%, significantly lower than the 28.4% ± 1.8% of the deionized water soaking in Comparative Example 2), thus ensuring the physiological integrity of the embryonic tissue.

[0025] (2) This invention accurately captures the critical physiological window when the seed water absorption reaches 60% to 70% saturation. By applying a synchronous induction signal during this window period, the metabolic pace within the seed population was successfully unified, overcoming the germination delay or inconsistency caused by individual seed differences, and achieving a high degree of convergence in the germination time distribution of the population (the seedling uniformity deviation in Example 3 was only 1.6±0.1h, significantly better than the 8.4±0.8h in the lightless induction group of Comparative Example 1), laying the foundation for modern mechanized precision sowing.

[0026] (3) This invention breaks through the limitations of decoupling induction factors in traditional processes, precisely aligning the dynamic switching of the osmotic pressure gradient with the red / far-red light pulses on the time axis. This "multi-field coupling" induction mode produces a stronger activation effect on the synthesis pathway of endogenous gibberellin (GA3) (Example 3: GA3 rise rate reaches 18.2%). The concentration was significantly higher than 4.2 ng / g·h for Comparative Example 1 with single osmotic pressure adjustment and 8.9 ng / g·h for Comparative Example 4 with decoupling treatment. Its induction efficiency is significantly better than that of single osmotic pressure regulation or single light treatment, achieving a dual acceleration of "repair" and "start-up" at the physiological and metabolic level.

[0027] (4) The seeds treated by this invention not only show a significantly improved germination rate (98.5% ± 0.5% in Example 3), but also, due to sufficient initial membrane repair, reduced nutrient leakage, allowing more nutrients inside the seed to be converted into seedling growth energy. Experimental data show that the dry weight of seedlings treated by this invention is significantly increased, and the seedling vigor index is greatly improved. This superior physiological state enables seedlings to exhibit stronger growth potential during the hypocotyl elongation and root development stages, providing a solid physiological foundation for stable and high yields in complex field environments.

[0028] In summary, this invention overcomes the problems of severe water absorption damage and poor germination synchronicity in traditional broad bean soaking processes by synergistically regulating the osmotic pressure gradient and light pulse signal during the soaking process. Within the induction window of the seed water absorption trigger point (60%–70%), solution switching time (20–30 min), and R / FR ratio (2.5–4.0), the germination synchronicity and the accumulation rate of the endogenous hormone GA3 are optimal. The photoosmotic synchronization coupling treatment introduced in this invention is beneficial for maintaining the cell membrane structure during osmotic pressure changes.

[0029] Integrity, accelerating the biosynthetic kinetics of endogenous gibberellins, is beneficial for seeds to maintain germination consistency in complex field environments. Detailed Implementation

[0030] To enable those skilled in the art to better understand the technical solutions of this invention, the present application will be further described in detail below with reference to embodiments.

[0031] In this invention, "saturated water absorption" refers to the maximum water absorption of broad bean seeds when they reach an equilibrium state of water absorption. It serves as the benchmark for calculating the trigger threshold of "seed water absorption reaching 60%–70% of its saturated water absorption." The specific measurement conditions and calculation methods are as follows: Measurements are conducted in deionized water at 25℃±1℃. The criterion for reaching the water absorption plateau (i.e., "saturation") is a difference of less than 0.5% between two consecutive weighings (2 hours apart) after 48 hours of soaking. The calculation is based on the average weight gain of a batch of at least 100 seeds, using the formula: Saturated water absorption (%) = ( ) / ×100%, of which The dry weight of the seeds is (g). Seed mass (g) required to reach the above saturation criteria. In actual cultivation, when the cumulative water absorption of the population (calculated in the same way as above) reaches 60% to 70% of the saturation water absorption measured according to this standard method, the synchronous induction program is triggered.

[0032] In both the embodiments and comparative examples of this invention, "Zhongcan No. 1" broad bean seeds were used as the test material, with an initial moisture content of 11.5% ± 0.3% and a thousand-seed weight of 1250g ± 25g. The experiment was conducted in a controlled chamber equipped with a precision peristaltic pump (flow rate accuracy ± 0.1 mL / min) and a narrow-band LED light source (half-width ≤ 15 nm). The seeds were placed in a stainless steel filter layer, and the solution was pumped in from the bottom and dynamically circulated. By precisely adjusting the pumping flow rate of the second permeation solution using the precision peristaltic pump, the solute concentration in the soaking chamber could change linearly with the displacement process, thereby guiding the water potential of the soaking environment to smoothly transition from the low water potential of the first permeation solution to the high water potential of the second permeation solution, avoiding osmotic shock caused by sudden changes in water potential. Water absorption was monitored by weighing the total mass of the soaking container in real time using a high-precision electronic balance, and calculated from the cumulative mass increment based on the above calculation method, with the real-time error controlled within ± 0.5%.

[0033] Unless otherwise specified, the raw materials used in this invention are all commercially available conventional products; the technical means used are all conventional means well known to those skilled in the art unless otherwise specified.

[0034] Example 1

[0035] 100g of broad bean seeds were placed in a soaking chamber equipped with a filter, and a 10% sorbitol solution was injected, along with 0.1% calcium chloride. The solution potential was adjusted to -1.5 MPa, and the ambient temperature to 12℃. Under this low potential (-1.5 MPa) environment, the seeds entered a slow imbibition state. The total mass of the soaking container was monitored in real time using a high-precision electronic balance. Based on the aforementioned calculation method for saturated water absorption (converted by cumulative mass increment), when the seed water absorption reached 60% of its saturated water absorption, the system automatically initiated a solution switching program: within 20 minutes, a second permeation solution was pumped in at a constant flow rate (2.0 mL / min) using a precision peristaltic pump, gradually replacing the original solution, so that the water potential of the soaking environment smoothly transitioned from -1.5 MPa to -0.3 MPa within 20 minutes. Simultaneously with the start of the replacement, an LED array was activated to apply pulsed light with an R / FR ratio of 2.5 (red light 660nm irradiation for 3 min, far-red light 730nm irradiation for 1 min), and this was induced in three cycles. After replacement, raise the temperature to 18℃ and maintain it until germination, and provide 50% oxygen after the radicle breaks through. Supplemental lighting treatment (lasting 1 hour).

[0036] Example 2

[0037] Except for the following parameter adjustments, the operating procedure is the same as in Example 1: In the first stage, a 15% sorbitol solution (0.2% calcium chloride) was used, with a water potential of -0.8 MPa and a temperature of 15°C. Switching was triggered when the seed's water absorption reached 70% of its saturation water absorption capacity. The solution replacement process was set to 30 minutes, pumped in at a flow rate of 0.5 mL / min, with the target solution being 5% mannitol (-0.1 MPa). Simultaneously, light pulses with an R / FR ratio of 4.0 were applied (red light for 5 minutes, far-red light for 2 minutes), repeated 5 times. In the later stages, the ambient temperature was set to 22°C, and the supplemental light intensity was 80 ppm. .

[0038] Example 3

[0039] Except for the following parameter adjustments, the operating procedure is the same as in Example 1: In the first stage, a 12.5% ​​sorbitol solution (0.15% calcium chloride) was used, with a water potential of -1.15 MPa and a temperature of 13.5℃. Switching was triggered when the seed water absorption reached 65% of its saturation water absorption. The solution replacement process was set to 25 minutes, pumped at a flow rate of 1.2 mL / min, with the target solution being 4% mannitol (-0.2 MPa). Simultaneously, light pulses with an R / FR ratio of 3.25 were applied (red light for 4 min, far-red light for 1.5 min), repeated 4 times. In the later stages, the ambient temperature was set to 20℃, and the supplemental light intensity was 65. .

[0040] Comparative Example 1

[0041] Similar to Example 3, except that the LED light source is not turned on during solution switching, and only the osmotic pressure gradient is adjusted.

[0042] Comparative Example 2

[0043] Same as Example 3, except that no sugar alcohol solution is used, and deionized water is used for seed soaking throughout the process, with light pulses applied when the water absorption reaches 65%.

[0044] Comparative Example 3

[0045] Same as Example 3, except that the solution switching and light pulse-induced triggering node are advanced to when the water absorption reaches 30% of the saturation value.

[0046] Comparative Example 4

[0047] Similar to Example 3, the difference lies in decoupling the two inducing factors: the solution switching is completed within 25 minutes, and the same intensity of light pulse irradiation is performed after the osmotic pressure stabilizes at -0.2 MPa.

[0048] Comparative Example 5

[0049] Similar to Example 3, except that the solution switching time is set to an extremely short 45 seconds (instantaneous replacement) to simulate an osmotic shock environment.

[0050] Comparative Example 6

[0051] Same as Example 3, except that the solution switching time is set to 120 minutes to simulate a very slow osmotic pressure transition.

[0052] Data Analysis and Comparison

[0053] To objectively evaluate the technical effects of the present invention, germination indices were measured on the seeds treated in Examples 1-3 and Comparative Examples 1-6.

[0054] 1. Germination rate and uniform seedling emergence deviation

[0055] The sand bed germination method was used in a 25℃ constant temperature artificial climate chamber. Each group was repeated 4 times, with 100 seeds per time. Germination was judged when the radicle broke through the seed coat by 2 mm. The number of germinated seeds was recorded every 2 hours. The uniformity deviation was defined as the time interval (h) required for the population to grow from a 10% germination rate to a 90% germination rate. The smaller the value, the higher the synchronicity.

[0056] 2. Cell solute extravasation rate

[0057] Twenty seeds were randomly selected from each of the treated seeds, and the process was repeated four times. 100 mL of deionized water was added, and the seeds were soaked at 25°C for 24 hours. The conductivity of the extract was measured using a conductivity meter (C1). Subsequently, the seeds were placed in a boiling water bath for 15 minutes, and after cooling to 25°C, the total conductivity was measured (C2). Percolation rate = (C1 / C2) × 100%.

[0058] 3. Seedling Vigor Index

[0059] On the 7th day of germination, 30 normal seedlings were randomly selected and the process was repeated 4 times. The hypocotyl and root system were cut off and blanched at 105℃ for 30 minutes. Then, they were dried at 80℃ to constant weight. The calculation formula is: Vigor Index = Germination Rate × Average Dry Weight of Seedlings (mg).

[0060] 4. Kinetic analysis of endogenous hormone GA3: 0.5 g of seed embryo tissue was collected at the solution switching start time (0 h) and 4 hours after the end of induction treatment (4 h). The tissue was quickly frozen in liquid nitrogen and ground. Extraction was performed at 4°C for 12 hours in cold methanol containing 1 mmol / L butylated hydroxytoluene (BHT) antioxidant, protected from light. The extract was concentrated under reduced pressure until an alcohol-free phase was obtained, and the pH was adjusted to 2.5-3.0 with 1 mol / L HCl. The extract was then extracted three times with an equal volume of ethyl acetate. The organic phases were combined and evaporated to dryness under reduced pressure. The residue was dissolved in chromatographic grade methanol and filtered through a 0.22 μm filter. GA3 content was determined using high-performance liquid chromatography (HPLC, C18 reversed-phase column, mobile phase: methanol:0.1% acetic acid aqueous solution, volume ratio 60:40, detection wavelength 254 nm). The assay was repeated four times. The GA3 rise rate was defined as the ratio of the difference in GA3 content in the embryo before and after induction treatment to the induction time. ).

[0061] Table 1 records the full sample statistical mean (Mean±SD) after each group was repeated 4 times.

[0062] Table 1

[0063] The results (Table 1) show that Examples 1-3 exhibited significant correlational advantages in core indicators such as germination rate, uniform emergence deviation, and vigor index. Compared to Comparative Examples 1-6, the Example groups achieved a narrower germination time distribution (uniform emergence deviation less than 3 hours) and a higher endogenous hormone metabolism rate while maintaining a lower solute extravasation rate. This overall trend reflects the synergistic contribution of initial hyperosmolar remediation, triggering of a specific water absorption window, and synchronous coupling of light and osmolarity to the broad bean seed germination process.

[0064] Comparative Example 1: In the absence of light pulse induction, the GA3 rise rate (4.2%) The germination synchronicity was significantly lower than in Example 3, and the germination deviation increased to 8.4 hours. This reflects that the activation intensity of the endogenous hormone synthesis pathway by single osmolarity regulation is limited, and the improvement of germination synchronicity depends to some extent on the further triggering of physiological metabolism by light signals.

[0065] Comparative Example 2 showed the highest solute extravasation rate (28.4%) and the lowest germination rate. This result is related to the physical damage to the cell membrane caused by the rapid imbibition rate of seeds in pure water, confirming the fundamental role of the initial high-osmolarity rate-limiting environment in ensuring the integrity of the membrane structure.

[0066] Comparative Example 3, with the trigger point advanced to 30% water absorption, showed a significantly higher solute extravasation rate (18.2%) than Example 3, and a lower viability index. Based on the seed's physiological state, it is speculated that the cell membrane had not yet completed a smooth transition from the gel phase to the liquid crystal phase during the initial imbibition stage. The osmotic pressure changes and signal stimulation at this time may have exacerbated the mechanical stress on the membrane structure.

[0067] Comparative Example 4 employed a decoupling treatment (switching osmotic pressure before irradiation with light pulses). Although the solute extravasation rate was controlled, the GA3 rise rate and uniformity deviation were inferior to those of Example 3, which was treated synchronously. Data trends indicate that the overlap between the dynamic decrease in osmotic pressure and the light pulses on the time axis has a non-linear promoting effect on improving the metabolic response rate.

[0068] In Comparative Example 5 (instantaneous switching), the solute extravasation rate surged to 22.8%, which can be attributed to the osmotic shock induced by the dramatic change in water potential gradient within a very short time, leading to secondary damage to the membrane structure. While Comparative Example 6 (120-minute switching) maintained better membrane integrity, the emergence deviation significantly increased (12.1 hours), and the GA3 rise rate was low. This phenomenon reflects a threshold effect in the rate of osmotic pressure decrease; excessively slow physical stimuli may dilute the induction signal, making it difficult to form an effective physiological resonance with the light pulse, thus affecting the convergence of population germination pace.

[0069] In summary, this invention overcomes the problems of severe imbibition damage and poor germination synchronicity in traditional seed soaking processes by synergistically regulating the osmotic pressure gradient and light pulse signal during broad bean seed soaking. The seed water absorption trigger point, solution switching time, and light pulse parameters are correlated, jointly influencing the physiological metabolic level of the seed embryo and the physicochemical state of the membrane structure. Within the induction window defined by this invention—60%–70% water absorption, 20–30 min solution switching time, and R / FR ratio of 2.5–4.0—seed germination synchronicity and the accumulation rate of endogenous hormone GA3 show good performance. The photoosmotic synchronization coupling treatment introduced in this invention helps maintain the structural integrity of the cell membrane during osmotic pressure changes. This treatment accelerates the biosynthetic kinetics of endogenous gibberellins, which is beneficial for maintaining germination consistency in complex field environments. Under the conditions of this embodiment, the three-stage induction effect based on initial membrane repair and photoosmotic coupling is superior to conventional constant osmotic pressure treatment or single light induction. This invention improves the germination performance and seedling vigor of broad bean seeds without altering their genetic characteristics by combining specific physical parameters with physiological window design.

Claims

1. A method for cultivating robust seedlings from broad bean seeds, characterized in that, Includes the following steps: (1) Initial repair seed soaking: Place broad bean seeds in the first osmosis solution and carry out imbibition repair at 12℃~15℃; (2) Induction node triggering: Real-time monitoring of seed water absorption. When the seed water absorption reaches 60% to 70% of its saturation water absorption, the synchronous induction program is started. (3) Photoosmosis synchronous induction: The first osmosis solution is replaced with the second osmosis solution within 20 to 30 minutes, and red / far-red light pulse induction is applied at the same time as the solution replacement; (4) Germination management: After replacement, adjust the ambient temperature to 18℃~22℃ and maintain it until germination, and provide supplemental light treatment after the radicle breaks through the seed coat.

2. The method for cultivating robust broad bean seedlings according to claim 1, characterized in that, In step (1), the first permeation solution is a sorbitol solution containing calcium chloride, wherein the mass concentration of the sorbitol is 10% to 15% and the mass concentration of the calcium chloride is 0.1% to 0.2%.

3. The method for cultivating robust broad bean seedlings according to claim 2, characterized in that, The water potential of the first permeation solution is -1.5 MPa to -0.8 MPa.

4. The method for cultivating robust seedlings from broad bean seeds according to claim 1, characterized in that, In step (3), the second permeation solution is a mannitol solution with a mass concentration of 3% to 5%.

5. The method for cultivating robust seedlings from broad bean seeds according to claim 4, characterized in that, The water potential of the second permeation solution is -0.3 MPa to -0.1 MPa.

6. The method for cultivating robust broad bean seedlings according to claim 1, characterized in that, In step (3), the red light / far-red light pulse induction uses a narrow-band LED light source with a red light wavelength of 660nm and a far-red light wavelength of 730nm.

7. The method for cultivating robust seedlings from broad bean seeds according to claim 6, characterized in that, The red light to far-red light energy ratio R / FR induced by the red light / far-red light pulse is 2.5 to 4.

0.

8. The method for cultivating robust seedlings from broad bean seeds according to claim 7, characterized in that, The operation of the red / far-red pulse induction is as follows: red light irradiation for 3-5 minutes, followed by far-red light irradiation for 1-2 minutes, and the induction is repeated 3-5 times.

9. The method for cultivating robust seedlings from broad bean seeds according to claim 1, characterized in that, In step (3), the first permeation solution is replaced with the second permeation solution by a peristaltic pump, and the pumping flow rate of the second permeation solution is controlled by the peristaltic pump to be 0.5 to 2.0 mL / min, so as to achieve a smooth transition of the water potential of the soaking environment from the water potential of the first permeation solution to the water potential of the second permeation solution.

10. The method for cultivating robust seedlings from broad bean seeds according to claim 1, characterized in that, The supplementary lighting treatment in step (4) has a supplementary lighting intensity of 50-80. The duration is 1 hour.

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