Precise positioning method for key decline nodes of aging activity of rice seeds

Abstract

The invention discloses a precise positioning method for key decline nodes of aging activity of rice seeds, and belongs to the technical field of agricultural science. The method specifically comprises the following steps: (1) performing continuous different-time-length artificial accelerated aging treatment on rice seeds with the same batch and the same activity level; (2) seed soaking and germination accelerating treatment; (3) indoor normal germination test; (4) measuring seed vigor indexes; (5) statistical comparison; and (6) defining the number of aging days corresponding to the first time when statistical significant decline occurs as the aging activity key decline node of the rice seed. According to the method, by simulating natural aging conditions, the nodes (namely critical points) of key decline of the activity of the rice seeds in the artificial accelerated aging process can be accurately positioned. According to the method, the mutation inflection point of the seed activity under the high-strength aging condition is quantified for the first time, and an accurate and reliable technical standard is provided for seed aging mechanism research, activity early warning and storability evaluation.

Description

Technical Field

[0001] This invention relates to the field of agricultural science and technology, and more specifically to a method for accurately locating key decline nodes in the aging vitality of rice seeds. Background Technology

[0002] Rice seeds are susceptible to aging during storage and distribution due to high temperature and humidity, leading to decreased seed viability and directly impacting germination rate and final yield. Therefore, accurately assessing the degree of seed aging and providing early warning of the aging process is crucial for ensuring agricultural production and minimizing economic losses.

[0003] Currently, accelerated aging experiments are a common method for rapidly assessing the storage tolerance and aging degree of rice seeds. However, traditional methods often focus on measuring the final vigor (e.g., germination rate) of rice seeds after a single, fixed, long-term aging treatment. This "endpoint" assessment fails to reveal that aging is a dynamic process, and even less can it accurately identify the critical turning point from stable maintenance to a sharp decline in rice seed vigor. The lack of a precise definition and measurement method for this critical point makes it difficult to quantitatively analyze and effectively intervene in the early stages of aging in fields such as rice seed physiology research, vigor monitoring and early warning, and germplasm resource evaluation.

[0004] Therefore, how to develop a method that can accurately and quantitatively locate the initial critical decline nodes in the aging process of rice seeds is a problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0005] In view of this, the purpose of the present invention is to provide a method for accurately locating key decline nodes of rice seed aging vitality, so as to overcome the shortcomings of the prior art.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] A method for accurately locating key decline nodes in the aging vigor of rice seeds, specifically including the following steps: (1) Rice seeds (gymnosperms) of the same batch and with the same vigor level were subjected to artificial accelerated aging treatment for different durations. (2) Soaking and germination treatments were carried out on rice seeds with different aging times; (3) Rice seeds with different aging times were used as aging treatment groups, and unaged rice seeds (gloves) were used as unaged control groups. Indoor normal germination tests were conducted on both groups. (4) Determine the seed vigor indicators of the aging treatment group and the non-aging control group; (5) The seed vigor index of the aging treatment group and the non-aging control group was statistically compared; (6) The number of days of aging when the seed vigor index of the aging treatment group first showed a statistically significant decrease compared with the non-aging control group was defined as the key decline node of the aging vigor of the rice seed.

[0008] Furthermore, in step (1) above, the device for artificial accelerated aging treatment is an artificial climate chamber; the temperature is 44.5-45.5℃, preferably 45℃; the relative humidity (RH) is 95%-99%, preferably 97%; and the duration includes at least 1 day, 2 days, 3 days, 4 days and 5 days.

[0009] Furthermore, in step (2) above, the temperature for soaking and germination treatment is room temperature 25℃, and the time is 24h for each.

[0010] Furthermore, in step (3) above, the temperature for normal germination testing indoors is a constant 25°C, during which water and light are supplemented.

[0011] Furthermore, in step (4) above, the measurement time is the 5th and 11th day of the normal indoor germination test.

[0012] Furthermore, in step (4) above, the seed vigor indicators include germination potential, germination rate and aboveground fresh weight of seedlings.

[0013] Furthermore, in step (5) above, the statistical comparison is performed using statistical significance level analysis software (…). P <0.05).

[0014] Furthermore, in step (6) above, the standard for a significant decrease is: P <0.05.

[0015] As can be seen from the above technical solution, compared with the prior art, the beneficial effects of the present invention are as follows: 1. For the first time, key mutation points in the aging process were quantified. Through precisely controlled time-series experiments, this invention reveals for the first time that under specific high-intensity aging conditions of 45℃ / 97%RH, the vitality of rice seeds does not decline at a uniform rate, but rather experiences a "cliff-like" drop from quantitative to qualitative change at a specific number of days (such as the 3rd day). This point is precisely defined as the "critical decline point of aging vitality".

[0016] 2. Provides accurate early warning tools This invention can issue an early warning at the starting point when irreversible and significant loss of rice seed vigor occurs, providing an unprecedentedly accurate basis for seed quality monitoring and the determination of safe storage period.

[0017] 3. The method has good standardization and reproducibility. The aging conditions (45℃ / 97%RH) and judgment criteria (statistically significant decrease) set by this invention are clear and specific, making the method easy to standardize and reproduce in different laboratories. Furthermore, this method can be used to test key nodes of vitality changes in multiple rice varieties, laying the foundation for the industry to establish a unified early evaluation standard for seed aging.

[0018] In summary, this invention, by simulating natural aging conditions, can precisely pinpoint the critical point (i.e., the point of decline) in the vigor of rice seeds during artificially accelerated aging. This invention is the first to quantify the inflection point of seed vigor mutation under high-intensity aging conditions, providing a precise and reliable technical standard for research on seed aging mechanisms, early warning of vigor, and evaluation of storage tolerance. Attached Figure Description

[0019] Figure 1 Germination potential and germination rate of 'Longping No. 5' rice in the non-aging control group and aging treatment group were tested in the room for normal germination. Figure 2 Fresh weight of aboveground parts of seedlings for indoor normal germination tests of 'Longping No. 5' rice in the non-aging control group and aging treatment group. Figure 3 Germination potential and germination rate of 'Longjing 31' rice in the non-aging control group and aging treatment group were tested in the room for normal germination. Figure 4 The aboveground fresh weight of seedlings for indoor normal germination tests of 'Longjing 31' rice in the non-aging control group and the aging treatment group. Detailed Implementation

[0020] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0021] Example 1 A method for accurately locating key decline points in the aging vigor of rice seeds includes the following steps: (1) In an artificial climate chamber with a temperature of 45℃ and a relative humidity of 97%, rice seeds of the same batch and with the same vigor level were subjected to artificial accelerated aging treatment for different durations, namely 1 day, 2 days, 3 days, 4 days and 5 days. (2) Rice seeds with different aging times were soaked and germinated at room temperature (25°C) for 24 hours each. (3) Rice seeds with different aging times were used as aging treatment groups, and unaged rice seeds were used as unaged control groups. Indoor normal germination tests were conducted. The required temperature for the test was a constant 25℃. Water and light were provided in time during the germination period to ensure the normal growth of rice plants. (4) On the 5th and 11th days of the normal germination test in the room, the germination potential, germination rate and fresh weight of the aboveground parts of the seedlings in the aging treatment group and the non-aging control group were recorded as plant phenotypic data. (5) Use statistical significance level analysis software ( P <0.05), the germination potential, germination rate and aboveground fresh weight of seeds in the aging treatment group and the non-aging control group were compared and analyzed; (6) The germination potential, germination rate, and aboveground fresh weight of seedlings in the aging treatment group showed a statistically significant decrease for the first time compared with the non-aging control group. P The number of aging days corresponding to a value <0.05 is defined as the critical decline node of the aging vigor of the rice seed.

[0022] Example 2 A method for accurately locating key decline points in the aging vigor of rice seeds includes the following steps: (1) In an artificial climate chamber with a temperature of 44.5℃ and a relative humidity of 95%, rice seeds of the same batch and with the same vigor level were subjected to artificial accelerated aging treatment for different durations, namely 1 day, 2 days, 3 days, 4 days and 5 days. (2) Rice seeds with different aging times were soaked and germinated at room temperature (25°C) for 24 hours each. (3) Rice seeds with different aging times were used as aging treatment groups, and unaged rice seeds were used as unaged control groups. Indoor normal germination tests were conducted. The required temperature for the test was a constant 25℃. Water and light were provided in time during the germination period to ensure the normal growth of rice plants. (4) On the 5th and 11th days of the normal germination test in the room, the germination potential, germination rate and fresh weight of the aboveground parts of the seedlings in the aging treatment group and the non-aging control group were recorded as plant phenotypic data. (5) Use statistical significance level analysis software ( P <0.05), the germination potential, germination rate and aboveground fresh weight of seeds in the aging treatment group and the non-aging control group were compared and analyzed; (6) The germination potential, germination rate, and aboveground fresh weight of seedlings in the aging treatment group showed a statistically significant decrease for the first time compared with the non-aging control group. P The number of aging days corresponding to a value <0.05 is defined as the critical decline node of the aging vigor of the rice seed.

[0023] Example 3 A method for accurately locating key decline points in the aging vigor of rice seeds includes the following steps: (1) In an artificial climate chamber with a temperature of 45.5℃ and a relative humidity of 99%, rice seeds of the same batch and with the same vigor level were subjected to artificial accelerated aging treatment for different durations, namely 1 day, 2 days, 3 days, 4 days and 5 days. (2) Rice seeds with different aging times were soaked and germinated at room temperature (25°C) for 24 hours each. (3) Rice seeds with different aging times were used as aging treatment groups, and unaged rice seeds were used as unaged control groups. Indoor normal germination tests were conducted. The required temperature for the test was a constant 25℃. Water and light were provided in time during the germination period to ensure the normal growth of rice plants. (4) On the 5th and 11th days of the normal germination test in the room, the germination potential, germination rate and fresh weight of the aboveground parts of the seedlings in the aging treatment group and the non-aging control group were recorded as plant phenotypic data. (5) Use statistical significance level analysis software ( P <0.05), the germination potential, germination rate and aboveground fresh weight of seeds in the aging treatment group and the non-aging control group were compared and analyzed; (6) The germination potential, germination rate, and aboveground fresh weight of seedlings in the aging treatment group showed a statistically significant decrease for the first time compared with the non-aging control group. P The number of aging days corresponding to a value <0.05 is defined as the critical decline node of the aging vigor of the rice seed.

[0024] Performance testing Based on the method in Example 1, the key decline nodes of aging vitality in rice varieties 'Longping 5' and 'Longjing 31' were determined respectively.

[0025] I. Precisely determining the key decline points of aging vitality in rice variety 'Longping 5' 1. Materials and Processing 'Longping No. 5' rice seeds with an initial germination rate ≥93% were selected. The artificial climate chamber was precisely set to 45℃ and 97%RH. There were two groups: a control group (CK) aged for 0 days (non-aged) and groups aged for 1, 2, 3, 4, and 5 days (aged treatment groups), with 50 seeds in each group and 4 replicates.

[0026] 2. Indicator Measurement After each treatment group completed its corresponding number of aging days, it underwent an indoor normal germination test together with the control group. At this time, the germination test started from indoor sowing, and the germination potential of each treatment was counted on the 5th day after sowing. The germination rate was counted on the 11th day, and the fresh weight of the aboveground parts of the seedlings was measured.

[0027] 3. Results and Analysis ( Figure 1-2 ) CK group (day 0): germination potential 91.33%, germination rate 93%, aboveground fresh weight 6.53 g.

[0028] The 1-day aging group had a germination potential of 87.33%, a germination rate of 91.33%, and a fresh weight of 6.38g above ground, which was not significantly different from the CK group (P>0.05).

[0029] Two-day aging group: germination potential 80.67%, germination rate 90.67%, and aboveground fresh weight 6.13 g, which were not significantly different from the CK group (P>0.05).

[0030] In the 3-day aging group, the germination potential dropped significantly to 44%, the germination rate dropped to 74%, and the fresh weight of the aboveground parts dropped to 5.09 g. Compared with the CK group, all indicators were significantly different (P<0.05), and the seed vigor showed a rapid decline during this period.

[0031] The 4-day aging group had a germination potential of 30.33%, a germination rate of 61.67%, and a fresh weight of the aboveground parts that decreased to 4.57g.

[0032] The aging group after 5 days had a germination potential of 18.67%, a germination rate of 42.33%, and a fresh weight of the aboveground parts that decreased to 3.94g.

[0033] 4. Conclusion According to the method of the present invention, analysis of the germination potential, germination rate, and aboveground fresh weight of seedlings after aging shows that, starting from day 3, all three indicators of 'Longping 5' rice were significantly lower than those of the control group. Therefore, it can be determined that the critical decline point of aging vigor of 'Longping 5' rice seeds under 45℃ / 97%RH conditions is day 3. That is, under these aging conditions, seed vigor enters a significant decline stage starting from day 3.

[0034] II. Precisely Determining the Key Decline Nodes of Aging Vitality in Rice Variety 'Longjing 31' 1. Materials and Processing 'Longjing 31' rice seeds with an initial germination rate ≥94.67% were selected. The artificial climate chamber was precisely set to 45℃ and 97%RH. There were two groups: a control group (CK) aged for 0 days (non-aged) and groups aged for 1, 2, 3, 4, and 5 days (aged treatment groups), with 50 seeds in each group and 4 replicates.

[0035] 2. Indicator Measurement After each treatment group completed its corresponding aging period, it underwent an indoor normal germination test together with the control group. Germination potential was recorded on day 5, germination rate was recorded on day 11, and the fresh weight of the aboveground parts of the seedlings was measured.

[0036] 3. Results and Analysis ( Figure 3-4 ) CK group (day 0): germination potential 84.67%, germination rate 94.67%, aboveground fresh weight 6.75 g.

[0037] The 1-day aging group had a germination potential of 78.33%, a germination rate of 88%, and a fresh weight of 6.55g above ground, which was not significantly different from the CK group (p>0.05).

[0038] Two-day aging group: germination potential 73.33%, germination rate 83.33%, and aboveground fresh weight 6.35 g, which were not significantly different from the CK group (p>0.05).

[0039] In the 3-day aging group, the germination potential decreased significantly to 49.14%, the germination rate decreased to 64%, and the fresh weight of the aboveground parts decreased to 5.11 g. Compared with the CK group, all indicators were significantly different (p<0.05).

[0040] Group aged for 4 days: germination potential 47.33%, germination rate 52%, and fresh weight of aboveground parts 4.65g.

[0041] Group aged for 5 days: germination potential 45%, germination rate 50.67%, and fresh weight of aboveground parts 4.15g.

[0042] 4. Conclusion According to the method of the present invention, analysis of the germination potential, germination rate, and aboveground fresh weight of seedlings after aging shows that, starting from day 3, all three indicators of 'Longjing 31' rice are significantly lower than those of the control group. Therefore, it can be determined that the critical point of decline in the aging vigor of 'Longjing 31' rice seeds under 45℃ / 97%RH conditions is day 3. That is, under these aging conditions, seed vigor enters a significant decline stage starting from day 3.

[0043] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for accurately locating key decline nodes in the aging vigor of rice seeds, characterized in that, Specifically, the following steps are included: (1) Rice seeds of the same batch and with the same vigor level were subjected to artificial accelerated aging treatment for different durations. (2) Soaking and germination treatments were carried out on rice seeds with different aging times; (3) Rice seeds with different aging times were used as aging treatment groups, and unaged rice seeds were used as unaged control groups. Indoor normal germination tests were conducted respectively. (4) Determine the seed vigor indicators of the aging treatment group and the non-aging control group; (5) The seed vigor index of the aging treatment group and the non-aging control group was statistically compared; (6) The number of days of aging when the seed vigor index of the aging treatment group first showed a statistically significant decrease compared with the non-aging control group was defined as the key decline node of the aging vigor of the rice seed.

2. The method for accurately locating key decline nodes in the aging vigor of rice seeds according to claim 1, characterized in that, In step (1), the device for the artificial accelerated aging treatment is an artificial climate chamber with a temperature of 44.5-45.5℃ and a relative humidity of 95%-99%, and the duration includes at least 1 day, 2 days, 3 days, 4 days and 5 days.

3. The method for accurately locating key decline nodes in the aging vigor of rice seeds according to claim 1, characterized in that, In step (2), the temperature for soaking and germination treatment is room temperature 25°C, and the time is 24h for each.

4. The method for accurately locating key decline nodes in the aging vigor of rice seeds according to claim 1, characterized in that, In step (3), the temperature for the indoor normal germination test is a constant 25°C, during which water and light are added.

5. The method for accurately locating key decline nodes in the aging vigor of rice seeds according to claim 1, characterized in that, In step (4), the measurement time is the 5th and 11th day of the indoor normal germination test.

6. The method for accurately locating key decline nodes in the aging vigor of rice seeds according to claim 1, characterized in that, In step (4), the seed vigor indicators include germination potential, germination rate and aboveground fresh weight of seedlings.

7. The method for accurately locating key decline nodes in the aging vigor of rice seeds according to claim 1, characterized in that, In step (5), the statistical comparison is performed using statistical significance level analysis software (...). P <0.05).

8. The method for accurately locating key decline nodes of rice seed aging vigor according to claim 1, characterized in that, In step (6), the criterion for a significant decrease is: P <0.05.

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