A nanoscale safe herbicide that alleviates butachlor damage to rice, its preparation method and application.

Abstract

This invention relates to the field of herbicide safeners, specifically to a nanoscale herbicide safener for mitigating butachlor phytotoxicity in rice, its preparation method, and its application. The preparation method of the nanoscale herbicide safener involves dissolving melatonin in water, adding trehalose, stirring until completely dissolved, then adding a reducing agent and reacting to obtain a mixed solution. Selenite is added to the mixed solution to conduct a redox reaction, yielding the nanoscale butachlor safener. The nanoscale butachlor safener of this invention has a particle size of 22 nm and exhibits good stability and in vitro antioxidant activity. Treating rice seeds with the nanoscale butachlor safener not only alleviates butachlor phytotoxicity by improving rice's resistance to stress and accelerating butachlor metabolism in the rice plant, but also increases the selenium content of the rice.

Description

Technical Field

[0001] This invention relates to the field of herbicide safeners, and more specifically, to a nanoscale herbicide safener for mitigating butachlor damage to rice, its preparation method, and its application. Background Technology

[0002] Rice is one of my country's main food crops. However, weeds such as barnyard grass and Echinochloa crus-galli threaten rice yield and quality by competing with rice for resources and spreading pests and diseases. Statistics show that weed damage causes approximately 15% of rice yield loss annually in my country. Butachlor is a selective chloroacetamide herbicide mainly used in direct-seeded and transplanted rice fields. It has good control effects on barnyard grass, Echinochloa crus-galli, and some broadleaf weeds during the budding and pre-two-leaf stages. However, its application in the field is easily affected by environmental factors such as temperature and humidity, as well as application techniques such as dosage and timing, leading to phytotoxicity symptoms in rice seedlings, such as stunting, chlorosis, and root necrosis, thus affecting rice yield and quality.

[0003] Herbicide safeners can improve crop tolerance to herbicides without reducing their effectiveness against target weeds, selectively protecting crops from herbicide damage. In agricultural production, glyphosate is often used in conjunction with pretilachlor to mitigate butachlor damage to rice. However, glyphosate primarily acts as a specific antidote for pretilachlor and lacks specificity for butachlor; furthermore, recent studies have shown that glyphosate is highly toxic to aquatic organisms. Therefore, developing novel environmentally friendly safeners to reduce the risk of butachlor damage is of significant practical importance for building a green and sustainable weed control system in rice paddies. Summary of the Invention

[0004] This invention provides a nanoscale safe herbicide that alleviates butachlor damage to rice, along with its preparation method and application. This safe herbicide not only alleviates butachlor damage to rice but also increases the selenium content of the plant.

[0005] The above-mentioned technical objective of the present invention is achieved through the following technical solution:

[0006] A method for preparing a nano-scale herbicide safener that alleviates butachlor damage to rice involves dissolving melatonin in water, adding trehalose, stirring until completely dissolved, then adding a reducing agent to react and obtain a mixed solution; then, adding selenite to the mixed solution to carry out a redox reaction to obtain a nano-scale butachlor safener (MT-SeNPs).

[0007] The redox reaction is carried out at 35~45℃ for 20~60 min.

[0008] The reaction time for melatonin, trehalose, and reducing agent is 10 minutes, and the reaction temperature is 40~65℃.

[0009] Preferably, the concentration of melatonin in the mixed solution is 0.1-0.3%.

[0010] Preferably, the concentration of trehalose in the mixed solution is 1.5-4%.

[0011] Preferably, the reducing agent is at least one of ascorbic acid, glutathione, and sodium dodecyl sulfate.

[0012] Preferably, the concentration of the reducing agent in the mixed solution is 20~80 mmol / L.

[0013] Preferably, the selenite is added to the mixed solution at a concentration of 5-20 mmol / L.

[0014] Another objective of this invention is to provide a safe herbicide that mitigates the herbicidal damage of butachlor to rice.

[0015] The nanoscale herbicide safener that alleviates butachlor phytotoxicity to rice is prepared by the above-described method.

[0016] Another object of the present invention is to provide the application of the above-mentioned nanoscale herbicide safener.

[0017] Specifically, the application of nanoscale herbicide safeners that alleviate butachlor damage to rice includes seed soaking in a herbicide safener (MT-SeNPs) solution. This involves transferring rice seeds into an MT-SeNPs solution, initiating the reaction for 48 hours, and then removing the seeds and removing any residual solution from the surface.

[0018] The MT-SeNPs solution soaking concentration was 2-12 mg / L. -1 Preferably, the soaking concentration is 6-8 mg / L. -1 The triggering condition is 28℃.

[0019] The beneficial effects of this invention are as follows: 1. Synergistic Enhancement Mechanism: Selenium is an important active component of glutathione peroxidase, which can scavenge reactive oxygen species; melatonin has anti-stress and growth-promoting functions; trehalose is a stress-protective substance that can form a protective film on the cell surface to protect the integrity of biomolecular structures when subjected to stress. The nanomaterial MT-SeNPs, prepared by modifying with trehalose and melatonin, significantly enhanced the antioxidant capacity of rice through synergistic effects.

[0020] 2. Mechanism of herbicide damage mitigation: The nano-level herbicide safener can effectively improve the rice's ability to resist herbicide stress by soaking rice seeds and accelerate the metabolism of butachlor in the rice, thereby significantly reducing herbicide damage.

[0021] 3. Quality Improvement: The nano-grade herbicide safe agent can alleviate herbicide damage while effectively increasing the selenium content of rice plants, thus achieving selenium-enriched rice production.

[0022] 4. Simple process: The preparation method of the present invention is simple to operate, mild in conditions, and suitable for large-scale promotion and application. Attached Figure Description

[0023] Figure 1 Characterization of MT-SeNPs; (a) TEM of MT-SeNPs, (b) Particle size of MT-SeNPs, (c) Zeta potential, (d) XPS of MT-SeNPs, melatonin, and trehalose, (e) FTIR of MT-SeNPs, melatonin, and trehalose, (f) DPPH scavenging rate of MT-SeNPs, melatonin, and trehalose, (g) ABTS of MT-SeNPs, melatonin, and trehalose. + Free radical scavenging rate.

[0024] Figure 2 Stability analysis of MT-SeNPs; (A) Changes in the transparency of MT-SeNPs over time at different temperatures, (B) Changes in the particle size of MT-SeNPs over time at different temperatures, (C) Changes in the transparency of MT-SeNPs over time at different pH levels, (D) Changes in the particle size of MT-SeNPs over time at different pH levels.

[0025] Figure 3 Effects of MT-SeNPs on rice growth, selenium content in different parts of rice, and butachlor residue under butachlor stress; (a) Root length, (b) Root fresh weight, (c) Selenium content in rice roots and aboveground parts, (d) Plant height, (e) Fresh weight of aboveground parts, (f) Butachlor residue in rice roots and aboveground parts, (g) Rice growth chart after 7 days of treatment, (h) Rice growth chart after 10 days of treatment.

[0026] Figure 4 The alleviating effect of MT-SeNPs on rice seedlings under butachlor stress; (a) Plant height, (b) Root length, (c) Fresh weight, (d) Rice growth chart after 7 days of treatment.

[0027] Figure 5 The alleviating effects of different substances on rice seedlings under butachlor stress; (a) Root length, (b) Plant height, (c) Fresh weight, (d) Rice growth chart after 7 days of treatment. Detailed Implementation

[0028] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0029] Unless otherwise specified in the examples, the conditions shall be performed according to the standard conditions or the conditions recommended by the manufacturer.

[0030] Example 1: Preparation, characterization, and stability analysis of a nanoscale herbicide safener Dissolve 0.15% melatonin and 2% trehalose in water sequentially, then add 40 mmol L. -1 Ascorbic acid was reacted with stirring at 40°C for 10 min to obtain a mixed solution. 10 mmol L⁻¹ was then added to the mixed solution. -1 Sodium selenite, reacted at 40℃ for 40 min, yields a nano-scale herbicide safener (MT-SeNPs).

[0031] The MT-SeNPs obtained in this example were characterized using transmission electron microscopy (TEM), laser particle size analyzer, X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared spectroscopy (FTIR). The results showed that the synthesized MT-SeNPs were spherical nanoparticles with a particle size of 22 nm and a zeta potential of -12.3 mV. Figure 1 ac), composed of four elements: C, N, O, and Se. Figure 1 d), and showed specific peak positions at 3004 (substituted benzene CH), 1608 (benzene ring C=C), and 1513 (amide NH). Figure 1 e), indicating that MT-SeNPs were successfully synthesized. Furthermore, in vitro antioxidant activity analysis was performed, using ascorbic acid as a positive control. The results showed that at a concentration of 100 mg / L... -1 Or 25 mg L -1 At that time, the corresponding DPPH clearance rate or ABTS + Clearance rate greater than 90% Figure 1 (f, g), indicating that it has good in vitro antioxidant activity.

[0032] MT-SeNPs were then placed at 4℃, 15℃, 25℃, 35℃, and 45℃ for 1 day, 7 days, 14 days, and 21 days, respectively. With increasing temperature and longer placement time, the transparency of MT-SeNPs decreased, and the particle size increased. When placed at 4℃, 15℃, 25℃, 35℃, and 45℃ for 1 day, the particle sizes were 21 nm, 24 nm, 32 nm, 43 nm, and 68 nm, respectively. After 21 days, the particle size at 4℃ was 28 nm, and the color was red and transparent; the particle sizes at 15℃ and 25℃ were 43-68 nm, and the color was red but slightly cloudy; the particle sizes at 35℃ and 45℃ were greater than 100 nm, and black precipitate was present at the bottom. Figure 2 A, B). Furthermore, MT-SeNPs were placed at pH 4, 5, 6, 7, and 8 for 1, 7, 14, and 21 days, respectively. The results showed that at pH 4, the transparency decreased and the particle size increased with prolonged storage time; the other pH values ​​had no significant effect on MT-SeNPs, and even after 21 days, the color remained red and transparent, with a particle size of 28-32 nm. Figure 2 (C, D). This indicates that MT-SeNPs are sensitive to high temperatures and strong acids.

[0033] Example 2: The alleviating effect of MT-SeNPs on butachlor stress in rice Select uniformly sized Huanghuazhan rice seeds, disinfect them with 5% (v / v) NaClO for 15 min, rinse them three times with deionized water, and then transfer them to a solution containing 0 mg L. -1 6 mg L -1 After initiation at 28°C for 48 h in an MT-SeNPs solution (the preparation method and dosage of the material are the same as in Example 1), excess solution was absorbed with filter paper. Rice seeds were arranged at 1.5 cm intervals in a solution moistened with 0.4 mg / mL... -1 Butachlor germination paper (25 cm x 38 cm, Anchor Paper Co., Saint Paul, MN, USA) with 20 seeds per sheet was placed vertically in a resealable bag, with a water-soaked control as a reference. The paper was placed in an incubator at a day / night temperature of 30℃ / 25℃ and a 14-hour photoperiod, with each treatment replicated three times. Seven days after treatment, some rice plants were harvested, and their agronomic traits (plant height, root length, and fresh weight), selenium content in different parts of the rice, and butachlor residue were measured. Ten days after treatment, agronomic traits were measured again. The following treatments were set up: water initiation (H2O), MT-SeNPs initiation (MT-SeNPs), water initiation + butachlor (H2O + BT), and MT-SeNPs initiation + butachlor (MT-SeNPs + BT).

[0034] Experimental results showed that under normal growth conditions, rice seedlings had bright green leaves and robust root systems. Furthermore, rice induced by MT-SeNPs exhibited superior root length, plant height, and fresh weight compared to that induced by water. When rice was subjected to butachlor stress, growth was stunted, plants were dwarfed, and plant height, root length, and fresh weight were significantly lower than in the normal treatment. p <0.05). However, after 7 days of butachlor stress treatment, the root length, plant height, root fresh weight, and aboveground fresh weight of rice treated with MT-SeNPs were increased by 33.68%, 16.72%, 7.56%, and 16.28%, respectively, compared with those treated with water. After 10 days of stress, the root length and plant height of rice treated with MT-SeNPs recovered to 31.54% and 28.97% of the normal values, respectively. Figure 3 (ab, de). Normal treatment refers to fenbutatin stress.

[0035] Furthermore, rice seedlings treated with MT-SeNPs exhibited selenium content ranging from 0.38 to 1.36 mg / kg. -1 Significantly higher than that caused by water ( Figure 3 c). Meanwhile, GC-MC analysis revealed that butachlor residue levels in the roots and aboveground parts of rice seedlings treated with MT-SeNPs were reduced by 21.72% and 50.55%, respectively, compared to those treated with water. Figure 3 f). This indicates that MT-SeNPs can be absorbed by rice through seed soaking and can effectively reduce the residue of butachlor in rice, thereby alleviating the toxic effects of butachlor on rice seedlings.

[0036] Example 3: The alleviating effect of MT-SeNPs on butachlor stress in rice 0.2% melatonin and 3% trehalose were dissolved sequentially in water, then 60 mmol / L ascorbic acid was added, and the mixture was stirred at 40°C for 10 min to obtain a mixed solution. 15 mmol / L sodium selenite was added to the mixed solution, and the mixture was reacted at 40°C for 40 min to obtain MT-SeNPs. Similarly, MT-SeNPs (0 mg / L) prepared using this method... -1 6 mg L -1 The treatment of yellow-flowered rice seeds was carried out using the same methods and settings as in Example 2. After 7 days of treatment, the agronomic traits were measured. The results showed that ( Figure 4 Under butachlor stress, compared with water initiation, the plant height, root length and fresh weight of Huanghuazhan rice treated with MT-SeNPs were increased by 10.07%, 62.90% and 9.43%, respectively.

[0037] Example 4: The alleviating effects of different substances on rice under butachlor stress After disinfection, uniformly sized Huanghuazhan rice seeds were transferred to clean water and 22.5 mg L of water, respectively. -1 Melatonin, 300 mg / L -1 Trehalose, 13.1 mg / L -1 Sodium selenite, 6 mg / L -1 Nano selenium (SeNPs), 6 mg L -1 Initiation was performed for 48 h in MT-SeNPs solution (the preparation method of the material is the same as in Example 1). The remaining treatment conditions were the same as in Example 2, and agronomic traits were measured after 7 days of treatment. The treatment settings were as follows: water initiation (H2O), water initiation + butachlor (H2O + BT), melatonin initiation + butachlor (Melatonin + BT), trehalose initiation + butachlor (Trehalose + BT), sodium selenite initiation + butachlor (Selenite + BT), nano-selenium + butachlor (SeNPs + BT), and MT-SeNPs initiation + butachlor (MT-SeNPs + BT). The results showed that under butachlor stress, the root length, plant height, and fresh weight of rice induced by melatonin, trehalose, sodium selenite, nano-selenium, and MT-SeNPs were all higher than those induced by water. Among them, the root length, plant height, and fresh weight of rice induced by MT-SeNPs were 1.30-1.72 times, 1.27-1.32 times, and 1.04-1.15 times that induced by other substances, respectively. Figure 5 This indicates that, under butachlor stress, MT-SeNPs are more effective than trehalose, melatonin, sodium selenite, and nano-selenium in alleviating herbicide damage to rice.

[0038] Example 5: The alleviating effect of MT-SeNPs on different rice varieties under butachlor stress Select rice seeds of uniform size, including Qingxiangyou 19, Hanyou 78, and Zhonghua 11. First, disinfect the seeds with 5% (v / v) NaClO for 15 min, then rinse them three times with deionized water. Transfer the seeds to a solution containing 0 mg / L... -1 6 mg L -1The rice seedlings were prepared using the same MT-SeNPs solution as in Example 1. All other culture conditions and treatment settings were the same as in Example 2. Rice seedlings treated for 7 and 10 days were harvested, and their agronomic traits were measured. The results are shown in Table 1. After 7 days of butachlor stress, the root length, plant height, and fresh weight of the three rice varieties (Qingxiangyou 19, Hanyou 78, and Zhonghua 11) induced by MT-SeNPs were all higher than those induced by water. After 10 days of stress, the root length, plant height, and fresh weight of the three rice varieties induced by MT-SeNPs recovered to 3.72-26.30%, 25.88-32.59%, and 19.57-39.47% of the normal treatment values, respectively. This indicates that the prepared nano-sized butachlor safener can alleviate the phytotoxicity of butachlor to rice, and there is no specific selectivity for the varieties to which it is alleviated.

[0039] Table 1. Alleviating effects of MT-SeNPs on different rice varieties under butachlor stress.

[0040] All data in the table are mean ± standard error (n=5), and different lowercase letters indicate significant differences between different treatments (p<0.05).

[0041] It should be particularly emphasized that the above description is merely illustrative of preferred embodiments of the present invention and is not intended to limit the scope of protection of the present invention. Those skilled in the art can make various changes and modifications to the present invention within the spirit and principles disclosed herein. Any modifications, equivalent substitutions, improvements, or other variations falling within this scope should be considered as included within the scope of protection claimed by the present invention.

Claims

1. A method for preparing a nanoscale herbicide safener that alleviates butachlor phytotoxicity to rice, characterized in that, Melatonin was dissolved in water, trehalose was added, and the mixture was stirred until completely dissolved. Then, a reducing agent was added, and the mixture was reacted to obtain a mixed solution. Selenite was then added to the mixed solution to carry out a redox reaction, resulting in nano-sized butachlor safener.

2. The method for preparing the nanoscale herbicide safener for mitigating butachlor phytotoxicity in rice according to claim 1, characterized in that, The redox reaction is carried out at 35~45℃ for 20~60 min.

3. The method for preparing the nanoscale herbicide safener for mitigating butachlor phytotoxicity in rice according to claim 1, characterized in that, The reaction time for melatonin, trehalose, and reducing agent is 10 min, and the reaction temperature is 40~65℃.

4. The method for preparing the herbicide safener for mitigating butachlor phytotoxicity in rice according to claim 1, characterized in that, In the mixed solution, the concentration of melatonin is 0.1-0.3%.

5. The method for preparing the nanoscale herbicide safener for mitigating butachlor phytotoxicity in rice according to claim 1, characterized in that, In the mixed solution, the concentration of trehalose is 1.5-4%.

6. The method for preparing the nanoscale herbicide safener for mitigating butachlor phytotoxicity in rice according to claim 1, characterized in that, The reducing agent is at least one of ascorbic acid, glutathione, and sodium dodecyl sulfate.

7. The method for preparing the nanoscale herbicide safener for mitigating butachlor phytotoxicity in rice according to claim 1, characterized in that, In the mixed solution, the concentration of the reducing agent is 20~80 mmol / L.

8. The method for preparing the nanoscale herbicide safener for mitigating butachlor phytotoxicity in rice according to claim 1, characterized in that, The selenite is added to the mixed solution at a concentration of 5-20 mmol / L.

9. The nanoscale herbicide safener prepared by the preparation method according to any one of claims 1-8.

10. The application of the nanoscale herbicide safener according to claim 9, characterized in that, The process includes soaking rice seeds in a herbicide-safe solution, transferring the rice seeds into the herbicide-safe solution, initiating them at 28°C for 48 hours, then removing the seeds and removing the solution from them.

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