Method for improving saline alkali soil through apple enzyme liquid and application

Abstract

The invention discloses a method for improving saline-alkali soil through apple enzyme liquid and application, belongs to the technical field of soil improvement and agricultural planting, and aims to solve the problem that existing saline-alkali soil improvement technologies are insufficient. The method comprises the steps that fallen apples serve as raw materials, fermentation liquid is prepared according to the mass ratio of brown sugar to apples to deionized water being 1: 3: 10, and the fermentation liquid is fermented for 90-100 d at the room temperature of 25 + / -2 DEG C in a dark place; the method comprises the following steps: adding a proper amount of enzyme liquid into a culture medium, uniformly mixing the enzyme liquid with soil until the pH value is stabilized at 3.17-3.24 and the electric conductivity fluctuation is less than 5% in a continuous week, filtering, sealing and storing at 4 DEG C, and applying the enzyme liquid: applying 200mL of the enzyme liquid with a specific concentration in a basal application and topdressing manner when rice is transplanted, then topdressing 100mL of the enzyme liquid every 7 days, and topdressing for 10 times in total, so that the improvement effect is remarkable; the plant height, stem length and root length of the rice diluted by 1: 25 are respectively increased by 70.7%, 111.4% and 72.9% compared with those of a control group; the dry matter accumulation amount is increased by 115.1%, salt and alkali stress is effectively relieved, and the method is green and sustainable, dropped apples serve as raw materials, agricultural waste resource utilization is achieved, the preparation process is simple, the cost is low, secondary pollution is avoided, and the requirement for green agriculture development is met.

Description

Technical Field

[0001] This invention relates to the field of soil improvement and agricultural planting technology, specifically to a method and application of apple enzyme liquid for improving saline-alkali soil. Background Technology

[0002] Saline-alkali soil is a significant obstacle to agricultural production. Globally, there are approximately 950 million hectares of saline-alkali land, with my country alone having 36 million hectares, and this area is expanding annually. Saline-alkali soils are characterized by high pH, ​​high salinity, nutrient deficiency, and poor structure. They inhibit crop growth through osmotic stress, ion toxicity, and oxidative damage, leading to reduced yields or even crop failure.

[0003] Traditional methods for improving saline-alkali soils include engineering measures, chemical amendments, and biological amendments. However, these methods generally suffer from high costs, long cycles, and unstable effects. Chemical amendments can also cause secondary soil pollution. In recent years, biological amendment technologies have gained attention due to their green, environmentally friendly, and sustainable advantages. Fruit and vegetable enzyme fermentation liquid, as a novel biological amendment, is rich in organic acids, amino acids, beneficial microorganisms, and other bioactive substances, and has been proven to regulate soil pH and improve soil physicochemical properties. However, current research on the microbiological mechanisms of fruit and vegetable enzyme liquid in improving saline-alkali soils and its comprehensive impact on crop growth is lacking, limiting the widespread application of this technology. There is an urgent need to clarify its mechanism of action and appropriate application parameters.

[0004] To address the above problems, a method and application of apple enzyme solution for improving saline-alkali soil are proposed. Summary of the Invention

[0005] The purpose of this invention is to provide a method and application for improving saline-alkali soil with apple enzyme liquid. By using this device, the shortcomings of existing saline-alkali soil improvement technologies mentioned above are solved.

[0006] To achieve the above objectives, the present invention provides the following technical solution: A method for improving saline-alkali soil with apple enzyme liquid includes the following steps: S1: Preparation of apple enzyme liquid: Using fallen apples as raw materials, prepare fermentation liquid according to the mass ratio of brown sugar: apple: deionized water 1:3:10. Ferment at room temperature 25±2℃ in the dark for 90-100 days until the pH stabilizes at 3.17-3.24 and the conductivity fluctuates by less than 5% for one week. After filtration, seal and store at 4℃. S2: Enzyme solution application: Use the "basal application and top dressing" method. When transplanting rice, apply 200mL of a specific concentration of enzyme solution as a base and mix it with the soil. Then, apply 100mL every 7 days for a total of 10 top dressings. S3: Concentration control: The application concentration of the apple enzyme solution is 1:25, 1:50, 1:100 or 1:200 dilution ratio.

[0007] Furthermore, in step 1, select disease-free and pest-free Hanfu apples, chop them into pieces with a diameter of 1-2cm, mix them with brown sugar and let them stand for 2 hours, then add deionized water. The amount of material should not exceed 80% of the container volume. Seal the container opening with 4 layers of sterile gauze and stir once a day.

[0008] Furthermore, the saline-alkali soil is a soda-type saline-alkali soil with a pH of 9.5-10.5, and the soil tillage layer depth is 0-20cm. Before application, plant residues and stones are removed, and the soil is naturally air-dried and then sieved through a 2mm sieve.

[0009] Furthermore, the rice variety used in the experiment was a japonica rice variety. The seedling transplanting method was adopted. After the seedlings were raised for 40 days until they reached the three-leaf and one-heart stage, they were transplanted. Five seedlings were transplanted into each pot, arranged in a plum blossom shape, with a spacing of about 8 cm between the seedlings.

[0010] Furthermore, during the rice growing season, flooding management is adopted, maintaining a water depth of 2-3 cm, controlling the growing environment temperature at 25-30℃, providing natural light, and rotating the pots once every 3 days to eliminate the position effect.

[0011] Furthermore, the bioactive substances include organic acids, amino acids, vitamins, active enzymes, and plant growth regulators, which can regulate soil pH and activate the expression of rice stress-resistance genes.

[0012] Another technical solution proposed in this invention is the application of the method of improving saline-alkali soil with apple enzyme liquid in the cultivation of rice in saline-alkali soil. The improvement of saline-alkali soil and the promotion of rice growth are achieved through multiple synergistic pathways, including improving soil physicochemical properties, reshaping the root microbial community, and activating the expression of plant stress resistance genes.

[0013] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. Significant improvement effect: The rice plant height, stem length and root length of the 1:25 dilution treatment increased by 70.7%, 111.4% and 72.9% respectively compared with the control, and the dry matter accumulation increased by 115.1%, effectively alleviating salt and alkali stress.

[0014] 2. Green and sustainable: Using fallen apples as raw materials, it realizes the resource utilization of agricultural waste. The preparation process is simple, low-cost, and has no secondary pollution, which meets the requirements of green agricultural development.

[0015] 3. Clear mechanism of action: The improvement mechanism is elucidated from multiple dimensions including physiology, microbiology and molecularity, providing theoretical support for the improvement of saline-alkali land. The clear appropriate concentration parameters (1:25-1:50) facilitate field promotion and application.

[0016] 4. Outstanding comprehensive benefits: It simultaneously improves soil physical and chemical properties, optimizes microbial communities, and enhances rice's stress resistance, providing new ideas for the efficient development and utilization of saline-alkali land and the guarantee of food security. Attached Figure Description

[0017] Figure 1 This is a flowchart illustrating the process of using apple enzyme solution to improve saline-alkali soil according to the present invention. Figure 2 This is a comparative diagram showing the effect of the apple enzyme solution of the present invention on the morphology of rice plants. Figure 3 This is a comparison diagram showing the effect of the apple enzyme solution of the present invention on the fresh weight of rice plants. Figure 4 This is a comparative diagram showing the effect of the apple enzyme solution of the present invention on the dry weight of rice plants. Detailed Implementation

[0018] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] like Figures 1-4 As shown, the following technical solutions are provided: 1. Preparation of Apple Enzyme Solution (1) Raw material selection: Select healthy fallen apples of good quality and free from pests and diseases (mainly Hanfu apples) as the core raw material for fermentation.

[0020] (2) Ingredient ratio: Weigh out brown sugar, apples and deionized water by mass ratio of 1:3:10.

[0021] (3) Pre-treatment: After washing the apples, remove the cores, chop them into pieces with a diameter of 1-2 cm, mix them thoroughly with brown sugar, and let them stand for 2 hours to allow the brown sugar to completely dissolve and fully contact the apple flesh.

[0022] (4) Fermentation process: The mixed materials and deionized water are put into a 5L food-grade polyethylene plastic bucket. The amount of material should not exceed 80% of the container volume. The container opening is sealed with 4 layers of sterile gauze and placed in a room temperature (25±2℃) environment to ferment in the dark. Stir once a day. The fermentation cycle is 90-100 days.

[0023] (5) Finished product judgment and processing: When the pH value of the fermentation liquid is stable at 3.17-3.24 and the conductivity fluctuates less than 5% for a week, the fermentation is judged to be complete; the supernatant is filtered through 4 layers of gauze, and then dispensed into brown glass bottles and sealed at 4°C for later use. Concentration control: the application concentration of the apple enzyme liquid is 1:25, 1:50, 1:100 or 1:200 dilution ratio, preferably 1:25-1:50 dilution concentration.

[0024] 2. Soil sample collection and pretreatment (1) Soil collection: Soil from the top 0-20cm layer was collected near Ailianbao Garden, Longfeng District, Daqing City, Heilongjiang Province (geographical coordinates 46°36'N, 125°08'E). The soil was a typical soda-type saline-alkali soil with a pH value of 10.08±0.02.

[0025] (2) Control soil collection: Ordinary planting soil in the top 0-20cm layer was collected from Boyi Ecological Park, Lindian County, Daqing City, Heilongjiang Province (geographical coordinates 47°11'N, 124°52'E), with a pH value of 7.88±0.02.

[0026] (3) Soil pretreatment: Remove plant residues and stones from the two types of soil, air dry them naturally, grind them, pass them through a 2mm sieve, and put them into self-sealing bags for storage.

[0027] 3. Rice varieties tested and seedling cultivation (1) Variety selection: The conventional japonica rice variety "Qijing No. 7" was selected as the test crop.

[0028] (2) Seed treatment: Disinfect rice seeds with 75% alcohol for 5 minutes, rinse with sterile water 3 times, and place them in an incubator to germinate for later use.

[0029] (3) Seedling raising operation: Raise seedlings in 32.5cm×24.5cm×4.5cm seedling trays and cultivate them for 40 days until the rice seedlings reach the three-leaf and one-heart stage, ready for transplanting.

[0030] 4. Experimental Group Setup (1) Treatment group design: A completely randomized design was adopted, with 5 treatment groups set up as follows: CK group: Deionized water was applied. JS200 group: Apple enzyme solution diluted at 1:200 (v / v); JS100 group: Apple enzyme solution diluted at 1:100 (v / v); JS50 group: Apple enzyme solution diluted at a ratio of 1:50 (v / v); JS25 group: Apple enzyme solution diluted at 1:25 (v / v).

[0031] (2) Replication setting: Each treatment group was set up with 3 replicates, for a total of 15 pots; the pots used for the experiment were plastic pots, and each pot was filled with 5 kg of pretreated saline-alkali soil.

[0032] 5. Rice transplanting and application of enzyme solution (1) Transplanting operation: Select healthy seedlings with uniform growth from the seedling tray, plant 5 seedlings in each tray, arranged in a plum blossom shape, with a spacing of about 8cm between seedlings; after transplanting, use flooding management to maintain a water depth of 2-3cm.

[0033] (2) Enzyme solution application method: The combination of "basal application + top dressing" was adopted. When transplanting, 200 mL of apple enzyme solution of the corresponding concentration was applied as a base and mixed thoroughly with the soil. After that, 100 mL was applied every 7 days, and a total of 10 top dressings were applied throughout the entire growth period. The CK group was given the same amount of deionized water, and the application time was the same as that of the treatment group.

[0034] (3) Environmental control: The experiment was conducted in a greenhouse with the temperature controlled at 25-30℃ and natural light. To reduce the position effect, the potted plants were randomly moved once every 3 days.

[0035] (4) Transcriptome analysis: JS100 treatment activated 3713 differentially expressed genes, and the 272 core genes shared by the three comparison groups constituted a key molecular network for resistance to salt and alkali stress.

[0036] 6. Sampling Time and Method (1) Sampling time: Samples were collected on the 70th day after rice transplanting (maturity period).

[0037] (2) Plant sample collection: Three plants were randomly selected from each treatment group for growth index determination. The plants were gently pulled up to separate the aboveground parts from the roots. The roots were rinsed clean with sterile deionized water and the surface moisture was absorbed with filter paper for later use.

[0038] (3) Rhizosphere soil collection: The soil tightly attached to the root surface was collected by shaking method, with about 50g collected from each pot; after removing visible root debris, it was divided into two parts. One part was air-dried and passed through a 2mm sieve for soil physicochemical property analysis; the other part was immediately packed into sterile cryovials, flash-frozen in liquid nitrogen and stored at -80℃ for microbial community analysis.

[0039] (4) Transcriptome sample collection: Select about 1g of tender root segments 0-3cm from the root tip from the root system, immediately place them in liquid nitrogen for quick freezing, and then transfer them to -80℃ for storage for transcriptome analysis; all sample collection processes are performed on ice.

[0040] 7. Measurement Indicators and Methods (1) Determination of physicochemical properties of apple enzyme solution: pH value was measured using a Leici PHS-3C pH meter; conductivity was measured using a Leici DDS-307 conductivity meter; total acid content was determined by sodium hydroxide titration (calculated as citric acid); organic matter content was determined by potassium dichromate oxidation method; all indicators were measured three times.

[0041] (2) Determination of soil physicochemical properties: Soil pH was determined by potentiometric method (soil-water ratio 1:2.5); organic matter content was determined by potassium dichromate titration method; total nitrogen content was determined by Kjeldahl method; total phosphorus content was determined by molybdenum-antimony spectrophotometric method; total potassium content was determined by sodium hydroxide fusion method; available nitrogen was determined by alkaline hydrolysis diffusion method; extraction-molybdenum-antimony colorimetric method; available potassium was determined by 1 mol / L NH4OAc extraction and flame photometry.

[0042] (3) Morphological indicators: Plant height was measured using a ruler, from the base of the root to the tip of the highest leaf; stem diameter was measured using a vernier caliper, from the diameter of the first internode at the base; root length was measured, from the maximum length of the main root. Biomass indicators: Fresh weight of the aboveground and underground parts of the plant was measured separately, and then the samples were blanched in an oven at 105℃ for 15 min, dried at 80℃ to constant weight, and the dry weight was measured.

[0043] (4) Determination of physiological and biochemical indicators of rice: chlorophyll content was determined by acetone extraction method; proline (Pro) content was determined by acidic ninhydrin method; superoxide dismutase (SOD) activity was determined by nitroblue tetrazolium (NBT) photochemical reduction method; catalase (CAT) activity was determined by ultraviolet spectrophotometry; malondialdehyde (MDA) content was determined by thiobarbituric acid method.

[0044] (5) Soil microbial community analysis: Total DNA of soil microorganisms was extracted using the FastDNA kit; using DNA as a template, the fungal ITS1 region was amplified by PCR using primers ITS1F and ITS2; the PCR products were excised, purified, and used to construct sequencing libraries, and paired-end sequencing was performed using the platform; the sequencing data were quality controlled, denoised, spliced, and dechimeric processed by QIIME2 software, and OTU clustering was performed at a similarity threshold of 97%, and species classification and annotation were performed by comparing with the UNITE database. The α diversity index and β diversity were calculated, and differential species analysis was performed using the LEfSe method.

[0045] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0046] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for improving saline-alkali soil with apple enzyme liquid, characterized in that, Includes the following steps: S1: Preparation of apple enzyme liquid: Using fallen apples as raw materials, prepare fermentation liquid according to the mass ratio of brown sugar, apples and deionized water of 1:3:

10. Ferment at room temperature (25±2℃) in the dark for 90-100 days until the pH stabilizes at 3.17-3.24 and the conductivity fluctuates by less than 5% for one week. After filtration, store in a sealed container at 4℃. S2: Enzyme solution application: Use the "basal application and top dressing" method. When transplanting rice, apply 200mL of a specific concentration of enzyme solution as a base and mix it with the soil. Then, apply 100mL every 7 days for a total of 10 top dressings. S3: Concentration control: The application concentration of the apple enzyme solution is 1:25, 1:50, 1:100 or 1:200 dilution ratio.

2. The method for improving saline-alkali soil with apple enzyme solution according to claim 1, characterized in that: In step 1, select disease-free and pest-free Hanfu apples, chop them into pieces with a diameter of 1-2cm, mix them with brown sugar and let them stand for 2 hours, then add deionized water. The amount of material should not exceed 80% of the container volume. Seal the container opening with 4 layers of sterile gauze and stir once a day.

3. The method for improving saline-alkali soil with apple enzyme liquid according to claim 2, characterized in that: The saline-alkali soil is a soda-type saline-alkali soil with a pH of 9.5-10.

5. The soil tillage layer depth is 0-20cm. Before application, remove plant residues and stones, and after natural air drying, pass through a 2mm sieve.

4. The method for improving saline-alkali soil with apple enzyme liquid according to claim 3, characterized in that: The rice variety used in the experiment was a japonica rice variety. The seedling transplanting method was adopted. After the seedlings were raised for 40 days until they reached the three-leaf and one-heart stage, they were transplanted. Five seedlings were planted in each pot, arranged in a plum blossom shape, with a spacing of about 8 cm between the seedlings.

5. The method for improving saline-alkali soil with apple enzyme solution according to claim 4, characterized in that: During the rice growing season, flooding management is adopted, maintaining a water depth of 2-3cm, controlling the growing environment temperature at 25-30℃, providing natural light, and rotating the pots once every 3 days to eliminate the position effect.

6. The method for improving saline-alkali soil with apple enzyme solution according to claim 5, characterized in that: The bioactive substances include organic acids, amino acids, vitamins, active enzymes, and plant growth regulators, which can regulate soil pH and activate the expression of rice stress-resistance genes.

7. The application of the method for improving saline-alkali soil with apple enzyme liquid according to any one of claims 1-6 in the cultivation of rice in saline-alkali soil, characterized in that: The improvement of saline-alkali soil and promotion of rice growth are achieved through a multi-pronged approach that improves soil physicochemical properties, reshapes the rhizosphere microbial community, and activates the expression of plant stress-resistance genes.

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