Application of nano sulfur fertilizer to improvement of plant salt tolerance and fruit quality
By preparing and applying nano-sulfur fertilizer, the Na+ efflux capacity of plants is enhanced, and the impact of salt stress on plants is solved, which significantly improves the salt tolerance and fruit quality of plants.
Patent Information
- Application Number
- CN202510201881.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-24
- Publication Date
- 2025-05-30
AI Technical Summary
Salt stress has had a significant impact on plant seed germination, growth and fruit quality, and existing nanomaterials still have room for improvement in improving plant salt tolerance.
Nanosulfur fertilizer prepared by hydrothermal reaction is used to alleviate salt stress by enhancing the plant's Na+ efflux capacity, improve the salt tolerance of the plant, and significantly improve the fruit quality.
Nanosulfur fertilizer significantly improves the germination rate and growth potential of plants under salt stress conditions, enhances the antioxidant ability of plants, and increases the single fruit weight and soluble solid content of the fruit.
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Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of nanoagriculture, and particularly relates to the application of a nano sulfur fertilizer in improving the salt tolerance of plants and the quality of fruits. Background Art
[0002] Salt stress can hinder processes such as seed germination, growth and development, flowering and fruiting, resulting in slowed plant growth, physiological imbalance and even death. The process from seed germination to seedling stage of plant seeds is a process sensitive to changes in seed moisture content and metabolism. Under salt stress conditions, the seed germination process is subject to osmotic stress, ionic stress and oxidative stress, reducing the seed germination rate. Excessive soluble salt content in the soil will weaken the ability of plants to passively and actively absorb water. The saline-alkali environment also indirectly affects the physiological and biochemical metabolic processes of plants, breaking the redox state and ionic balance state of plants, limiting the productivity of plants, and causing a reduction in yield and the quality of harvested products in agricultural production.
[0003] Nanomaterials refer to materials with at least one dimension between 1 - 100 nm. The large specific surface area of nanomaterials endows them with special interfacial effects, such as special surface structures, fluorescence characteristics, etc. At present, there have been some reports on using nanomaterials to improve the salt tolerance of plants. For example: Mn 3 O 4 nanomimetic enzymes enhanced the salt tolerance of cucumbers (Lu et al, 2020), and Ce 2 O nanoparticles improved the performance of crops such as rape, cotton, Arabidopsis thaliana, and rice under salt stress. However, nanomaterials that can effectively improve the salt tolerance of plants still need to be further developed and studied. Summary of the Invention
[0004] Aiming at the above-mentioned existing technologies, the purpose of the present invention is to provide the application of a nano sulfur fertilizer in improving the salt tolerance of plants and the quality of fruits.
[0005] To achieve the above purpose, the present invention adopts the following technical solutions:
[0006] In the first aspect of the present invention, there is provided the application of a nano sulfur fertilizer in improving the germination rate of plant seeds under salt stress conditions; the nano sulfur fertilizer is prepared by the following method:
[0007] Dissolve N-acetyl-L-cysteine and citric acid in water, and carry out a hydrothermal reaction at 160 - 200 °C for 3 - 5 h.
[0008] Preferably, the plant is Arabidopsis thaliana.
[0009] Preferably, the salt stress condition is 150 mM NaCl.
[0010] In a second aspect of the present invention, there is provided the use of the above-mentioned nano sulfur fertilizer in improving the salt tolerance of plants.
[0011] Preferably, the plant is Arabidopsis thaliana, apple rootstock and / or tomato.
[0012] In the above application, the nano sulfur fertilizer relieves plant salt stress by enhancing the Na + efflux ability of plants, thereby improving the salt tolerance of plants.
[0013] In a third aspect of the present invention, there is provided the use of the above-mentioned nano sulfur fertilizer in improving the quality of apple fruits.
[0014] In the above application, the fruit quality includes: single fruit weight and soluble solid content.
[0015] In a fourth aspect of the present invention, there is provided a method for improving the salt tolerance and fruit quality of plants, comprising the following steps:
[0016] Treat the plants with the above-mentioned nano sulfur fertilizer by irrigation.
[0017] Advantages of the present invention:
[0018] The nano sulfur fertilizer of the present invention can relieve salt stress during seed priming and at the same time improve the salt tolerance of plants during growth. Experiments show that the nano sulfur fertilizer of the present invention can scavenge hydroxyl radicals, reduce the content of MDA, H 2 O 2 , reduce the relative conductivity and increase the antioxidant enzyme activity.
[0019] The nano sulfur fertilizer of the present invention can also significantly improve the growth potential of plants under salt stress conditions, increase the Na + efflux flow rate, and relieve plant salt stress by enhancing the Na + efflux ability of plants. Description of the drawings
[0020] Figure 1 Chemical characterization of the nano sulfur fertilizer prepared in Example 1.
[0021] Figure 2 Distribution and localization of the nano sulfur fertilizer prepared in Example 1 in apple rootstock G935.
[0022] Figure 3 Phenotype and germination rate of Arabidopsis thaliana seeds primed with nano sulfur fertilizer under salt stress.
[0023] Figure 4 Effects of different dosages of nano sulfur fertilizer on Arabidopsis thaliana seedlings under salt stress.
[0024] Figure 5Effect of adding N-acetyl-L-cysteine alone on Arabidopsis thaliana seedlings under salt stress.
[0025] Figure 6 Effect of adding citric acid alone on Arabidopsis thaliana seedlings under salt stress.
[0026] Figure 7 Effect of nano-sulfur fertilizer on sodium ion efflux.
[0027] Figure 8 Effect of nano-sulfur fertilizer on salt tolerance of apple rootstocks.
[0028] Figure 9 Effect of nano-sulfur fertilizer on salt tolerance of tomato seedlings.
[0029] Figure 10 Effect of nano-sulfur fertilizer on apple fruit quality. Detailed implementation manners
[0030] It should be noted that the following detailed descriptions are all illustrative and are intended to provide further explanations of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which the present application belongs.
[0031] In order to enable those skilled in the art to more clearly understand the technical solutions of the present application, the technical solutions of the present application will be described in detail below with reference to specific embodiments.
[0032] The test materials used in the embodiments of the present invention are all conventional test materials in the art and can be obtained through commercial channels. The experimental methods without detailed conditions are carried out according to conventional test methods or according to the operation manuals recommended by the suppliers. Among them:
[0033] CAS number of N-acetyl-L-cysteine: 616-91-1; CAS number of citric acid: 77-92-9.
[0034] Example 1: Preparation of nano-sulfur fertilizer
[0035] Take 3.21 g of N-acetyl-L-cysteine and 1.92 g of citric acid and dissolve them in 30 ml of distilled water. Place the obtained solution in a high-pressure reaction kettle with a polytetrafluoroethylene lining and heat it at 180 °C for 4 hours. After the reaction, cool it naturally to obtain a crude nano-sulfur fertilizer product; add acetone to the crude nano-sulfur fertilizer product, and the addition amount of acetone is 90% of the weight of the crude nano-sulfur fertilizer product. Let it stand and precipitate naturally, and use a freeze dryer to remove the excess water in the precipitate to prepare a brownish-yellow powdery nano-sulfur fertilizer (S-CD).
[0036] Perform structural characterization on the nano-sulfur fertilizer prepared in this example, such as Figure 1As shown in A, the results show that the nano-sulfur fertilizer is spherical particles with an average particle size of 2.3 nm and a lattice spacing of 0.21 nm. As Figure 1 As shown in B, the nano-sulfur fertilizer has obvious infrared absorption. The absorption peaks at 3166, 2931 and 2520 cm -1 correspond to the stretching vibrations of OH / N-H, C-H and S-H bonds respectively. The absorption peaks at 1714 and 1622 cm -1 belong to C=O / C=N and C=C bonds respectively. The absorption peaks at 1400 cm -1 and 1115 cm -1 are the bending vibration absorptions of C-H and C-O bonds. As Figure 1 As shown in C, the nano-sulfur fertilizer has fluorescence properties and shows a strong absorption peak at 350 nm; under ultraviolet light excitation at about 360 nm, it can emit obvious blue light (near 431 nm). As Figure 1 As shown in D, the nano-sulfur fertilizer contains C, N, O, and S elements. The peaks at 174.9, 297.9, 409.9 and 545.1 eV in the X-ray photoelectron spectroscopy diagram of the nano-sulfur fertilizer correspond to S2p (2.69%), C 1s (44.25%), N 1s (7.31%) and O1s (43.11%) respectively.
[0037] Example 2: Investigation on the transport ability of nano-sulfur fertilizer in plants
[0038] 1. Test method:
[0039] The nano-sulfur fertilizer prepared in Example 1 was formulated into an aqueous solution of nano-sulfur fertilizer with a concentration of 200 mg / L. Using the leafy rootstock seedlings of apple rootstock G935 with a height of 3-5 cm, under the environmental conditions of simulated sunlight, the rootstock seedlings were hydroponically treated in the aqueous solution of nano-sulfur fertilizer. After 4 days of cultivation, the distribution of nano-sulfur fertilizer in apple rootstock G935 was photographed using a high-resolution laser confocal microscope.
[0040] 2. Test results:
[0041] The results are as Figure 2 shown. Strong blue fluorescence exists in both the leaves and roots of apple rootstock G935, indicating that the nano-sulfur fertilizer can be absorbed from the roots, transported upward, and finally reach the leaves.
[0042] Example 3: Effect of nano-sulfur fertilizer on seed germination under salt stress
[0043] 1. Test method:
[0044] Two treatments were set in the test, namely:
[0045] Treatment 1 (150 mM NaCl + S-CDs): Arabidopsis thaliana seeds disinfected with sodium hypochlorite were evenly sown on a 1 / 2 MS plate containing 100 mg / L of the nano-sulfur fertilizer prepared in Example 1 and 150 mM NaCl;
[0046] Treatment 2 (150 mM NaCl): Arabidopsis thaliana seeds disinfected with sodium hypochlorite were evenly sown on a 1 / 2 MS plate containing 150 mM NaCl without nano-sulfur fertilizer.
[0047] The 1 / 2 MS plates of Treatment 1 and Treatment 2 were placed at 4 °C in the dark for 3 days, then transferred to a light incubator for continued vernalization for 4 days. On the 8th day, the number of germinated seeds was counted, and the germination rate was calculated for 5 days.
[0048] 2. Test results:
[0049] The observation results of the seed germination on the 5th day are as Figure 3 shown in A, and the statistical results of the germination rate within 5 days are as Figure 3 shown in B. Compared with the addition of no nano-sulfur fertilizer, the addition of nano-sulfur fertilizer significantly promoted the germination of Arabidopsis thaliana seeds. This indicates that the nano-sulfur fertilizer can improve the salt tolerance of Arabidopsis thaliana seeds and promote germination.
[0050] Example 4: Effect of nano-sulfur fertilizer on salt tolerance of Arabidopsis thaliana seedlings
[0051] 1. Test method:
[0052] The following treatments were set up for the test, namely:
[0053] Treatment 1: Arabidopsis thaliana seedlings vernalized for 6 days were transferred to a 1 / 2 MS plate containing 0 mg / L, 100 mg / L, 200 mg / L, 300 mg / L of nano-sulfur fertilizer (prepared in Example 1) and 150 mM NaCl.
[0054] Treatment 2: Arabidopsis thaliana seedlings vernalized for 6 days were transferred to a 1 / 2 MS plate containing 300 mg / L of N-acetyl-L-cysteine and 150 mM NaCl.
[0055] Treatment 3: Arabidopsis thaliana seedlings vernalized for 6 days were transferred to a 1 / 2 MS plate containing 300 mg / L of citric acid and 150 mM NaCl.
[0056] The 1 / 2 MS plates of Treatment 1 - Treatment 3 were placed in a light incubator for 7 days. After 7 days, the phenotypes were observed and indicators such as fresh weight and relative conductivity were measured.
[0057] 2. Test results:
[0058] The effects of different addition amounts of nano-sulfur fertilizer in Treatment 1 on Arabidopsis thaliana seedlings under salt stress are asFigure 4 As shown, the use of nano-sulfur fertilizer can significantly promote the growth of Arabidopsis seedlings. After treatment with nano-sulfur fertilizer, root length and fresh weight increased significantly, and relative conductivity decreased significantly.
[0059] Effects of single addition of N-acetyl-L-cysteine on Arabidopsis seedlings under salt stress Figure 5 The effect of adding citric acid alone on Arabidopsis seedlings under salt stress is shown in Figure 6 The results showed that compared with the use of N-acetyl-L-cysteine alone and citric acid alone, the nano-sulfur fertilizer prepared by combining N-acetyl-L-cysteine and citric acid had a synergistic effect in improving the resistance of plants to salt stress.
[0060] Example 5: Effect of Nano-sulfur Fertilizer on Sodium Ion Efflux
[0061] 1. Test method:
[0062] In order to explore the effect of nano-sulfur fertilizer on sodium ion efflux, Arabidopsis seedlings with similar growth were used as experimental objects. The experiment was set up as follows:
[0063] Treatment 1: Arabidopsis seedlings were cultured in 1 / 2MS medium;
[0064] Treatment 2: Arabidopsis seedlings were cultured in 1 / 2MS medium containing 150 mM NaCl;
[0065] Treatment 3: Arabidopsis seedlings were cultured in 1 / 2MS medium containing 300 mg / L of the nano-sulfur fertilizer prepared in Example 1 and 150 mM NaCl.
[0066] The other culture conditions of each treatment remained the same. After 14 days of culture, the flux of sodium ions at the root apex was measured by the NMT method.
[0067] 2. Test results:
[0068] The results are as follows Figure 7 The results showed that under normal conditions (treatment 1), Arabidopsis seedlings showed lower Na + Under salt stress conditions (treatment 2), the Na + The addition of nano-sulfur fertilizer can significantly increase the Na excretion of Arabidopsis seedlings under salt stress conditions. + Therefore, the nano-sulfur fertilizer of the present invention can enhance the Na + Efflux capacity to alleviate plant salt stress.
[0069] Example 6: Effect of Nano-sulfur Fertilizer on Salt Tolerance of Apple Stock
[0070] 1. Test method:
[0071] Using apple rootstock G935 seedlings with similar growth vigor as the experimental objects, the following treatments were set in the experiment, namely:
[0072] Treatment 1: Irrigate the roots with 150 mM NaCl solution, once every 7 days, and the amount of irrigation per plant is 500 ml / plant;
[0073] Treatment 2: Irrigate the roots with 150 mM NaCl solution containing 200 mg / L of the nano-sulfur fertilizer prepared in Example 1, once every 7 days, and the amount of irrigation per plant is 500 ml / plant;
[0074] Treatment 3: Irrigate the roots with 150 mM NaCl solution containing 300 mg / L of the nano-sulfur fertilizer prepared in Example 1, once every 7 days, and the amount of irrigation per plant is 500 ml / plant;
[0075] Treatment 4: Irrigate the roots with 150 mM NaCl solution containing 400 mg / L of the nano-sulfur fertilizer prepared in Example 1, once every 7 days, and the amount of irrigation per plant is 500 ml / plant.
[0076] All treatments were cultured for 30 days under simulated sunlight conditions (from 8 am to 22 pm is simulated daytime, using white light (20.43 μmol·m -2 ·s -1 ) and UV-A light (λmax 365 nm, 0.83 μmol·m -2 ·s -1 ); from 22 pm to 8 am the next day is simulated night, without light).
[0077] Observe the phenotypes of apple rootstock G935 seedlings in each treatment group, and measure the fresh weight, relative conductivity, MDA content, O 2- activity, H 2 O 2 content, POD activity, CAT activity and SOD activity of apple rootstock seedlings in each treatment group.
[0078] 2. Test results:
[0079] The results are as Figure 8 shown: Under salt stress conditions, apple rootstock seedlings showed obvious growth inhibition, including leaf yellowing and even leaf margin scorching ( Figure 8 the leftmost pot in A), while after adding different concentrations of nano-sulfur fertilizer, the growth inhibition caused by salt stress on apple rootstock seedlings could be significantly alleviated ( Figure 8 the second pot to the fourth pot from the left in A are respectively added with 200 mg / L, 300 mg / L and 400 mg / L of nano-sulfur fertilizer).
[0080] Compared with Treatment 1, after adding nano-sulfur fertilizer at different concentrations, the fresh weight of apple rootstock seedlings can be significantly increased ( Figure 8 B), and the activities of SOD, POD and CAT ( Figure 8 G-I); the relative conductivity can be reduced ( Figure 8 C), the MDA content ( Figure 8 D), the O 2- activity ( Figure 8 E) and the H 2 O 2 content ( Figure 8 F).
[0081] The above results show that the nano-sulfur fertilizer of the present invention has a significant effect on alleviating the salt stress of apple rootstocks.
[0082] Example 7: Effect of nano-sulfur fertilizer on salt tolerance of tomatoes
[0083] 1. Test method:
[0084] Using tomato seedlings with similar growth as the experimental objects, the following treatments were set in the test, respectively:
[0085] Treatment 1: Irrigate the roots with 150 mM NaCl solution, once every 7 days, and the irrigation amount per plant is 500 ml / plant;
[0086] Treatment 2: Irrigate the roots with 150 mM NaCl solution containing 200 mg / L of the nano-sulfur fertilizer prepared in Example 1, once every 7 days, and the irrigation amount per plant is 500 ml / plant;
[0087] Treatment 3: Irrigate the roots with 150 mM NaCl solution containing 300 mg / L of the nano-sulfur fertilizer prepared in Example 1, once every 7 days, and the irrigation amount per plant is 500 ml / plant;
[0088] Treatment 4: Irrigate the roots with 150 mM NaCl solution containing 400 mg / L of the nano-sulfur fertilizer prepared in Example 1, once every 7 days, and the irrigation amount per plant is 500 ml / plant.
[0089] All treatments were cultured for 30 days under simulated sunlight conditions (from 8:00 am to 22:00 pm is simulated daytime, using white light (20.43 μmol·m -2 ·s -1 ) and UV-A light (λmax 365 nm, 0.83 μmol·m -2 ·s -1 ); from 22:00 pm to 8:00 am the next day is simulated night, without light).
[0090] Observe the phenotypes of tomato seedlings in each treatment group, and measure the fresh weight, relative conductivity, and MDA content of tomato seedlings in each treatment group.
[0091] 2. Test results:
[0092] The results are as Figure 9 shown: Under salt stress conditions, tomato seedlings showed obvious growth inhibition ( Figure 9 the leftmost pot in A), while after adding nano-sulfur fertilizers at different concentrations, the growth inhibition caused by salt stress on tomato seedlings could be significantly alleviated ( Figure 9 the second to fourth pots from the left in A are the pots added with 200 mg / L, 300 mg / L, and 400 mg / L nano-sulfur fertilizers respectively).
[0093] Compared with the treatment group 1, after adding nano-sulfur fertilizers at different concentrations, the fresh weight of tomato seedlings could be significantly increased ( Figure 9 B); the relative conductivity ( Figure 9 C) and MDA content ( Figure 9 D) were decreased.
[0094] The above results indicate that the nano-sulfur fertilizer of the present invention has a significant effect on alleviating salt stress of tomato seedlings.
[0095] Example 8: Effect of nano-sulfur fertilizer on apple fruit quality
[0096] 1. Test method:
[0097] Use four-year-old apple trees with similar growth vigor and growing in saline-alkali environments as experimental objects. The following treatments are set in the test, which are respectively:
[0098] Treatment group 1 (T1): On June 3 (day 0), June 23 (day 20), July 15 (day 41), and July 31 (day 57), irrigate the apple trees with clear water by root irrigation, and the irrigation amount is 2 L / tree.
[0099] Treatment group 2 (T2): On June 3 (day 0), June 23 (day 20), July 15 (day 41), and July 31 (day 57), irrigate the apple trees with the nano-sulfur fertilizer aqueous solution prepared in Example 1 at 1000 mg / L by root irrigation, and the irrigation amount is 2 L / tree.
[0100] Treatment group 3 (T3): On June 3 (day 0), June 23 (day 20), July 15 (day 41), and July 31 (day 57), irrigate the apple trees with the nano-sulfur fertilizer aqueous solution prepared in Example 1 at 2000 mg / L by root irrigation, and the irrigation amount is 2 L / tree.
[0101] Treatment group 4 (T4): On June 3 (day 0), June 23 (day 20), July 15 (day 41), and July 31 (day 57), apple trees were treated by root irrigation with the aqueous solution of nano sulfur fertilizer prepared in Example 1 at a concentration of 3000 mg / L, and the irrigation amount was 2 L per tree.
[0102] Samples were taken for quality determination after each treatment.
[0103] Fruit quality determination:
[0104] Single fruit weight: Weigh and record the weight of each fruit with an electronic balance accurate to one ten-thousandth.
[0105] Soluble solids: Measured using a hand-held refractometer.
[0106] Titratable acid: Determined by acid-base neutralization method.
[0107] Solid-acid ratio: The ratio of soluble solids to titratable acid.
[0108] Fruit hardness: Measured using a hand-held hardness tester.
[0109] Fruit glossiness: Measured using a hand-held glossmeter.
[0110] 2. Test results:
[0111] Data statistical analysis found that compared with group T1, after applying nano sulfur fertilizer (T2, T3, and T4), the soluble sugar content of the fruit increased significantly, by 1.43%, 7.8%, and 14.28% respectively; the content of titratable acid in the fruit decreased significantly by 6.25%, 30.76%, and 30.76%; the hardness of the fruit pulp increased by 4.8%, 36.5%, and 64.3% respectively, and the single fruit weight of the fruit increased by 33.9%, 37.6%, and 82.5% ( Figure 10 ).
[0112] Therefore, the treatment with nano sulfur fertilizer can significantly improve fruit hardness, soluble content, single fruit weight, and solid-acid ratio, and significantly reduce the content of titratable acid.
[0113] The above are only the preferred embodiments of the present application and are not used to limit the present application. For those skilled in the art, the present application can have various changes and modifications. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims
1. Application of nano-sulfur fertilizer in improving the germination rate of plant seeds under salt stress conditions; characterized in that: The nano-sulfur fertilizer is prepared by the following method: Dissolve N-acetyl-L-cysteine and citric acid in water and perform hydrothermal reaction at 160-200°C for 3-5 hours.
2. The use according to claim 1, characterized in that: The plant is Arabidopsis thaliana.
3. The use according to claim 1, characterized in that: The salt stress condition was 150 mM NaCl.
4. Application of nano-sulfur fertilizer in improving plant salt tolerance; characterized in that, The nano-sulfur fertilizer is prepared by the following method: Dissolve N-acetyl-L-cysteine and citric acid in water and perform hydrothermal reaction at 160-200°C for 3-5 hours.
5. The use according to claim 4, characterized in that: The plants are Arabidopsis thaliana, apple and / or tomato.
6. The use according to claim 4, characterized in that: The nano-sulfur fertilizer can enhance the Na + Efflux capacity to alleviate plant salt stress.
7. The application of nano-sulfur fertilizer in improving the quality of apple fruit is characterized by: The nano-sulfur fertilizer is prepared by the following method: Dissolve N-acetyl-L-cysteine and citric acid in water and perform hydrothermal reaction at 160-200°C for 3-5 hours.
8. The use according to claim 7, characterized in that: The fruit quality includes: single fruit weight and soluble solid content.
9. A method for improving plant salt tolerance and fruit quality, characterized in that: The following steps are involved: Treat plants with nano-sulfur fertilizer by irrigation; The nano-sulfur fertilizer is prepared by the following method: Dissolve N-acetyl-L-cysteine and citric acid in water and perform hydrothermal reaction at 160-200°C for 3-5 hours.