Use of mnpses3 nanoparticles in improving salt stress resistance of plants

Abstract

The present application relates to the field of economic crop cultivation, and more particularly to application of MnPSe3 nanoparticles in improving salt stress resistance of plants. The present application uniformly mixes MnPSe3 nanoparticles with water to prepare a nano-solution, and then sprays the nano-solution on the surface of plant leaves. Spraying this MnPSe3 nanoparticles on the leaves of ice plants under salt stress environment can significantly alleviate the salt stress of the ice plants, and at the same time, enhance the antioxidant capacity and sugar resistance of the ice plants. The method of the present application not only improves the yield of the ice plants under salt stress, but also improves the alpha-amylase inhibitory activity of the ice plants.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of economic crop cultivation and planting, and particularly relates to application of MnPSe3 nanoparticles in improving salt stress resistance of plants. BACKGROUND

[0002] The formation of saline-alkali soil is mainly due to the combined action of natural factors and human activities. Natural factors include rainfall less than evaporation, high groundwater level, and high salt content in soil parent material. Human factors such as excessive irrigation, unreasonable land use, poor drainage, etc. can also exacerbate salinization. Saline-alkali soil has a serious impact on agricultural production and the ecological environment, leading to reduced crop yields or even failure to grow, disrupting the balance of the ecological system, and affecting biodiversity.

[0003] Saline-alkali soil causes multiple hazards to vegetable growth. First, high salt content leads to an increase in soil solution osmotic pressure, making it difficult for vegetable roots to absorb water, and even causing water to flow in reverse, causing dehydration and physiological drought in vegetables. Second, excessive sodium and chloride ions are toxic, interfering with the activity of enzymes in vegetable cells, affecting the absorption and metabolism of nutrients, causing leaf burns and growth inhibition. In addition, the high salt content of saline-alkali soil also changes the soil structure, making the soil harden, and the aeration and water permeability worse, further affecting the growth and development of the roots. These factors together severely limit the growth and development of vegetables, reduce crop yield and quality, and even cause vegetable death, posing a major challenge to agricultural production.

[0004] Ice lettuce is a vegetable with both nutritional and medicinal value. From a nutritional perspective, it is rich in vitamin C, vitamin A, and dietary fiber, which helps to enhance immunity, promote digestion, and maintain skin health, and also contains important minerals such as calcium, iron, and potassium. In terms of medicinal value, ice lettuce has the effects of clearing heat and detoxifying, diuresis and swelling relief, and can be used to relieve hot diseases such as fever and sore throat. In addition, its active ingredients help to delay aging and reduce blood sugar, making it a functional vegetable with high nutritional value. However, saline-alkali soil significantly affects the growth of ice lettuce due to high salt content and alkaline substances, making it difficult for the roots to absorb water and nutrients, and changing the soil properties, inhibiting the absorption of trace elements by ice lettuce, causing physiological diseases, and ultimately significantly reducing yield.

[0005] Currently, the main methods to alleviate salt stress include soil improvement, irrigation management and biotechnology. Soil improvement reduces the content of soil salt and improves soil structure by applying organic fertilizer or chemical modifier; irrigation management reduces the accumulation of salt on the soil surface by using drip irrigation or underground drip irrigation technology; biotechnology focuses on using salt-tolerant plants or genetic engineering to breed salt-tolerant crop varieties to improve the adaptability of plants to salt stress. In recent years, nanomaterials have shown unique advantages in alleviating salt stress due to their large specific surface area and high reactivity. Nanomaterials can more efficiently adsorb soil salt and promote the absorption of nutrients by plant roots, thereby enhancing the salt tolerance of plants. Therefore, in the face of the increasingly serious problem of saline-alkali land, nanotechnology as a new solution has attracted widespread attention. SUMMARY

[0006] The application provides application of MnPSe3 nanoparticles in improving salt stress resistance of plants. The application discloses a nanomaterial named MnPSe3 which is rich in manganese (Mn), phosphorus (P) and selenium (Se) elements beneficial to the growth of vegetables. The nanomaterial is applied to the leaves of ice plants under salt stress environment, can significantly alleviate the salt stress of the ice plants, and can enhance the antioxidant capacity and sugar activity of the ice plants. The method not only improves the yield of the ice plants under salt stress, but also improves the alpha-amylase inhibitory activity of the ice plants. By spraying the nanometer MnPSe3 on the leaves, the salt stress resistance and nutritional value of the ice plants can be improved under the stress of saline-alkali land.

[0007] In order to achieve the above application purposes, the application provides the following technical solutions.

[0008] The application provides application of MnPSe3 nanoparticles in improving salt stress resistance of plants.

[0009] Preferably, the application method is as follows.

[0010] The MnPSe3 nanoparticles are uniformly mixed with water to prepare a nanosolution, and the nanosolution is sprayed on the surface of the leaves of the plants.

[0011] Preferably, the mass-volume ratio of the MnPSe3 nanoparticles to water is 50-150 mg / L.

[0012] Preferably, the interval time of spraying is 4-7 days, and the spraying frequency is 1-2 times.

[0013] Preferably, the plants are ice plants.

[0014] The application also provides application of the MnPSe3 nanoparticles in improving the antioxidant capacity of plants under salt stress, and the application method is: spraying the plant leaf surface with the MnPSe3 nanoparticles after the MnPSe3 nanoparticles are dissolved to a concentration of 50-150 mg / L.

[0015] The application also provides application of the MnPSe3 nanoparticles in improving the antioxidant capacity of plants under salt stress, and the application method is: spraying the plant leaf surface with the MnPSe3 nanoparticles after the MnPSe3 nanoparticles are dissolved to a concentration of 50-150 mg / L.

[0016] Compared with the prior art, the application has the following beneficial effects:

[0017] The application shows that the nano MnPSe3 relieves the damage of salt stress to the ice vegetables and improves the sugar resistance of the ice vegetables. By applying the nano MnPSe3 to the ice vegetables under the salt stress, it is observed that the growth condition of the ice vegetables is significantly improved, specifically, the leaf area is increased, and the contents of malondialdehyde and proline are reduced. The improvement of the above physiological indexes not only relieves the adverse effects of the salt stress on the ice vegetables, but also enhances the sugar resistance of the ice vegetables by inhibiting the activity of alpha-amylase, and improves the nutritional value and medicinal value of the ice vegetables.

[0018] The preparation method of the MnPSe3 nano solution is simple, the application process is convenient, and the environment is friendly, so the MnPSe3 nano solution has wide application prospect and market potential. BRIEF DESCRIPTION OF DRAWINGS

[0019] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or the prior art description. Obviously, the drawings in the following description only illustrate the embodiments of the application, and for those skilled in the art, other drawings can also be obtained without creative labor on the basis of the provided drawings.

[0020] Figure 1 It is a transmission electron microscope image of the MnPSe3 nanoparticles, and the scale is 50 nm;

[0021] Figure 2 It is a growth schematic diagram of the ice vegetables under different treatment conditions;

[0022] Figure 3Figures of leaf area, proline content and malondialdehyde content; wherein, A is leaf area, B is proline content; C is malondialdehyde content; CK is a 0 mM NaCl treatment group, Salt is a 200 mM NaCl treatment group, Mn50 is a 200 mM NaCl stress with 50 mg / L MnPSe3 treatment group, Mn100 is a 200 mM NaCl stress with 100 mg / L MnPSe3 treatment group, Mn150 is a 200 mM NaCl stress with 150 mg / L MnPSe3 treatment group;

[0023] Figure 4 Figures of photosynthetic indicators of plant leaves before and after nano-material treatment; wherein, A is photosynthetic efficiency; B is intercellular CO2 concentration; C is transpiration rate; CK is a 0 mM NaCl treatment group, Salt is a 200 mM NaCl treatment group, Mn50 is a 200 mM NaCl stress with 50 mg / L MnPSe3 treatment group, Mn100 is a 200 mM NaCl stress with 100 mg / L MnPSe3 treatment group, Mn150 is a 200 mM NaCl stress with 150 mg / L MnPSe3 treatment group;

[0024] Figure 5 Figure of superoxide anion confocal fluorescence imaging, the scale is 10 μm;

[0025] Figure 6 Figure of hydrogen peroxide confocal fluorescence imaging, the scale is 10 μm;

[0026] Figure 7 Figure of MnPSe3 nanoparticle metabolism, the scale is 10 μm;

[0027] Figure 8 Figure of detection results of soluble sugar and soluble protein content; wherein, A is soluble sugar content; B is soluble protein content;

[0028] Figure 9 Figure of detection results of antioxidant activity of crude extract of Brassica campestris leaves under different treatments; wherein, CK is a 0 mM NaCl treatment group, Salt is a 200 mM NaCl treatment group, Mn50 is a 200 mM NaCl stress with 50 mg / L MnPSe3 treatment group, Mn100 is a 200 mM NaCl stress with 100 mg / L MnPSe3 treatment group, Mn150 is a 200 mM NaCl stress with 150 mg / L MnPSe3 treatment group;

[0029] Figure 10Figure of the inhibition of α-amylase activity by the crude extract of the leaves of I. cibotium under different treatments, wherein CK is a 0 mM NaCl treatment group, Salt is a 200 mM NaCl treatment group, Mn50 is a 200 mM NaCl stress treatment group to which 50 mg / L of MnPSe3 is added, Mn100 is a 200 mM NaCl stress treatment group to which 100 mg / L of MnPSe3 is added, and Mn150 is a 200 mM NaCl stress treatment group to which 150 mg / L of MnPSe3 is added. DETAILED DESCRIPTION

[0030] The technical solutions provided by the present application are described in detail below in conjunction with the examples, but they should not be understood as limiting the scope of protection of the present application.

[0031] In the following examples, the experimental methods are all conventional methods unless otherwise specified. In the following examples, the materials, reagents, etc. used are all commercially available unless otherwise specified.

[0032] In the following examples, the MnPSe3 (abbreviated as "NPs") nanoparticles are purchased from Mokonano (particle size 5-20 μm, purity ≥ 99.95%), the MnPSe3 crystals are dissolved in N-methyl-2-pyrrolidone solution (2 mg / mL), the ultrasonic disrupter (600 W ice bath for 24 h, 5 s on / 3 s off) is used for crushing, and then gradient centrifugation is combined, 4000 rpm is used to remove large particles that are not crushed, and 14000 rpm is used to collect small particles, so that nanoparticles with a particle size of about 30 nm are obtained.

[0033] Example 1 Application of Nanometer MnPSe3 in Improving the Salt Stress Resistance of I. cibotium

[0034] 1. Planting of I. cibotium seedlings

[0035] Uniform-sized and plump I. cibotium seeds are selected and placed in 16 cm x 16 cm flowerpots, and a mixed substrate with a volume ratio of nutrient soil to perlite of 2:1 is used for planting at opposite corners to ensure sufficient space. The I. cibotium is cultivated at 23°C and a humidity of 45%, and each group has 6 treatments. Regular watering is performed until the I. cibotium grows 3 pairs of leaves, and then salt treatment is performed.

[0036] 2. Salt solution treatment

[0037] A 200 mM NaCl solution is prepared, and the I. cibotium is irrigated once a week, and the soil is watered every two weeks to wash away excess salt to maintain an appropriate salt concentration and prevent salt accumulation.

[0038] 3. Nanometer solution treatment

[0039] Preparation of nanosolutions: Weigh 50 mg, 100 mg and 150 mg of MnPSe3 nanoparticles respectively, and treat them with 180 W ultrasonic waves for 30 min at room temperature to ensure that the nanoparticles are uniformly dispersed in ultrapure water, so as to obtain nanosolutions with concentrations of 50 mg / L, 100 mg / L and 150 mg / L respectively. Figure 1 The transmission electron microscope image of the nano-MnPSe3 shows that the nano-MnPSe3 has good dispersion, uniform particle size, and appears to be spherical with a diameter of about 30 nm.

[0040] Application of nano solution: Use a spray bottle to evenly spray the ice plant leaves twice a week, ensuring that each leaf surface is fully covered with nano solution for effective experiments.

[0041] 4. Experimental Grouping

[0042] This study investigated the effects of spraying different concentrations of nanoparticles on the growth of *Ilex crenata* under salt stress, aiming to optimize the spraying concentration and improve the salt tolerance of *Ilex crenata*. *Ilex crenata* seedlings with uniform growth were divided into 5 groups of 6 seedlings each. The seedlings were first subjected to salt stress for one week, followed by spraying with the nanoparticle solution. The specific grouping method is as follows:

[0043] Control group (CK group): Sprayed with 0mM NaCl + water;

[0044] Salt treatment group: Spray with 200mM NaCl once a week, and water with clean water every four weeks to rinse off the salt and prevent the salt from accumulating.

[0045] Salt + Nanosolution Treatment Group:

[0046] Mn50: Spray 50mg / L MnPSe3 nano solution twice a week on the basis of salt treatment;

[0047] Mn100: Spray with 100 mg / L MnPSe3 nano solution twice a week on the basis of salt treatment. Mn150: Spray with 150 mg / L MnPSe3 nano solution twice a week on the basis of salt treatment.

[0048] After 30 days of cultivation, the first and second true leaves were taken to measure various indicators.

[0049] 5. Determination of leaf area, proline content, and malondialdehyde content.

[0050] (1) Measurement of leaf area

[0051] Two weeks after treatment with nanomaterials, leaf photographs were taken when significant differences in ice plant growth were observed (see [link]). Figure 2 The leaf area was measured using ImageJ software to quantitatively analyze the effects of different treatments on the growth of ice plant.

[0052] (2) Detection of proline and malondialdehyde content

[0053] Select 0.1 g of fresh leaves at the same leaf position of the plant under different treatments and place them in a mortar. Then add 1.5 mL of 100 mM, pH = 7.8 PBS buffer (NaH2PO4 and Na2HPO4 stock solution, PBS needs to be prepared and stored at room temperature) for homogenization. The entire operation process is carried out on ice. Then centrifuge at 4°C, 10000g for 20 min. After centrifugation, transfer the supernatant to a new centrifuge tube and place it on ice. The crude extract is ready for subsequent determination of proline and malondialdehyde content. Finally, the concentration of the crude extract is determined by the formula protein concentration (mg / mL) = 1.55 x A 280 -0.76 x A 260 The results show that the concentration of the CK group is 1.249 mg / mL, the concentration of the Salt group is 1.328 mg / mL, the concentration of the Mn50 group is 1.263 mg / mL, the concentration of the Mn100 group is 1.177 mg / mL, and the concentration of the Mn150 group is 1.230 mg / mL.

[0054] Proline content determination: Prepare 100 mM PBS (pH = 7.0), 2.5% acid ninhydrin, and 3% sulfosalicylic acid. Prepare working solution (10 mL 3% sulfosalicylic acid, 10 mL acetic acid, and 20 mL 2.5% acid ninhydrin). Prepare 1.5 mL centrifuge tube, add 100 μL of the above crude extract to 1 mL of working solution, and add 100 μL of 100 mM PBS (pH = 7.8) to 1 mL of working solution for the control group. Boil the mixture in a water bath for 15 min, then cool the mixture on ice for 5 min. Take 200 μL of the mixture and measure the absorbance at 520 nm.

[0055] Malondialdehyde content determination: Prepare 10% trichloroacetic acid (TCA) and 0.25% thiobarbituric acid (TBA). The working solution is 0.25% TBA. Prepare 1.5 mL centrifuge tube, add 100 μL of crude extract of ice plant under different treatments to 1 mL of working solution. Add 100 μL of 100 mM PBS (pH = 7.8) to 1 mL of working solution for the control group. Boil the mixture in a water bath for 15 min, then cool the mixture on ice for 5 min. Take 200 μL of the mixture and measure the absorbance at 532 nm and 600 nm.

[0056] The growth results of ice plants under different treatment conditions are shown in Table 1. Figure 2 Figure 2 ​The growth of ice vegetables under five different treatment conditions is shown in the schematic diagram. It can be seen that the ice vegetable plants subjected to salt stress exhibit a clear phenotype of weak and small plants. After spraying the nanomaterial, the growth of the ice vegetable plants gradually tends to the control group, and when the spraying concentration of the nanomaterial is 100 mg / L, the growth of the ice vegetables is the best. When the spraying concentration of MnPSe3 is 150 mg / L, the leaves curl, indicating that the spraying of the nanomaterial is excessive.

[0057] The results of the determination of leaf area, proline content and malondialdehyde content are shown in Figure 3 When the ice vegetables are subjected to salt stress, the plant growth is short, and the plants undergo oxidative stress. The content of malondialdehyde as an oxidative stress marker is significantly increased, and the content of proline plays a role in maintaining water balance and is significantly increased. The leaf area of the ice vegetables is the largest under the treatment of 100 mg / L of the nanosolution compared to the maximum salt stress Figure 3 A), the proline content is the lowest under the treatment of 100 mg / L of the nanosolution Figure 3 B), and the malondialdehyde content is also the lowest under the treatment of 100 mg / L of the nanosolution Figure 3 C); the above results show that the spraying of the nanomaterial has a significant effect, and the best concentration is 100 mg / L. However, when the concentration is too high, it may have an inhibitory effect. Plants undergo oxidative stress to produce superoxide anions (O 2- ) after being subjected to salt stress.

[0058] 6. Measurement of photosynthetic indicators

[0059] The photosynthetic efficiency, transpiration rate and intercellular CO2 concentration of plant leaves were determined using a plant photosynthesis instrument. The results are shown in Figure 4 , wherein Figure 4 A is the change in photosynthetic efficiency before and after the treatment of the nanomaterial. It can be seen that the photosynthetic efficiency of the plants is significantly reduced after being subjected to salt stress, and is alleviated after spraying 50 mg / L and 150 mg / L of the nanosolution; Figure 4 B is the change in intercellular CO2 concentration before and after the treatment of the nanomaterial. It can be seen that the intercellular CO2 concentration of the plants is significantly increased after being subjected to salt stress, and is alleviated after spraying the nanosolution; Figure 4 C is the change in transpiration rate before and after the treatment of the nanomaterial. It can be seen that the transpiration rate of the plants is significantly reduced after being subjected to salt stress, and is alleviated after spraying the nanosolution.

[0060] 7. In vivo imaging of reactive oxygen species

[0061] After two weeks of ice-leaf treatment, small round pieces of 5 mm in diameter were taken from the first and second true leaves, respectively. After 3-5 small holes were made by sharp tweezers, the round pieces were incubated in 25 μM of 2',7'-Dichlorodihydrofluorescein Diacetate (H2DCFDA) and 10 μM of Dihydroethidium (DHE), respectively. After 30 min of incubation, the round pieces were washed with TES buffer for 3 times, and were placed on glass slides with the front side up and pressed.

[0062] The results are shown in Figure 5 After the plants were subjected to salt stress, oxidative stress occurred to produce superoxide anion (O 2- ). Figure 5 The figure is a superoxide anion confocal fluorescence imaging figure. DHE can stain the superoxide anion in the leaf to show green fluorescence. The chloroplast autofluorescence is red fluorescence. The combination of the two can locate and quantify the content of superoxide anion. As can be seen from the figure, the fluorescence content of the 200 mM NaCl treatment group is significantly greater than that of the watering control group, and the fluorescence content detected in the ice-leaf treated with 100 mg / L nano solution is less than that of the 200 mM NaCl treatment group, indicating that under salt stress, spraying 100 mg / L MnPSe3 nano solution reduces the content of O 2- in the leaf.

[0063] The results are shown in Figure 6 After the plants were subjected to salt stress, oxidative stress occurred to produce hydrogen peroxide (H2O2). Figure 6 The figure is a hydrogen peroxide confocal fluorescence imaging figure. H2DCFDA can stain the hydrogen peroxide in the leaf to show green fluorescence. The chloroplast autofluorescence is red fluorescence. The combination of the two can locate and quantify the content of H2O2. As can be seen from the figure, the fluorescence content of the 200 mM NaCl treatment group is significantly greater than that of the watering control group, and the fluorescence content detected in the ice-leaf treated with 100 mg / L nano solution is less than that of the 200 mM NaCl treatment group, indicating that under salt stress, spraying 100 mg / L MnPSe3 nano solution reduces the content of H2O2 in the leaf.

[0064] 8. Metabolism of nanoparticles

[0065] MnPSe3 nano material was incubated with Dil dye to form Dil-MnPSe3. After the leaf was applied with Dil-MnPSe3, small round pieces of 5 mm in diameter were taken at 0 h, 3 h, 6 h, 9 h, 12 h, and 24 h, respectively. The round pieces were placed on glass slides with the front side up and pressed.

[0066] The samples were imaged using a Leica SP8 laser confocal scanning microscope. Dil dye can make nano MnPSe3 show green fluorescence; chloroplast autofluorescence shows red; the combination of the two can show that as the treatment time of the material is prolonged, nano MnPSe3 is metabolized. The results are shown in Figure 7 As the time after spraying MnPSe3 on the leaves is prolonged, the fluorescence content gradually decreases, which indicates that the nanoparticles may have been degraded, diffused or absorbed by plant cells on the leaf surface, thereby causing the fluorescence signal to weaken, suggesting that the residual amount of MnPSe3 in the ice vegetables decreases as the time is prolonged, and therefore does not cause pollution of the ice vegetables by MnPSe3 nanomaterials.

[0067] 9. Detection of soluble sugar and soluble protein content

[0068] The detection of the soluble sugar content of the sample used a soluble sugar content kit (Suzhou Gaisi Biological, soluble sugar content (SS) kit (G0501F). Under the action of concentrated sulfuric acid, sugars are dehydrated to generate furfural or hydroxymethyl glyoxal, which reacts with anthrone to form a blue-green derivative, which has a maximum absorption at 620 nm, and the light absorption value is proportional to the sugar content. The detection of the soluble protein content of the sample used a BCA method protein content determination kit (Suzhou Gaisi Biological, protein content (SP) kit (G0418W). The BCA method is to use the peptide bond in the protein to reduce copper ions to cuprous ions under alkaline conditions, and then form a purple-blue complex, which has a maximum absorption peak at 562 nm, and the color depth is proportional to the protein concentration, so as to determine the protein content by colorimetry.

[0069] Plants will increase the content of soluble sugars under salt stress, which is used to maintain osmotic balance, protect cells from oxidative damage, provide energy, and regulate signal transduction; plants will increase the content of soluble proteins under salt stress, which is used to regulate osmotic balance, protect enzyme activity, signal transduction, protein synthesis and metabolic regulation, and maintain the stability of cell structure. These mechanisms can help plants survive and grow in salt stress environments. From Figure 8 As can be seen from the figure, salt stress will increase the content of soluble sugars and soluble proteins, and the use of nano solution can restore it to normal level. It can be seen that after spraying nano MnPSe3 solution, the content of soluble sugars and soluble proteins has no obvious difference from the control group, further confirming that nano MnPSe3 can alleviate the salt stress of ice vegetables.

[0070] Example 2 Verification of the effect of spraying nano MnPSe3 on the hypoglycemic and anti-aging functions of ice vegetables under salt stress

[0071] 1. Preparation of ice vegetable crude extract

[0072] The dried and ground ice plant of different treatments in Example 1 was taken 10 g, and added with ethyl acetate or 70% ethanol (w:v = 1:20) to extract for 2 h at room temperature. After standing and filtration, the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain the crude extract. The crude extract was dissolved in a methanol and water solution (9:1 by volume) by ultrasonic, and extracted with petroleum ether to remove the lipid components. The crude extract was continuously concentrated under reduced pressure and stored in a 4°C refrigerator for subsequent experiments.

[0073] 2. Detection of antioxidant activity

[0074] The antioxidant activity of the sample was detected by 1,1-diphenyl-2-trinitrobenzene hydrazine (DPPH) method. 3.5 mg of DPPH was dissolved in anhydrous ethanol and transferred into a 10 mL volumetric flask to obtain a working solution which was stored at 4°C in the dark for standby. The ice plant sample was dissolved in methanol to obtain a concentration of 10 mg / mL. A 96-well plate was used, 200 μL of the above working solution + 50 μL of the crude extract was added to each well, and the absorbance was measured at 517 nm by an enzyme marker for 30 min, with a detection interval of 1 min. The control group used 200 μL of working solution + 50 μL of methanol.

[0075] The results are shown in Figure 9 Figure 9 The results of the DPPH free radical scavenging rate experiment of the crude extract of ice plant leaves are shown. In this embodiment 2, ethyl acetate was used as the extraction reagent to compare the antioxidant activity of the crude extract of ice plant leaves under different treatments. As can be seen from the figure, the ethyl acetate extraction method has a significant advantage in improving the DPPH free radical scavenging rate. The crude extract of ice plant leaves obtained by ethyl acetate extraction can more effectively extract the antioxidant active ingredients therein, and significantly enhance the DPPH free radical scavenging capacity.

[0076] 3. Detection of α-amylase inhibitory activity

[0077] In a 96-well plate, 20 μL of the sample to be tested (DMSO as the solvent), 80 μL of sodium phosphate buffer (pH 6.0) and 0.125 U / mL a-amylase solution were added and mixed. After shaking for 2 min, 20 μL of 10 mM CNP-G3 solution was added after incubation at 37°C for 10 min, and the absorbance of the product was measured at 405 nm. The reaction time was 30 min, the determination was 11 times, and 10% DMSO was used as a blank control.

[0078] The results are shown in Figure 10 Figure 10 ​​The results of the experiment of the inhibition rate of the crude extract of the leaves of the ice plant on α-amylase are presented. In this embodiment 2, 70% ethanol is used as the extraction reagent, and the inhibition of the α-amylase activity by the extracts of the leaves of the ice plant under different treatments is compared. As can be seen from the figure, the 70% ethanol extraction method has a significant advantage in improving the inhibition rate of α-amylase. It is shown that 70% ethanol as the extraction reagent can more effectively extract the components in the leaves of the ice plant with anti-amylase activity, thereby significantly enhancing the inhibition of α-amylase.

[0079] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. Use of MnPSe3 nanoparticles in alleviating ice lettuce salt stress, characterized in that, The concentration of the MnPSe3 nanoparticles is 50-150 mg / L.

2. Use according to claim 1, characterized in that, The application method is: The MnPSe3 nanoparticles are mixed with water to form a nano-solution, and the nano-solution is sprayed on the surface of the plant leaves.

3. Use according to claim 2, characterized in that, The interval time of spraying is 4-7 days, and the spraying frequency is 1-2 times.

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