A method of applying l-alpha-aspartyl-l-phenylalanine to enhance salt tolerance in bermudagrass
By applying L-α-aspartic-L-phenylalanine solution to Bermuda grass, the content of H2O2, MDA and proline in Bermuda grass under salt stress is reduced, which solves the problem of growth damage caused by salt stress and achieves a simple and low-cost effect of improving saline soil. It is suitable for lawn establishment and ecological restoration.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- NANTONG UNIV
- Filing Date
- 2026-03-11
- Publication Date
- 2026-06-12
AI Technical Summary
In existing technologies, salt stress causes serious damage to plant growth and development. Traditional improvement techniques are costly and difficult to promote on a large scale. There is an urgent need for a simple and environmentally friendly saline soil improvement technology to improve the salt tolerance of plants.
Applying L-α-aspartic-L-phenylalanine solution to Bermuda grass alleviates salt stress and reduces the content of H2O2, MDA, and proline during the cultivation process, thereby enhancing the salt tolerance of Bermuda grass.
It significantly reduces oxidative damage and osmotic stress of bermudagrass under salt stress, improves its growth in high-salt environments, and is suitable for turf establishment and ecological restoration in saline soils.
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Figure CN122181531A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of plant stress resistance technology, and more particularly to a method for enhancing the salt tolerance of Cynodon dactylon by applying L-α-aspartic-L-phenylalanine. Background Technology
[0002] Soil salinity severely impairs plant growth and development, causing enormous losses in crop quality and yield worldwide. Excessive sodium (Na+) in the soil leads to ionic, osmotic, and oxidative stress in plants, resulting in growth inhibition, developmental changes, and subsequent metabolic adjustments. Salt stress, as a typical abiotic stress, negatively impacts plant growth and development through a triple mechanism of ion toxicity, osmotic imbalance, and oxidative burst. The main ion causing salt stress is Na+. + and Cl - Excessive accumulation of K in the cytoplasm destroys K + / Na + Homeostasis inhibits photosynthetic enzyme activity, leading to a decrease in osmotic potential and difficulty in water absorption by plant roots, resulting in wilting and yellowing of the above-ground parts. A surge in reactive oxygen species triggers membrane lipid peroxidation (increased MDA), protein oxidation, and DNA damage, ultimately manifesting as yellowing and wilting of plant leaves, and even death. Traditional soil improvement techniques suffer from high costs, repetitive degradation, and difficulty in large-scale implementation; therefore, there is an urgent need for a simple, environmentally friendly saline soil improvement technology to ensure the efficient, rational, and sustainable development and utilization of saline soils.
[0003] During long-term adaptation to salt stress, plants have gradually evolved a series of strategies and response mechanisms to address it. In previous salt stress experiments on Bermuda grass, the applicant discovered that gradient-acclimated Bermuda grass exhibited stronger salt stress tolerance compared to direct salt treatment. Metabolomics analysis revealed that gradient-acclimated Bermuda grass significantly accumulated amino acid metabolites. Amino acids, as important physiologically active substances in plants, play multiple crucial roles in responding to salt stress. Under salt stress, plants actively accumulate osmotic-regulating amino acids and their derivatives, such as proline and glycine betaine, to alleviate osmotic stress damage by reducing cell osmotic potential and maintaining turgor pressure balance. Simultaneously, amino acids, as precursors for the synthesis of polyamines and γ-aminobutyric acid (GABA), can activate antioxidant defense systems, activate antioxidant enzyme systems, scavenge excess reactive oxygen species, and reduce membrane lipid peroxidation. Many amino acids have been used in applications and development to improve crop salt tolerance, but research reports on L-α-aspartic-L-phenylalanine are lacking. Summary of the Invention
[0004] The purpose of this invention is to overcome the shortcomings of existing technologies by proposing a method for enhancing the salt tolerance of Bermuda grass by applying L-α-aspartic-L-phenylalanine, which can improve the growth of Bermuda grass in high-salt environments. This method is simple to operate, low in cost, and suitable for turf establishment and ecological restoration in saline soils, showing great application potential.
[0005] To achieve the above objectives, the present invention employs the following technical solution: a method for enhancing the salt tolerance of bermudagrass roots by applying L-α-aspartic-L-phenylalanine, comprising the following steps:
[0006] Step 1: Prepare L-α-aspartic-L-phenylalanine solutions with concentrations of 0.03-0.1 mM using 1 / 2 Hoagland modified nutrient solution and 300 mM NaCl solution, respectively. Specifically, these concentrations are 0.03, 0.05, and 0.1 mM, and are recorded as 0.03 AP, 0.05 AP, 0.1 AP, D300+0.03 AP, D300+0.05 AP, and D300+0.1 AP, respectively.
[0007] Step 2: Select bermudagrass rhizome segments of approximately 15 cm in length and similar growth, and culture them in 1 / 2 Hogland modified nutrient solution for 20 days. The culture conditions are: 28 ℃ / 26 ℃ day and night temperature, 70% relative humidity, 12 h light / 12 h dark; pH of 1 / 2 Hogland modified nutrient solution 5.9-6.0.
[0008] Step 3: After culturing the bermudagrass root and stem segments for 20 days, first culture the bermudagrass roots in Hogland nutrient solution containing L-α-aspartic-L-phenylalanine for 3 days, and then add a salt solution containing L-α-aspartic-L-phenylalanine, keeping the culture conditions unchanged.
[0009] By adopting the above technical solution, the contents of H2O2, MDA and proline (Pro) in the leaves and roots of Bermuda grass were significantly reduced after applying L-α-aspartic-L-phenylalanine, indicating that the salt tolerance of Bermuda grass was enhanced.
[0010] Preferably, in step 2, the treatment method is to pretreat with L-α-aspartic-L-phenylalanine prepared with 1 / 2 Hogland modified nutrient solution for 3 days before treatment with 300 mM NaCl.
[0011] The base of the bermudagrass root segment is fixed with a sponge, which is about 5cm wide and used for fixation and attachment of new roots.
[0012] Preferably, in step 3, the salt tolerance of the plant is tested by verifying the content of H2O2, MDA and Pro. The test and phenotypic record are performed after 4 days of treatment to evaluate the differences in salt tolerance of bermudagrass.
[0013] Preferably, the external application concentration of L-α-aspartic-L-phenylalanine is 0.03-0.1 mM, and all of these concentrations are effective.
[0014] Preferably, the nutrient solution for bermudagrass should be changed every 4 days.
[0015] By employing the above-mentioned technical solution—applying 0.03-0.1 mM L-α-aspartic-L-phenylalanine under 300 mM NaCl stress—the levels of H2O2, MDA, and proline in the leaves and roots of Bermuda grass can be significantly reduced, alleviating oxidative damage, membrane lipid peroxidation, and osmotic balance, thereby improving the growth status of Bermuda grass in high-salt environments. This method is simple to operate, low in cost, and suitable for turf establishment and ecological restoration in saline soils, showing great application potential.
[0016] Compared with the prior art, the present invention has the following beneficial effects:
[0017] Compared with salt treatment alone, the application of L-α-aspartic-L-phenylalanine in this invention significantly reduced the contents of H2O2, MDA and Pro in bermudagrass under salt stress, indicating that L-α-aspartic-L-phenylalanine can effectively alleviate oxidative damage, membrane lipid peroxidation and osmotic balance. Compared to the D300 group, the D300+0.03AP group showed a 19.0% decrease in H2O2, a 56.5% decrease in MDA, and a 90.4% decrease in Pro in leaves, and a 6.8% decrease in H2O2, an 86.6% decrease in MDA, and a 73.2% decrease in Pro in roots; the D300+0.05AP group showed a 37.7% decrease in H2O2, a 43.2% decrease in MDA, and an 80.4% decrease in Pro in leaves, and a 58.0% decrease in H2O2, an 83.8% decrease in MDA, and a 80.6% decrease in Pro in roots; the D300+0.1AP group showed a 20.0% decrease in H2O2, a 64.3% decrease in MDA, and an 86.8% decrease in Pro in leaves, and a 48.6% decrease in H2O2, an 85.2% decrease in MDA, and a 72.3% decrease in Pro in roots, indicating that L-α-aspartic-L-phenylalanine application can reduce the damage of salt stress to plants. Attached Figure Description
[0018] Figure 1Phenotypic images of plants subjected to salt stress after applying different concentrations of L-α-aspartic-L-phenylalanine according to this invention are shown. a) is the control group, denoted as CK (cultured in 1 / 2 Hogland modified nutrient solution for 4 days); b) is the salt stress group, denoted as D300 (cultured in 300 mM NaCl for 4 days, prepared with 1 / 2 Hogland modified nutrient solution); c) the experimental group subjected to exogenous application of 0.03 mM L-α-aspartic-L-phenylalanine, denoted as D300+0.03AP (cultured in 0.03 mM L-α-aspartic-L-phenylalanine solution prepared in 1 / 2 Hogland modified nutrient solution for 3 days, followed by 0.03 mM L-α-aspartic-L-phenylalanine solution prepared in 300 mM NaCl for 4 days); d) the group subjected to exogenous application of 0.05 mM L-α-aspartic-L-phenylalanine. The L-α-aspartic-L-phenylalanine experimental group was denoted as D300+0.05AP (cultured for 3 days in 0.05 mM L-α-aspartic-L-phenylalanine solution prepared with 1 / 2 Hogland modified nutrient solution, followed by 4 days in 0.05 mM L-α-aspartic-L-phenylalanine solution prepared with 300 mM NaCl). e) The experimental group with exogenous application of 0.1 mM L-α-aspartic-L-phenylalanine was denoted as D300+0.1AP (cultured for 3 days in 0.1 mM L-α-aspartic-L-phenylalanine solution prepared with 1 / 2 Hogland modified nutrient solution, followed by 4 days in 0.1 mM L-α-aspartic-L-phenylalanine solution prepared with 300 mM NaCl).
[0019] Figure 2This diagram illustrates the changes in the content of damage indicators in the leaves of *Cynodon dactylon* after exogenous application of L-α-aspartic-L-phenylalanine according to the present invention. CK represents the control group (cultured in 1 / 2 Hogland modified nutrient solution for 4 days), D300 represents the salt stress treatment group (cultured in 300 mM NaCl for 4 days, prepared with 1 / 2 Hogland modified nutrient solution), D300+0.03AP represents the experimental group with exogenous application of 0.03 mM L-α-aspartic-L-phenylalanine (cultured in 0.03 mM L-α-aspartic-L-phenylalanine solution prepared in 1 / 2 Hogland modified nutrient solution for 3 days, followed by 4 days of culture in 0.03 mM L-α-aspartic-L-phenylalanine solution prepared in 300 mM NaCl), and D300+0.05AP represents the group with exogenous application of 0.05 mM L-α-aspartic-L-phenylalanine. The L-α-aspartic-L-phenylalanine experimental group (cultured for 3 days in 0.05 mM L-α-aspartic-L-phenylalanine solution prepared with 1 / 2 Hogland modified nutrient solution, followed by 4 days in 0.05 mM L-α-aspartic-L-phenylalanine solution prepared with 300 mM NaCl), and the D300+0.1AP experimental group with exogenous 0.1 mM L-α-aspartic-L-phenylalanine (cultured for 3 days in 0.1 mM L-α-aspartic-L-phenylalanine solution prepared with 1 / 2 Hogland modified nutrient solution, followed by 4 days in 0.1 mM L-α-aspartic-L-phenylalanine solution prepared with 300 mM NaCl).
[0020] Figure 3This diagram illustrates the changes in the content of damage indicators in *Cynodon dactylon* roots after exogenous application of L-α-aspartic-L-phenylalanine according to the present invention. CK represents the control group (cultured in 1 / 2 Hogland modified nutrient solution for 4 days), D300 represents the salt stress treatment group (cultured in 300 mM NaCl for 4 days, prepared with 1 / 2 Hogland modified nutrient solution), D300+0.03AP represents the experimental group with exogenous application of 0.03 mM L-α-aspartic-L-phenylalanine (cultured in 0.03 mM L-α-aspartic-L-phenylalanine solution prepared in 1 / 2 Hogland modified nutrient solution for 3 days, followed by 4 days of culture in 0.03 mM L-α-aspartic-L-phenylalanine solution prepared in 300 mM NaCl), and D300+0.05AP represents the group with exogenous application of 0.05 mM L-α-aspartic-L-phenylalanine. The L-α-aspartic-L-phenylalanine experimental group (cultured for 3 days in 0.05 mM L-α-aspartic-L-phenylalanine solution prepared with 1 / 2 Hogland modified nutrient solution, followed by 4 days in 0.05 mM L-α-aspartic-L-phenylalanine solution prepared with 300 mM NaCl), and the D300+0.1AP experimental group with exogenous 0.1 mM L-α-aspartic-L-phenylalanine (cultured for 3 days in 0.1 mM L-α-aspartic-L-phenylalanine solution prepared with 1 / 2 Hogland modified nutrient solution, followed by 4 days in 0.1 mM L-α-aspartic-L-phenylalanine solution prepared with 300 mM NaCl). Detailed Implementation
[0021] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings, so that those skilled in the art can better understand the advantages and features of the present invention, thereby making a clearer definition of the scope of protection of the present invention. The embodiments described in this invention are only some embodiments of the present invention, not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.
[0022] illustrate:
[0023] In the examples described in this specification, the Bermuda grass root used was from Yangjiang. L-α-aspartic-L-phenylalanine was provided by Shanghai Maclean Biochemical Technology Co., Ltd., with a content >98%. NaCl was a conventional chemical reagent.
[0024] Example 1: (Applying 0.03 mM AP)
[0025] processing method
[0026] Step 1: (1) Select a bermudagrass rhizome segment about 15cm in length with good growth, remove the bottom leaves, fix the bottom of the bermudagrass rhizome segment with a sponge, and place it in a culture box to be cultured in 1 / 2 Hogland modified nutrient solution for 20 days under the following conditions: 28°C / 26°C day and night temperature cycle, 70% relative humidity and 12 hours light / 12 hours dark light cycle; change the nutrient solution once every 4 days.
[0027] (2) After the seedlings have recovered and rooted, bermudagrass materials with similar growth were selected for experimental treatment. Before treatment, the D300+0.03AP group was pretreated with 0.03AP prepared by 1 / 2 Hogland modified nutrient solution for 3 days, and then treated with L-α-aspartic-L-phenylalanine solution prepared by 300 mM NaCl.
[0028] Step 2: Four days after treatment, the changes in the contents of H2O2, MDA, and Pro, indicators of root damage in canine teeth, were detected.
[0029] Results: During salt stress treatment, 300 mM NaCl treatment significantly affected the growth of Bermuda grass, causing yellowing of leaves, basal wilting, and water loss between stems. Application of 0.03 mM L-α-aspartic-L-phenylalanine significantly improved the yellowing and wilting caused by salt stress, indicating that exogenous L-α-aspartic-L-phenylalanine can improve the salt stress tolerance of Bermuda grass (e.g., ...). Figure 1 (As shown). Compared to the direct salt stress treatment group, the contents of H2O2, MDA, and Pro in the D300+0.03AP group were significantly reduced, demonstrating at the physiological level that L-α-aspartic-L-phenylalanine has the ability to alleviate salt damage and improve the salt stress tolerance of Bermuda grass. Using D300 as a reference, the H2O2 in the leaves of the D300+0.03AP group decreased by 19.0%, MDA by 56.5%, and Pro by 90.4% (as shown). Figure 2 As shown), H2O2 in the roots decreased by 6.8%, MDA decreased by 86.6%, and Pro decreased by 73.2%. This clearly demonstrates that 0.03 mM L-α-aspartic-L-phenylalanine can effectively alleviate salt stress-induced membrane lipid peroxidation damage, excess reactive oxygen species production, and osmotic imbalance, thereby improving the salt tolerance of Bermuda grass roots (e.g., ...). Figure 3 (As shown).
[0030] Example 2: (Applying 0.05mM AP)
[0031] 0.05 mM L-α-aspartic-L-phenylalanine was treated with 300 mM NaCl and recorded as D300+0.05AP. The treatment conditions were the same as in Example 1.
[0032] Results: MDA in leaves decreased by 43.2%, H2O2 decreased by 37.7%, and Pro decreased by 80.4% (e.g. Figure 2 As shown); in the root, MDA decreased by 83.8%, H2O2 decreased by 58.0%, and Pro decreased by 80.6% (as shown). Figure 3 (As shown).
[0033] Example 3: (Applying 0.1 mM AP)
[0034] 0.1 mM L-α-aspartic-L-phenylalanine was treated with 300 mM NaCl and recorded as D300+0.1AP. The treatment conditions were the same as in Example 1.
[0035] Results: MDA in leaves decreased by 64.3%, H2O2 decreased by 20.0%, and Pro decreased by 86.8% (e.g. Figure 2 As shown); in the root, MDA decreased by 85.2%, H2O2 decreased by 48.6%, and Pro decreased by 72.3% (as shown). Figure 3 (As shown).
[0036] Compared with the salt-treated group alone, the contents of hydrogen peroxide (H2O2), MDA, and Pro in Bermuda grass treated with exogenous L-α-aspartic-L-phenylalanine showed a significant decreasing trend. The significant decrease in H2O2 indicates that this substance can effectively scavenge excess reactive oxygen species (ROS) accumulated in plant cells under salt stress, inhibit oxidative stress, and reduce oxidative damage to cell membranes, proteins, and nucleic acids. The decrease in MDA, a core marker product of membrane lipid peroxidation, directly proves that L-α-aspartic-L-phenylalanine can effectively maintain the structural integrity and functional stability of the cell membrane, alleviate membrane lipid peroxidation damage induced by salt stress, reduce cell membrane permeability, and ensure the normal transport of substances and signal transduction between intracellular and extracellular spaces. The significant decrease in Pro content indicates that under the regulatory effect of this substance, the degree of salt stress suffered by Bermuda grass was greatly reduced, and the osmotic pressure imbalance in cells was effectively improved. Normal physiological metabolic balance could be maintained without the need for large accumulations of osmotic regulators such as Pro, indirectly reflecting a significant relief of osmotic stress. The above results fully demonstrate that exogenous application of L-α-aspartic-L-phenylalanine can alleviate the physiological damage caused by salt stress to bermudagrass at three levels: scavenging reactive oxygen species, protecting the cell membrane system, and improving cell osmotic balance, thereby enhancing the plant's adaptability to salt stress and significantly improving the plant's salt tolerance.
[0037] In conclusion, the exogenous L-α-aspartic-L-phenylalanine of this invention can effectively alleviate the damage of salt stress to bermudagrass roots in the range of 0.03-0.1 mM, with 0.05 mM being the optimal concentration, and has application value.
[0038] In summary, this invention demonstrates that applying 0.03-0.1 mM L-α-aspartic-L-phenylalanine under 300 mM NaCl stress significantly reduces the levels of H2O2, MDA, and proline in the leaves and roots of Bermuda grass, alleviating oxidative damage, membrane lipid peroxidation, and osmotic balance, thereby improving the growth of Bermuda grass in high-salt environments. This method is simple to operate, low in cost, and suitable for turf establishment and ecological restoration in saline soils, showing great application potential.
[0039] The descriptions and practices disclosed in this invention are readily apparent and understandable to those skilled in the art, and various modifications and refinements can be made without departing from the principles of this invention. Therefore, any modifications or improvements made without departing from the spirit of this invention should also be considered within the scope of protection of this invention.
Claims
1. A method for enhancing the salt tolerance of bermudagrass roots by applying L-α-aspartic-L-phenylalanine, characterized in that, Includes the following steps: Step 1: Prepare L-α-aspartic-L-phenylalanine solution with a concentration of 0.03-0.1 mM using 1 / 2 Hogland modified nutrient solution and 300 mM NaCl solution as solvents respectively; Step 2: Select bermudagrass rhizome segments of approximately 15 cm in length and similar growth, and culture them in 1 / 2 Hogland modified nutrient solution for 20 days. The culture conditions are: 28 ℃ / 26 ℃ day and night temperature, 70% relative humidity, 12 h light / 12 h dark; pH of 1 / 2 Hogland modified nutrient solution 5.9-6.
0. Step 3: After culturing the bermudagrass root and stem segments for 20 days, first culture the bermudagrass roots in Hogland nutrient solution containing L-α-aspartic-L-phenylalanine for 3 days, and then add a salt solution containing L-α-aspartic-L-phenylalanine, keeping the culture conditions unchanged.
2. The method for enhancing the salt tolerance of bermudagrass roots by applying L-α-aspartic-L-phenylalanine according to claim 1, characterized in that, In step 2, the treatment method is to pretreat with L-α-aspartic-L-phenylalanine prepared by 1 / 2 Hogland modified nutrient solution for 3 days before treatment with 300 mM NaCl.
3. The method for enhancing the salt tolerance of bermudagrass roots by applying L-α-aspartic-L-phenylalanine according to claim 1, characterized in that, In step 3, the salt tolerance of the plant is tested by verifying the content of H2O2, MDA and Pro. The test and phenotypic record are performed after 4 days of treatment.
4. The method for enhancing the salt tolerance of bermudagrass roots by applying L-α-aspartic-L-phenylalanine according to claim 1, characterized in that, The external application concentration of L-α-aspartic-L-phenylalanine is 0.03-0.1 mM.
5. The method for enhancing the salt tolerance of bermudagrass roots by applying L-α-aspartic-L-phenylalanine according to claim 1, characterized in that, Change the nutrient solution for bermudagrass every 4 days.