Culture medium for enhancing nutrition absorption of potato tissue culture seedlings, preparation method and application thereof

Abstract

The present application relates to the technical field of plant tissue culture, and discloses a culture medium for enhancing the nutrient absorption of potato tissue culture seedlings, a preparation method and application thereof, which is prepared from graphene oxide, an anionic high-molecular surfactant, trace element components, macroelement components, organic auxiliary material components, sucrose, plant tissue culture grade agar powder and deionized water; the anionic high-molecular surfactant adsorbs the graphene oxide to form a physical steric hindrance layer, separates the trace element components from the macroelement components, and blocks the precipitation reaction; the preparation method of the culture medium for enhancing the nutrient absorption of potato tissue culture seedlings adopts a stepwise feeding process; and the root system of the potato tissue culture seedlings secretes organic acid to trigger protonation competition substitution, and promote the dissociation of trace metal elements. The present application can improve the effective state retention rate of trace metal elements, prevent heavy metal toxicity and nutrient deficiency of potato tissue culture seedlings, and promote the root system development of the potato tissue culture seedlings.

Description

Technical Field

[0001] This invention relates to the field of plant tissue culture technology, specifically to a culture medium for enhancing nutrient absorption in potato tissue culture seedlings, its preparation method, and its application. Background Technology

[0002] Potatoes are an important crop. Plant tissue culture is a common method for achieving virus-free propagation and rapid seedling propagation of potatoes. In plant tissue culture, the plant tissue culture medium provides all the nutrients for the growth of potato tissue culture seedlings.

[0003] Existing plant tissue culture media generally contain macro-element components, micro-element components, and organic excipient components. Macro-element components provide abundant anions such as phosphate and sulfate, while micro-element components provide essential metal cations such as zinc ions. During the preparation of existing plant tissue culture media and the mandatory autoclaving process, the free micro-metal cations readily react with high concentrations of anions, forming water-insoluble precipitates. The formation of these insoluble precipitates reduces the effective concentration of micro-metal elements within the existing plant tissue culture media, failing to meet the normal growth requirements of potato tissue culture seedlings.

[0004] Existing plant tissue culture media are static nutrient supply systems. The concentration of free metal cations is relatively high in the early stages of potato explant inoculation, posing a risk of inhibiting cell division and causing heavy metal toxicity. As the culture process progresses and precipitation reactions continue, micronutrient deficiency gradually becomes apparent in the later stages. This deficiency limits the accumulation and downward transport of photosynthetic substances in potato tissue culture seedlings, ultimately resulting in slow root development and low overall biomass. Therefore, this invention proposes a culture medium, its preparation method, and its application to enhance nutrient absorption in potato tissue culture seedlings, addressing the shortcomings of existing technologies. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a culture medium, its preparation method, and its application for enhancing nutrient absorption in potato tissue culture seedlings. This solves the problem that in existing plant tissue culture media, during preparation and high-pressure steam sterilization, trace metal elements readily react with anions in macro-inorganic salts to produce insoluble precipitates, leading to a decrease in the effective concentration of trace elements. This, in turn, limits the accumulation of photosynthetic substances in potato tissue culture seedlings, resulting in delayed growth and development.

[0006] To achieve the above objectives, the present invention provides the following technical solution: Firstly, the present invention provides a culture medium that enhances nutrient absorption in potato tissue culture seedlings, employing the following technical solution: A culture medium that enhances nutrient absorption in potato tissue culture seedlings is made from raw materials comprising the following parts by weight: 0.1 to 0.3 parts by weight of graphene oxide and 1.0 to 2.0 parts by weight of anionic polymeric surfactant; Trace element composition: ferrous sulfate heptahydrate 25.0–30.0 parts by weight, disodium ethylenediaminetetraacetate dihydrate 35.0–40.0 parts by weight, manganese sulfate tetrahydrate 20.0–25.0 parts by weight, zinc sulfate heptahydrate 7.0–10.0 parts by weight, boric acid 5.0–7.0 parts by weight, potassium iodide 0.7–1.0 parts by weight, sodium molybdate dihydrate 0.2–0.3 parts by weight, copper sulfate pentahydrate 0.02–0.03 parts by weight, cobalt chloride hexahydrate 0.02–0.03 parts by weight; Macro-element components: ammonium nitrate 1550-1750 parts by weight, potassium nitrate 1800-2000 parts by weight, calcium chloride dihydrate 400-480 parts by weight, magnesium sulfate heptahydrate 350-390 parts by weight, potassium dihydrogen phosphate 150-190 parts by weight. Organic excipients: 90-110 parts by weight of inositol, 1.8-2.2 parts by weight of glycine, 0.09-0.15 parts by weight of thiamine hydrochloride, 0.4-0.6 parts by weight of pyridoxine hydrochloride, 0.4-0.6 parts by weight of nicotinic acid; 28,000-32,000 parts by weight of sucrose; 6500-7500 parts by weight of plant tissue culture grade agar powder; Including the remaining deionized water, the total volume of the mixed system is brought to a final volume of 900,000 to 1,100,000 parts by volume.

[0007] By adopting the above technical solution, the release of trace elements is controlled by using anionic polymeric surfactants to encapsulate graphene oxide and utilizing a non-covalent assembly mechanism, thereby improving the retention rate of effective trace metal elements and promoting root development of potato tissue culture seedlings.

[0008] The specific mechanism is as follows: First, the polymeric framework structure of the anionic polymeric surfactant is adsorbed onto the surface of graphene oxide through intermolecular forces, forming a physical steric hindrance layer. Second, the anionic functional groups contained in the anionic polymeric surfactant undergo dynamic coordination bonding with free zinc ions in the trace element components at the solid-liquid interface. Subsequently, when macro-element components are introduced and high-pressure steam sterilization heating is performed, the physical steric hindrance layer physically isolates the coordinated zinc ions from phosphate and sulfate ions in the external aqueous phase, blocking the reaction pathway of inorganic salt crystallization and precipitation. Finally, after inoculation with potato tissue culture seedlings, the organic small molecule acids secreted by the roots of the potato tissue culture seedlings reduce the local pH, and free hydrogen ions in the environment undergo protonation competitive substitution, promoting the dissociation of zinc ions from the anionic functional groups and their continuous diffusion into the aqueous medium for absorption by the potato tissue culture seedlings.

[0009] Preferably, the anionic polymeric surfactant is sodium lignosulfonate.

[0010] By adopting the above technical solution, sodium lignosulfonate comprises a rigid phenylpropane framework and sulfonic acid groups. The rigid phenylpropane framework of sodium lignosulfonate is anchored to the two-dimensional carbon surface of graphene oxide through physical conjugation, providing strong physical steric hindrance. The sulfonic acid groups of sodium lignosulfonate preferentially coordinate with trace element components at the solid-liquid interface, avoiding the co-precipitation loss of trace metal elements from a chemical reaction kinetics perspective.

[0011] Preferably, anionic polymeric surfactants and graphene oxide are assembled into a mesoscopic micro-reservoir structure through non-covalent assembly, and the hydrodynamic radius of the mesoscopic micro-reservoir structure is distributed between 340 nm and 410 nm.

[0012] By adopting the above technical solution, the mesoscopic micro-reservoir structure with a hydrodynamic radius distribution between 340nm and 410nm ensures that the active components in the culture medium that enhances the nutrient absorption of potato tissue culture seedlings are in a highly uniform suspension state, preventing irreversible layering and precipitation of graphene oxide.

[0013] Preferably, the culture medium for enhancing nutrient absorption of potato tissue culture seedlings before the addition of plant tissue culture-grade agar powder has a pH of 5.6–6.0 and a Zeta potential maintained between -26.5 mV and -22.3 mV.

[0014] By adopting the above technical solution, the Zeta potential value within the specified range provides sufficient electrostatic repulsion to resist the compression effect of cations in high-concentration macro-element components on the double electric layer of colloidal particles, thus maintaining the colloidal thermodynamic stability of the mixed system.

[0015] Secondly, the present invention provides a method for preparing a culture medium that enhances nutrient absorption in potato tissue culture seedlings, using the following technical solution: A method for preparing a culture medium that enhances nutrient absorption in potato tissue culture seedlings includes the following steps: An anionic polymeric surfactant was dissolved in deionized water, and an aqueous dispersion containing graphene oxide was slowly added dropwise. The mixture was then stirred under heating and constant temperature conditions to obtain an intermediate composite precursor solution. Lower the temperature of the intermediate composite precursor solution, add trace element components to form the first mixture, adjust the pH value of the first mixture with hydrochloric acid solution, and stir under constant temperature conditions. In a separate container, macro-element components, organic auxiliary components, and sucrose are dissolved in deionized water to form a transparent homogeneous solution; The transparent homogeneous solution was mixed with the first mixture to form the final mixture system. Room temperature deionized water was added, and the pH value of the final mixture system was adjusted using sodium hydroxide solution. Plant tissue culture grade agar powder was added to the final mixture at a uniform rate, and high-shear stirring was maintained to obtain a liquid suspension culture medium. The liquid suspension culture medium was then sterilized by autoclaving and cooled to solidify, thus obtaining a culture medium that enhances nutrient absorption of potato tissue culture seedlings.

[0016] By employing the above technical solution, a step-by-step feeding process is used. Anionic polymeric surfactants and graphene oxide are initially mixed in a pure water environment, ensuring sufficient time for the anionic polymeric surfactants to fully coat the graphene oxide. Subsequently, trace element components are added separately, allowing the metal trace elements to preferentially enter the coordination sites provided by the anionic polymeric surfactants. Finally, a transparent homogeneous solution containing a high concentration of inorganic salts is added. This step-by-step feeding process effectively blocks the colloidal instability pathways caused by inorganic salts, ensuring efficient anti-precipitation retention of trace element components within the culture medium that enhances nutrient absorption in potato tissue culture seedlings.

[0017] Preferably, the heating temperature is controlled at 65℃~75℃, the stirring speed is 300rpm~500rpm, and the stirring time is 45min~75min.

[0018] By adopting the above technical solution, the specific heating and constant temperature stirring steps promote the high expansion of the polymer chain segments of the anionic polymeric surfactant in a low concentration state, and the hydrophobic skeleton is stably adsorbed on the surface of graphene oxide, thus constructing a thick polymeric adsorption layer.

[0019] Preferably, the temperature of the intermediate composite precursor solution is lowered to 35℃~45℃, the concentration of the hydrochloric acid solution is 0.1mol / L~0.5mol / L, the pH value of the first mixture is adjusted to 4.3~4.8 using hydrochloric acid solution, the stirring temperature is controlled at 35℃~45℃ under constant temperature conditions, and the stirring time under constant temperature conditions is 25min~40min.

[0020] By adopting the above technical solution, a slightly acidic environment combined with medium-temperature constant-temperature stirring promotes the full coordination reaction between metal ions and the functional groups inside the anionic polymer surfactant, preventing the metal ions from undergoing premature hydroxide precipitation reaction in an alkaline environment.

[0021] Preferably, the concentration of the sodium hydroxide solution is 0.8 mol / L to 1.2 mol / L, and the pH of the final mixed system is adjusted to 5.6 to 6.0 using the sodium hydroxide solution.

[0022] By adopting the above technical solution, a standard and suitable pH environment is provided for the growth of potato stem explants.

[0023] Preferably, the working pressure of high-pressure steam sterilization is 0.10MPa~0.11MPa, the temperature of high-pressure steam sterilization is 115℃~125℃, and the time of high-pressure steam sterilization is 15min~20min.

[0024] By adopting the above technical solution, constant temperature and high pressure steam can effectively kill the microorganisms inside the culture medium that enhances the nutrient absorption of potato tissue culture seedlings. At the same time, it can cause plant tissue culture grade agar powder to melt at high temperature and solidify into a uniform solid gel that supports the growth of explants after natural cooling.

[0025] Thirdly, the present invention provides an application of a culture medium that enhances nutrient absorption in potato tissue culture seedlings, employing the following technical solution: Application of a culture medium that enhances nutrient absorption in potato tissue culture seedlings, comprising the following steps: Potato stem segments with a length of 0.8cm to 1.2cm and containing axillary buds were cut as explants; The morphological lower end of a potato stem segment is vertically inserted into a culture medium that enhances nutrient absorption in potato tissue culture seedlings. The culture medium for enhancing nutrient absorption of potato tissue culture seedlings inoculated with potato stem segments was transferred into an environmentally controlled culture room and cultured continuously for 25–35 days. The temperature of the environmentally controlled culture room was set at 18℃–22℃, the relative humidity was set at 30%–40%, and LED plant grow lights were turned on to maintain a light flux of 1000 lm–1400 lm. The photocycle was set to alternate between a light duration of 16h–20h and a dark duration of 4h–8h.

[0026] By employing the above technical solution, the slow-release efficiency of the culture medium that enhances nutrient absorption in potato tissue culture seedlings is matched with controlled physical environmental conditions. Specific ranges of light flux and photoperiod parameters stimulate photosynthesis in potato tissue culture seedlings, and the downward transport of photosynthetic products promotes root development. The organic acids secreted by the roots of potato tissue culture seedlings continuously trigger the nutrient release mechanism within the culture medium that enhances nutrient absorption, achieving on-demand supply of trace metal elements and preventing heavy metal toxicity in the early stages of cultivation and nutrient deficiencies in the later stages.

[0027] This invention provides a culture medium for enhancing nutrient absorption in potato tissue culture seedlings, its preparation method, and its application. It has the following beneficial effects: 1. This invention utilizes anionic polymeric surfactants to adsorb graphene oxide, forming a physical steric hindrance layer. This layer physically isolates free zinc ions in the trace element component from phosphate and sulfate ions in the macro element component. This physical isolation blocks the reaction pathway of inorganic salts crystallizing and precipitating during the high-pressure steam sterilization heating stage, thereby improving the retention rate of effective trace metal elements in the culture medium that enhances nutrient absorption in potato tissue culture seedlings.

[0028] 2. This invention utilizes the organic small-molecule acids secreted by the roots of potato tissue culture seedlings inoculated on a culture medium that enhances nutrient absorption. These acids lower the local pH. The increased free hydrogen ions in the environment undergo protonation substitution, prompting the coordinated zinc ions to dissociate from the anionic functional groups of the anionic polymeric surfactant and diffuse into the aqueous medium. This protonation competitive substitution mechanism ensures a continuous and stable supply of trace metal elements, preventing heavy metal toxicity in the early stages of potato tissue culture and nutrient deficiency in the later stages, thereby promoting normal root development.

[0029] 3. This invention employs a step-by-step feeding process, initially mixing anionic polymeric surfactants and graphene oxide in deionized water, then separately adding trace element components to achieve coordination bonding, and finally incorporating macro element components dissolved with a high concentration of inorganic salts. This step-by-step feeding process, combined with pH adjustment, maintains the hydrodynamic radius and Zeta potential of the mesoscopic micro-sink structure within the culture medium that enhances nutrient absorption in potato tissue culture seedlings within a specific range. These numerical parameters within this range provide sufficient electrostatic repulsion, maintaining the colloidal thermodynamic stability of the overall suspension system of the culture medium that enhances nutrient absorption in potato tissue culture seedlings. Attached Figure Description

[0030] Figure 1 This is a schematic diagram comparing the retention rates of the effective state obtained from the conversion of the absolute mass concentration of free and soluble zinc elements in this invention. Figure 2 This is a schematic diagram comparing the rooting rates of explants in this invention; Figure 3 This is a schematic diagram comparing root length and plant height in this invention; Figure 4 This is a schematic diagram comparing the fresh weight and dry weight of a single plant according to the present invention. Detailed Implementation

[0031] The technical solutions in 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.

[0032] Raw materials: The main raw materials and reagents used in the following examples and comparative examples have the following sources and specifications. Reagents not specifically mentioned are all commercially available analytical grade or higher grade products.

[0033] Sodium lignosulfonate (CAS No. 8061-51-6) is an anionic polymeric surfactant. Its main structure is a three-dimensional network polymer formed by phenylpropane repeating units with sulfonic acid groups crosslinked by ether bonds and carbon-carbon bonds. Its weight-average molecular weight is 8,000 to 15,000 Daltons and its degree of sulfonation is 1.2 to 1.8 mmol / g.

[0034] Preparation Examples 1-3: Preparation Example 1: This preparation example provides a method for preparing an intermediate complex precursor liquid, including the following steps: Add 300 mL of deionized water to a reaction vessel equipped with a temperature control and stirring device, add 1.5 mg of sodium lignosulfonate to the deionized water, and turn on the stirring device to completely dissolve the sodium lignosulfonate. Subsequently, an aqueous dispersion of graphene oxide containing 0.2 mg of graphene oxide was slowly added dropwise to the reaction vessel; Turn on the heating jacket of the reactor to raise the temperature inside the reactor to 70°C, and keep it under constant temperature and closed stirring at 400 rpm for 60 minutes to obtain the intermediate composite precursor liquid.

[0035] Preparation Example 2: This preparation example provides a method for preparing an intermediate complex precursor liquid, including the following steps: Add 250 mL of deionized water to a reaction vessel equipped with a temperature control and stirring device, add 1.0 mg of sodium lignosulfonate to the deionized water, and turn on the stirring device to completely dissolve the sodium lignosulfonate. Subsequently, an aqueous dispersion of graphene oxide containing 0.2 mg of graphene oxide was slowly added dropwise to the reaction vessel; Turn on the heating jacket of the reactor to raise the temperature inside the reactor to 65°C, and keep it under constant temperature and closed stirring at 300 rpm for 45 minutes to obtain the intermediate composite precursor liquid.

[0036] Preparation Example 3: This preparation example provides a method for preparing an intermediate complex precursor liquid, including the following steps: Add 350 mL of deionized water to a reaction vessel equipped with a temperature control and stirring device, add 2.0 mg of sodium lignosulfonate to the deionized water, and turn on the stirring device to completely dissolve the sodium lignosulfonate. Subsequently, an aqueous dispersion of graphene oxide containing 0.3 mg of graphene oxide was slowly added dropwise to the reaction vessel; Turn on the heating jacket of the reactor to raise the temperature inside the reactor to 75°C, and keep it under constant temperature and closed stirring at 500 rpm for 75 minutes to obtain the intermediate composite precursor liquid.

[0037] Examples 1-3: Example 1: This embodiment provides a culture medium for enhancing nutrient absorption in potato tissue culture seedlings, its preparation method, and its application, including the following steps: The temperature of the reactor containing the intermediate composite precursor solution prepared in Example 1 was lowered to 40°C. Trace element components were added to the reactor, including 27.8 parts by weight of ferrous sulfate heptahydrate, 37.3 parts by weight of disodium ethylenediaminetetraacetate dihydrate, 22.3 parts by weight of manganese sulfate tetrahydrate, 8.6 parts by weight of zinc sulfate heptahydrate, 6.2 parts by weight of boric acid, 0.83 parts by weight of potassium iodide, 0.25 parts by weight of sodium molybdate dihydrate, 0.025 parts by weight of copper sulfate pentahydrate, and 0.025 parts by weight of cobalt chloride hexahydrate. The pH of the mixture in the reactor was adjusted to 4.5 using a 0.3 mol / L hydrochloric acid solution, and the mixture was stirred at a constant temperature of 40°C for 30 min.

[0038] Add 400,000 parts by volume of room temperature deionized water to another separate container. Add macro-element components, organic auxiliary components, and 30,000 parts by weight of sucrose to the separate container. The macro-element components include 1,650 parts by weight of ammonium nitrate, 1,900 parts by weight of potassium nitrate, 440 parts by weight of calcium chloride dihydrate, 370 parts by weight of magnesium sulfate heptahydrate, and 170 parts by weight of potassium dihydrogen phosphate. The organic auxiliary components include 100 parts by weight of inositol, 2.0 parts by weight of glycine, 0.12 parts by weight of thiamine hydrochloride, 0.5 parts by weight of pyridoxine hydrochloride, and 0.5 parts by weight of nicotinic acid. Start stirring to completely dissolve all soluble solids at room temperature to form a transparent homogeneous solution.

[0039] The transparent homogeneous solution in an independent container was pumped into a reaction vessel containing trace element components for mixing. Room temperature deionized water was added to the reaction vessel to bring the total volume of the reaction vessel to a constant 1,000,000 parts by volume. The final pH value of the mixture in the reaction vessel was adjusted to 5.8 using a 1.0 mol / L sodium hydroxide solution. Then, 7,000 parts by weight of plant tissue culture grade agar powder were added to the reaction vessel at a uniform rate, and high shear stirring was maintained to ensure that the agar powder was uniformly suspended in the mixture to obtain a liquid suspension culture medium.

[0040] Liquid suspension culture medium was continuously filled into glass tissue culture bottles, with 40,000 volume portions per bottle. After sealing the bottle mouths, the bottles were placed in an industrial high-pressure steam sterilizer and sterilized for 17 minutes at a working pressure of 0.105 MPa and a temperature of 121°C. After sterilization, the bottles were allowed to cool naturally to room temperature. The agar powder in the liquid suspension culture medium melted at high temperature and solidified upon cooling into a homogeneous solid gel, thus obtaining a culture medium that enhances nutrient absorption in potato tissue culture seedlings.

[0041] Aseptic operation was performed in a clean bench. Virus-free potato tissue culture seedlings were selected, and potato stem segments with a length of 1.0 cm and containing axillary buds were cut using a sterile scalpel as explants. The morphological lower end of the potato stem segment was vertically inserted into a culture medium that enhances nutrient absorption of potato tissue culture seedlings.

[0042] The glass tissue culture bottles inoculated with potato stem segments were transferred to an environmentally controlled culture room. The temperature of the environmentally controlled culture room was maintained at 20°C, the relative humidity was controlled at 35%, and the LED plant grow lights were turned on to maintain the light flux at 1200lm. The photocycle was set to alternate between 18 hours of light and 6 hours of darkness. The culture was carried out continuously in the environmentally controlled culture room for 30 days.

[0043] Example 2: This embodiment describes a culture medium for enhancing nutrient absorption in potato tissue culture seedlings, its preparation method, and its application, including the following steps: The temperature of the reactor containing the intermediate composite precursor solution prepared in Example 2 was lowered to 35°C. Trace element components were added to the reactor, including 25.0 parts by weight of ferrous sulfate heptahydrate, 35.0 parts by weight of disodium ethylenediaminetetraacetate dihydrate, 20.0 parts by weight of manganese sulfate tetrahydrate, 7.0 parts by weight of zinc sulfate heptahydrate, 5.0 parts by weight of boric acid, 0.7 parts by weight of potassium iodide, 0.2 parts by weight of sodium molybdate dihydrate, 0.02 parts by weight of copper sulfate pentahydrate, and 0.02 parts by weight of cobalt chloride hexahydrate. The pH of the mixture in the reactor was adjusted to 4.3 using a 0.1 mol / L hydrochloric acid solution, and the mixture was stirred at a constant temperature of 35°C for 25 min.

[0044] Add 400,000 parts by volume of room temperature deionized water to another separate container. Add macro-element components, organic auxiliary components, and 28,000 parts by weight of sucrose to the separate container. The macro-element components include 1,550 parts by weight of ammonium nitrate, 1,800 parts by weight of potassium nitrate, 400 parts by weight of calcium chloride dihydrate, 350 parts by weight of magnesium sulfate heptahydrate, and 150 parts by weight of potassium dihydrogen phosphate. The organic auxiliary components include 90 parts by weight of inositol, 1.8 parts by weight of glycine, 0.09 parts by weight of thiamine hydrochloride, 0.4 parts by weight of pyridoxine hydrochloride, and 0.4 parts by weight of nicotinic acid. Start stirring to completely dissolve all soluble solids at room temperature to form a transparent homogeneous solution.

[0045] The transparent homogeneous solution in an independent container was pumped into a reaction vessel containing trace element components for mixing. Room temperature deionized water was added to the reaction vessel to bring the total volume to a constant 900,000 parts by volume. The final pH of the mixture in the reaction vessel was adjusted to 5.6 using a 0.8 mol / L sodium hydroxide solution. Subsequently, 6,500 parts by weight of plant tissue culture grade agar powder were added to the reaction vessel at a uniform rate, and high shear stirring was maintained to ensure that the agar powder was uniformly suspended in the mixture, thus obtaining a liquid suspension culture medium.

[0046] Liquid suspension culture medium was continuously filled into glass tissue culture bottles, with 30,000 volume portions per bottle. After sealing the bottle mouths, the bottles were placed in an industrial high-pressure steam sterilizer and sterilized for 15 minutes at a working pressure of 0.10 MPa and a temperature of 115°C. After sterilization, the bottles were allowed to cool naturally to room temperature. The agar powder in the liquid suspension culture medium melted at high temperature and solidified into a homogeneous solid gel, thus obtaining a culture medium that enhances nutrient absorption in potato tissue culture seedlings.

[0047] Aseptic operation was performed in a clean bench. Virus-free potato tissue culture seedlings were selected, and potato stem segments with a length of 0.8 cm and containing axillary buds were cut using a sterile scalpel as explants. The morphological lower end of the potato stem segment was vertically inserted into a culture medium that enhances nutrient absorption of potato tissue culture seedlings.

[0048] The glass tissue culture bottles inoculated with potato stem segments were transferred to an environmentally controlled culture room. The temperature of the environmentally controlled culture room was maintained at 18℃, the relative humidity was controlled at 30%, and the LED plant grow lights were turned on to maintain the light flux at 1000lm. The photocycle was set to alternate between 16 hours of light and 8 hours of darkness. The culture was carried out continuously in the environmentally controlled culture room for 25 days.

[0049] Example 3: This embodiment describes a culture medium for enhancing nutrient absorption in potato tissue culture seedlings, its preparation method, and its application, including the following steps: The temperature of the reactor containing the intermediate composite precursor solution prepared in Example 3 was lowered to 45°C. Trace element components were added to the reactor, including 30.0 parts by weight of ferrous sulfate heptahydrate, 40.0 parts by weight of disodium ethylenediaminetetraacetate dihydrate, 25.0 parts by weight of manganese sulfate tetrahydrate, 10.0 parts by weight of zinc sulfate heptahydrate, 7.0 parts by weight of boric acid, 1.0 part by weight of potassium iodide, 0.3 parts by weight of sodium molybdate dihydrate, 0.03 parts by weight of copper sulfate pentahydrate, and 0.03 parts by weight of cobalt chloride hexahydrate. The pH of the mixture in the reactor was adjusted to 4.8 using a 0.5 mol / L hydrochloric acid solution, and the mixture was stirred at a constant temperature of 45°C for 40 min.

[0050] Add 400,000 parts by volume of room temperature deionized water to another separate container. Add macro-element components, organic auxiliary components, and 32,000 parts by weight of sucrose to the separate container. The macro-element components include 1,750 parts by weight of ammonium nitrate, 2,000 parts by weight of potassium nitrate, 480 parts by weight of calcium chloride dihydrate, 390 parts by weight of magnesium sulfate heptahydrate, and 190 parts by weight of potassium dihydrogen phosphate. The organic auxiliary components include 110 parts by weight of inositol, 2.2 parts by weight of glycine, 0.15 parts by weight of thiamine hydrochloride, 0.6 parts by weight of pyridoxine hydrochloride, and 0.6 parts by weight of nicotinic acid. Start stirring to completely dissolve all soluble solids at room temperature to form a transparent homogeneous solution.

[0051] The transparent homogeneous solution in an independent container was pumped into a reaction vessel containing trace element components for mixing. Room temperature deionized water was added to the reaction vessel to bring the total volume of the reaction vessel to a constant 1,100,000 parts by volume. The final pH value of the mixture in the reaction vessel was adjusted to 6.0 using a 1.2 mol / L sodium hydroxide solution. Then, 7,500 parts by weight of plant tissue culture grade agar powder were added to the reaction vessel at a uniform rate, and high shear stirring was maintained to ensure that the agar powder was uniformly suspended in the mixture to obtain a liquid suspension culture medium.

[0052] Liquid suspension culture medium was continuously filled into glass tissue culture bottles, with 50,000 volume portions per bottle. After sealing the bottle mouths, the bottles were placed in an industrial high-pressure steam sterilizer and sterilized for 20 minutes at a working pressure of 0.11 MPa and a temperature of 125°C. After sterilization, the bottles were allowed to cool naturally to room temperature. The agar powder in the liquid suspension culture medium melted at high temperature and solidified into a homogeneous solid gel, thus obtaining a culture medium that enhances nutrient absorption in potato tissue culture seedlings.

[0053] Aseptic operation was performed in a clean bench. Virus-free potato tissue culture seedlings were selected, and potato stem segments with a length of 1.2 cm and containing axillary buds were cut using a sterile scalpel as explants. The morphological lower end of the potato stem segment was vertically inserted into a culture medium that enhances nutrient absorption of potato tissue culture seedlings.

[0054] The glass tissue culture bottles inoculated with potato stem segments were transferred to an environmentally controlled culture room. The temperature of the environmentally controlled culture room was maintained at 22℃, the relative humidity was controlled at 40%, and the LED plant grow lights were turned on to maintain the light flux at 1400lm. The photocycle was set to alternate between 20h light duration and 4h dark duration. The culture was carried out continuously in the environmentally controlled culture room for 35 days.

[0055] Comparative Examples 1-5: Comparative Example 1: Compared with Example 1, the difference is that the culture medium formula does not contain graphene oxide and sodium lignosulfonate, and the steps of Preparation Example 1 are not performed. The trace element components, macro element components, organic auxiliary material components, sucrose and plant tissue culture grade agar powder are directly mixed, adjusted to a certain volume, and then autoclaved. All other aspects are the same.

[0056] Comparative Example 2: Compared with Example 1, the difference is that the culture medium formula does not contain sodium lignosulfonate. The aqueous dispersion of graphene oxide is directly mixed with trace element components, macro element components, organic auxiliary components, sucrose and plant tissue culture grade agar powder in one go, and then sterilized by high pressure steam. All other aspects are the same.

[0057] Comparative Example 3: Compared with Example 1, the difference is that the sequential split feeding process is not used. The aqueous dispersion of graphene oxide, sodium lignosulfonate, trace element components, macro element components, organic auxiliary material components and sucrose are mixed and brought to a constant volume in room temperature deionized water. Then, plant tissue culture grade agar powder is added and autoclaved. All other aspects are the same.

[0058] Comparative Example 4: Compared with Example 1, the difference lies in the disordered feeding order. The trace element components and macro element components are first dissolved in deionized water, and then the aqueous dispersion of graphene oxide and sodium lignosulfonate are added. The rest are the same.

[0059] Comparative Example 5: Compared with Example 1, the difference is that sodium lignosulfonate is replaced by a conventional polymeric dispersant, polyethylene glycol, in equal weight; otherwise, they are the same.

[0060] Test Examples 1-4: Test Example 1: Dimensional Evolution Test of Colloid Electrodynamics and Hydrodynamics in Hybrid Systems In a cleanroom environment, 2.0 mL liquid samples were extracted using a pipette from the reactors at the following stages: after the intermediate composite precursor solutions were prepared in Examples 1, 2, and 3 (defined as Stage 1); after the addition of trace element components and constant-temperature stirring (defined as Stage 2); and after the addition of macro element components and constant-volume mixing, but before the addition of plant tissue culture-grade agar powder (defined as Stage 3). Simultaneously, 2.0 mL liquid samples were extracted from Comparative Example 2 at the same process nodes as Stage 1, Stage 2, and Stage 3, and from Comparative Example 3 after a single mixing and volume adjustment, but before the addition of plant tissue culture-grade agar powder.

[0061] The extracted liquid sample was transferred to a clean polystyrene cuvette and a collapsible capillary electrolysis cell. The Stage 3 liquid sample, which contained a high concentration of inorganic salts, was diluted tenfold with room temperature deionized water to ensure that the optical transmittance of the liquid sample met the detection threshold of the dynamic light scattering instrument.

[0062] Start the dynamic light scattering instrument and its associated electrophoretic light scattering test module, and maintain the temperature of the detection chamber of the dynamic light scattering instrument at 25.0℃. Input the refractive index and viscosity parameters of the dispersion medium, pure water, into the control software of the dynamic light scattering instrument. Perform three consecutive scans of the zeta potential and hydrodynamic radius of the liquid samples in the polystyrene cuvette and the foldable capillary electrolysis cell, respectively. Record the absolute value of the zeta potential and the distribution value of the hydrodynamic radius output by the dynamic light scattering instrument, and calculate the arithmetic mean of the three consecutive scans.

[0063] Table 1. Data on the dimensional evolution of colloidal electrodynamics and hydrodynamics at each preparation stage in the examples and comparative examples.

[0064] Conclusions and Analysis: According to the data in Table 1, the phase one test results of Examples 1 to 3 show that the mixed system has a negative potential of less than -40mV and a hydrodynamic radius distributed in the range of 340nm to 380nm, which proves that sodium lignosulfonate in an extremely dilute state and after heat treatment has highly extended polymer chain segments. The hydrophobic benzene ring skeleton has been firmly bonded to the surface of graphene oxide through van der Waals forces and π-π conjugation, forming a thick polymer adsorption layer.

[0065] Comparing the data from Stage 1 of Comparative Example 2, the Zeta potential of the standalone aqueous dispersion of graphene oxide was -36.4 mV, and the hydrodynamic radius was 302.8 nm, demonstrating that the introduction of sodium lignosulfonate contributed additional sulfonate negative charge and a significant steric hindrance layer thickness to the graphene oxide. Upon entering Stages 2 and 3, with the addition of trace amounts of metal cations and high concentrations of inorganic salts, the Zeta potential of the mixed systems from Examples 1 to 3 exhibited a regular decay, indicating that the large number of inorganic cations exerted a compression effect on the electrical double layer of the colloidal particles.

[0066] In Examples 1 to 3, the hydrodynamic radius of the mixture under the stage 3 environment only increased slightly, with the highest not exceeding 410 nm, and the mixture remained in a highly homogeneous mesoscopic suspension state.

[0067] Comparative Example 2, lacking sodium lignosulfonate and thus lacking a high-strength steric hindrance layer, experienced a rapid increase in hydrodynamic radius to 845.6 nm and initial agglomeration when trace metal ions were introduced in stage two. When Comparative Example 2 entered stage three and high concentrations of nitrate, sulfate, calcium, magnesium, and potassium ions were introduced, the double layer was completely destroyed, the Zeta potential dropped to -2.8 mV, resulting in the complete loss of electrostatic repulsion. The hydrodynamic radius exceeded 2100 nm, leading to lamellar precipitation and salting-out flocculation.

[0068] Comparative Example 3 did not employ a sequential splitting feeding process, resulting in a lack of high-temperature expansion and conformational rearrangement of sodium lignosulfonate in a pure water environment. Consequently, the polymeric surfactant failed to adequately encapsulate the graphene oxide, leading to a hydrodynamic radius of 1435.9 nm in the mixed system of Comparative Example 3, thus disrupting the colloidal thermodynamic stability. The test results of Examples 1 to 3 demonstrate that the sequential splitting process and the non-covalent assembly mechanism successfully blocked the colloidal instability pathway induced by inorganic salts, ensuring the efficient construction and uniform distribution of the mesoscopic reservoir of trace elements in the culture medium.

[0069] Test Example 2: Quantitative Test of Retention Rate of Effective Metal Ions under Extreme Thermodynamic Conditions of High-Pressure Steam Sterilization In a dust-free experimental environment, using a pipette, 50.0 mL of mixed liquid samples were extracted from each of the reaction vessels of Examples 1, 2, 3, and Comparative Examples 1, 4, and 5 after macro-element components were added and mixed to a constant volume, but before the addition of plant tissue culture grade agar powder. The samples were then transferred to high-pressure resistant quartz glass tubes and sealed.

[0070] The high-pressure resistant quartz glass tube containing the mixed liquid sample was transferred into an industrial high-pressure steam sterilizer and sterilized under constant temperature and high pressure at 121°C. The constant temperature and high pressure steam sterilization time for Examples 1, 1, 4 and 5 was 17 min, for Example 2 it was 15 min, and for Example 3 it was 20 min. After sterilization, the tube was naturally vented and cooled to room temperature (25°C).

[0071] The cooled mixed liquid sample was transferred to a polycarbonate centrifuge tube and placed in a high-speed refrigerated centrifuge. The centrifugation temperature was set to 20°C and the speed to 12,500 rpm. The centrifugation was carried out continuously for 25 minutes to promote the physical separation of macroscopic inorganic salt precipitates that may be generated during the high-pressure steam sterilization process from the liquid phase.

[0072] Use a micropipette to transfer 5.0 mL of the supernatant from the centrifuge tube to a polytetrafluoroethylene (PTFE) digestion vessel. Add 8.0 mL of 65% concentrated nitric acid solution to the PTFE digestion vessel. Place the PTFE digestion vessel in a microwave digester and heat it to 180°C for 30 minutes to completely release the non-covalently bound trace metal elements in the supernatant into free ions.

[0073] After the digestion solution has cooled, transfer it to a 50 mL volumetric flask and dilute to the mark with room temperature deionized water. Start the inductively coupled plasma atomic emission spectrometer (ICP-AES) and use the standard curve method to determine the absolute mass concentration of zinc in the digestion solution. Divide the mass concentration values ​​output by the ICP-AES by the theoretical initial mass concentration values ​​of each component to obtain the percentage of effective retention.

[0074] Table 2. Retention rate of trace metal elements in liquid culture medium before and after sterilization

[0075] Conclusions and Analysis: According to the data in Table 2, the effective zinc retention rates in Examples 1, 2, and 3 are all higher than 95.0%. The test results verify that the mesoscopic micro-reservoir constructed from sodium lignosulfonate and graphene oxide maintains extremely high structural stability even under extreme thermodynamic conditions of high-pressure steam sterilization at 121℃. The rigid phenylpropane skeleton of sodium lignosulfonate is firmly anchored to the two-dimensional carbon matrix of graphene oxide through π-π conjugation. The sulfonic acid groups of sodium lignosulfonate facing the aqueous phase preferentially undergo solid-liquid interfacial dynamic coordination with trace metal ions (such as zinc ions). When easily precipitating anions such as phosphate are added and subjected to high-temperature heating, the high-strength physical steric hindrance layer effectively blocks the diffusion and penetration of external precipitating anions into the inner layer of free trace metal ions, cutting off the reaction path of inorganic salt crystallization and precipitation from a chemical reaction kinetics perspective, and avoiding the co-precipitation loss of trace metal elements.

[0076] In contrast, Comparative Example 1 did not introduce graphene oxide or sodium lignosulfonate. The trace metal ions in the mixed liquid sample were completely exposed to a macro-element aqueous environment containing high concentrations of phosphate and sulfate. Driven by high energy at 121°C, the free metal ions rapidly exceeded the solubility product limit, undergoing violent side reactions to form insoluble precipitates. This resulted in significant loss of trace elements (for example, the retention rate of zinc was only 61.2%), failing to meet the effective supply concentration required for the normal metabolism of potato tissue culture seedlings.

[0077] In Comparative Example 4, due to the disruption of the feeding sequence, sodium lignosulfonate failed to form a complete steric hindrance layer, allowing anions to still penetrate and contact the underlying metal ions. This resulted in poor anti-precipitation effect of trace metal elements (zinc retention rate was only 79.4%), confirming the irreplaceable role of the time-separated feeding process in constructing the mesoscopic anti-precipitation barrier.

[0078] In Comparative Example 5, sodium lignosulfonate was replaced with polyethylene glycol. The loose polyethylene glycol molecular chains were prone to thermal desorption and adsorption. Anions easily penetrated the polyethylene glycol layer and captured the metal ions in the bottom layer, resulting in a large amount of micronutrient elements being precipitated and lost (the zinc element retention rate was only 65.8%).

[0079] The difference in absolute concentration measurements between the examples and the comparative examples demonstrates that the present invention can achieve the retention of micronutrients without precipitation in the high-salt and high-temperature environment of plant tissue culture media.

[0080] Test Example 3: Comparative Evaluation of Core Biological and Agronomic Growth and Development Indicators of Potato Tissue Culture Seedlings After the continuous culture cycle in the controlled environment was completed, all glass tissue culture bottles inoculated with potato stem segments were transferred to a clean bench. The number of potato stem segments with visible roots in each glass tissue culture bottle in the same test group was counted. The number of potato stem segments with visible roots was divided by the total number of potato stem segments initially inoculated, and the rooting rate of the explants was calculated and recorded.

[0081] Break the glass culture flask and use sterile dissecting forceps to completely remove the potato tissue culture seedlings from the culture medium that enhances nutrient absorption. Gently rinse the removed seedlings in a beaker of room temperature deionized water to thoroughly wash away any solid gel adhering to the roots.

[0082] Place the washed potato tissue culture seedlings flat on a black rubber mat with millimeter graduations. Use a ruler to measure the absolute vertical distance from the lower morphological point of the seedling to the growth point of the highest leaf, and record this as the plant height. Use a ruler to measure the absolute length of the longest taproot of the seedling, and record this as the average root length. All length measurements should be accurate to 0.1 cm.

[0083] Use highly absorbent filter paper to absorb the room temperature deionized water from the surface of potato tissue culture seedlings. Place the dried potato tissue culture seedlings on the weighing pan of a precision analytical balance with an accuracy of 0.1 mg, weigh them, and record the fresh weight of each seedling.

[0084] After weighing the fresh weight of each potato tissue culture seedling, place them in kraft paper envelopes and transfer them to an electric blast drying oven. Set the temperature of the electric blast drying oven to 105℃ and heat for 15 minutes to sterilize the plant tissue. Then, reduce the temperature of the electric blast drying oven to 75℃ and continue heating to dry until the difference in weight between two consecutive weighings of the plant tissue in a single kraft paper envelope is less than 0.002g. Place the dried potato tissue culture seedlings in a desiccator containing silica gel to cool to 25℃, and weigh them again using a precision analytical balance, recording the dry weight of each seedling.

[0085] Table 3. Data on core growth and development indicators of potato tissue culture seedlings in biology and agronomy.

[0086] Conclusions and Analysis: According to Table 3 and in conjunction with Figure 2 , Figure 3 and Figure 4The data show that the potato tissue culture seedlings of Examples 1, 2 and 3 all exhibited significant advantages in five core biological and agronomic indicators: percentage of explant rooting rate, average root length, plant height, fresh weight per plant, and dry weight per plant.

[0087] The rooting rate of explants in Examples 1 to 3 all exceeded 95.0%, the plant height exceeded 11.0 cm, and the fresh weight of a single plant exceeded 500.0 mg. The test results demonstrate that the mesoscopic micro-sink constructed from sodium lignin sulfonate and graphene oxide effectively maintained the homogeneous suspension of trace elements in the culture medium. The mesoscopic micro-sink can dynamically and steadily release trace nutrients based on the decrease in microenvironment pH caused by organic acids secreted by the roots of potato tissue culture seedlings. This dynamic release process matches the nutrient absorption kinetics of potato tissue culture seedlings in the closed tissue culture environment, avoiding nutrient toxicity in the early stages of culture and nutrient deficiency in the later stages, and promoting the accumulation of photosynthetic substances and cell division and elongation in potato tissue culture seedlings.

[0088] In Comparative Example 1, no graphene oxide or sodium lignosulfonate was added. The precipitation and loss of trace metal elements within the culture medium led to nutrient deficiency, resulting in stunted growth. This nutrient deficiency resulted in a rooting rate of only 52.4% for potato tissue culture seedlings in Comparative Example 1, and a single plant dry weight of only 19.3 mg, indicating severe stunted growth.

[0089] In Comparative Example 2, due to the lack of steric hindrance protection from sodium lignin sulfonate, the exposed graphene oxide underwent irreversible lamellar aggregation and precipitation in an environment rich in inorganic macro-cations. The aggregated graphene oxide hindered the development of capillary roots and water metabolism in the potato tissue culture seedlings, resulting in an average root length of only 3.1 cm for Comparative Example 2.

[0090] Comparative Example 3 did not employ a sequential feeding process, and Comparative Example 4 disrupted the feeding sequence, both resulting in sodium lignosulfonate failing to completely coat the two-dimensional carbon substrate of graphene oxide. This incomplete coating led to the continued presence of some metal salt precipitation side reactions in the culture medium system, hindering the effective supply of trace elements and causing a significant decline in various growth indicators of the potato tissue culture seedlings.

[0091] In Comparative Example 5, sodium lignosulfonate was replaced with polyethylene glycol. Polyethylene glycol could not provide dynamic coordination anchoring and strong physical steric hindrance for trace metal ions, thus failing to block the high-temperature crystallization and precipitation side reaction of trace metal elements. This resulted in a lower effective supply rate of micronutrients, leading to a significantly smaller final biomass accumulation in the potato tissue culture seedlings of Comparative Example 5 compared to Examples 1 to 3. This verified the effectiveness of the preparation methods provided in Examples 1 to 3 in real plant tissue culture applications.

Claims

1. A culture medium for enhancing nutrient absorption in potato tissue culture seedlings, characterized in that, It is made from raw materials comprising the following parts by weight: 0.1 to 0.3 parts by weight of graphene oxide and 1.0 to 2.0 parts by weight of anionic polymeric surfactant; Trace element composition: ferrous sulfate heptahydrate 25.0–30.0 parts by weight, disodium ethylenediaminetetraacetate dihydrate 35.0–40.0 parts by weight, manganese sulfate tetrahydrate 20.0–25.0 parts by weight, zinc sulfate heptahydrate 7.0–10.0 parts by weight, boric acid 5.0–7.0 parts by weight, potassium iodide 0.7–1.0 parts by weight, sodium molybdate dihydrate 0.2–0.3 parts by weight, copper sulfate pentahydrate 0.02–0.03 parts by weight, cobalt chloride hexahydrate 0.02–0.03 parts by weight; Macro-element components: ammonium nitrate 1550-1750 parts by weight, potassium nitrate 1800-2000 parts by weight, calcium chloride dihydrate 400-480 parts by weight, magnesium sulfate heptahydrate 350-390 parts by weight, potassium dihydrogen phosphate 150-190 parts by weight. Organic excipients: 90-110 parts by weight of inositol, 1.8-2.2 parts by weight of glycine, 0.09-0.15 parts by weight of thiamine hydrochloride, 0.4-0.6 parts by weight of pyridoxine hydrochloride, 0.4-0.6 parts by weight of nicotinic acid; 28,000-32,000 parts by weight of sucrose; 6500-7500 parts by weight of plant tissue culture grade agar powder; Including the remaining deionized water, the total volume of the mixed system is brought to a final volume of 900,000 to 1,100,000 parts by volume.

2. The culture medium for enhancing nutrient absorption in potato tissue culture seedlings according to claim 1, characterized in that, The anionic polymeric surfactant is sodium lignosulfonate.

3. The culture medium for enhancing nutrient absorption in potato tissue culture seedlings according to claim 1, characterized in that, The anionic polymeric surfactant and the graphene oxide are assembled non-covalently to form a mesoscopic micro-reservoir structure, and the hydrodynamic radius of the mesoscopic micro-reservoir structure is distributed between 340 nm and 410 nm.

4. The culture medium for enhancing nutrient absorption in potato tissue culture seedlings according to claim 1, characterized in that, Before the addition of the plant tissue culture-grade agar powder, the pH of the culture medium for enhancing nutrient absorption in potato tissue culture seedlings was 5.6–6.0, and the Zeta potential was maintained between -26.5 mV and -22.3 mV.

5. A method for preparing a culture medium that enhances nutrient absorption in potato tissue culture seedlings, characterized in that, The method for preparing the culture medium for enhancing nutrient absorption of potato tissue culture seedlings as described in any one of claims 1-4 comprises the following steps: An anionic polymeric surfactant was dissolved in deionized water, and an aqueous dispersion containing graphene oxide was slowly added dropwise. The mixture was heated and stirred under constant temperature conditions to obtain an intermediate composite precursor solution. The temperature of the intermediate composite precursor solution is lowered, trace element components are added to form a first mixture, the pH value of the first mixture is adjusted using hydrochloric acid solution, and the mixture is stirred under constant temperature conditions. In a separate container, macro-element components, organic auxiliary components, and sucrose are dissolved in deionized water to form a transparent homogeneous solution; The transparent homogeneous solution is mixed with the first mixture to form a final mixture system, room temperature deionized water is added, and the pH value of the final mixture system is adjusted using sodium hydroxide solution. Plant tissue culture grade agar powder is added to the final mixture at a uniform rate, and high-shear stirring is maintained to obtain a liquid suspension culture medium; the liquid suspension culture medium is sterilized by high-pressure steam and cooled to solidify, thus obtaining the culture medium that enhances nutrient absorption of potato tissue culture seedlings.

6. The method for preparing the culture medium for enhancing nutrient absorption in potato tissue culture seedlings according to claim 5, characterized in that, The process of heating and stirring under constant temperature conditions to prepare the intermediate composite precursor liquid specifically includes: The heating temperature is controlled at 65℃~75℃, the stirring speed is 300rpm~500rpm, and the stirring time is 45min~75min.

7. The method for preparing the culture medium for enhancing nutrient absorption in potato tissue culture seedlings according to claim 5, characterized in that, Lowering the temperature of the intermediate composite precursor solution, adding trace element components to form a first mixture, adjusting the pH of the first mixture using hydrochloric acid solution, and stirring under constant temperature conditions specifically include: The temperature of the intermediate composite precursor solution is lowered to 35℃~45℃, the concentration of the hydrochloric acid solution is 0.1mol / L~0.5mol / L, the pH value of the first mixture is adjusted to 4.3~4.8 using the hydrochloric acid solution, the stirring temperature under constant temperature conditions is 35℃~45℃, and the stirring time under constant temperature conditions is 25min~40min.

8. The method for preparing the culture medium for enhancing nutrient absorption in potato tissue culture seedlings according to claim 5, characterized in that, The specific steps of adjusting the pH of the final mixture using sodium hydroxide solution include: The concentration of the sodium hydroxide solution is 0.8 mol / L to 1.2 mol / L, and the pH of the final mixture is adjusted to 5.6 to 6.0 using the sodium hydroxide solution.

9. The method for preparing the culture medium for enhancing nutrient absorption in potato tissue culture seedlings according to claim 5, characterized in that, The process of autoclaving and cooling the liquid suspension culture medium specifically includes: The working pressure of the high-pressure steam sterilization is 0.10MPa to 0.11MPa, the temperature of the high-pressure steam sterilization is 115℃ to 125℃, and the time of the high-pressure steam sterilization is 15min to 20min.

10. The application of a culture medium that enhances nutrient absorption in potato tissue culture seedlings, characterized in that, The application of the culture medium for enhancing nutrient absorption in potato tissue culture seedlings as described in any one of claims 1-4, the application comprising the following steps: Potato stem segments with a length of 0.8cm to 1.2cm and containing axillary buds were cut as explants; The morphological lower end of the potato stem segment is vertically inserted into the culture medium that enhances nutrient absorption of potato tissue culture seedlings; The culture medium for enhancing nutrient absorption of potato tissue culture seedlings inoculated with the potato stem segments was transferred into an environmentally controlled culture room and cultured continuously for 25 to 35 days. The temperature of the environmentally controlled culture room was set to 18℃ to 22℃, the relative humidity was set to 30% to 40%, and the LED plant grow lights were turned on to maintain a light flux of 1000lm to 1400lm. The photocycle was set to alternate between a light duration of 16h to 20h and a dark duration of 4h to 8h.

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