Application of N-cis-feruloyltyramine in improvement of rice alkali stress tolerance

Abstract

The invention belongs to the technical field of crop planting, and discloses application of N-cis-feruloyltyramine in improvement of rice alkali stress tolerance. According to the method, N-cis-feruloyl tyramine (the CAS number is 80510-09-4) serves as a regulating agent and is prepared into a working solution with the concentration of 10 <-8 >-10 <-4 > g / L, and spraying treatment is conducted on leaves in the one-leaf one-core period of rice seedlings. The chlorophyll content of the rice in the alkali stress environment can be remarkably increased, chlorophyll degradation is relieved, growth of overground parts is promoted, and the alkali stress tolerance of the rice in the seedling stage is effectively enhanced only through simple foliar spraying operation. The method is easy and convenient to operate, low in cost, environmentally friendly and efficient, and has important agricultural application value for stable and yield increasing of saline-alkali soil rice and utilization of marginal land resources.

Description

Technical Field

[0001] This invention belongs to the field of agricultural product planting technology, specifically relating to the application of N-cis-feruloyltyramine in enhancing the tolerance of rice to alkali stress. Background Technology

[0002] Soil alkalization is a major environmental stressor facing global agricultural production, posing a serious threat to the growth and yield of food crops such as rice. Alkaline soils have a high pH value (typically >8.5), which not only leads to soil structure deterioration and reduced aeration, but also significantly reduces the availability of key nutrients such as Fe and P in the soil, causing plant yellowing and stunted growth.

[0003] In soda-type alkaline soil environments (mainly NaCO3 / NaHCO3), high concentrations of Na⁺ and OH⁻ can disrupt the stability of rice cell membranes, triggering ion toxicity and oxidative stress, leading to decreased photosynthetic efficiency and inhibited enzyme activity. Studies have shown that under such alkaline stress, rice seed germination rates can drop below 20%, seedling biomass can decrease by more than 40%, ultimately resulting in a significant reduction in seed setting rate.

[0004] Existing technologies for alleviating alkaline stress mainly include traditional agronomic measures (such as leaching saline-alkali land with irrigation water) and alkali-tolerant breeding. However, the former suffers from high costs, while the latter faces problems such as long breeding cycles, limited genetic resources, and unstable resistance traits. In light of this, chemical regulation has become an important research direction for improving crop stress resistance in recent years. However, most research in this field remains at the level of superficial descriptions of stress phenomena, such as observing a decrease in chlorophyll content and biomass under stress conditions, without delving into the underlying physiological regulatory mechanisms. Simply supplementing chlorophyll exogenously based on the discovery of chlorophyll degradation under alkaline stress is not only technically difficult to implement (chlorophyll molecules are difficult for leaves to absorb and utilize), but also only a temporary solution, failing to address the core regulatory nodes of the stress response. A true technological breakthrough lies in identifying signaling molecules or metabolites that play a core regulatory role in the stress response and developing methods to effectively activate the plant's own stress resistance mechanisms through exogenous application.

[0005] Therefore, developing an efficient, environmentally friendly, and easy-to-apply alkali stress relief technology based on specific signaling molecules is of great significance for ensuring stable and increased rice yields in saline-alkali land. Summary of the Invention

[0006] Through systematic research, the inventors of this invention have discovered for the first time the crucial role of N-cis-feruloyltyramine in the alkali stress response: its decreased content is not simply a result of stress, but rather a significant cause of weakened antioxidant defense systems and photosynthetic structural protection mechanisms. Exogenous supplementation with appropriate concentrations of N-cis-feruloyltyramine can effectively activate downstream stress responses, enhancing the plant's tolerance from the root cause. This discovery provides a novel and innovative chemical regulatory strategy.

[0007] The purpose of this invention is to provide a method for enhancing the tolerance of rice to alkali stress using N-cis-feruloyltyramine. By exogenously spraying a specific concentration of this compound, the inhibitory effect of alkali stress on rice seedling growth is significantly alleviated, and chlorophyll content and plant biomass are increased. The specific technical solution is as follows: Step 1: After soaking and germinating the rice seeds, place the sprouting seeds evenly in the germination device and cultivate them in a greenhouse using hydroponic nutrient solution. The greenhouse temperature is controlled at 28±2 ℃, and the light duration is controlled at 12 hours during the day and 12 hours at night. The hydroponic nutrient solution formula refers to the standard recommended by the International Rice Research Institute. Step 2: When rice seedlings reach the one-leaf-one-heart stage, N-cis-feruloyltyramine is used as a regulator to prepare a working solution, which is then sprayed onto the seedlings at this stage. The concentration of N-cis-feruloyltyramine in the working solution is 10%. -8 ~10 -4 g / L.

[0008] In the above method, the working solution is an aqueous solution of N-cis-feruloyltyramine, and the preferred concentration of N-cis-feruloyltyramine is 10. -6 g / L.

[0009] In the above method, the target of the spraying treatment is the leaves of rice seedlings, and the amount of spraying should be such that the liquid droplets are evenly attached to the leaf surface without dripping.

[0010] In the above method, the alkali stress includes soda-type alkaline earth stress mainly composed of NaCO3 and NaHCO3, and alkaline environmental stress with pH ≥ 8.8.

[0011] The beneficial effects of this invention are: (1) High efficiency: in 10 -6 At a concentration of g / L, spraying can increase the total chlorophyll content of rice under alkali stress (pH=9.2) by 47.5% compared with the untreated group and increase the plant height by 6.8% compared with the untreated group, significantly alleviating the inhibition of rice growth by alkali stress.

[0012] (2) Easy to operate: It can be effective simply by foliar spraying, without the need for complicated equipment and professional operation, and is suitable for large-scale agricultural production.

[0013] (3) Low cost: The amount of N-cis-feruloyltyramine used is extremely small (effective concentration as low as 10). -6 (g / L), low application cost per unit area, and easy to promote.

[0014] (4) Environmentally friendly: The regulatory substances used are naturally occurring small molecule compounds, with no residues or environmental pollution risks, which meet the needs of green agricultural development.

[0015] (5) Wide applicability: It is effective for different rice varieties such as Zhonghua 11 and Longgeng 31, and can be used in various alkaline environments with pH 8.8~9.2. Detailed Implementation

[0016] The specific embodiments of the present invention are described below to enable those skilled in the art to understand the present invention. However, it should be understood that the present invention is not limited to the scope of the specific embodiments. For those skilled in the art, various changes are obvious as long as they are within the spirit and scope of the present invention as defined and determined by the appended claims. All inventions utilizing the concept of the present invention are protected.

[0017] The hydroponic nutrient solution formulations used in the following examples are based on the standards recommended by the International Rice Research Institute (IRRI), as shown in Table 1: Table 1. Nutrient solution formulas from the International Rice Research Institute Example 1

[0018] 1. Experimental material: Zhonghua 11.

[0019] 2. The experimental treatment is as follows: (1) The culture was carried out in a greenhouse using hydroponic culture medium. The greenhouse temperature was controlled at 28±2 ℃ and the daytime and nighttime hours were 12 h and 12 h respectively. The hydroponic nutrient solution formula was based on that of the International Rice Research Institute (IRRI).

[0020] (2) After soaking and germinating the seeds, the seeds with white sprouts are evenly placed in the germination device and cultivated in a greenhouse.

[0021] (3) After the seedlings have grown for 10 days, change the nutrient solution. Use the normal nutrient solution (CK) as the control group, and use hydroponic nutrient solutions with different pH values ​​to cultivate the seedlings in the greenhouse for another 3 days (without changing the nutrient solution).

[0022] (4) Investigation of traits and evaluation methods: After 3 days of alkali stress, the chlorophyll content in the aboveground parts and the N-cis-feruloyltyramine content in the underground parts of each treatment group were measured.

[0023] 3. Results Analysis The results showed that, as shown in Table 2, the chlorophyll content of plants decreased significantly with increasing alkali stress intensity (pH increasing from 9.0 to 9.2). As shown in Table 3, unlike the chlorophyll trend, the N-cis-feruloyltyramine content exhibited the following characteristics: when treated with 70 mM NaCl alone, the N-cis-feruloyltyramine content increased significantly by 15.6% compared to the control (CK). Using 70 mM NaCl alone as a control, the N-cis-feruloyltyramine content gradually decreased with increasing alkali stress intensity (pH 8.8-9.2).

[0024] Table 2 Chlorophyll content of plants under different alkaline treatments

[0025] Note: Different letters indicate significant differences between different groups (p<0.05). Table 3. N-cis-feruloyltyramine content in the underground parts of plants under different alkaline treatments.

[0026] Note: Different letters indicate significant differences between different groups (p<0.05). Example 2

[0027] 1. Experimental material: Zhonghua 11 2. The experimental treatment is as follows: (1) Dilute N-cis-feruloyltyramine to 10 -4 g / L, 10 -6 g / L, 10 -8 Three working solutions with concentrations of g / L.

[0028] (2) The culture was carried out in a greenhouse using hydroponic culture medium. The greenhouse temperature was controlled at 28±2 ℃ and the daytime and nighttime hours were 12 h and 12 h respectively. The hydroponic nutrient solution formula was based on that of the International Rice Research Institute (IRRI).

[0029] (3) After soaking and germinating the seeds, place the white seeds evenly in the germination device and cultivate them in the greenhouse.

[0030] (4) When the seedlings have grown to 10 days, change the nutrient solution. Use the normal nutrient solution (CK) as the control group, spray the leaves with different concentrations of N-cis-feruloyltyramine working solution, and cultivate in the greenhouse for 3 days with an alkaline stress nutrient solution of pH=9.2 (without changing the nutrient solution in between).

[0031] (5) Investigation of traits and evaluation methods: After 3 days of alkali stress, the chlorophyll content in the aboveground parts and the N-cis-feruloyltyramine content in the underground parts of each treatment group were measured.

[0032] 3. Results Analysis The results, as shown in Table 4, indicate that under alkaline stress, exogenous application of N-cis-feruloyltyramine working solution significantly increased the content in the aboveground parts, and even foliar application alone could increase the content in the roots. (The text abruptly ends here, so the translation also ends here.) -6 Treatment with g / L N-cis-feruloyltyramine working solution resulted in the highest N-cis-feruloyltyramine content in the plants, and this concentration can be used as the recommended concentration.

[0033] As shown in Table 5, compared with the control (CK), alkali stress significantly reduced chlorophyll content, with total chlorophyll decreasing by 62.4%, and chlorophyll a and b decreasing by 52.3% and 62.7%, respectively. Under alkali stress, spraying different concentrations of N-cis-feruloyltyramine resulted in both increases and decreases in chlorophyll content. The spraying concentration was 10... -6 The g / L N-cis-feruloyltyramine working solution is the most effective, as it can increase chlorophyll content and alleviate chlorophyll degradation under alkaline stress.

[0034] Table 4. N-cis-feruloyltyramine content in the aboveground and underground parts of plants under different treatments.

[0035] Note: Different letters indicate significant differences between different groups (p<0.05). Table 5 Chlorophyll content of plants under different treatments

[0036] Note: Different letters indicate significant differences between different groups (p<0.05). Example 3

[0037] 1. Experimental material: Dragon's Root 31 2. The experimental treatment is as follows: (1) Dilute N-cis-feruloyltyramine to 10 -6 Working solution with a concentration of g / L.

[0038] (2) The culture was carried out in a greenhouse using hydroponic culture medium. The greenhouse temperature was controlled at 28±2 ℃ and the daytime and nighttime hours were 12 h and 12 h respectively. The hydroponic nutrient solution formula was based on that of the International Rice Research Institute (IRRI).

[0039] (3) After soaking and germinating the seeds, place the white seeds evenly in the germination device and cultivate them in the greenhouse.

[0040] (4) Cultivate the seedlings until they grow to 10 days, then... -6 The leaves were sprayed with g / L N-cis-feruloyltyramine working solution, with water spraying as a control group. The leaves were cultured in a greenhouse for 10 days with an alkaline stress nutrient solution of pH=8.8 and 65mM NaHCO3 (without changing the nutrient solution).

[0041] (5) Investigation of traits and evaluation methods: After 10 days of alkali stress, plant height, root length and chlorophyll content of each treatment group were measured.

[0042] 3. Results Analysis The results showed that, as shown in Table 6, compared with the control (CK), alkali stress significantly reduced the chlorophyll content of the plants, while spraying with 10... -6 The working solution of g / L N-cis-feruloyltyramine significantly reduced the degradation of chlorophyll content in plants. As shown in Table 7, compared with the control (CK), alkali stress significantly inhibited plant growth, especially the aboveground parts, while external application of 10 g / L N-cis-feruloyltyramine significantly reduced chlorophyll content degradation in plants. -6 The g / L N-cis-feruloyltyramine working solution can alleviate the inhibition of alkaline stress on the aboveground parts of plants, but its effect on the underground parts is not obvious.

[0043] Table 6 Chlorophyll content of plants under different treatments

[0044] Note: Different letters indicate significant differences between different groups (p<0.05). Table 7 Plant height and root length under different treatments

[0045] Note: Different letters indicate significant differences between different groups (p<0.05). Example 4

[0046] 1. Experimental material: Zhonghua 11 2. The experimental treatment is as follows: (1) Dilute N-cis-feruloyltyramine and N-trans-feruloyltyramine (CisAK1) to 10. -6 g / L working solution.

[0047] (2) The culture was carried out in a greenhouse using hydroponic culture medium. The greenhouse temperature was controlled at 28±2 ℃ and the daytime and nighttime hours were 12 h and 12 h respectively. The hydroponic nutrient solution formula was based on that of the International Rice Research Institute (IRRI).

[0048] (3) After soaking and germinating the seeds, place the white seeds evenly in the germination device and cultivate them in the greenhouse.

[0049] (4) When the seedlings have grown to 10 days, change the nutrient solution. Spray the leaves with N-cis-feruloyltyramine working solution of different concentrations, and cultivate them in the greenhouse for 3 days with alkaline stress nutrient solution of pH=9.2 (without changing the nutrient solution in between).

[0050] (5) Investigation of traits and evaluation methods: After 3 days of alkali stress, the chlorophyll content of the aboveground parts of each treatment group was measured.

[0051] 3. Results Analysis The results showed that, as shown in Table 8, under salt-alkali stress, external application of N-cis-feruloyltyramine and N-trans-feruloyltyramine could increase chlorophyll content and alleviate chlorophyll degradation under alkaline stress.

[0052] Table 8 Chlorophyll content of plants under different alkaline treatments

[0053] Note: Different letters indicate significant differences between different groups (p<0.05). In summary, the intensity of alkali stress is negatively correlated with the content of N-cis-feruloyltyramine in rice: as the pH value increases from 8.8 to 9.2, the content of N-cis-feruloyltyramine in the underground parts of rice gradually decreases, while the chlorophyll content decreases significantly. Exogenous application of N-cis-feruloyltyramine can supplement the deficiency of this substance in the plant, thereby enhancing the rice's tolerance to alkali stress by maintaining cell membrane stability, inhibiting oxidative stress, reducing chlorophyll degradation, and improving photosynthetic efficiency. N-trans-feruloyltyramine has similar effects.

[0054] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. The application of a chemical substance, N-cis-feruloyltyramine, in improving the alkali tolerance of rice seedlings under alkali stress, characterized in that... The application involves applying a concentration of 10 to the leaves of rice at the one-leaf-one-heart stage. -8 ~10 -4 g / L N-cis-feruloyltyramine.

2. The application according to claim 1, characterized in that, The concentration of the N-cis-feruloyltyramine is 10. -6 g / L.

3. A method for enhancing rice seedling tolerance to alkali stress using N-cis-feruloyltyramine, characterized in that, Includes the following steps: Step 1: After soaking and germinating the rice seeds, place the sprouting seeds evenly in the germination device and cultivate them in a greenhouse using hydroponic nutrient solution. The hydroponic nutrient solution formula refers to the standard recommended by the International Rice Institute. Step 2: When rice seedlings reach the one-leaf-one-heart stage, N-cis-feruloyltyramine is used as a regulator to prepare a working solution, which is then sprayed onto the seedlings at this stage. The concentration of N-cis-feruloyltyramine in the working solution is 10%. -8 ~10 -4 g / L.

4. The method according to claim 3, characterized in that, The concentration of N-cis-feruloyltyramine in the working solution is 10. -6 g / L.

5. The method according to claim 3, characterized in that, The spraying treatment targets rice seedling leaves, and the spraying amount is such that the liquid droplets are evenly attached to the leaf surface without any droplets falling.

6. The method according to claim 3, characterized in that, The rice varieties mentioned include Zhonghua 11 and Longgeng 31.

7. The method according to claim 3, characterized in that, The alkali stress includes soda-type alkaline earth stress composed of NaCO3 and NaHCO3; the pH of the alkali stress is between 8.8 and 9.

2.

8. The method according to claim 3, characterized in that, The working solution is an aqueous solution of N-cis-feruloyltyramine.

9. The method according to claim 3, characterized in that, In step one, the greenhouse temperature is controlled at 28±2 ℃, and the light duration is controlled at 12 hours during the day and 12 hours at night.

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