Wheat seed vigor detection method based on oxygen sensing

Abstract

The invention provides a wheat seed vigor detection method based on oxygen sensing, and the wheat seed vigor detection method based on oxygen sensing comprises the steps of pretreatment, detection and statistics, and specifically comprises the following steps: selecting full, consistent, insect-hole-free and mildew-free wheat seeds, and carrying out aging treatment to obtain pre-detected wheat seeds a; the method comprises the following steps: adding a single seed of a to-be-detected wheat seed a into a sample tube, transferring into a seed vitality instrument, monitoring an oxygen consumption curve in real time, and screening out five parameters including an initial metabolic rate SMR, an oxygen metabolic rate OMR, time R50 for 50% of seeds to reach 50% of oxygen concentration, area AUC50 under the 50% oxygen concentration curve and relative germination time RGT; the indexes of SMR, OMR, R50, AUC50 and RGT are used for determining the seed vigor. Compared with the prior art, the method has the advantages that breakthrough progress is realized in detection efficiency, parameter value and variety suitability, and particularly, the technical blank of wheat seed oxygen sensing activity detection is filled through a three-in-one innovative design of'medium-parameter-algorithm '.

Description

Technical Field

[0001] The present invention relates to the field of detection technology, and in particular to a wheat seed vitality detection method based on oxygen sensing. Background Art

[0002] Seed vigor refers to the potential for seeds to germinate under actual field conditions and its correlation with yield and related agronomic indicators. It is not only closely linked to crop production but also to physiological and biochemical processes such as seed development, maturation, germination, storage life, and deterioration. As a key assessment criterion for seed quality, seed vigor plays a central role in seed quality assessment and seed trading. Previously, one of the most commonly used methods for testing seed vigor was the seed germination test. However, traditional germination tests are time-consuming and labor-intensive, making them difficult to meet the current demand for rapid, efficient, and accurate seed viability assessment in modern agricultural production. Therefore, the development of a product that can address these technical issues is crucial. Summary of the Invention

[0003] The purpose of the present invention is to provide a wheat seed vitality detection method based on oxygen sensing.

[0004] The object of the present invention is achieved in this way. The wheat vitality detection method based on oxygen sensing includes pre-processing, detection and statistical steps, specifically including: A. Pre-treatment: Select plump, uniform, insect-free and mildew-free wheat seeds and perform aging treatment to obtain pre-test wheat seeds a; B. Testing: 1) Add agar to distilled water to prepare agar solution b with a concentration of 0.4-0.6%; 2) Add 900-1100 µL of agar solution b to a sample tube. After solidification, add a single seed from pre-tested wheat seed a to the sample tube. Transfer the seed to a seed viability meter and monitor the oxygen consumption curve in real time. Five parameters are screened: initial metabolic rate (SMR), oxygen metabolic rate (OMR), time for 50% of seeds to reach 50% oxygen concentration (R50), area under the 50% oxygen concentration curve (AUC50), and relative germination time (RGT); C. Statistics: SMR, OMR, R50, AUC50 and RGT indicators were used to determine seed vigor.

[0005] The application is directed to the construction of a set of innovative systems based on oxygen sensing technology for wheat seed vigor detection, the core technical features of which include: the use of 0.5% concentration agar medium (0.15g agar + 150ml distilled water) and 1000ul volume of special combination, real-time monitoring of oxygen consumption curve by seed vigor instrument under constant temperature 20℃ conditions, and screening of five core parameters of initial metabolic rate (SMR), oxygen metabolic rate (OMR), 50% seed reaching 50% oxygen concentration time (R50), 50% oxygen concentration curve area (AUC50) and relative germination time (RGT) as the vigor determination index. The system realizes the accurate detection of conventional wheat variety "Yunmai 56" by optimizing the physical properties of the medium (avoiding seed sinking or suspension, ensuring 100% anti-S-shaped oxygen consumption curve) and the equipment adaptation method (40-hole plate regional scanning). The technical contribution lies in significantly improving the detection efficiency and sensitivity: on the one hand, the 7-day cycle of traditional germination test is shortened to a few hours (vigor is predicted by the initial oxygen consumption curve shape), solving the time efficiency problem that has existed in the industry for a long time; on the other hand, the high sensitivity of R50 parameter to seed aging is first found (R50 value changes from 59.35 to 106.93 after 30 days of aging, with a change rate of 80%, which is 5.7 times more sensitive than the germination rate index), and a multi-parameter cross-validation model is established combining the strong stability of SMR / OMR (no significant difference under volume fluctuation), with an error rate of less than 5%. In addition, the system provides a standardized vigor monitoring scheme for seed banks through modular design (such as adjusting the medium volume to adapt to different crops), supporting the safety preservation of germplasm, for example, SMR<0.002 can quickly identify deteriorated seeds. Compared with existing technologies, the application has made breakthrough progress in detection efficiency, parameter value and variety adaptability, especially through the "medium-parameter-algorithm" trinity innovation design, filling the technical gap in wheat seed oxygen sensing vigor detection.

[0006] (I) Special detection system parameter combination, detection medium: 0.5% agar (specific ratio: 0.15g agar + 150ml distilled water boiling). Medium volume: 1000ul (optimal volume for wheat variety "Yunmai 56"). Environmental conditions: constant temperature 20℃ (adapted to the physiological characteristics of wheat). Detection index: five core oxygen sensing parameters are selected - initial metabolic rate (SMR), oxygen metabolic rate (OMR), 50% seed reaching 50% oxygen concentration time (R50), 50% oxygen concentration curve area (AUC50), relative germination time (RGT), and equipment adaptation method. Sample arrangement: different concentrations / volume combinations are arranged in different regions of a 40-hole plate, and scanned by column (A1→F1→A2→F2). Automated process: monitor every half hour for 7 days (337 cycles in total), and automatically transfer data to analysis software.

[0007] (II) Vitality determination model: Establishing a quantitative correlation between parameters and changes in vitality: High vitality characteristics: SMR↑, OMR↑, R50↓, AUC50↓, RGT↓. Aging patterns: With aging time, SMR / OMR significantly decreases, while R50 / AUC50 / RGT significantly increases.

[0008] Advantages of the present invention: 1. This technology addresses industry pain points, replacing the traditional germination test (which takes seven days) with rapid testing within hours (determining viability through the initial morphology of the oxygen consumption curve). This technology overcomes the lack of universality of existing technologies: For the first time, a dedicated testing system has been developed for a conventional wheat variety (Yunmai 56), addressing errors caused by embedding and suspension in the medium.

[0009] 2. Parameter screening breakthroughs revealed the sensitivity of R50, AUC50, and RGT in wheat aging detection (Table 3-2). After 30 days of aging, the R50 value increased from 59.35 to 106.93 (an 80% increase), significantly outperforming the germination rate indicator (which only decreased by 14%). SMR and OMR were established as core vitality indicators: they showed a strong negative correlation with aging degree (p<0.05) and demonstrated high assay stability (no significant fluctuations despite volume changes).

[0010] 3. Technical Verification and Accuracy: Highly consistent with standard germination test results (germination potential decreased from 90.7% to 31.0% after 30 days of aging, with a synchronous response from oxygen sensor indicators). Reproducibility: Under the conditions of 0.5% agar + 1000μl, the oxygen consumption curve exhibited a 100% inverted S-shape, eliminating false negatives. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 This is a schematic diagram of the oxygen consumption curve of Yunmai 56 in Example 1 of the present invention; Figure 2 This is a schematic diagram of the seed growth in Example 1 of the present invention; Figure 3 This is a bar chart of various indicators of Yunmai 56 in Example 1 of the present invention. DETAILED DESCRIPTION

[0012] The present invention is further described below with reference to the embodiments, but the present invention is not limited in any way. Any changes or substitutions made based on the teachings of the present invention fall within the scope of protection of the present invention.

[0013] The wheat vitality detection method based on oxygen sensing of the present invention includes pre-processing, detection and statistical steps, specifically including: A. Pre-treatment: Select plump, uniform, insect-free and mildew-free wheat seeds and perform aging treatment to obtain pre-test wheat seeds a; B. Testing: 1) Add agar to distilled water to prepare agar solution b with a concentration of 0.4-0.6%; 2) Add 900-1100 µL of agar solution b to a sample tube. After solidification, add a single seed from pre-tested wheat seed a to the sample tube. Transfer the seed to a seed viability meter and monitor the oxygen consumption curve in real time. Five parameters are screened: initial metabolic rate (SMR), oxygen metabolic rate (OMR), time for 50% of seeds to reach 50% oxygen concentration (R50), area under the 50% oxygen concentration curve (AUC50), and relative germination time (RGT); C. Statistics: SMR, OMR, R50, AUC50 and RGT indicators were used to determine seed vigor.

[0014] The aging treatment described in step A is to place the wheat seeds under the conditions of a temperature of 30-50° C. and a humidity of 70-80% for aging treatment.

[0015] The aging treatment is to place the wheat seeds at a temperature of 35-45° C. and a humidity of 74-76% for aging treatment.

[0016] The aging treatment time is 7 to 30 days.

[0017] The temperature for real-time monitoring of the oxygen consumption curve in step B2) is 15-25°C.

[0018] The present invention will be further described below with specific embodiments: Example 1

[0019] 1. Wheat seed vitality analysis 1.1 Analysis of oxygen consumption by vigor meter in unaged wheat seeds Figure 1 This is a schematic diagram of the oxygen consumption curve of Yunmai 56 under different germination concentration and volume conditions; in, Figure 1 a, b, c, and d are oxygen consumption curves of 0.1% agar with different volumes. Figure 1 e, f, g, and h are oxygen consumption curves of 0.3% agar with different volumes. Figure 1 Figures i, j, k, and l are oxygen consumption curves of different volumes of 0.5% agar. It was observed that the oxygen consumption curves of 0.1% concentration did not show an inverse "S" shape. Under the same concentration conditions, as the volume increased, more oxygen consumption curves showed an inverse "S" shape. Under the same volume conditions, as the concentration increased, more and more oxygen consumption curves showed an inverse "S" shape. Figure 1 J, Figure 1 k and Figure 1The l all showed a good inverse "S" curve, indicating that when the conventional wheat variety Yunmai 56 was tested using the seed vigor instrument, the germination conditions with an agar concentration of 0.5% and a volume of 600, 800, and 1000 μl were more suitable.

[0020] Based on the oxygen consumption curve, which identified an optimal agar concentration of 0.5%, further comparisons of oxygen consumption indicators in different agar volumes revealed no significant differences in SMR, OMR, R50, AUC50, and RGT for volumes of 600, 800, and 1000 μl. While IMT values ​​differed between 600 μl, 800 μl, and 1000 μl, there was no significant difference between 800 μl and 1000 μl (Table 1). This suggests that an agar concentration of 0.5% and volumes of 800 and 1000 μl are suitable for seed viability analysis.

[0021] Table 1 Yunmai 56 vitality detection parameters Note: The difference is significant at the p = 0.05 level. At the same time, combined with the germination performance of seeds in different media, it can be seen that ( Figure 2 ). In 0.1% agar, the vast majority of Yunmai 56 seeds sank to the bottom of the tube and essentially failed to germinate. In 0.3% agar, most seeds remained suspended in the agar. In 0.5% agar, the seeds were completely above the agar and not embedded in it. Seedlings grown in 800 and 1000 μl agar volumes showed good growth. This further demonstrates that 0.5% agar concentration and 800 and 1000 μl agar volumes are suitable for seed viability testing.

[0022] 1.2 Analysis of oxygen consumption by vigor meter after wheat seed aging treatment Figure 3 Figure 1 shows the statistical results of different oxygen consumption indicators after aging treatment. Generally speaking, the stronger the seed vigor, the greater the SMR and OMR values, and the smaller the IMT, R50, AUC50, and RGT values. With increasing aging time, seed vigor decreases, with SMR and OMR values ​​showing a downward trend, while IMT, R50, AUC50, and RGT values ​​show an upward trend. Overall, the trends in oxygen consumption indicators after aging as measured by the seed vigor meter are consistent with the observed decrease in vigor after aging of wheat seeds.

[0023] A specific comparison of the changes in different oxygen consumption indicators during the aging stage revealed: When the germination medium concentration was 0.1%, the oxygen consumption indicators SMR, IMT, OMR, R50, AUC50, and RGT showed an overall trend consistent with declining seed vigor at different volumes. However, there were fluctuations at specific time points. For example, the SMR (0.1%, 800μl) showed some fluctuations at different time points when the germination medium concentration was 0.3%. For example, the OMR (0.3%, various volumes) showed some fluctuations at specific time points when the germination medium concentration was 0.5%. At 400μl, 600μl, and 800μl, the oxygen consumption indicators also showed some fluctuations at specific time points. However, at 1000μl, the oxygen consumption indicators showed consistent trends, with no fluctuations. This suggests that the 0.5% and 1000μl volumes are more suitable for seed viability testing.

[0024] Table 2 Viability parameter values ​​of Yunmai 56 after different aging treatments in germination medium with an agar concentration of 0.5% and a volume of 1000 μl Note: The difference is significant at the p = 0.05 level. Generally speaking, the stronger the seed vigor, the larger the SMR and OMR values, and the smaller the IMT, R50, AUC50, and RGT values. With increasing aging time, seed vigor decreases, with SMR and OMR values ​​showing a decreasing trend, while IMT, R50, AUC50, and RGT values ​​show an increasing trend. Table 2 shows the various parameters of Yunmai 56 aged in 0.5% 1000μl agar. Table 2 shows that the SMR and OMR values ​​show a decreasing trend at 0, 7, 14, and 30 days of aging, with significant differences observed at the first three aging time points, consistent with the pattern of seed vigor variation. IMT values ​​showed no significant differences at 0, 7, 14, and 30 days of aging, but showed a significant difference at 30 days. The IMT value at 14 days of aging was lower than that at 7 and 0 days, which is inconsistent with the trend that stronger vigor corresponds to smaller IMT values. R50, AUC50, and RGT all increase with aging time, consistent with the trend that lower seed vigor corresponds to larger values. In summary, SMR, OMR, R50, AUC50, and RGT indicators can be used to determine seed vigor.

[0025] 1.3 Wheat Seed Standard Germination Test Analysis Table 3 shows the statistical results of germination rate and germination potential of Yunmai 56 seeds after aging for different time periods. The results show that both germination potential and germination rate decrease with increasing aging time, and germination potential decreases faster.

[0026] Comparing the oxygen consumption index test results of the vitality meter with those of the standard germination experiment, it was found that the two test results were consistent, which shows that the seed vitality meter can be used in wheat seed vitality testing, and like the standard germination experiment, it has reliable and accurate test results.

[0027] Table 3. Germination potential (rate) of Yunmai 56 As can be seen from Table 4, the germination potential of Yunmai 56 gradually decreased with the increase of aging time. There was no significant difference between aging for 0 days and aging for 7 days, but there was a significant difference after aging for 14 days and 30 days. There was no significant difference in germination rate among aging for 0 days, 7 days and 14 days, but it decreased significantly after aging for 30 days.

[0028] Table 4 Variance analysis of germination potential (rate) of Yunmai 56 Note: The difference is significant at the p = 0.05 level.

Claims

1. A wheat seed vitality detection method based on oxygen sensing, characterized in that: The wheat vitality detection method based on oxygen sensing includes pre-processing, detection and statistical steps, specifically including: A. Pre-treatment: Select plump, uniform, insect-free and mildew-free wheat seeds and perform aging treatment to obtain pre-test wheat seeds a; B. Testing: 1) Add agar to distilled water to prepare agar solution b with a concentration of 0.4-0.6%; 2) Add 900-1100 µL of agar solution b to a sample tube. After solidification, add a single seed from pre-tested wheat seed a to the sample tube. Transfer the seed to a seed viability meter and monitor the oxygen consumption curve in real time. Five parameters are screened: initial metabolic rate (SMR), oxygen metabolic rate (OMR), time for 50% of seeds to reach 50% oxygen concentration (R50), area under the 50% oxygen concentration curve (AUC50), and relative germination time (RGT); C. Statistics: SMR, OMR, R50, AUC50 and RGT indicators were used to determine seed vigor.

2. The wheat seed vitality detection method based on oxygen sensing according to claim 1, characterized in that: The aging treatment described in step A is to place the wheat seeds under the conditions of a temperature of 30-50° C. and a humidity of 70-80% for aging treatment.

3. The wheat seed vitality detection method based on oxygen sensing according to claim 2, characterized in that: The aging treatment is to place the wheat seeds at a temperature of 35-45° C. and a humidity of 74-76% for aging treatment.

4. The wheat seed vigor detection method based on oxygen sensing according to claim 2 or 3, characterized in that: The aging treatment time is 7 to 30 days.

5. The wheat seed vitality detection method based on oxygen sensing according to claim 1, characterized in that: The temperature for real-time monitoring of the oxygen consumption curve in step B2) is 15-25°C.

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