Comprehensive evaluation and application of the mitigation effect of microplastic toxicity in maize seedlings

Abstract

This invention presents a comprehensive evaluation method and application for mitigating microplastic toxicity in maize seedlings. It utilizes the biological concepts of relative mitigation effect index and absolute mitigation intensity coefficient for individual traits in maize seedlings affected by polystyrene (PS) microplastics, derived from the application of different concentrations of nano-zinc oxide (ZnONPs) / monolayer graphene (GO) solutions. This method scientifically, objectively, accurately, and comprehensively evaluates the overall mitigation effect and intensity of different concentrations of ZnONPs / GO solutions on PS microplastic toxicity in maize seedlings after 7 or 21 days of growth. Through comprehensive comparison, a scientific and efficient application method for mitigating PS microplastic toxicity in maize seedlings using ZnONPs / GO solution treatment is proposed. This invention is simple to operate, requires small amounts of ZnONPs / GO, is economical and practical, does not pollute the environment, and has a good overall mitigation effect on microplastic toxicity in maize seedlings, showing great potential for application in green, efficient, and clean maize production.

Description

Technical Field

[0001] This invention belongs to the field of efficient and safe corn production under microplastic (MP) exposure environments. Specifically, it relates to a comprehensive evaluation method for mitigating the toxicity of polystyrene (PS) microplastics in corn seedlings, and an application method for mitigating the toxicity of PS microplastics in corn seedlings using zinc oxide nanoparticales (ZnONPs) and graphene oxide (GO). Background Technology

[0002] Since the first synthetic synthesis of phenolic plastics (PF) in 1909, they have been widely used in various fields such as agriculture, food packaging, medical and health care, and industrial production due to their excellent properties such as light weight, low cost, practicality, high plasticity, and ease of processing. By the early 1940s, the production and use of plastic products began to increase rapidly, gradually developing into various types and functions of plastics such as polyvinyl chloride (PVC), polyethylene (PE), polypropylene (PP), and polystyrene (PS). According to statistics from the United Nations Environment Programme (UNEP; https: / / www.unep.org / ), the cumulative global production of plastic products from 1950 to 2017 was 9.2 billion tons, and it is projected to reach 33 billion tons by 2050 (Nong Daiqian et al., 2024). Typically, most of these plastic products are not recycled after use and are directly discharged into the environment, forming plastic waste. Under long-term exposure to sunlight, ultraviolet radiation, weathering, mechanical wear, animals, and microorganisms, they will gradually decompose into microplastics (MPs) with particles smaller than 5 mm (Thompson et al. 2004). Soil is a major source of microplastic enrichment (Geyer et al. 2017). Increased microplastic concentration in soil not only significantly increases soil pH, ammonium nitrogen, and dissolved organic carbon content, and decreases soil nitrate nitrogen content (Gao et al. 2021), thereby altering soil structure and physicochemical properties, but also affects the structure and function of microbial communities (Aralappanavar et al. 2024). Furthermore, microplastics significantly inhibit seed germination, seedling growth, and biomass accumulation in maize (Zea mays L.) (Liu et al., 2022; Wang et al. 2020), reduce the germination rate and shoot height of ryegrass (Lolium perenne L.) (Boots et al. 2019), and have toxic effects on onion (Allium cepa L.) cells, thus posing a serious threat to the ecosystem.

[0003] Zinc (Zinc; Zn) is an essential micronutrient for crop growth and development. Although the required amount is extremely small, Zn deficiency in crops is a global problem. In fact, "hidden hunger" caused by Zn deficiency affects one-third of the global population, seriously impacting human health (Mcclung 2019). In crops, Zn primarily functions as a metal activator of various enzymes, playing a crucial role in the biosynthesis and metabolism of carbohydrates, proteins, and auxins, thus influencing crop resistance and growth. Soil Zn content <20 mg / kg -1 At that time, it was believed that there was a deficiency of Zn. Although traditional Zn fertilizers can supplement crop growth with Zn through root or foliar application, the root application of these traditional Zn fertilizers will cause a large amount of Zn loss, and the crop's utilization efficiency of Zn is low when applying Zn by leaves, and may cause adverse reactions such as leaf burn.

[0004] Nanomaterials / particles (NPs) refer to ultrafine particles with solid particle sizes down to the nanometer scale (1~100 nm) and materials with special properties with crystal sizes down to the nanometer scale (1~100 nm). In agriculture, NPs can improve the efficacy of agricultural inputs through targeted delivery, controlled release, enhanced solubility, and improved adhesion, thereby improving soil quality and enhancing crop resistance, ultimately stimulating crop growth and increasing agricultural productivity (Raza et al. 2023). In recent years, zinc oxide nanoparticles (ZnONPs) have become the most effective alternative to traditional Zn fertilizers and are widely used in stress-resistant and high-efficiency production of various crops. For example, Guo et al. (2024) found that under heat wave stress, foliar application of 100 mg L... -1 ZnONPs-treated rice ( Oryza sativa L . The yield, protein content, and nutritional quality content (such as amino acids) of the product increased by 22.1%, 11.8%, and 77.5%, respectively. Ghani et al. (2022) reported that applying 100 mg L... -1 ZnONPs can alleviate the effects of cucumber (by promoting antioxidant defense systems and osmotic accumulation) Cucumis sativus Drought-induced water shortage damage to L. seedlings. Hussain et al. (2024) showed that application of 25 mg L. -1ZnONPs can improve the toxic effects of cadmium stress on maize plants by regulating primary metabolites and antioxidant activity. Furthermore, graphene oxide (GO), known as "black gold," is currently the thinnest, strongest, and most electrically and thermally conductive nanomaterial discovered. In agricultural applications, Yan et al. (2024) revealed that 400 mg L... -1 GO soaked peanuts ( Arachis hypogaea L. seeds can significantly improve seed germination rate. Applying GO during the seedling stage can alleviate NaCl stress damage in peanut seedlings by enhancing photosynthesis, maintaining plasma membrane integrity, and promoting nutrient accumulation. Xue et al. (2020) found that adding 7.5 mg L. seeds significantly improved seed germination rate. -1 GO can improve European aspen (Populus tremula) The survival rate and rooting rate of L. under NaCl stress were improved, but the roots were relatively thin, weak, and short. However, there are no reports on the research and application of these nanomaterials in alleviating microplastic stress in crops. Summary of the Invention

[0005] The applicant, in its application for "Comprehensive Evaluation of Microplastic Toxicity Resistance of Maize Seedlings and Method for Identifying Microplastic-Resistant Maize Genotypes" submitted on the same day, found that 0.50% concentration (ratio of polystyrene (PS) microplastic mass to cultivation substrate mass) of PS microplastics caused the most severe toxicity to maize seedlings at 7 days and 21 days of growth. Based on this, the technical problem to be solved by the present invention is to provide a comprehensive evaluation method for mitigating the toxicity of polystyrene (PS) microplastics in maize seedlings. Specifically, the present invention systematically studies the effects of soaking seeds or spraying leaves with different concentrations of ZnONPs / GO solution on the growth and development, leaf chlorophyll accumulation, and 12 root traits of maize seedlings grown for 7 or 21 days under 0.50% PS microplastic toxicity. Furthermore, it comprehensively and systematically reveals the mitigation mechanism of ZnONPs / GO on maize seedlings under PS microplastic toxicity from the perspectives of aboveground and underground growth, biomass accumulation, leaf chlorophyll accumulation, and root activity and physiological metabolism at multiple growth stages of maize seedlings. This lays a theoretical foundation for the future scientific application of soaking seeds or spraying leaves with ZnONPs / GO to mitigate the toxic effects of PS microplastics on maize seedlings. Meanwhile, in order to scientifically, objectively, simply, accurately, and quantitatively evaluate the comprehensive mitigation effect / intensity of different concentrations of ZnONPs / GO solution soaking in seeds or foliar spraying on maize seedlings grown for 7 or 21 days under 0.50% PS microplastic toxicity, this invention also innovatively proposes a relative mitigation effect index of ZnONPs / GO (RMEI) for individual traits of maize seedlings under 0.50% PS microplastic toxicity.ZnONPS and RMEI GO The absolute mitigation intensity coefficient of ZnONPS / GO (AMIC) for individual traits of maize seedlings under 0.50% PS microplastic toxicity was determined by soaking seeds or foliar spraying with different concentrations of ZnONPS / GO solution. ZnONPs and AMIC GO This invention utilizes two biological concepts, characterized by a comprehensive consideration of the effects of control treatment, PS microplastic stress treatment, and seed soaking or foliar spraying with corresponding concentrations of ZnONPs / GO solution on each trait of maize seedlings at different growth stages. This effectively avoids the influence of specific treatments on individual traits of maize seedlings. It scientifically, accurately, and simply quantifies the relative mitigation effect index or absolute mitigation intensity coefficient of different concentrations of ZnONPs / GO solution on each trait of maize seedlings at different growth stages under PS microplastic toxicity. Furthermore, this invention also utilizes RMEI... Z n ONPS / RMEI GO and AMIC Z n ONP s / AMIC GO The values ​​were used as the comprehensive mitigation effect of ZnONPs / GO (CME) solution on maize seedlings cultured on day 7 or 21 of 0.50% PS microplastic toxicity, respectively, by soaking seeds or foliar spraying. ZnONPs and CME GO ) and comprehensive mitigation intensity of ZnONPs / GO, CMI ZnONPs and CMI GOThis provides a scientific evaluation method for the comprehensive effects of applying different substances to alleviate microplastic toxicity or other abiotic stresses in maize seedlings, and its application potential is enormous. Furthermore, through comprehensive standard gel analysis, this invention also proposes two methods to alleviate 0.50% PS microplastic toxicity in maize seedlings: soaking seeds in a 75 μmol / L ZnONPs solution or foliar spraying; and soaking seeds in a 12.5 mg / L GO solution or foliar spraying with a 5.0 mg / L GO solution. This provides an effective strategy for mitigating PS microplastic toxicity in maize seedlings during production, offers technical reference for the comprehensive management of microplastics in farmland ecosystems, and ensures the safe production of maize seedlings under PS microplastic toxicity conditions. Simultaneously, it achieves comprehensive management of microplastic pollution, demonstrating significant economic, ecological, and social benefits, and possessing immense application value.

[0006] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0007] 1. A comprehensive evaluation method for mitigating the toxicity of polystyrene microplastics in corn seedlings, the specific steps of which are as follows:

[0008] (1) Preparation of high-quality corn seeds: Prepare high-quality new corn genotype seeds that are uniform in size, plump, vigorous and pure, harvested from the field in the current year, for later use.

[0009] (2) Preparation of different types of solutions: A 0.50% PS microplastic solution and a 150 μmol / L nano-sized zinc oxide (ZnONPs) stock solution were prepared, and then diluted to prepare six ZnONPs solutions of different concentrations: 25 μmol / L, 50 μmol / L, 75 μmol / L, 100 μmol / L, 125 μmol / L, and 150 μmol / L. 0.50 g of PS microplastic was added to 100 mL of each ZnONPs solution to dissolve the microplastics, resulting in 0.50% PS + 25 μmol / L ZnONPs, 0.50% PS + 50 μmol / L ZnONPs, 0.50% PS + 75 μmol / L ZnONPs, 0.50% PS + 100 μmol / L ZnONPs, 0.50% PS + 125 μmol / L ZnONPs, and 0.50% PS + 150 μmol / L ZnONPs. Six PS and ZnONPs mixed solutions with ZnONPs concentrations were prepared fresh for each use. In addition, six monolayer graphene oxide (GO) solutions with concentrations of 2.5 mg / L, 5.0 mg / L, 7.5 mg / L, 10.0 mg / L, 12.5 mg / L, and 15.0 mg / L were prepared. 0.50 g of PS microplastics was added to 100 mL of each GO solution of the corresponding concentration to dissolve them, resulting in six PS and GO mixed solutions with concentrations of 0.50% PS + 2.5 mg / L GO, 0.50% PS + 5.0 mg / L GO, 0.50% PS + 7.5 mg / L GO, 0.50% PS + 10.0 mg / L GO, 0.50% PS + 12.5 mg / L GO, and 0.50% PS + 15.0 mg / L GO, prepared fresh for each use.

[0010] (3) Experiment on the 7-day growth of corn seedlings treated with different types of ZnONPs / GO solutions to alleviate polystyrene (PS) microplastic toxicity: High-quality corn genotype seeds were disinfected with 70% ethanol (v / v) for 10 min, rinsed with ddH2O water 5 times, and the water adhering to the seed surface was dried with sterile filter paper to obtain the corresponding disinfected corn genotype seeds. The disinfected corn genotype seeds were then soaked in ddH2O water from step (2); 0.50% PS microplastic solution; 6 kinds of PS and ZnONPs mixtures; 6 kinds of PS and GO mixtures, for a total of 14 types of solutions for 24 h. Simultaneously, two concentrations of PS microplastics were prepared in the soil culture medium: a 0.00% PS microplastic soil culture medium (i.e., 0.00g PS microplastics thoroughly mixed with 300.00g nutrient soil, then 250mL ddH2O water was added and stirred thoroughly), and a 0.50% PS microplastic soil culture medium (i.e., 1.50g PS microplastics thoroughly mixed with 300.00g nutrient soil, then 250mL ddH2O water was added and stirred thoroughly). Ten maize seeds of each of the 14 different solutions were then sown in pots containing the corresponding concentrations of PS microplastics in the soil culture medium and placed in an artificial climate chamber for cultivation. During cultivation, the relative humidity was set at 65%, the temperature was set at 25±0.5℃ / 20±0.5℃ alternating for 12 hours each, the photoperiod was set at 16 / 8 hours of light / dark, and the light intensity was set at 300μM m². -2 s -1The CO2 concentration was 450 PPM. In total, there were 14 treatments in the experiment: CK control (soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate), 0.50% PS treatment (soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate), 0.50% PS + 25 μmol / L ZnONPs treatment (soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate), 0.50% PS + 50 μmol / L ZnONPs treatment (soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate), 0.50% PS + 75 μmol / L ZnONPs treatment, and 0.50% PS + 75 μmol / L ZnONPs treatment. ZnONPs treatment (seed soaking in a mixture of 0.50% PS and 75 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate), 0.50% PS + 100 μmol / L ZnONPs treatment (seed soaking in a mixture of 0.50% PS and 100 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate), 0.50% PS + 125 μmol / L ZnONPs treatment (seed soaking in a mixture of 0.50% PS and 125 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate), 0.50% PS + 150 μmol / L ZnONPs treatment (seed soaking in a mixture of 0.50% PS and 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate), 0.50% PS + 2.5 mg / L GO treatment (soaking in a mixture of 0.50% PS and 2.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate), 0.50% PS + 5.0 mg / L GO treatment (soaking in a mixture of 0.50% PS and 5.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate), 0.50% PS + 7.5 mg / L GO treatment (soaking in a mixture of 0.50% PS and 7.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate), 0.50% PS + 10.0 mg / L GO treatment (soaking in a mixture of 0.50% PS and 10.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate), 0.50% PS + 12.5 mg / L GO treatment (0.50% PS + 12.5 mg / L GO). The treatments included: GO mixture soaking + 0.50% PS microplastic soil substrate culture; 0.50% PS + 15.0 mg / L GO treatment (0.50% PS + 15.0 mg / L GO mixture soaking + 0.50% PS microplastic soil substrate culture), with 4 biological replicates for each treatment.During the cultivation period, 50 mL of ddH2O water was evenly applied to each pot every 3 days. On the 7th day of cultivation, 12 traits of maize seedlings under 14 treatments were measured, including seedling length (SL), seedling fresh weight (SFW), seedling dry weight (SDW), root length (RL), root fresh weight (RFW), root dry weight (RDW), stem diameter (SD), root diameter (RD), total plant dry biomass (PDW), leaf chlorophyll SPAD value (SPAD), root activity (RV), and seedling vigor index (SSI). SSI was calculated according to formula (1): SSI = (SD / SL) × PDW (1). Where SSI is the seedling vigor index, SD is the stem diameter, SL is the seedling length, and PDW is the total plant dry biomass.

[0011] (4) Leaf spraying of different types of ZnONPs / GO solutions to alleviate polystyrene (PS) microplastic toxicity in corn seedlings for 21 days: Corn genotype seedlings cultured for 7 days under the corresponding 14 treatments in step (3) were cultured in an artificial climate chamber with the same environment for another 14 days. Every 2 days, the leaves of the corn genotype seedlings under the corresponding treatments were sprayed with 10 mL of each of the 14 types of solutions of the corresponding concentration in the morning, for a total of 7 leaf sprayings. In summary, a total of 14 treatments were tested: CK control treatment (seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water), 0.50% PS treatment (seed soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water), 0.50% PS + 25 μmol / L ZnONPs treatment (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 25 μmol / L ZnONPs solution), and 0.50% PS + 50 μmol / L ZnONPs treatment (seed soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 25 μmol / L ZnONPs solution). Treatments included: foliar spraying of seedlings with ZnONPs solution; treatment with 0.50% PS + 75 μmol / L ZnONPs solution (seed soaking in a mixture of 0.50% PS + 75 μmol / L ZnONPs + culture in a microplastic soil substrate + foliar spraying of seedlings with 75 μmol / L ZnONPs solution); treatment with 0.50% PS + 100 μmol / L ZnONPs solution (seed soaking in a mixture of 0.50% PS + 100 μmol / L ZnONPs + culture in a microplastic soil substrate + foliar spraying of seedlings with 100 μmol / L ZnONPs solution); and treatment with 0.50% PS + 125 μmol / L ZnONPs solution (seed soaking in a mixture of 0.50% PS + 125 μmol / L ZnONPs + culture in a microplastic soil substrate + 125 μmol / L ZnONPs solution). Treatments included: foliar spraying of seedlings with ZnONPs solution; treatment with 0.50% PS + 150 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + culture in a microplastic soil substrate + foliar spraying of seedlings with 150 μmol / L ZnONPs solution); treatment with 0.50% PS + 2.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 2.5 mg / L GO + culture in a microplastic soil substrate + foliar spraying of seedlings with 2.5 mg / L GO solution); and treatment with 0.50% PS + 5.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 5.0 mg / L GO + 0.50% PS + 5.0 mg / L GO solution).Treatments included: 50% PS microplastic soil substrate culture + foliar spraying of seedlings with 5.0 mg / L GO solution; 0.50% PS + 7.5 mg / L GO treatment (seed soaking in a mixture of 0.50% PS + 7.5 mg / L GO + foliar spraying of seedlings with 0.50% PS microplastic soil substrate culture + foliar spraying of seedlings with 7.5 mg / L GO solution); 0.50% PS + 10.0 mg / L GO treatment (seed soaking in a mixture of 0.50% PS + 10.0 mg / L GO + foliar spraying of seedlings with 0.50% PS microplastic soil substrate culture + foliar spraying of seedlings with 10.0 mg / L GO solution); 0.50% PS + 15.0 mg / L GO treatment (seed soaking in a mixture of 0.50% PS + 12.5 mg / L GO + foliar spraying of seedlings with 0.50% PS microplastic soil substrate culture + foliar spraying of seedlings with 12.5 mg / L GO solution); 0.50% PS + 15.0 mg / L GO treatment. GO treatment (seed soaking in a mixture of 0.50% PS and 15.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 15.0 mg / L GO solution), with four biological replicates for each treatment. During culture, the relative humidity was set at 65%, the temperature was set at alternating cycles of 25±0.5℃ and 20±0.5℃ for 12 h each, the photoperiod was set at 16 / 8 h light / dark, and the light intensity was set at 300 μM m. -2 s -1 The CO2 concentration was 450 PPM. During the cultivation period, 50 mL of ddH2O water was evenly poured into each pot every 3 days. On the 21st day of seedling cultivation, 12 traits of maize seedlings under these 14 treatments were measured.

[0012] (5) Statistical analysis of different experimental data: Using ExCel software, the mean and standard deviation of 12 traits of maize seedlings in step (3) soaking maize seeds in different types of ZnONPs / GO solutions to alleviate PS microplastic toxicity for 7 days and step (4) foliar spraying of different types of ZnONPs / GO solutions to alleviate PS microplastic toxicity for 21 days were determined. IBM-SPSSStatistics 19 data statistical analysis software was used to perform joint variance analysis on all traits of maize seedlings in different types of ZnONPs / GO solutions soaking seeds and foliar spraying to alleviate PS microplastic toxicity for 7 days and 21 days. Using online GENESCLOUD software, correlation coefficient diagrams of the corresponding 12 traits of maize seedlings in different types of ZnONPs / GO solutions soaking seeds and foliar spraying to alleviate PS microplastic toxicity for 7 days and 21 days were plotted. The IMLOG2 function was used to further standardize the mean values ​​of 12 traits in maize genotype seedlings treated with different types of ZnONPs / GO solutions for seed soaking and foliar spraying to alleviate PS microplastic toxicity in 7-day and 21-day growth trials. IBM-SPSS Statistics 16 data analysis software was used to perform inter-group clustering in all treatments of maize genotype seedlings treated with different types of ZnONPs / GO solutions for seed soaking and foliar spraying to alleviate PS microplastic toxicity in 7-day and 21-day growth trials. This evaluated the overall growth status of maize genotype seedlings at different cultivation stages and treatments, and thus scientifically and qualitatively identified the mitigation effect of different concentrations of ZnONPs / GO solutions on maize genotype seedlings at different cultivation stages under PS microplastic toxicity.

[0013] (6) Relative mitigation effect index of different concentrations of ZnONPs / GO solution soaking or foliar spraying on individual traits of maize seedlings under 0.50% polystyrene (PS) microplastic toxicity: In order to more scientifically, objectively and accurately measure the mitigation effect of different concentrations of ZnONPs / GO solution soaking or foliar spraying on individual traits of maize genotype seedlings under 0.50% PS microplastic toxicity, this invention newly defines the relative mitigation effect index (RMEI) of different concentrations of ZnONPs / GO solution soaking or foliar spraying on individual traits of maize seedlings under 0.50% polystyrene (PS) microplastic toxicity. ZnONPS and RMEI GO ). Formula (2): (2). In the formula The relative mitigation index of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. The values ​​for the j-th trait in maize seedlings treated with a 0.50% PS microplastic toxicity solution (i-ZnONPs) at concentration i-ZnONPs concentration, measured on day 7 or day 21 of cultivation. The ΣT represents the phenotypic value of genotype j in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity treatment. j / m represents the average value of the j-th trait in maize seedlings cultured on day 7 or day 21 after treatment with m types of solutions (m=8) for seed soaking or foliar spraying. Similarly, formula (3): and In the formula The relative mitigation index of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. The value of the jth trait in maize seedlings cultured on day 7 or day 21 after treatment with i-GO concentration solution (immersion in 0.50% PS microplastic toxicity solution) is the result of the i-GO concentration solution. The ΣT represents the phenotypic value of genotype j in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity treatment. j / n represents the average value of the jth trait in maize seedlings cultured on day 7 or day 21 after treatment with n (n=8) types of solutions for seed soaking or foliar spraying. The higher the value, the stronger the mitigating effect of soaking seeds or spraying leaves with the i-ZnONPs concentration solution on the j-th genotype of maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. The higher the value, the stronger the mitigating effect of soaking seeds or spraying leaves with the i-GO concentration solution on the j-th trait of maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity.

[0014] (7) Absolute mitigation strength coefficients (AMICs) of different concentrations of ZnONPs / GO solution for seed soaking or foliar spraying on individual traits of maize seedlings under 0.50% polystyrene (PS) microplastic toxicity: To facilitate a more scientific, simple, and objective evaluation of the mitigation strength of different concentrations of ZnONPs / GO solution for seed soaking or foliar spraying on individual traits of maize seedlings under 0.50% polystyrene (PS) microplastic toxicity, this invention also newly defines the absolute mitigation strength coefficients (AMICs) of different concentrations of ZnONPs / GO solution for seed soaking or foliar spraying on individual traits of maize seedlings under 0.50% polystyrene (PS) microplastic toxicity. ZnONPs and AMIC GO ). Formula (4): In the formula The absolute mitigation strength coefficient of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. The values ​​for the j-th trait in maize seedlings treated with a 0.50% PS microplastic toxicity solution (i-ZnONPs) at concentration i-ZnONPs concentration, measured on day 7 or day 21 of cultivation. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity treatment. The value of the j-th genotype in maize seedlings cultured on day 7 or day 21 under CK treatment. Similarly, formula (5): In the formula The absolute mitigation strength coefficient of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. The value of the jth trait in maize seedlings cultured on day 7 or day 21 after treatment with i-GO concentration solution (immersion in 0.50% PS microplastic toxicity solution) is the result of the i-GO concentration solution. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity treatment. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under CK treatment. The higher the value, the greater the mitigation effect of soaking seeds or spraying leaves with the i-ZnONPs concentration solution on the j-th genotype of maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. The higher the value, the greater the mitigation effect of soaking seeds in i-GO concentration solution or spraying leaves on the j-th phenotype of maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity.

[0015] (8) Comprehensive evaluation of the mitigating effect of different concentrations of ZnONPs / GO solution soaking seeds or foliar spraying on the toxicity of 0.50% polystyrene (PS) microplastics in maize seedlings: Further, based on the RMEI of different concentrations of ZnONPs / GO solution soaking seeds or foliar spraying on the toxicity of 0.50% PS microplastics in maize seedlings cultured for 7 days or 21 days, calculated in step (6), the RMEI of 12 traits of maize seedlings with different genotypes cultured under 0.50% PS microplastic toxicity was obtained. ZnONPS and RMEI GOThe values ​​were used as evaluation indicators for the mitigating effects of different concentrations of ZnONPs / GO solution on seed soaking or foliar spraying on maize genotype seedlings under 0.50% PS microplastic toxicity. The membership function method was employed to scientifically, objectively, comprehensively, and quantitatively evaluate the comprehensive mitigating effects of different concentrations of ZnONPs / GO solution on maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity (CME). ZnONPs and CME GO ). Formula (6): In the formula: Membership value for mitigation effect of soaking seeds or spraying leaves with i-ZnONPs concentration solution on the j-th trait of maize seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity. RMEI is the relative mitigation index of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity, calculated by soaking seeds or spraying leaves with i-ZnONPs concentration solution. j-max RMEI is the maximum relative mitigation index of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity, measured by soaking seeds or foliar spraying with ZnONPs solutions at all concentrations. j-min The minimum relative mitigation index for the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity, obtained by soaking seeds or foliar spraying with ZnONPs solutions at all concentrations. Formula (7): In the formula, CME i-ZnONPs The overall mitigation effect of soaking seeds or spraying leaves with i-ZnONPs concentration solution on maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity is shown, where k (k=12) represents the number of traits measured. The membership value of the mitigation effect of soaking seeds or spraying leaves with i-ZnONPs concentration solution on the j-th trait of maize seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity. Formula (8): In the formula: Membership value for mitigation effect of soaking seeds or foliar spraying with i-GO concentration solution on the j-th trait of maize seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity. The RMEI (Relative Mitigation Index) is the index of the relative mitigation effect of soaking seeds or foliar spraying with i-GO concentration solution on the j-th trait of maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. j-maxRMEI is the maximum relative mitigation index of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity, measured by soaking seeds or foliar spraying with GO solution at all concentrations. j-min The minimum relative mitigation index for the j-th genotype of maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity, obtained by soaking seeds or foliar spraying with GO solution at all concentrations. Formula (9): In the formula, CME i-GO The overall mitigation strength of soaking seeds or foliar spraying with i-GO concentration solution on maize genotype seedlings cultured at 0.50% PS microplastic toxicity on day 7 or day 21 is given, where k (k=12) represents the number of traits measured. Membership value for mitigation effect of soaking seeds or foliar spraying with i-GO concentration solution on the j-th trait of maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity. CME i -ZnONPs The higher the value, the stronger the overall mitigating effect of soaking seeds or spraying leaves with the i-ZnONPs concentration solution on maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity.

[0016] (9) Comprehensive evaluation of the mitigation strength of different concentrations of ZnONPs / GO solution soaking or foliar spraying on maize seedlings under 0.50% polystyrene (PS) microplastic toxicity: Further, based on the AMIC of different concentrations of ZnONPs / GO solution soaking or foliar spraying on maize seedlings under 0.50% PS microplastic toxicity on day 7 or day 21 of culture, the AMIC of 12 traits of maize genotype seedlings under 0.50% PS microplastic toxicity was calculated in step (7). ZnONPS and AMIC GO The values ​​were used as evaluation indicators for the mitigation intensity of different concentrations of ZnONPs / GO solution applied to seeds or leaves to maize genotype seedlings under 0.50% PS microplastic toxicity. The membership function method was employed to comprehensively, systematically, objectively, and quantitatively evaluate the overall mitigation intensity (CMI) of different concentrations of ZnONPs / GO solution applied to seeds or leaves to maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity. ZnONPs and CMI GO ). Formula (10): In the formula: The membership value of the mitigation strength of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. The absolute mitigation strength coefficient (AMIC) of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. j-max The maximum absolute mitigation strength coefficient (AMIC) for the j-th trait in maize genotype seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with ZnONPs solutions at all concentrations. j-min The minimum absolute mitigation strength coefficient for the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with ZnONPs solution at all concentrations. Formula (11): In the formula, CMI i-ZnONPs The overall mitigation strength of soaking seeds or foliar spraying with i-ZnONPs concentration solution on maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity is determined, where k (k=12) represents the number of traits measured. The membership value of the mitigation strength of the jth trait in maize seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. Formula (12): In the formula: The membership value of the mitigation strength of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. The absolute mitigation strength coefficient (AMIC) of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or foliar spraying with i-GO concentration solution. j-max The maximum absolute mitigation strength coefficient (AMIC) for seed or foliar spraying with GO solution at all concentrations against the j-th genotype of maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. j-min The minimum absolute mitigation strength coefficient for the j-th genotype of maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with GO solution of all concentrations. Formula (13): In the formula, CMI i-GO The overall mitigation strength of soaking seeds or foliar spraying with i-GO concentration solution on maize genotype seedlings cultured at 0.50% PS microplastic toxicity on day 7 or day 21 is given, where k (k=12) represents the number of traits measured. CMI (Combined Minerals Index) is the membership value for mitigation strength of the j-th trait in maize seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity after soaking seeds or spraying leaves with i-GO concentration solution. i-ZnONPs The higher the value, the greater the overall mitigation effect of soaking seeds or foliar spraying with the i-ZnONPs concentration solution on maize genotype seedlings cultured for 7 or 21 days under 0.50% PS microplastic toxicity. CMI i-GO The higher the value, the greater the overall mitigation effect of soaking seeds or spraying leaves with the i-GO concentration solution on maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity.

[0017] (10) Optimal ZnONPs / GO solution concentration for best mitigation effect / maximum mitigation intensity of 0.50% polystyrene (PS) microplastic toxicity in maize seedlings: Further evaluation was conducted by comparing the intergroup clustering results of the growth status of maize genotype seedlings in all treatments during the 7-day and 21-day growth trials of different types of ZnONPs / GO solutions for mitigating PS microplastic toxicity, and by comparing the mitigation effect of different concentrations of ZnONPs / GO solutions for mitigating 0.50% PS microplastic toxicity in maize seedlings. The comprehensive evaluation results of the effects and the comprehensive evaluation results of the mitigation intensity of different concentrations of ZnONPs / GO solution soaking seeds or foliar spraying on maize genotype seedlings with 0.50% concentration of PS microplastics were comprehensively compared and analyzed. The mitigation effects of different concentrations of ZnONPs / GO solution soaking seeds or foliar spraying on maize genotype seedlings with 0.50% concentration of PS microplastics at different cultivation periods were finally determined. The optimal concentration of ZnONPs / GO solution for seed soaking or foliar spraying was determined as the best mitigation effect / maximum mitigation intensity of 0.50% concentration of PS microplastics in maize genotype seedlings at different cultivation periods.

[0018] 2. The application of nano-grade zinc oxide (ZnONPs) in mitigating the toxicity of PS microplastics in corn seedlings is as follows:

[0019] Spraying corn seedlings with 10 mL of 75 μmol / L ZnONPs solution every 2 days for 7 consecutive days, after 7 days of microplastic PS toxicity, can significantly reduce the toxicity of microplastics to corn seedlings.

[0020] Forty high-quality maize seeds sterilized with 70% ethanol (v / v) were soaked in 100 mL of a mixed solution of 0.50% PS and 75 μmol / L ZnONPs for 24 h. Ten of these soaked seeds were then sown in pots containing 0.50% PS microplastic substrate and cultured in an artificial climate chamber. On day 7 of seedling culture, every two days, the leaves of each pot were sprayed with 10 mL of a 75 μmol / L ZnONPs solution in the morning. This process was continued for 14 days, for a total of 7 foliar sprays. During the 21-day culture period, the relative humidity was set at 65%, the temperature was alternated between 25±0.5℃ and 20±0.5℃ for 12 h, the photoperiod was 16 / 8 h light / dark, and the light intensity was 300 μM m². -2 s -1 The CO2 concentration was 450 PPM; simultaneously, 50 mL of ddH2O water was evenly applied to each pot every 3 days. Then, 12 traits of maize genotype seedlings were measured on days 7 and 21 of culture. The experiment was performed in four biological replicates.

[0021] 3. The application of single-layer graphene oxide (GO) in mitigating microplastic toxicity in corn seedlings is as follows:

[0022] Spraying corn seedlings with 1 mL of 5.0 mg / L monolayer graphene oxide solution every 2 days for 14 consecutive days, for a total of 7 sprays, can significantly reduce the toxicity of microplastics to corn seedlings.

[0023] Forty high-quality new maize seeds of the corresponding genotype, sterilized with 70% ethanol (v / v), were soaked in 100 mL of a mixed solution of 0.50% PS and 12.5 mg / L GO for 24 hours. Ten of these soaked seeds were then sown in pots containing 0.50% PS microplastic substrate and cultured in an artificial climate chamber. On day 7 of seedling culture, the leaves of each pot were sprayed with 10 mL of a 5.0 mg / L GO solution every two days in the morning. This process was continued for 14 days, for a total of 7 foliar sprays. During the 21-day culture period, the relative humidity was set at 65%, the temperature was alternated between 25±0.5℃ and 20±0.5℃ for 12 hours each, the photoperiod was 16 / 8 hours of light / dark, and the light intensity was 300 μMm. -2 s -1 The CO2 concentration was 450 PPM; simultaneously, 50 mL of ddH2O water was evenly applied to each pot every 3 days. Then, 12 traits of maize genotype seedlings were measured on days 7 and 21 of culture. The experiment was performed in four biological replicates.

[0024] The beneficial effects of this invention are:

[0025] PS microplastics are among the most significant microplastic pollutants in the environment, and the application of nanomaterials is expected to create a new type of productivity in the agricultural field. This invention systematically studies the effects of different concentrations of ZnONPs / GO solution soaking in seeds or foliar spraying on 12 traits, including growth phenotype, leaf chlorophyll accumulation, and root activity, in maize seedlings grown for 7 or 21 days under 0.50% PS microplastic toxicity. Furthermore, it systematically reveals the mechanism by which ZnONPs / GO drives maize seedlings to overcome PS microplastic toxicity and scientifically and innovatively proposes a relative mitigation effect index (RMEI) for individual traits in maize seedlings under 0.50% PS microplastic toxicity caused by different concentrations of ZnONPs / GO solution soaking in seeds or foliar spraying. ZnONPS and RMEI GO The absolute mitigation intensity coefficient (AMIC) of individual traits in maize seedlings under 0.50% PS microplastic toxicity was determined by soaking seeds or foliar spraying with different concentrations of ZnONPs / GO solutions. ZnONPs and AMIC GO The biological concepts and calculation formulas of ZnONPs / GO solution were used as evaluation indicators to assess the mitigating effects and intensity of different concentrations of ZnONPs / GO solution on maize seedlings exposed to 0.50% PS microplastic toxicity. The membership function method was employed to scientifically, objectively, comprehensively, and quantitatively evaluate the comprehensive mitigating effects of different concentrations of ZnONPs / GO solution on maize seedlings cultured for 7 or 21 days under 0.50% PS microplastic toxicity (CME). ZnONPs and CME GO ) and Comprehensive Mitigation Intensity (CMI) ZnONPs and CMI GO This invention, through comprehensive comparison, ultimately yielded an application method for mitigating the toxicity of 0.50% PS microplastics in maize seedlings by soaking seeds in an optimal concentration of ZnONPs / GO solution or by foliar spraying. This invention not only provides a scientific, objective, and comprehensive evaluation method for mitigating PS microplastics in maize seedlings, but also offers an effective strategy for mitigating PS microplastic toxicity in maize seedlings using nano-ZnONPs / GO in production. The method is simple to operate, has good repeatability, requires a small dosage of nano-ZnONPs / GO, and is economical. In production, nano-ZnONPs / GO shows good mitigation effects on PS microplastic toxicity in maize seedlings, and is also environmentally friendly. It can effectively solve microplastic pollution in farmland ecosystems, ensure national food security, and has many other beneficial effects, possessing significant production application value and generating substantial ecological, social, and economic benefits. Attached Figure Description

[0026] Figure 1Formula (1) for calculating the seedling vigor index (SSI) of maize genotype seedlings: The relative mitigation effect index of the jth trait of maize genotype seedlings cultured on the 7th or 21st day under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with i-ZnONPs concentration solution. Formula (2) is used to calculate the relative mitigation index of the jth trait in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with i-GO concentration solution. Formula (3) is used to calculate the absolute mitigation strength coefficient of the jth trait in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with i-ZnONPs concentration solution. Formula (4) is used to calculate the absolute mitigation strength coefficient of the jth trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with i-GO concentration solution. Formula (5) is used to calculate the membership value of the mitigation effect of soaking seeds or spraying leaves with the i-th ZnONPs concentration solution on the j-th trait of maize seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity. Calculation formula (6) shows the comprehensive mitigation effect of soaking seeds or spraying leaves with i-ZnONPs concentration solution on maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity (CME). i-ZnONPs Formula (7) is used to calculate the membership value of the mitigation effect of soaking seeds or spraying leaves with i-GO concentration solution on the j-th trait of maize genotype seedlings cultured on the 7th or 21st day of culture under 0.50% PS microplastic toxicity. Formula (8) is used to calculate the overall mitigation strength (CME) of seed soaking or foliar spraying with i-GO concentration solution on maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. i-GO Formula (9) is used to calculate the mitigation strength membership value of the jth trait in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with i-ZnONPs concentration solution. Formula (10) is used to calculate the overall mitigation intensity (CMI) of i-ZnONPs concentration solution applied to seeds or leaves during foliar spraying against maize seedlings cultured at 0.50% PS microplastic toxicity on day 7 or day 21. i-ZnONPs Formula (11) is used to calculate the mitigation strength membership value of the jth trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with i-GO concentration solution. Formula (12) is used to calculate the overall mitigation strength (CMI) of i-GO concentration solution soaking seeds or foliar spraying against maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. i-GO ) Calculation formula (13).

[0027] Figure 2 Seedling growth of TS163 maize genotype seedlings was investigated during a 7-day experiment to mitigate the toxicity of 0.50% PS microplastics by soaking seeds in ZnONPs solutions of different concentrations.

[0028] The control treatment (CK) in the 7-day seedling growth experiment (seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate), the treatment with 0.50% PS in the 7-day seedling growth experiment (seed soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate), the treatment with 0.50% PS + 25 μmol / L ZnONPs in the 7-day seedling growth experiment (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate), and the treatment with 0.50% PS + 50 μmol / L ZnONPs in the 7-day seedling growth experiment (0.50% PS + 50 μmol / L ZnONPs) Treatments for 7-day seedling growth were as follows: 0.50% PS + 75 μmol / L ZnONPs (soil-based culture), 0.50% PS + 100 μmol / L ZnONPs (soil-based culture), and 0.50% PS + 125 μmol / L ZnONPs. ZnONPs treatment (seed soaking in a mixture of 0.50% PS and 125 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate), and 0.50% PS + 150 μmol / L ZnONPs treatment for seedling growth 7 days (seed soaking in a mixture of 0.50% PS and 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate).

[0029] Figure 3 Leaf spraying of ZnONPs solutions of different concentrations to alleviate the toxicity of 0.50% PS microplastics during the 21-day growth of TS163 maize genotype seedlings.

[0030] The control treatment (CK) for the 21-day seedling growth experiment was seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water. The treatments included: 0.50% PS (seed soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water); 0.50% PS + 25 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 25 μmol / L ZnONPs solution); and 0.50% PS + 50 μmol / L ZnONPs. ZnONPs were used in a 21-day seedling growth experiment with treatments including: 0.50% PS + 50 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 50 μmol / L ZnONPs solution); 0.50% PS + 75 μmol / L ZnONPs were used in a 21-day seedling growth experiment with treatments including: 0.50% PS + 100 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 75 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 75 μmol / L ZnONPs solution); and 0.50% PS + 100 μmol / L ZnONPs were used in a 21-day seedling growth experiment with treatments including: 0.50% PS + 100 μmol / L ZnONPs. ZnONPs treatments (seed soaking in a mixture of 0.50% PS and 100 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 100 μmol / L ZnONPs solution), 0.50% PS + 125 μmol / L ZnONPs treatments for a 21-day seedling growth experiment, 0.50% PS + 150 ... Seedlings were soaked in a mixture of ZnONPs, cultured in a soil substrate of 0.50% PS microplastics, and foliar sprayed with a 150 μmol / L ZnONPs solution.

[0031] Figure 4Analysis of 12 phenotypic changes in TS163 maize genotype seedlings during 7-day or 21-day seedling growth trials of seedlings treated with different concentrations of ZnONPs solution soaking or foliar spraying to alleviate the toxicity of 0.50% PS microplastics.

[0032] CK represents the control treatment in the 7-day seedling growth experiment (seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate) or the control treatment in the 21-day seedling growth experiment (seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water); 0.50% PS represents the 0.50% PS treatment in the 7-day seedling growth experiment (seed soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate) or the 0.50% PS treatment in the 21-day seedling growth experiment (seed soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water); 0.50% PS + 25 μmol / L ZnONPs represents the 0.50% PS + 25 μmol / L ZnONPs treatment in the 7-day seedling growth experiment. ZnONPs treatment (seed soaking in a mixture of 0.50% PS and 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate) or foliar spraying of seedlings with a 25 μmol / L ZnONPs solution in a 21-day seedling growth test; 0.50% PS + 50 μmol / L ZnONPs treatment (seed soaking in a mixture of 0.50% PS and 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 25 μmol / L ZnONPs solution); 0.50% PS + 50 μmol / L ZnONPs treatment in a 7-day seedling growth test (seed soaking in a mixture of 0.50% PS and 50 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate) or foliar spraying of seedlings with a 25 μmol / L ZnONPs solution in a 21-day seedling growth test. ZnONPs treatment (seed soaking in a mixture of 0.50% PS and 50 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 50 μmol / L ZnONPs solution), 0.50% PS + 75 μmol / L ZnONPs treatment for 7-day seedling growth (seed soaking in a mixture of 0.50% PS and 75 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate) or 0.50% PS + 100 μmol / L ZnONPs treatment for 21-day seedling growth (seed soaking in a mixture of 0.50% PS and 75 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 75 μmol / L ZnONPs solution), 0.50% PS + 100 μmol / L ZnONPs treatment ZnONPs were used in the 7-day seedling growth test with 0.50% PS + 100 μmol / L ZnONPs treatment (seed soaking in a mixture of 0.50% PS + 100 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate) or in the 21-day seedling growth test with 0.Treatment with 50% PS + 100 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 100 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 100 μmol / L ZnONPs solution), 0.50% PS + 125 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 125 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate) for 7-day seedling growth trials, or 0.50% PS + 125 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 125 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + 125 μmol / L ZnONPs solution) (for 21-day seedling growth trials) (ZnONPs solution foliar spraying of seedlings), 0.50% PS + 150 μmol / L ZnONPs treatment for 7-day seedling growth (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate) or 0.50% PS + 150 μmol / L ZnONPs treatment for 21-day seedling growth (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 150 μmol / L ZnONPs solution). SL represents seedling length, SFW represents seedling fresh weight, SDW represents seedling dry weight, RL represents root length, RFW represents root fresh weight, RDW represents root dry weight, SD represents stem diameter, RD represents root diameter, PDW represents total plant dry biomass, SPAD represents leaf chlorophyll SPAD value, SSI represents seedling vigor index, and RV represents root activity. Different lowercase letters indicate that the difference in a single trait between different treatments was statistically significant at the P<0.05 level in the 7-day or 21-day seedling growth experiment.

[0033] Figure 5 Seedling growth of TS163 maize genotype seedlings was investigated during a 7-day experiment to mitigate the toxicity of 0.50% PS microplastics by soaking seeds in GO solutions of different concentrations.

[0034] The control treatment (CK) for the 7-day seedling growth experiment was seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate. The treatments included: 0.50% PS (0.50% PS microplastic solution soaking + 0.50% PS microplastic soil substrate); 0.50% PS + 2.5 mg / L GO (0.50% PS + 2.5 mg / L GO mixture soaking + 0.50% PS microplastic soil substrate); 0.50% PS + 5.0 mg / L GO (0.50% PS + 5.0 mg / L GO mixture soaking + 0.50% PS microplastic soil substrate); and 0.50% PS + 7.5 mg / L GO. GO was used in the 7-day seedling growth experiment for treatments including: 0.50% PS + 7.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 7.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 10.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 10.0 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 12.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 12.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); and 0.50% PS + 15.0 mg / L GO. GO was used in a 7-day seedling growth experiment, consisting of a 0.50% PS + 15.0 mg / L GO treatment (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate).

[0035] Figure 6 Foliar spraying of different concentrations of GO solution to alleviate the toxicity of 0.50% PS microplastics during the 21-day growth of TS163 maize genotype seedlings.

[0036] The control treatment (CK) for the 21-day seedling growth experiment consisted of: seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water; the treatment with 0.50% PS for the 21-day seedling growth experiment consisted of: seed soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water; the treatment with 0.50% PS + 2.5 mg / L GO for the 21-day seedling growth experiment consisted of: seed soaking in a mixture of 0.50% PS + 2.5 mg / L GO + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 2.5 mg / L GO solution; and the treatment with 0.50% PS + 5.0 mg / L GO for the 21-day seedling growth experiment consisted of: 0.50% PS + 5.0 mg / L GO. GO treatment (seed soaking in a mixture of 0.50% PS and 5.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 5.0 mg / L GO solution), 0.50% PS + 7.5 mg / L GO treatment for 21-day seedling growth, 0.50% PS + 10.0 mg / L GO treatment for 21-day seedling growth, 0.50% PS + 10.0 mg / L GO treatment for 21-day seedling growth, 0.50% PS + 10.0 mg / L GO treatment for 21-day seedling growth, 0.50% PS + 10.0 mg / L GO treatment, 0.50% PS microplastic soil substrate + 10.0 mg / L GO solution. The treatments included: foliar spraying of seedlings with GO solution; 0.50% PS + 12.5 mg / L GO treatment for 21-day seedling growth (seed soaking in a mixture of 0.50% PS + 12.5 mg / L GO + culturing in a microplastic soil substrate with 0.50% PS + foliar spraying of seedlings with 12.5 mg / L GO solution); and 0.50% PS + 15.0 mg / L GO treatment for 21-day seedling growth (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culturing in a microplastic soil substrate with 0.50% PS + foliar spraying of seedlings with 15.0 mg / L GO solution).

[0037] Figure 7 Analysis of 12 phenotypic changes in TS163 maize genotype seedlings during 7-day or 21-day seedling growth trials of seedlings treated with different concentrations of GO solution for seed soaking or foliar spraying to alleviate the toxicity of 0.50% PS microplastics.

[0038] CK represents the control treatment in the 7-day seedling growth experiment (seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate) or the control treatment in the 21-day seedling growth experiment (seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water); 0.50% PS represents the 0.50% PS treatment in the 7-day seedling growth experiment (seed soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate) or the 0.50% PS treatment in the 21-day seedling growth experiment (seed soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water); 0.50% PS + 2.5 mg / L GO represents the 0.50% PS + 2.5 mg / L GO treatment in the 7-day seedling growth experiment (0.50% PS + 2.5 mg / L GO). The following treatments were used in a 21-day seedling growth trial: a 0.50% PS + 2.5 mg / L GO mixture (seed soaking in a 0.50% PS + 2.5 mg / L GO mixture + 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 2.5 mg / L GO solution); a 7-day seedling growth trial using a 0.50% PS + 5.0 mg / L GO mixture (seed soaking in a 0.50% PS + 5.0 mg / L GO mixture + 0.50% PS microplastic soil substrate); or a 21-day seedling growth trial using a 0.50% PS + 5.0 mg / L GO mixture (seed soaking in a 0.50% PS + 5.0 mg / L GO mixture + 0.50% PS microplastic soil substrate + 5.0 mg / L GO solution). The treatments included: foliar spraying of seedlings with GO solution; 0.50% PS + 7.5 mg / L GO treatment for 7-day seedling growth (seed soaking in a mixture of 0.50% PS + 7.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate); foliar spraying of seedlings with 7.5 mg / L GO solution for 21-day seedling growth; and 0.50% PS + 10.0 mg / L GO treatment for 7-day seedling growth (0.50% PS + 10.0 mg / L GO + 0.0 mg / L GO + 0.50% PS + 1 ... The treatments included: soaking seeds in a GO mixture followed by cultivation in a 0.50% PS microplastic soil substrate; foliar spraying of seedlings with 0.50% PS + 10.0 mg / L GO solution for 21 days of seedling growth; and foliar spraying of seedlings with 0.50% PS + 10.0 mg / L GO + 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 10.0 mg / L GO solution for 7 days of seedling growth.Treatment with 50% PS + 12.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 12.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or foliar spraying of seedlings with 12.5 mg / L GO solution for 21-day seedling growth trials; treatment with 0.50% PS + 15.0 mg / L GO for 7-day seedling growth trials (seed soaking in a mixture of 0.50% PS + 12.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or foliar spraying of seedlings with 12.5 mg / L GO solution for 21-day seedling growth trials (0.50% PS + 12.5 mg / L GO ... Seeds were soaked in a mixture of GO and cultured in a 0.50% PS microplastic soil substrate, followed by foliar spraying of seedlings with a 15.0 mg / L GO solution. SL represents seedling length, SFW represents seedling fresh weight, SDW represents seedling dry weight, RL represents root length, RFW represents root fresh weight, RDW represents root dry weight, SD represents stem diameter, RD represents root diameter, PDW represents total plant dry biomass, SPAD represents leaf chlorophyll SPAD value, SSI represents seedling vigor index, and RV represents root activity. Different lowercase letters indicate significant differences (P < 0.05) in individual traits between different treatments in 7-day or 21-day seedling growth trials.

[0039] Figure 8 Pearson correlation analysis of 12 traits in a 7-day growth experiment of TS163 maize genotype seedlings, using ZnONPs solutions of different concentrations to alleviate the toxicity of 0.50% PS microplastics.

[0040] Wherein SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. "*" indicates that the two traits are significantly correlated at the P<0.05 level.

[0041] Figure 9 Pearson correlation analysis of 12 traits in a 21-day growth experiment of TS163 maize genotype seedlings to alleviate the toxicity of 0.50% PS microplastics by foliar spraying with different concentrations of ZnONPs solutions.

[0042] Wherein SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. "*" indicates that the two traits are significantly correlated at the P<0.05 level.

[0043] Figure 10 Pearson correlation analysis of 12 traits in a 7-day growth experiment of TS163 maize genotype seedlings, using different concentrations of GO solution to alleviate the toxicity of 0.50% PS microplastics.

[0044] Wherein SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. "*" indicates that the two traits are significantly correlated at the P<0.05 level.

[0045] Figure 11 Pearson correlation analysis of 12 traits in TS163 maize genotype seedlings during a 21-day growth experiment of foliar spraying with different concentrations of GO solution to alleviate the toxicity of 0.50% PS microplastics.

[0046] Wherein SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. "*" indicates that the two traits are significantly correlated at the P<0.05 level.

[0047] Figure 12 A comprehensive evaluation of cluster analysis among all treatment groups in a 7-day growth experiment of TS163 maize genotype seedlings was conducted by soaking seeds in ZnONPs solutions of different concentrations to mitigate the toxicity of 0.50% PS microplastics.

[0048] CK was the control treatment in the 7-day seedling growth experiment (seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate); 0.50% PS was the 0.50% PS treatment in the 7-day seedling growth experiment (seed soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 25 μmol / L ZnONPs was the 0.50% PS + 25 μmol / L ZnONPs treatment in the 7-day seedling growth experiment (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate); and 0.50% PS + 50 μmol / L ZnONPs was the 0.50% PS + 50 μmol / L ZnONPs treatment in the 7-day seedling growth experiment (0.50% PS + 50 μmol / L ZnONPs). Treatments included: a mixture of ZnONPs for seed soaking + 0.50% PS microplastic soil substrate; 0.50% PS + 75 μmol / L ZnONPs for seedling growth (7-day seedling growth test); 0.50% PS + 100 μmol / L ZnONPs for seedling growth (7-day seedling growth test); and 0.50% PS + 125 μmol / L ZnONPs. ZnONPs were used in a 7-day seedling growth experiment with two treatments: 0.50% PS + 125 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 125 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate) and 0.50% PS + 150 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate). Treatment I represents good corn seedling growth, treatment II represents weak corn seedling growth, and treatment III represents poor corn seedling growth.

[0049] Figure 13 Foliar spraying of different concentrations of ZnONPs solutions to alleviate microplastic toxicity of 0.50% PS at TS163 maize genotype for 21 days was evaluated by cluster analysis among all treatment groups.

[0050] The control treatment (CK) for the 21-day seedling growth experiment was seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water. The treatments included: 0.50% PS (seed soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water); 0.50% PS + 25 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 25 μmol / L ZnONPs solution); and 0.50% PS + 50 μmol / L ZnONPs. ZnONPs were used in a 21-day seedling growth experiment with treatments including: 0.50% PS + 50 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 50 μmol / L ZnONPs solution); 0.50% PS + 75 μmol / L ZnONPs were used in a 21-day seedling growth experiment with treatments including: 0.50% PS + 100 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 75 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 75 μmol / L ZnONPs solution); and 0.50% PS + 100 μmol / L ZnONPs were used in a 21-day seedling growth experiment with treatments including: 0.50% PS + 100 μmol / L ZnONPs. ZnONPs treatments (seed soaking in a mixture of 0.50% PS and 100 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 100 μmol / L ZnONPs solution), 0.50% PS + 125 μmol / L ZnONPs treatments for a 21-day seedling growth experiment, 0.50% PS + 150 ... The treatment regimen consisted of soaking seeds in a ZnONPs mixture, culturing in a 0.50% PS microplastic soil substrate, and foliar spraying of seedlings with a 150 μmol / L ZnONPs solution. Treatment I represented good seedling growth, treatment II represented weak seedling growth, and treatment III represented poor seedling growth.

[0051] Figure 14 A comprehensive evaluation of inter-treatment clusters was conducted on the 7-day growth of TS163 maize genotype seedlings by soaking seeds in GO solutions of different concentrations to mitigate the toxicity of 0.50% PS microplastics.

[0052] The control treatment (CK) for the 7-day seedling growth experiment was seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate. The treatments included: 0.50% PS (0.50% PS microplastic solution soaking + 0.50% PS microplastic soil substrate); 0.50% PS + 2.5 mg / L GO (0.50% PS + 2.5 mg / L GO mixture soaking + 0.50% PS microplastic soil substrate); 0.50% PS + 5.0 mg / L GO (0.50% PS + 5.0 mg / L GO mixture soaking + 0.50% PS microplastic soil substrate); and 0.50% PS + 7.5 mg / L GO. GO was used in the 7-day seedling growth experiment for treatments including: 0.50% PS + 7.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 7.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 10.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 10.0 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 12.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 12.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); and 0.50% PS + 15.0 mg / L GO. GO represents the treatment of 0.50% PS + 15.0 mg / L GO in a 7-day seedling growth experiment (seeds soaked in a mixture of 0.50% PS and 15.0 mg / L GO + cultured in a 0.50% PS microplastic soil substrate). Treatment I represents the type of maize seedling with good growth, treatment II represents the type of maize seedling with weak growth, and treatment III represents the type of maize seedling with poor growth.

[0053] Figure 15 Foliar spraying of different concentrations of GO solution to alleviate the toxicity of 0.50% PS microplastics in TS163 maize genotype seedlings for 21 days: comprehensive evaluation of clustering among all treatment groups.

[0054] The control treatment (CK) for the 21-day seedling growth experiment consisted of: seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water; the treatment with 0.50% PS for the 21-day seedling growth experiment consisted of: seed soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water; the treatment with 0.50% PS + 2.5 mg / L GO for the 21-day seedling growth experiment consisted of: seed soaking in a mixture of 0.50% PS + 2.5 mg / L GO + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 2.5 mg / L GO solution; and the treatment with 0.50% PS + 5.0 mg / L GO for the 21-day seedling growth experiment consisted of: 0.50% PS + 5.0 mg / L GO. GO treatment (seed soaking in a mixture of 0.50% PS and 5.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 5.0 mg / L GO solution), 0.50% PS + 7.5 mg / L GO treatment for 21-day seedling growth, 0.50% PS + 10.0 mg / L GO treatment for 21-day seedling growth, 0.50% PS + 10.0 mg / L GO treatment for 21-day seedling growth, 0.50% PS + 10.0 mg / L GO treatment for 21-day seedling growth, 0.50% PS + 10.0 mg / L GO treatment, 0.50% PS microplastic soil substrate + 10.0 mg / L GO solution. Treatments included: foliar spraying of seedlings with GO solution; 0.50% PS + 12.5 mg / L GO treatment for 21-day seedling growth (seed soaking in a mixture of 0.50% PS + 12.5 mg / L GO + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 12.5 mg / L GO solution); and 0.50% PS + 15.0 mg / L GO treatment for 21-day seedling growth (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 15.0 mg / L GO solution). Treatment I represented good seedling growth, treatment II represented weak seedling growth, and treatment III represented poor seedling growth.

[0055] Figure 16 The overall mitigation effect of different concentrations of ZnONPs solution soaking in seeds or foliar spraying on TS163 maize genotype seedlings on day 7 or day 21 of growth of 0.50% PS microplastic toxicity (CME)ZnONPs )analyze.

[0056] 7d refers to the 7th day of growth of the maize genotype seedlings, and 21d refers to the 21st day of growth of the maize genotype seedlings. 0.50% PS + 25 μmol / L ZnONPs was used for seedling growth trials at 7 days (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate) or for seedling growth trials at 21 days (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 25 μmol / L ZnONPs solution), and 0.50% PS + 50 μmol / L ZnONPs was used for seedling growth trials at 7 days (0.50% PS + 50 μmol / L ZnONPs + foliar spraying of seedlings). For seedling growth trials, treatments included: a mixed solution of ZnONPs (seed soaking + 0.50% PS microplastic soil substrate culture) or foliar spraying of seedlings with a 0.50% PS + 50 μmol / L ZnONPs solution (seed soaking + 0.50% PS microplastic soil substrate culture + 50 μmol / L ZnONPs solution); and foliar spraying of seedlings with a 0.50% PS + 75 μmol / L ZnONPs solution (seed soaking + 0.50% PS microplastic soil substrate culture) or ... ZnONPs treatment (seed soaking in a mixture of 0.50% PS and 75 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 75 μmol / L ZnONPs solution), 0.50% PS + 100 μmol / L ZnONPs treatment for 7-day seedling growth (seed soaking in a mixture of 0.50% PS and 100 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate), or 0.50% PS + 100 μmol / L ZnONPs treatment for 21-day seedling growth (seed soaking in a mixture of 0.50% PS and 100 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + 100 μmol / L ZnONPs solution). ZnONPs solution foliar spraying of seedlings), 0.50% PS + 125 μmol / L ZnONPs was used for seedling growth 7 days test 0.50% PS + 125 μmol / L ZnONPs treatment (0.50% PS + 125 μmol / L ZnONPs mixture seed soaking + 0.Treatments included: 0.50% PS + 125 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 125 μmol / L ZnONPs + 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 125 μmol / L ZnONPs solution) for seedlings grown in a 21-day culture trial; and 0.50% PS + 150 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + 0.50% PS microplastic soil substrate) for seedlings grown in a 7-day culture trial; and 0.50% PS + 150 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + 0.50% PS microplastic soil substrate) for seedlings grown in a 21-day culture trial. Seedlings were soaked in a mixture of ZnONPs, cultured in a 0.50% PS microplastic soil substrate, and then foliar-sprayed with a 150 μmol / L ZnONPs solution.

[0057] Figure 17 The overall mitigation effect of different concentrations of GO solution soaking in seeds or foliar spraying on TS163 maize genotype seedlings on day 7 or day 21 of growth of 0.50% PS microplastic toxicity (CME) GO )analyze.

[0058] 7d refers to the 7th day of growth of the maize genotype seedlings, and 21d refers to the 21st day of growth of the maize genotype seedlings. 0.50% PS + 2.5 mg / L GO was used for seedling growth trials at 7 days (seed soaking in a mixture of 0.50% PS + 2.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or at 21 days (seed soaking in a mixture of 0.50% PS + 2.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 2.5 mg / L GO solution). Alternatively, 0.50% PS + 5.0 mg / L GO was used for seedling growth trials at 7 days (seed soaking in a mixture of 0.50% PS + 5.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or at 21 days (seed foliar spraying of seedlings with 2.5 mg / L GO solution). GO treatment (seed soaking in a mixture of 0.50% PS and 5.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 5.0 mg / L GO solution), 0.50% PS + 7.5 mg / L GO treatment for 7 days of seedling growth (seed soaking in a mixture of 0.50% PS and 7.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or 0.50% PS + 7.5 mg / L GO treatment for 21 days of seedling growth (seed soaking in a mixture of 0.50% PS and 7.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 7.5 mg / L GO solution), 0.50% PS + 10.0 mg / L GO treatment for 7 days of seedling growth (0.50% PS + 10.0 mg / L GO). GO treatment (seed soaking in a mixture of 0.50% PS and 10.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or 0.50% PS + 10.0 mg / L GO treatment (seed soaking in a mixture of 0.50% PS and 10.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 10.0 mg / L GO solution) for 21 days of seedling growth; 0.50% PS + 12.5 mg / L GO treatment (seed soaking in a mixture of 0.50% PS and 12.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or 0.50% PS + 12.5 mg / L GO treatment (seed soaking in a mixture of 0.50% PS and 12.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or 0.50% PS + 12.5 mg / L GO treatment (seed soaking in a mixture of 0.50% PS and 12.5 mg / L GO + foliar spraying of seedlings) for 21 days of seedling growth. Seeds were soaked in a mixture of GO and cultured in a 0.50% PS microplastic soil substrate, with seedlings foliar sprayed with a 12.5 mg / L GO solution. 0.50% PS + 15.0 mg / L GO was used as the 0.50% PS + 15.0 mg / L GO solution in a 7-day seedling growth experiment.Treatment with 50% PS + 15.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or treatment with 0.50% PS + 15.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 15.0 mg / L GO solution) for a 21-day seedling growth trial.

[0059] Figure 18 The overall mitigation intensity (CMI) of TS163 maize genotype seedlings at 7 days or 21 days of growth was compared by soaking seeds or foliar spraying with ZnONPs solutions of different concentrations to alleviate microplastic toxicity caused by 0.50% PS. ZnONPs )analyze.

[0060] 7d refers to the 7th day of growth of the maize genotype seedlings, and 21d refers to the 21st day of growth of the maize genotype seedlings. 0.50% PS + 25 μmol / L ZnONPs was used for seedling growth trials at 7 days (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate) or for seedling growth trials at 21 days (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 25 μmol / L ZnONPs solution), and 0.50% PS + 50 μmol / L ZnONPs was used for seedling growth trials at 7 days (0.50% PS + 50 μmol / L ZnONPs + foliar spraying of seedlings). For seedling growth trials, treatments included: a mixed solution of ZnONPs (seed soaking + 0.50% PS microplastic soil substrate culture) or foliar spraying of seedlings with a 0.50% PS + 50 μmol / L ZnONPs solution (seed soaking + 0.50% PS microplastic soil substrate culture + 50 μmol / L ZnONPs solution); and foliar spraying of seedlings with a 0.50% PS + 75 μmol / L ZnONPs solution (seed soaking + 0.50% PS microplastic soil substrate culture) or ... ZnONPs treatment (seed soaking in a mixture of 0.50% PS and 75 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 75 μmol / L ZnONPs solution), 0.50% PS + 100 μmol / L ZnONPs treatment for 7-day seedling growth (seed soaking in a mixture of 0.50% PS and 100 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate), or 0.50% PS + 100 μmol / L ZnONPs treatment for 21-day seedling growth (seed soaking in a mixture of 0.50% PS and 100 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + 100 μmol / L ZnONPs solution). ZnONPs solution foliar spraying of seedlings), 0.50% PS + 125 μmol / L ZnONPs was used for seedling growth 7 days test 0.50% PS + 125 μmol / L ZnONPs treatment (0.50% PS + 125 μmol / L ZnONPs mixture seed soaking + 0.Treatments included: 0.50% PS + 125 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 125 μmol / L ZnONPs + 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 125 μmol / L ZnONPs solution) for seedlings grown in a 21-day culture trial; and 0.50% PS + 150 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + 0.50% PS microplastic soil substrate) for seedlings grown in a 7-day culture trial; and 0.50% PS + 150 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + 0.50% PS microplastic soil substrate) for seedlings grown in a 21-day culture trial. Seedlings were soaked in a mixture of ZnONPs, cultured in a 0.50% PS microplastic soil substrate, and then foliar-sprayed with a 150 μmol / L ZnONPs solution.

[0061] Figure 19 The overall mitigation intensity (CMI) of TS163 maize genotype seedlings at 7 days or 21 days of growth was compared by soaking seeds or foliar spraying with different concentrations of GO solution to alleviate the toxicity of 0.50% PS microplastics. GO )analyze.

[0062] 7d refers to the 7th day of growth of the maize genotype seedlings, and 21d refers to the 21st day of growth of the maize genotype seedlings. 0.50% PS + 2.5 mg / L GO was used for seedling growth trials at 7 days (seed soaking in a mixture of 0.50% PS + 2.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or at 21 days (seed soaking in a mixture of 0.50% PS + 2.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 2.5 mg / L GO solution). Alternatively, 0.50% PS + 5.0 mg / L GO was used for seedling growth trials at 7 days (seed soaking in a mixture of 0.50% PS + 5.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or at 21 days (seed foliar spraying of seedlings with 2.5 mg / L GO solution). GO treatment (seed soaking in a mixture of 0.50% PS and 5.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 5.0 mg / L GO solution), 0.50% PS + 7.5 mg / L GO treatment for 7 days of seedling growth (seed soaking in a mixture of 0.50% PS and 7.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or 0.50% PS + 7.5 mg / L GO treatment for 21 days of seedling growth (seed soaking in a mixture of 0.50% PS and 7.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 7.5 mg / L GO solution), 0.50% PS + 10.0 mg / L GO treatment for 7 days of seedling growth (0.50% PS + 10.0 mg / L GO). GO treatment (seed soaking in a mixture of 0.50% PS and 10.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or 0.50% PS + 10.0 mg / L GO treatment (seed soaking in a mixture of 0.50% PS and 10.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 10.0 mg / L GO solution) for 21 days of seedling growth; 0.50% PS + 12.5 mg / L GO treatment (seed soaking in a mixture of 0.50% PS and 12.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or 0.50% PS + 12.5 mg / L GO treatment (seed soaking in a mixture of 0.50% PS and 12.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or 0.50% PS + 12.5 mg / L GO treatment (seed soaking in a mixture of 0.50% PS and 12.5 mg / L GO + foliar spraying of seedlings) for 21 days of seedling growth. Seeds were soaked in a mixture of GO and cultured in a 0.50% PS microplastic soil substrate, with seedlings foliar sprayed with a 12.5 mg / L GO solution. 0.50% PS + 15.0 mg / L GO was used as the 0.50% PS + 15.0 mg / L GO solution in a 7-day seedling growth experiment.Treatment with 50% PS + 15.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate) or treatment with 0.50% PS + 15.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 15.0 mg / L GO solution) for a 21-day seedling growth trial. Specific implementation methods

[0063] Unless otherwise specified, the methods used in the following embodiments of the present invention are conventional methods; the equipment and reagents used are all conventional equipment and reagents purchased from reagent companies. It should also be noted that, in order to avoid obscuring the technical solution of the present invention due to unnecessary details, only technical solutions and / or processing steps closely related to the solution according to the present invention are shown in the embodiments, while other details that are not closely related are omitted.

[0064] Furthermore, those skilled in the art will understand that the maize genotype test material is not limited to one sample. This invention uses only one sample of maize genotype test material as an example in order to clearly describe the content of the technical solution of this invention.

[0065] Example 1

[0066] This invention provides a comprehensive evaluation method for mitigating microplastic toxicity in corn seedlings. The specific evaluation method is carried out according to the following steps:

[0067] 1. Preparation of high-quality maize seeds: Prepare high-quality TS163 maize genotype new seeds that were strictly bagged and self-pollinated in the same year, harvested in 2024 at the Longxi Maize Experiment Station (34.97°N, 104.40°E, altitude 2074m), and harvested when physiologically mature, uniform in size, plump, vigorous, and high in purity.

[0068] 2. Preparation of different types of solutions: 0.50 g of PS microplastics (purchased from Dongguan Ruixiang Plastic Raw Materials Co., Ltd.; PS specifications: 1000 mesh, particle size 13 μm) was accurately weighed using an electronic balance and dissolved in 100 mL of ddH2O to prepare a 0.50% concentration PS microplastic solution (PS mass to ddH2O mass ratio). The solution was prepared fresh for each use. 12.0 mg of nano-sized zinc oxide (ZnONPs; purchased from Shanghai Yuanye Biotechnology Co., Ltd.; ZnONPs specifications: CAS number 1314-13-2, molecular weight 81.39, purity 99.9%, particle size 30±10 nm) was accurately weighed using an electronic balance and dissolved in 1 L of ddH2O water. The ZnONPs were dispersed at room temperature using a Tianjin Taist SH-2 / SH-3 type magnetic stirrer. After 30 minutes, the ZnONPs were ultrasonically vibrated for another 30 minutes using a 40Hz SB-5200DT ultrasonic cleaner from Ningbo Xinzhi Biotechnology Co., Ltd., to ensure thorough and uniform dispersion of ZnONPs in ddH2O water, resulting in a 150 μmol / L ZnONPs stock solution. This stock solution was then diluted to obtain six concentrations of ZnONPs: 25 μmol / L, 50 μmol / L, 75 μmol / L, 100 μmol / L, 125 μmol / L, and 150 μmol / L. Simultaneously, 100 mL of each of these six concentrations of ZnONPs solution was taken, and 0.50 g of PS microplastics was added to each solution for dissolution, resulting in 0.50% PS + 25 μmol / L ZnONPs, 0.50% PS + 50 μmol / L ZnONPs, 0.50% PS + 75 μmol / L ZnONPs, and 0.50% PS + 100 μmol / L ZnONPs. Six PS and ZnONPs mixtures with concentrations of ZnONPs, 0.50% PS + 125 μmol / L ZnONPs, and 0.50% PS + 150 μmol / L ZnONPs were prepared fresh for each use. In addition, 1% (1g / 100mL ddH2O water) monolayer graphene oxide (GO; purchased from Shanghai Yuanye Biotechnology Co., Ltd.; ZnONPs specifications: purity 98%, carbon content <46%, oxygen content >45%, sulfur content <1.5%, sheet diameter 0.2-10μm) was pipetted into volumetric flasks at concentrations of 25μL, 50μL, 75μL, 100μL, 125μL, and 150μL, respectively, and then diluted to 100mL with ddH2O water to obtain six GO solutions of concentrations of 2.5mg / L, 5.0mg / L, 7.5mg / L, 10.0mg / L, 12.5mg / L, and 15.0mg / L, which were prepared fresh each time.Meanwhile, 100 mL of each of the six GO solutions of the above-mentioned concentrations was taken, and 0.50 g of PS microplastics was added to each solution to dissolve them, thereby obtaining six PS and GO mixtures with concentrations of 0.50% PS + 2.5 mg / L GO, 0.50% PS + 5.0 mg / L GO, 0.50% PS + 7.5 mg / L GO, 0.50% PS + 10.0 mg / L GO, 0.50% PS + 12.5 mg / L GO, and 0.50% PS + 15.0 mg / L GO. These mixtures were prepared fresh for each use. In summary, a total of 26 types of solutions were prepared, namely: ddH2O in water; 0.50% PS microplastic solution; six ZnONP solutions with concentrations of 25 μmol / L, 50 μmol / L, 75 μmol / L, 100 μmol / L, 125 μmol / L, and 150 μmol / L; and 0.50% PS + 25 μmol / L ZnONPs, 0.50% PS + 50 μmol / L ZnONPs, 0.50% PS + 75 μmol / L ZnONPs, 0.50% PS + 100 μmol / L ZnONPs, 0.50% PS + 125 μmol / L ZnONPs, and 0.50% PS + 150 μmol / L ZnONPs. Six PS and ZnONPs mixed solutions at ZnONPs concentrations; six GO solutions at concentrations of 2.5 mg / L, 5.0 mg / L, 7.5 mg / L, 10.0 mg / L, 12.5 mg / L, and 15.0 mg / L; and six PS and GO mixed solutions at concentrations of 0.50% PS + 2.5 mg / L GO, 0.50% PS + 5.0 mg / L GO, 0.50% PS + 7.5 mg / L GO, 0.50% PS + 10.0 mg / L GO, 0.50% PS + 12.5 mg / L GO, and 0.50% PS + 15.0 mg / L GO, all prepared fresh for use.

[0069] 3. Experiment on the 7-day growth of maize seedlings to alleviate PS microplastic toxicity by soaking maize seeds in different types of ZnONPs / GO solutions: Forty high-quality TS163 maize genotype seeds prepared in advance were placed in Erlenmeyer flasks, 100 mL of 70% ethanol (v / v) was added, and the seeds were placed on a TS-2 horizontal shaker of Shenzhen Sanli Technology Co., Ltd. to disinfect the seeds for 10 min. The seeds were then rinsed 5 times with 100 mL of ddH2O water to remove residual ethanol from the seed surface. The water adhering to the seed surface was absorbed by sterile filter paper to obtain the corresponding disinfected TS163 maize genotype seeds. The sterilized TS163 maize genotype seeds were then placed into corresponding Erlenmeyer flasks, and 100 mL of each of the 14 types of solutions prepared beforehand were added, including ddH2O water; 0.50% PS microplastic solution; six mixtures of PS and ZnONPs with concentrations of 0.50% PS + 25 μmol / L ZnONPs, 0.50% PS + 50 μmol / L ZnONPs, 0.50% PS + 75 μmol / L ZnONPs, 0.50% PS + 100 μmol / L ZnONPs, 0.50% PS + 125 μmol / L ZnONPs, and 0.50% PS + 150 μmol / L ZnONPs; 0.50% PS + 2.5 mg / L GO, 0.50% PS + 5.0 mg / L GO, and 0.50% PS + 7.5 mg / L GO. Six mixtures of PS and GO at concentrations of GO, 0.50% PS + 10.0 mg / L GO, 0.50% PS + 12.5 mg / L GO, and 0.50% PS + 15.0 mg / L GO were prepared and the solutions were placed in a dark indoor environment on a TS-2 horizontal shaker manufactured by Shenzhen Sanli Technology Co., Ltd., for 24 hours to soak the seeds. Simultaneously, two soil substrates with different concentrations (PS mass to nutrient soil mass ratio) of PS microplastics were prepared: a 0.00% PS microplastic soil substrate (i.e., 0.00 g PS microplastics thoroughly mixed with 300.00 g nutrient soil, then 250 mL ddH2O water was added and stirred thoroughly) and a 0.50% PS microplastic soil substrate (i.e., 1.50 g PS microplastics thoroughly mixed with 300.00 g nutrient soil, then 250 mL ddH2O water was added and stirred thoroughly). Ten seeds of each of the 14 types of solutions used for soaking were then sown in pots containing the corresponding concentration of PS microplastic soil substrate and placed in an artificial climate chamber for cultivation. During cultivation, the relative humidity was set at 65%, the temperature was alternated between 25±0.5℃ and 20±0.5℃ for 12 hours, the photoperiod was set at 16 / 8 hours of light / dark, and the light intensity was set at 300 μMm. -2 s -1The CO2 concentration was 450 PPM. In total, there were 14 treatments in the experiment: CK control (soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate), 0.50% PS treatment (soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate), 0.50% PS + 25 μmol / L ZnONPs treatment (soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate), 0.50% PS + 50 μmol / L ZnONPs treatment (soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate), and 0.50% PS + 75 μmol / L ZnONPs treatment (…). Treatments included: a mixture of ZnONPs soaking in a solution of 0.50% PS microplastic soil substrate; a mixture of ZnONPs soaking in a solution of 0.50% PS + 100 μmol / L ZnONPs in a solution of 0.50% PS + 125 μmol / L ZnONPs in a solution of 0.50% PS + 150 μmol / L ZnONPs in a solution of 0.50% PS + 2.5 mg / L GO in a solution of 0.50% PS + 2.5 mg / L GO. Treatments included: GO mixture soaking + 0.50% PS microplastic soil substrate culture; 0.50% PS + 5.0 mg / L GO treatment (0.50% PS + 5.0 mg / L GO mixture soaking + 0.50% PS microplastic soil substrate culture); 0.50% PS + 7.5 mg / L GO treatment (0.50% PS + 7.5 mg / L GO mixture soaking + 0.50% PS microplastic soil substrate culture); 0.50% PS + 10.0 mg / L GO treatment (0.50% PS + 10.0 mg / L GO mixture soaking + 0.50% PS microplastic soil substrate culture); 0.50% PS + 12.5 mg / L GO treatment (0.50% PS + 12.5 mg / L GO mixture soaking + 0.50% PS microplastic soil substrate culture); 0.50% PS + 15.0 mg / L GO treatment. GO treatment (soaking in a mixture of 0.50% PS and 15.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate), with 4 biological replicates for each treatment.During the cultivation period, 50 mL of ddH2O water was evenly applied to each pot every 3 days to replenish the seedlings with water in a timely manner. On the 7th day of cultivation, 12 traits of TS163 maize genotype seedlings under these 14 treatments were measured, including seedling length (SL), seedling fresh weight (SFW), seedling dry weight (SDW), root length (RL), root fresh weight (RFW), root dry weight (RDW), stem diameter (SD), root diameter (RD), total plant dry biomass (PDW), leaf chlorophyll SPAD value (SPAD), root activity (RV), and seedling vigor index (SSI). Among them, the SSI value was calculated according to Appendix. Figure 1 The formula (1) is used for calculation. In the formula, SSI is the seedling strength index, SD is the stem diameter, SL is the seedling length, and PDW is the total dry biomass of the plant.

[0070] 4. 21-day experiment on mitigating PS microplastic toxicity in maize seedlings by foliar spraying of different types of ZnONPs / GO solutions: TS163 maize genotype seedlings cultured for 7 days under the corresponding 14 treatments were further cultured in an artificial climate chamber under the same environment for 14 days. Every 2 days, the leaves of the corresponding TS163 maize genotype seedlings under each treatment were sprayed with 10 mL of each of the 14 types of solutions at the corresponding concentration in the morning. That is, 1 mL of each of the 14 types of solutions was sprayed on the leaves of each maize genotype seedling under each corresponding treatment, for a total of 7 foliar sprays. In summary, a total of 14 treatments were tested: CK control treatment (seed soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water), 0.50% PS treatment (seed soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with ddH2O water), 0.50% PS + 25 μmol / L ZnONPs treatment (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 25 μmol / L ZnONPs solution), and 0.50% PS + 50 μmol / L ZnONPs treatment (seed soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 25 μmol / L ZnONPs solution). Foliar application of ZnONPs solution to seedlings), treatment with 0.50% PS + 75 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 75 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar application of 75 μmol / L ZnONPs solution to seedlings), treatment with 0.50% PS + 100 μmol / L ZnONPs solution (… Treatment methods included: seed soaking in a ZnONPs mixture + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 100 μmol / L ZnONPs solution; treatment with 0.50% PS + 125 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 125 μmol / L ZnONPs + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 125 μmol / L ZnONPs solution); treatment with 0.50% PS + 150 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 150 μmol / L ZnONPs solution); and treatment with 0.50% PS + 2.5 mg / L GO (0.50% PS + 2.5 mg / L GO). Seedlings were soaked in a mixture of GO and cultured in a soil substrate containing 0.50% PS microplastics, followed by foliar spraying of seedlings with a 2.5 mg / L GO solution. Seedlings were also treated with a mixture of 0.50% PS and 5.0 mg / L GO.Seedlings were treated with a mixture of 50% PS and 5.0 mg / L GO (soaked in a solution of 50% PS + 5.0 mg / L GO, cultured in a microplastic soil substrate with 0.50% PS, and foliar sprayed with a 5.0 mg / L GO solution). Other treatments included: 0.50% PS + 7.5 mg / L GO (soaked in a solution of 0.50% PS + 7.5 mg / L GO, cultured in a microplastic soil substrate with 0.50% PS, and foliar sprayed with a 7.5 mg / L GO solution); 0.50% PS + 10.0 mg / L GO (soaked in a solution of 0.50% PS + 10.0 mg / L GO, cultured in a microplastic soil substrate with 0.50% PS, and foliar sprayed with a 10.0 mg / L GO solution); and 0.50% PS + 12.5 mg / L GO (…). Seedlings were treated with a mixture of GO (Glycogen peroxide) for soaking, followed by cultivation in a 0.50% PS microplastic soil substrate, and then foliar spraying with a 12.5 mg / L GO solution. Four biological replicates were used for each treatment: (1) seed soaking in a mixture of 0.50% PS and 15.0 mg / L GO, followed by cultivation in a 0.50% PS microplastic soil substrate, and then foliar spraying with a 15.0 mg / L GO solution). Relative humidity was 65%, temperature was 25 ± 0.5 °C / 20 ± 0.5 °C alternating for 12 h, photoperiod was 16 / 8 h light / dark, and light intensity was 300 μMm. -2 s -1 The CO2 concentration was 450 PPM. During the cultivation period, 50 mL of ddH2O water was evenly poured into each pot every 3 days to replenish the seedlings in time. On the 21st day of seedling cultivation, 12 traits of TS163 maize genotype seedlings under these 14 treatments were measured, including seedling length (SL), seedling fresh weight (SFW), seedling dry weight (SDW), root length (RL), root fresh weight (RFW), root dry weight (RDW), stem diameter (SD), root diameter (RD), total plant dry biomass (PDW), leaf chlorophyll SPAD value (SPAD), root activity (RV), and seedling vigor index (SSI).

[0071] 5. Statistical Analysis of Different Experimental Data: Using Excel software, the mean and standard deviation (SD) of 12 traits of TS163 maize genotype seedlings were analyzed in the 7-day and 21-day experiments of seedling growth of TS163 maize genotype seedlings treated with different types of ZnONPs / GO solutions to alleviate PS microplastic toxicity. IBM-SPSS Statistics was also used. 19. Using the statistical analysis software (https: / / www.ibm.com / products / spss-statistics / data-visualization), a joint analysis of variance was performed on all traits of TS163 maize genotype seedlings in 7-day and 21-day growth trials after seed soaking and foliar spraying with different types of ZnONPs / GO solutions to alleviate PS microplastic toxicity. This included the corrected model (CM), the PS microplastic concentration treatments (PS), ZnONPs concentration treatments (ZnONPs), GO concentration treatments (GO), the culture period treatments (S), and the relationship between PS microplastic concentration treatments and ZnONPs / GO concentration treatments. The degrees of freedom (DF), F-values, and significance of differences (SIG) were analyzed for the following interactions: interaction between ONP concentration treatments (PS×ZnONPs), interaction between PS microplastic concentration treatments and GO concentration treatments (PS×GO), interaction between PS microplastic concentration treatments and culture period treatments (PS×S), interaction between ZnONP concentration treatments and culture period (ZnONPs×S), interaction between GO concentration treatments and culture period (GO×S), interaction among PS microplastic concentration treatments, ZnONP concentration treatments, and culture period (PS×ZnONPs×S), and interaction among PS microplastic concentration treatments, GO concentration treatments, and culture period (PS×GO×S). The online analysis and plotting software GENESCLOUD (https: / / www.genescloud.cn / chart / CircleCorPlot) was used to plot the correlation coefficients of 12 traits in 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different types of ZnONPs / GO solutions to alleviate PS microplastic toxicity. The IMLOG2 function was used to further standardize the mean values ​​of 12 traits in the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different types of ZnONPs / GO solutions for seed soaking and leaf spraying to alleviate PS microplastic toxicity.IBM SPSS Statistics 16 (https: / / www.ibm.com / products / spss-statistics / data-visualization) software was used to perform between-group linkage analysis on all treatments in 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different types of ZnONPs / GO solutions to alleviate PS microplastic toxicity. This was to evaluate the overall growth status of TS163 maize genotype seedlings at different cultivation stages and treatments, and to scientifically and qualitatively identify the mitigation effects of different concentrations of ZnONPs / GO solution on TS163 maize genotype seedlings at different cultivation stages under PS microplastic toxicity.

[0072] 6. Relative mitigation effect index of different concentrations of ZnONPs / GO solution soaking or foliar spraying on individual traits of maize seedlings under 0.50% PS microplastic toxicity: To more scientifically, objectively, and accurately measure the mitigating effect of different concentrations of ZnONPs / GO solution soaking or foliar spraying on individual traits of maize genotype seedlings under 0.50% PS microplastic toxicity, we newly defined the relative mitigation effect index (RMEI) of different concentrations of ZnONPs / GO solution soaking or foliar spraying on individual traits of maize seedlings under 0.50% PS microplastic toxicity. ZnONPS and RMEI GO (See attached document for details) Figure 1 Formulas (2) and (3). In formula (2) The relative mitigation index of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. The values ​​for the j-th trait in maize seedlings treated with a 0.50% PS microplastic toxicity solution (i-ZnONPs) at concentration i-ZnONPs concentration, measured on day 7 or day 21 of cultivation. The ΣT represents the phenotypic value of genotype j in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity treatment. j / m represents the average value of the j-th trait in maize seedlings cultured on day 7 or day 21 after treatment with m (m=8) types of solutions for seed soaking or foliar spraying. In formula (3) The relative mitigation index of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. The value of the jth trait in maize seedlings cultured on day 7 or day 21 after treatment with i-GO concentration solution (immersion in 0.50% PS microplastic toxicity solution) is the result of the i-GO concentration solution. The ΣT represents the phenotypic value of genotype j in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity treatment. j / n represents the average value of the jth trait in maize seedlings cultured on day 7 or day 21 after treatment with n (n=8) types of solutions for seed soaking or foliar spraying. The higher the value, the stronger the mitigating effect of soaking seeds or spraying leaves with the i-ZnONPs concentration solution on the j-th genotype of maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. The higher the value, the stronger the mitigating effect of soaking seeds or spraying leaves with the i-GO concentration solution on the j-th trait of maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity.

[0073] 7. Absolute mitigation strength coefficient of different concentrations of ZnONPs / GO solution for seed soaking or foliar spraying on individual traits of maize seedlings under 0.50% PS microplastic toxicity: To facilitate a more scientific, simple, and objective evaluation of the mitigation strength of different concentrations of ZnONPs / GO solution for seed soaking or foliar spraying on individual traits of maize genotype seedlings under 0.50% PS microplastic toxicity, this invention also newly defines the absolute mitigation strength coefficient (AMIC) of different concentrations of ZnONPs / GO solution for seed soaking or foliar spraying on individual traits of maize seedlings under 0.50% PS microplastic toxicity. ZnONPs and AMIC GO (See attached document for details) Figure 1 Formulas (4) and (5). In formula (4) The absolute mitigation strength coefficient of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. The values ​​for the j-th trait in maize seedlings treated with a 0.50% PS microplastic toxicity solution (i-ZnONPs) at concentration i-ZnONPs concentration, measured on day 7 or day 21 of cultivation. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity treatment. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under CK treatment. In formula (5) The absolute mitigation strength coefficient of the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. The value of the jth trait in maize seedlings cultured on day 7 or day 21 after treatment with i-GO concentration solution (immersion in 0.50% PS microplastic toxicity solution) is the result of the i-GO concentration solution. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity treatment. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under CK treatment. The higher the value, the greater the mitigation effect of soaking seeds or spraying leaves with the i-ZnONPs concentration solution on the j-th genotype of maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. The higher the value, the greater the mitigation effect of soaking seeds in i-GO concentration solution or spraying leaves on the j-th phenotype of maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity.

[0074] 8. Comprehensive evaluation of the mitigating effect of different concentrations of ZnONPs / GO solution soaking or foliar spraying on the toxicity of 0.50% PS microplastics in maize seedlings: Further, based on the above calculations, the RMEI of 12 traits in TS163 maize genotype seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was obtained from soaking seeds or spraying them with different concentrations of ZnONPs / GO solution. ZnONPS and RMEI GO The values ​​were used as evaluation indicators for the mitigating effects of different concentrations of ZnONPs / GO solution soaking or foliar spraying on TS163 maize genotype seedlings under 0.50% PS microplastic toxicity. The membership function method was employed to scientifically, objectively, comprehensively, and quantitatively evaluate the comprehensive mitigating effects of different concentrations of ZnONPs / GO solution soaking or foliar spraying on TS163 maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity (CME). ZnONPs and CME GO See attached for details. Figure 1 Formulas (6), (7), (8), and (9). In formula (6): The membership value of the mitigation effect of soaking seeds or spraying leaves with i-ZnONPs concentration solution on the j-th trait of TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined. RMEI is the relative mitigation index of the jth trait in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity, calculated by soaking seeds or spraying leaves with i-ZnONPs concentration solution. j-max RMEI is the maximum relative mitigation index for the j-th trait in TS163 maize genotype seedlings cultured on day 7 or 21 of 0.50% PS microplastic toxicity after seed soaking or foliar spraying with ZnONPs solution at all concentrations. j-min The minimum relative mitigation index for the j-th trait in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity, obtained by soaking seeds or foliar spraying with ZnONPs solution at all concentrations. In formula (7), CME... i-ZnONPs The overall mitigation effect of soaking seeds or spraying leaves with i-ZnONPs concentration solution on TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity is shown, where k (k=12) represents the number of traits measured. The membership value of the mitigation effect of soaking seeds or spraying leaves with i-ZnONPs concentration solution on the j-th trait of TS163 maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity. In formula (8): The membership value of the mitigation effect of soaking seeds or spraying leaves with i-GO concentration solution on the j-th trait of TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by the effect of such treatment on seeds. The RMEI (Relative Mitigation Index) was used to measure the relative mitigation effect of soaking seeds or foliar spraying with i-GO concentration solution on the j-th trait in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. j-max The RMEI (Relative Remission Index) represents the maximum relative mitigation effect on the jth trait in TS163 maize genotype seedlings cultured on day 7 or 21 of 0.50% PS microplastic toxicity after seed soaking or foliar spraying with GO solution at all concentrations. j-min The minimum relative mitigation index for the j-th trait in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity, obtained by soaking seeds or foliar spraying with GO solution at all concentrations. In formula (9), CME... i-GO The overall mitigation strength of TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. k (k=12) represents the number of traits measured. Membership value for mitigating the j-th trait in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with i-GO concentration solution. i-ZnONPs The higher the value, the stronger the overall mitigation effect of soaking seeds or spraying leaves with the i-ZnONPs concentration solution on TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity.

[0075] 9. Comprehensive evaluation of the mitigation strength of different concentrations of ZnONPs / GO solution soaking or foliar spraying on maize seedlings under 0.50% PS microplastic toxicity: Further, based on the above calculations, the AMIC of 12 traits in TS163 maize genotype seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was evaluated using different concentrations of ZnONPs / GO solution soaking or foliar spraying. ZnONPS and AMIC GO The values ​​were used as evaluation indicators for the mitigation intensity of different concentrations of ZnONPs / GO solution soaking or foliar spraying on TS163 maize genotype seedlings under 0.50% PS microplastic toxicity. The membership function method was used to comprehensively, systematically, objectively, and quantitatively evaluate the overall mitigation intensity (CMI) of different concentrations of ZnONPs / GO solution soaking or foliar spraying on TS163 maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity. ZnONPs and CMI GO See attached for details. Figure 1 Formulas (10), (11), (12), and (13). In formula (10): The membership value of the mitigation strength of the jth trait in TS163 maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. The absolute mitigation strength coefficient (AMIC) of the j-th trait in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or foliar spraying with i-ZnONPs concentration solution. j-max The maximum absolute mitigation strength coefficient (AMIC) of the j-th trait in TS163 maize genotype seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or foliar spraying with ZnONPs solutions at all concentrations. j-min The minimum absolute mitigation strength coefficient for the j-th trait in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with ZnONPs solution at all concentrations. In formula (11), CMI... i-ZnONPs The overall mitigation strength of soaking seeds or foliar spraying with i-ZnONPs concentration solution on TS163 maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity was determined, where k (k=12) represents the number of traits measured. The membership value of the mitigation strength of the jth trait in TS163 maize seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. In formula (12): The membership value of the mitigation strength of the j-th trait in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. The absolute mitigation strength coefficient (AMIC) of the j-th trait in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or foliar spraying with i-GO concentration solution. j-max The maximum absolute mitigation strength coefficient (AMIC) of the j-th trait in TS163 maize genotype seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or foliar spraying with GO solution at all concentrations. j-min The minimum absolute mitigation strength coefficient for the j-th trait in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with GO solution of all concentrations. In formula (13), CMI... i-GO The overall mitigation strength of TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. k (k=12) represents the number of traits measured. CMI (Combined Minerals Index) is the membership value for mitigation strength of the j-th trait in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity, obtained by soaking seeds or spraying leaves with i-GO concentration solution. i-ZnONPs The higher the value, the greater the overall mitigation effect of soaking seeds or foliar spraying with the i-ZnONPs concentration solution on TS163 maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity. CMI i-GO The higher the value, the greater the overall mitigation effect of soaking seeds or spraying leaves with the i-GO concentration solution on TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity.

[0076] 10. Optimal ZnONPs / GO solution concentration for achieving the best mitigation effect / maximum mitigation intensity of 0.50% concentration of PS microplastic toxicity in maize seedlings: Further evaluation was conducted by comparing the intergroup clustering results of the growth status of TS163 maize genotype seedlings under all treatments in the 7-day and 21-day growth trials of seed soaking and foliar spraying with different types of ZnONPs / GO solutions to mitigate PS microplastic toxicity, the comprehensive evaluation results of the mitigation effects of seed soaking or foliar spraying with different concentrations of ZnONPs / GO solutions on 0.50% concentration of PS microplastic toxicity in TS163 maize genotype seedlings, and the above... The comprehensive evaluation results of the mitigation intensity of different concentrations of ZnONPs / GO solution soaking in seeds or foliar spraying on TS163 maize genotype seedlings with 0.50% PS microplastic toxicity were studied. Multiple analytical methods were used to comprehensively compare and analyze the mitigation effects of different concentrations of ZnONPs / GO solution soaking in seeds or foliar spraying on TS163 maize genotype seedlings with 0.50% PS microplastic toxicity at different cultivation stages. Finally, the optimal concentration of ZnONPs / GO solution for seed soaking or foliar spraying was determined to achieve the best mitigation effect / maximum mitigation intensity of 0.50% PS microplastic toxicity in TS163 maize genotype seedlings at different cultivation stages.

[0077] Example 2

[0078] This invention provides a comprehensive evaluation result of the effect of mitigating microplastic toxicity in corn seedlings. The specific evaluation results are as follows:

[0079] 1. Analysis of joint variance of 12 traits in the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of ZnONPs solution for seed soaking and foliar spraying to alleviate PS microplastic toxicity: As shown in Table 1, the corrected model (CM) for the joint variance analysis of the 12 traits was significant at the P<0.001 level. This indicates that the overall differences among these 12 traits were significant in the joint variance analysis. Therefore, it is valid to conduct ANOVA on these 12 traits for the interaction between PS microplastic concentration treatment (PS), ZnONPs concentration treatment (ZnONPs), culture period treatment (S), the interaction between PS microplastic concentration treatment and culture period treatment (PS×S), the interaction between ZnONPs concentration treatment and culture period (ZnONPs×S), the interaction between PS microplastic concentration treatment and ZnONPs concentration treatment (PS×ZnONPs), and the interaction between PS microplastic concentration treatment, ZnONPs concentration treatment, and culture period (PS×ZnONPs×S). However, since the degrees of freedom for the interaction between PS microplastic concentration treatment and ZnONPs concentration treatment, and the interaction between PS microplastic concentration treatment, ZnONPs concentration treatment, and culture period are 0, we can only further conduct ANOVA on the remaining 5 factors. Interestingly, the F-values ​​of the analysis of variance for these 12 traits among PS microplastic concentration treatments ranged from 5.160 (seedling vigor index, P<0.05) to 425.983 (root activity, P<0.001); the F-values ​​of the analysis of variance for these 12 traits among ZnONPs concentration treatments ranged from 2.828 (root diameter, P<0.05) to 194.268 (total plant dry biomass, P<0.001). The F-values ​​of the ANOVA among the 12 traits during the cultivation period ranged from 23.138 (root diameter, P<0.001) to 6028.824 (total plant dry biomass, P<0.001); the F-values ​​of the ANOVA among the interactions between ZnONPs concentration treatments and cultivation period ranged from 3.534 (root diameter, P<0.01) to 159.307 (total plant dry biomass, P<0.001). This indicates that the expression of these 12 traits in maize seedlings was significantly regulated by the interaction between PS microplastic concentration treatments, ZnONPs concentration treatments, cultivation period treatments, and the interaction between ZnONPs concentration treatments and cultivation period. In addition, only four traits—fresh seedling weight (F-value = 53.340, P < 0.001), root length (F-value = 4.142, P < 0.05), stem diameter (F-value = 5.688, P < 0.05), and root activity (F-value = 90.903, P < 0.001)—showed significant differences between the PS microplastic concentration treatment and the culture period treatment. This indicates that these four traits were also influenced by the interaction between PS microplastic concentration treatment and the culture period treatment. Therefore, it is necessary to analyze the performance of these 12 traits in TS163 maize genotype seedlings under different treatments.

[0080] Table 1. F-values, significance of differences, and degrees of freedom of the joint ANOVA analysis of 21 traits in the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of ZnONPs solution and sprayed with PS microplastics.

[0081]

[0082]

[0083] Note: CM represents the calibration model; PS represents the PS microplastic concentration treatment interval; ZnONPs represents the ZnONPs concentration treatment interval; S represents the culture period treatment interval; PS×S represents the interaction between PS microplastic concentration treatment and culture period treatment; ZnONPs×S represents the interaction between ZnONPs concentration treatment and culture period; PS×ZnONPs represents the interaction between PS microplastic concentration treatment and ZnONPs concentration treatment; PS×ZnONPs×S represents the interaction between PS microplastic concentration treatment, ZnONPs concentration treatment, and culture period; DF represents degrees of freedom; SL represents seedling length; SFW represents seedling fresh weight; SDW represents seedling dry weight; RL represents root length; RFW represents root fresh weight; RDW represents root dry weight; SD represents stem diameter; RD represents root diameter; PDW represents total plant dry biomass; SPAD represents leaf chlorophyll SPAD value; SSI represents seedling vigor index; RV represents root activity. "-" indicates missing information. "*", "**", or "***" represent significant differences at the P<0.05, P<0.01, or P<0.001 levels, respectively.

[0084] 2. Analysis of joint variance of 12 traits in the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of GO solution for mitigating PS microplastic toxicity: Similarly, as shown in Table 2, the corrected model (CM) for the joint variance analysis of the 12 traits was significant at the P<0.001 level. This indicates that the overall differences among these 12 traits were significant in the joint variance analysis. Therefore, it is valid to conduct ANOVA on these 12 traits for the interaction between PS microplastic concentration treatments (PS), GO concentration treatments (GO), culture period treatments (S), the interaction between PS microplastic concentration treatments and culture period treatments (PS×S), the interaction between GO concentration treatments and culture period treatments (GO×S), the interaction between PS microplastic concentration treatments and GO concentration treatments (PS×GO), and the interaction between PS microplastic concentration treatments, GO concentration treatments, and culture period (PS×GO×S). However, since the degrees of freedom for the interaction between PS microplastic concentration treatments and GO concentration treatments, and the interaction between PS microplastic concentration treatments, GO concentration treatments, and culture period is 0, we can only further conduct ANOVA on the remaining 5 factors. Interestingly, the F-values ​​of the analysis of variance for these 12 traits among PS microplastic concentration treatments ranged from 15.263 (seedling vigor index, P<0.001) to 806.862 (root activity, P<0.001); the F-values ​​of the analysis of variance for these 12 traits among treatments during the culture period ranged from 98.661 (root diameter, P<0.001) to 6746.794 (seedling length, P<0.001); and the F-values ​​of the analysis of variance for the interaction between GO concentration treatment and culture period ranged from 3.757 (root diameter, P<0.01) to 118.648 (seedling fresh weight, P<0.001). This indicates that the performance of these 12 traits in maize seedlings was significantly regulated by the interaction between PS microplastic concentration treatments, the interaction between treatments during the culture period, and the interaction between GO concentration treatment and the culture period. In addition, except for root diameter (F-value = 1.306, P>0.05), which was not significant in the ANOVA among GO concentration treatments, the F-values ​​of the other 11 traits ranged from 6.914 (leaf chlorophyll SPAD value, P<0.001) to 86.384 (root activity, P<0.001) in the ANOVA among GO concentration treatments. At the same time, five traits, namely seedling fresh weight (F-value = 50.338, P<0.001), root length (F-value = 4.586, P<0.05), root dry weight (F-value = 6.170, P<0.05), stem diameter (F-value = 11.358, P<0.001), and root activity (F-value = 172.182, P<0.001), showed significant differences between the PS microplastic concentration treatment and the culture period treatment. This indicates that these traits are also influenced by the interactions between GO concentration treatments, PS microplastic concentration treatments, and culture period treatments. Therefore, it is necessary to analyze the expression of these 12 traits in TS163 maize genotype seedlings under different treatments.

[0085] Table 2. F-values, significance of differences, and degrees of freedom of the joint ANOVA analysis of 21 traits in the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of GO solution and sprayed with PS microplastics.

[0086]

[0087]

[0088] Note: CM represents the calibration model; PS represents the interaction between PS microplastic concentration treatments; GO represents the interaction between GO concentration treatments; S represents the interaction between culture period treatments; PS×S represents the interaction between PS microplastic concentration treatments and culture period treatments; GO×S represents the interaction between GO concentration treatments and culture period treatments; PS×GO represents the interaction between PS microplastic concentration treatments and GO concentration treatments; PS×GO×S represents the interaction between PS microplastic concentration treatments, GO concentration treatments, and culture period treatments; DF represents degrees of freedom; SL represents seedling length; SFW represents seedling fresh weight; SDW represents seedling dry weight; RL represents root length; RFW represents root fresh weight; RDW represents root dry weight; SD represents stem diameter; RD represents root diameter; PDW represents total plant dry biomass; SPAD represents leaf chlorophyll SPAD value; SSI represents seedling vigor index; RV represents root activity. "-" indicates missing information. "*", "**", or "***" represent significant differences at the P < 0.05, P < 0.01, or P < 0.001 levels, respectively.

[0089] 3. Analysis of 12 phenotypic characteristics in 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of ZnONPs solution for seed soaking and foliar spraying to alleviate PS microplastic toxicity: The effects of different concentrations of ZnONPs solution soaking on the 7-day growth of TS163 maize genotype seedlings treated with 0.50% PS microplastic toxicity were analyzed. In the d-test, compared with the CK control treatment, the seedling length, seedling fresh weight, seedling dry weight, root length, root fresh weight, root dry weight, stem diameter, root diameter, total plant dry biomass, seedling vigor index, leaf chlorophyll SPAD value, and root activity of TS163 maize seedlings under 0.50% PS stress decreased by 26.13%, 26.91%, 35.73%, 14.32%, 38.50%, 20.59%, 7.51%, 13.22%, 31.00%, 13.81%, 8.84%, and 77.72%, respectively. (See appendix) Figure 2 and 4Different concentrations of ZnONPs soaking in seedlings showed a certain degree of improvement on 12 traits of TS163 maize seedlings under 0.50% PS stress for 7 days. Furthermore, the improvement effects of different concentrations of ZnONPs soaking on these 12 traits varied (see attached). Figure 2 and 4 Compared with the 0.50% PS stress treatment, the TS163 maize genotype seedlings showed the greatest increase in seedling length (61.08%) under the 0.50% PS + 100 μmol / L ZnONPs treatment; the greatest increase in seedling fresh weight (68.90%) under the 0.50% PS + 75 μmol / L ZnONPs treatment; the greatest increase in seedling dry weight (69.95%) under the 0.50% PS + 100 μmol / L ZnONPs treatment; the greatest increase in root length (35.99%) under the 0.50% PS + 75 μmol / L ZnONPs treatment; and the greatest increase in root fresh weight (70.94%) under the 0.50% PS + 75 μmol / L ZnONPs treatment. The largest increase in root dry weight of TS163 maize seedlings was 13.61% under ZnONPs treatment; the largest increase in stem diameter was 36.06% under 0.50% PS + 75 μmol / L ZnONPs treatment; the largest increase in root diameter was 6.36% under 0.50% PS + 100 μmol / L ZnONPs treatment; the largest increase in total dry biomass was 57.56% under 0.50% PS + 75 μmol / L ZnONPs treatment; the largest increase in seedling vigor index was 43.32% under 0.50% PS + 125 μmol / L ZnONPs treatment; and the largest increase in seedling vigor index was 43.32% under 0.50% PS + 125 μmol / L ZnONPs treatment. The chlorophyll SPAD value of leaves from TS163 maize genotype seedlings treated with ZnONPs increased by the largest amount (17.92%); the root activity of TS163 maize genotype seedlings treated with 0.50% PS + 25 μmol / L ZnONPs increased by the largest amount (395.69%). (See attached image) Figure 2 and 4Similarly, in a 21-day experiment on the foliar spraying of different concentrations of ZnONPs solutions to alleviate the microplastic toxicity of 0.50% PS in TS163 maize genotype seedlings, compared with the CK control treatment, the seedling length, seedling fresh weight, seedling dry weight, root length, root fresh weight, root dry weight, stem diameter, root diameter, total plant dry biomass, seedling vigor index, leaf chlorophyll SPAD value, and root activity of TS163 maize genotype seedlings under 0.50% PS stress treatment decreased by 11.11%, 30.35%, 11.21%, 22.94%, 16.52%, 28.17%, 13.22%, 13.35%, 15.75%, 17.75%, 21.19%, and 62.21%, respectively (see appendix). Figure 3 and 4 Foliar application of different concentrations of ZnONPs improved 12 traits of TS163 maize seedlings under 0.50% PS stress for 21 days. The improvement effects of different concentrations of ZnONPs on these traits varied (see attached figure). Figure 3 and 4Compared with the 0.50% PS stress treatment, the TS163 maize genotype seedlings under the 0.50% PS + 75 μmol / L ZnONPs treatment showed the largest increase in seedling length (21.49%), seedling fresh weight (37.62%), and seedling dry weight (33.47%) under the 0.50% PS + 125 μmol / L ZnONPs treatment. The root length of the TS163 maize genotype seedlings under the 0.50% PS + 25 μmol / L ZnONPs treatment also showed the largest increase. The largest increase in fresh root weight of TS163 maize seedlings was 66.78% under ZnONPs treatment; the largest increase in dry root weight was 66.96% under 0.50% PS + 75 μmol / L ZnONPs treatment; the largest increase in stem diameter was 18.55% under 0.50% PS + 75 μmol / L ZnONPs treatment; the largest increase in root diameter was 6.29% under 0.50% PS + 50 μmol / L ZnONPs treatment; and the largest increase in total dry biomass was 262.92% under 0.50% PS + 125 μmol / L ZnONPs treatment. The ZnONPs treatment resulted in the largest increase in seedling vigor index of TS163 maize genotype seedlings (275.11%); the 0.50% PS + 75 μmol / L ZnONPs treatment resulted in the largest increase in chlorophyll SPAD value of TS163 maize genotype seedlings (62.29%); and the 0.50% PS + 50 μmol / L ZnONPs treatment resulted in the largest increase in root activity of TS163 maize genotype seedlings (135.50%). (See appendix) Figure 3 and 4 The results indicate that the effects of different concentrations of ZnONPs solution soaking in seeds and foliar spraying on mitigating PS microplastic toxicity in TS163 maize genotype seedlings were not entirely the same in 7-day and 21-day growth trials.

[0090] 4. Analysis of 12 phenotypic characteristics in 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of GO solution to alleviate PS microplastic toxicity: In the 7-day growth trial of TS163 maize genotype seedlings treated with different concentrations of GO solution to alleviate 0.50% PS microplastic toxicity, compared with the CK control treatment, the 12 traits of TS163 maize genotype seedlings under 0.50% PS stress treatment showed varying degrees of reduction, ranging from 7.51% (stem diameter) to 77.72% (root activity) (see appendix). Figure 5 and 7 Different concentrations of GO soaking in seedlings showed a certain degree of improvement on 12 traits of TS163 maize seedlings under 0.50% PS stress for 7 days. Furthermore, the improvement effects of different concentrations of GO soaking on these 12 traits varied (see attached). Figure 5 and 7 Compared with the 0.50% PS stress treatment, the TS163 maize genotype seedlings showed the greatest increase in seedling length (29.12%) under the 0.50% PS + 12.5 mg / L GO treatment; the greatest increase in seedling fresh weight (68.33%) under the 0.50% PS + 12.5 mg / L GO treatment; the greatest increase in seedling dry weight (35.99%) under the 0.50% PS + 12.5 mg / L GO treatment; and the greatest increase in root length (35.99%) under the 0.50% PS + 7.5 mg / L GO treatment. The greatest increase in root fresh weight (82.64%) was observed under the 0.50% PS + 12.5 mg / L GO treatment. The largest increase in root dry weight of TS163 maize seedlings was 41.14% under GO treatment; the largest increase in stem diameter was 23.21% under 0.50% PS + 10.0 mg / L GO treatment; the largest increase in root diameter was 10.25% under 0.50% PS + 15.0 mg / L GO treatment; the largest increase in total dry biomass was 90.78% under 0.50% PS + 12.5 mg / L GO treatment; the largest increase in seedling vigor index was 82.31% under 0.50% PS + 5.0 mg / L GO treatment; and the largest increase in leaf chlorophyll SPAD value was 12.84% under 0.50% PS + 12.5 mg / L GO treatment. The GO treatment resulted in the largest increase in root vigor of TS163 maize seedlings, at 216.12% (see appendix). Figure 5 and 7 Similarly, in a 21-day experiment on the foliar spraying of different concentrations of GO solution to alleviate the microplastic toxicity of 0.50% PS, the traits of TS163 maize genotype seedlings under 0.50% PS stress were significantly reduced compared to the CK control treatment, with reductions ranging from 13.22% (stem diameter) to 62.21% (root activity) (see appendix). Figure 6 and 7Different concentrations of GO foliar spraying improved 12 traits of TS163 maize seedlings under 0.50% PS stress for 21 days to some extent, and the improvement effects of different concentrations of GO foliar spraying on the 12 traits of TS163 maize seedlings under 0.50% PS stress varied (see appendix). Figure 6 and 7 Compared with the 0.50% PS stress treatment, the 0.50% PS + 2.5 mg / L GO treatment resulted in the largest increase in seedling length (10.59%), seedling fresh weight (51.34%), and seedling dry weight (57.05%) of the TS163 maize genotype seedlings; the 0.50% PS + 5.0 mg / L GO treatment resulted in the largest increase in root length (36.45%), and the 0.50% PS + 2.5 mg / L GO treatment resulted in the largest increase in root fresh weight (104.72%). The largest increase in root dry weight of TS163 maize seedlings was 113.38% under GO treatment; the largest increase in stem diameter was 12.24% under 0.50% PS + 5.0 mg / L GO treatment; the largest increase in root diameter was 12.89% under 0.50% PS + 2.5 mg / L GO treatment; the largest increase in total dry biomass was 66.33% under 0.50% PS + 5.0 mg / L GO treatment; the largest increase in seedling vigor index was 70.32% under 0.50% PS + 5.0 mg / L GO treatment; and the largest increase in leaf chlorophyll SPAD value was 31.84% under 0.50% PS + 15.0 mg / L GO treatment. The GO treatment resulted in the largest increase in root vigor in maize seedlings with the TS163 genotype, at 167.93% (see appendix). Figure 6 and 7 The results indicate that different concentrations of GO solution applied to seeds and leaves have varying effects on mitigating PS microplastic toxicity in TS163 maize genotype seedlings during 7-day and 21-day growth trials.

[0091] 5. Pearson correlation analysis of 12 traits in 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of ZnONPs solution for mitigating PS microplastic toxicity: In the 7-day growth trial of TS163 maize genotype seedlings treated with different concentrations of ZnONPs solution for mitigating 0.50% PS microplastic toxicity, a total of 31 pairs of significant positive / negative correlations were detected among the 12 traits in the 8 treatments (P<0.05). (See Appendix) Figure 8 Regarding the 21-day growth experiment of TS163 maize genotype seedlings treated with foliar spraying of different concentrations of ZnONPs solutions to alleviate the toxicity of 0.50% PS microplastics, a total of 25 pairs of traits showed significant positive / negative correlations (P<0.05) among the 12 traits and 8 treatments (see appendix). Figure 9 This indicates that these 12 traits of maize seedlings have close synergistic or antagonistic negative effects on each other, jointly influencing the growth and development of maize seedlings under different treatments.

[0092] 6. Pearson correlation analysis of 12 traits in 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of GO solution to alleviate PS microplastic toxicity: Consistently, in the 7-day growth trials of TS163 maize genotype seedlings treated with different concentrations of GO solution to alleviate 0.50% PS microplastic toxicity, a total of 39 pairs of significant positive / negative correlations were found among all treatments of the 12 traits (P<0.05). (See Appendix) Figure 10 Regarding the 21-day trial of foliar spraying of different concentrations of GO solution to alleviate the toxicity of 0.50% PS microplastics in TS163 maize genotype seedlings, a total of 44 pairs of significant positive / negative correlations were identified among these traits across all treatments (P<0.05). (See Appendix) Figure 11 This indicates that these traits of maize seedlings work closely together, with positive or antagonistic negative effects jointly regulating the growth and development of maize seedlings under different treatments.

[0093] 7. Cluster analysis of all treatments in the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of ZnONPs solution for seed soaking and foliar spraying to alleviate PS microplastic toxicity: We further used the IMLOG2 function to standardize the means of the corresponding 12 traits among all treatments in the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of ZnONPs solution for seed soaking and foliar spraying to alleviate 0.50% PS microplastic toxicity; then IBM-SPSS Statistics were used. 16. Data statistical analysis software was used to conduct inter-group cluster comprehensive evaluation of all treatments in the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of ZnONPs solution for seed soaking and foliar spraying to alleviate 0.50% PS microplastic toxicity. The evaluation qualitatively assessed the overall PS microplastic toxicity mitigation status and growth condition of TS163 maize genotype seedlings in the corresponding treatments of the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of ZnONPs solution for seed soaking and foliar spraying to alleviate 0.50% PS microplastic toxicity. In a 7-day experiment on the growth of TS163 maize genotype seedlings, seed soaking in ZnONPs solutions of different concentrations alleviated the toxicity of 0.50% PS microplastics, based on the performance of 12 traits of TS163 maize genotype seedlings among 8 treatments, at an Euclidean distance of 6, the treatments of 0.50% PS + 25 μmol / L ZnONPs, 0.50% PS + 50 μmol / L ZnONPs, the control (CK) treatment, and the 0.50% PS + 75 μmol / L ZnONPs treatment were clustered into type I. All 12 traits of TS163 maize genotype seedlings under these four treatments were well-developed, and this type was named the treatment with good seedling growth. The treatments of 0.50% PS + 100 μmol / L ZnONPs, 0.50% PS + 150 μmol / L ZnONPs, and 0.50% PS + 125 μmol / L ZnONPs were also clustered. ZnONPs treatments clustered into type II, where all 12 traits of TS163 maize genotype seedlings under the three treatments were weak; this type was named the treatment type with weak maize seedling growth. Meanwhile, 0.50% PS treatments clustered into type III, where TS163 maize genotype seedlings under this type showed poor performance in all 12 traits due to the toxic effects of 0.50% PS microplastics; this type was named the treatment type with poor maize seedling growth. (See appendix) Figure 12In addition, in a 21-day foliar spraying experiment of different concentrations of ZnONPs solutions to alleviate the toxicity of 0.50% PS microplastics in TS163 maize genotype seedlings, based on the performance of 12 traits of TS163 maize genotype seedlings among 8 treatments, at an Euclidean distance of 5.5, the CK control treatment clustered into type I. Seedlings of the TS163 maize genotype under this type of treatment showed good performance in all 12 traits and were not affected by PS microplastic toxicity; this type was named the treatment with good maize seedling growth. The treatments included 0.50% PS + 50 μmol / L ZnONPs, 0.50% PS + 75 μmol / L ZnONPs, 0.50% PS + 125 μmol / L ZnONPs, 0.50% PS + 25 μmol / L ZnONPs, 0.50% PS + 100 μmol / L ZnONPs, and 0.50% PS + 150 μmol / L ZnONPs. ZnONPs treatments clustered into type II, where all 12 traits of TS163 maize genotype seedlings under the six treatments in this type were weak, and this type was named the weak maize seedling growth treatment type. Meanwhile, 0.50% PS treatments clustered into type III, where TS163 maize genotype seedlings under this type of treatment showed poor performance in all 12 traits due to the toxicity of 0.50% PS microplastics, and this type was named the poor maize seedling growth treatment type. (See appendix) Figure 13 These results indicate that the intergroup clustering comprehensive evaluation method can only qualitatively evaluate the growth status of maize seedlings under all treatments in the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of ZnONPs solution for seed soaking or foliar spraying, and the mitigation effect of corresponding concentrations of ZnONPs solution on maize seedlings under 0.50% concentration of PS microplastic toxicity. However, it cannot clearly and quantitatively distinguish the overall mitigation effect of different concentrations of ZnONPs solution for seed soaking or foliar spraying on maize seedlings under 0.50% concentration of PS microplastic toxicity, which seriously limits the application of suitable concentrations of ZnONPs solution for seed soaking or foliar spraying to mitigate the toxicity effect of 0.50% concentration of PS microplastic on maize seedlings.

[0094] 8. Comprehensive evaluation of inter-group clustering among all treatments in the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of GO solution for mitigating PS microplastic toxicity: Similarly, the mean values ​​of the corresponding 12 traits among all treatments in the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of GO solution for mitigating 0.50% PS microplastic toxicity were further standardized using IMLOG2. Then, a comprehensive inter-group clustering evaluation was performed among all treatments in the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of GO solution for mitigating 0.50% PS microplastic toxicity. This qualitatively evaluated the overall PS microplastic toxicity mitigation status and growth condition of TS163 maize genotype seedlings in the corresponding treatments of the 7-day and 21-day growth trials of TS163 maize genotype seedlings treated with different concentrations of GO solution for mitigating 0.50% PS microplastic toxicity. In a 7-day experiment on the growth of TS163 maize genotype seedlings, seed soaking in different concentrations of GO solution mitigated the toxicity of 0.50% PS microplastics. Based on the performance of 12 traits of TS163 maize genotype seedlings among 8 treatments, at an Euclidean distance of 6.5, the treatments of 0.50% PS + 7.5 mg / L GO, 0.50% PS + 10.0 mg / L GO, 0.50% PS + 12.5 mg / L GO, and the control (CK) treatment were clustered into type I. All 12 traits of TS163 maize genotype seedlings under these four treatments were well-developed, and this type was named the treatment with good seedling growth. The treatments of 0.50% PS + 2.5 mg / L GO, 0.50% PS + 5.0 mg / L GO, and 0.50% PS + 15.0 mg / L GO were also clustered. The GO treatment clustered into type II, where all 12 traits of TS163 maize genotype seedlings under the three treatments were weak, and this type was named the weak maize seedling growth treatment type. The 0.50% PS treatment clustered into type III, where TS163 maize genotype seedlings under this type of treatment showed poor performance in all 12 traits due to the toxicity of 0.50% PS microplastics, and this type was named the poor maize seedling growth treatment type. (See appendix) Figure 14In addition, in a 21-day experiment on the foliar spraying of 0.50% PS microplastic toxicity in TS163 maize genotype seedlings to alleviate the toxicity of 0.50% PS microplastics, based on the performance of 12 traits of TS163 maize genotype seedlings among 8 treatments, at an Euclidean distance of 13, the 0.50% PS + 2.5 mg / L GO treatment, 0.50% PS + 5.0 mg / L GO treatment, CK control treatment, 0.50% PS + 7.5 mg / L GO treatment, and 0.50% PS + 15.0 mg / L GO treatment clustered into type I. All 12 traits of TS163 maize genotype seedlings under these 5 treatments were well-developed, and this type was named the treatment with good seedling growth. The 0.50% PS + 12.5 mg / L GO treatment... The GO treatment clustered into type II, where all 12 traits of the TS163 maize genotype seedlings were weak, and this type was named the weak maize seedling growth treatment type. The 0.50% PS treatment and the 0.50% PS + 10.0 mg / L GO treatment clustered into type III, where all 12 traits of the TS163 maize genotype seedlings were poor, and this type was named the poor maize seedling growth treatment type. (See appendix) Figure 15 These analyses show that the intergroup clustering comprehensive evaluation method can only provide a vague qualitative assessment of the growth status of maize seedlings in all treatments of the TS163 maize genotype seedling growth trials (7-day and 21-day) using different concentrations of GO solution for seed soaking or foliar spraying to alleviate PS microplastic toxicity, as well as the mitigation effect of corresponding concentrations of GO solution for seed soaking or foliar spraying on maize seedlings under 0.50% PS microplastic toxicity. However, it cannot accurately and objectively quantitatively identify the overall mitigation effect of different concentrations of GO solution for seed soaking or foliar spraying on maize seedlings under 0.50% PS microplastic toxicity, which seriously affects the application of appropriate concentrations of GO solution for seed soaking or foliar spraying to alleviate the toxicity of 0.50% PS microplastics on maize seedlings.

[0095] 9. Comprehensive evaluation of the mitigating effect of different concentrations of ZnONPs solution soaking or foliar spraying on 0.50% PS microplastic toxicity in maize seedlings: We further evaluated the mitigating effect of different concentrations of ZnONPs solution soaking or foliar spraying on 12 traits of TS163 maize genotype seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity. ZnONPS The values ​​(Tables 3 and 4) were used as evaluation indicators for the mitigating effect of different concentrations of ZnONPs solution soaking in seeds or foliar spraying on TS163 maize genotype seedlings under 0.50% PS microplastic toxicity. The membership function method was employed, based on the attached values... Figure 1Formula (6) was used to calculate the mitigation effect of different concentrations of ZnONPs solution soaking or foliar spraying on 12 traits of TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. ZnONPs (Tables 5 and 6), then according to the appendix Figure 1 Formula (7) was used to evaluate the overall mitigation effect of different concentrations of ZnONPs solution soaking or foliar spraying on TS163 maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity (CME). ZnONPs The results are attached. Figure 16 The results showed that soaking seeds in six concentrations of ZnONPs solutions affected the CME of TS163 maize genotype seedlings cultured on day 7 under 0.50% PS microplastic toxicity. ZnONPs The concentrations ranged from 0.222 (0.50% PS + 125 μmol / L ZnONPs treatment) to 0.703 (0.50% PS + 75 μmol / L ZnONPs treatment); similarly, foliar spraying of six ZnONPs solutions had an effect on the CME of TS163 maize genotype seedlings cultured for 21 days under 0.50% PS microplastic toxicity. ZnONPs The values ​​ranged from 0.152 (0.50% PS + 100 μmol / L ZnONPs treatment) to 0.764 (0.50% PS + 75 μmol / L ZnONPs treatment). It is speculated that soaking seeds in a 75 μmol / L ZnONPs solution and foliar spraying provided the best overall mitigation effect on maize seedlings cultured for 7 days and 21 days under 0.50% PS microplastic toxicity.

[0096] Table 3. Relative mitigation index (RMEI) of seeds soaked in ZnONPs solution of different concentrations on 12 traits of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity during the 7-day growth experiment. ZnONPS )value

[0097]

[0098]

[0099] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. The following treatments were used in a 7-day seedling growth experiment: 0.50% PS + 25 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate); 0.50% PS + 50 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate); and 0.50% PS + 75 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 75 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate). Treatments for 7-day seedling growth were as follows: 0.50% PS + 100 μmol / L ZnONPs (soil-based culture), 0.50% PS + 125 μmol / L ZnONPs (soil-based culture), and 0.50% PS + 150 μmol / L ZnONPs (soil-based culture). ZnONPs treatment (soaking in a mixture of 0.50% PS and 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate).

[0100] Table 4. Relative mitigation index (RMEI) of foliar spraying with different concentrations of ZnONPs solution on 12 traits of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity during a 21-day growth experiment. ZnONPS )value

[0101]

[0102] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. Treatments for 21-day seedling growth were as follows: 0.50% PS + 25 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 25 μmol / L ZnONPs solution); 0.50% PS + 50 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 50 μmol / L ZnONPs solution); and 0.50% PS + 75 μmol / L ZnONPs (seed treatment for 21-day seedling growth). ZnONPs treatments (seed soaking in a mixture of 0.50% PS and 75 μmol / L ZnONPs + culturing in a microplastic soil substrate with 0.50% PS + foliar spraying of seedlings with 75 μmol / L ZnONPs solution), 0.50% PS + 100 μmol / L ZnONPs treatments for a 21-day seedling growth experiment, 0.50% PS + 125 μmol / L ZnONPs treatments for a 21-day seedling growth experiment, 0.50% PS + 125 μmol / L ZnONPs treatments for a 21-day seedling growth experiment. The treatment consisted of: seed soaking in a mixture of ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 125 μmol / L ZnONPs solution; and a 0.50% PS + 150 μmol / L ZnONPs treatment for a 21-day seedling growth test (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 150 μmol / L ZnONPs solution).

[0103] Table 5. Membership values ​​(U(RMEI)) of seed soaking in ZnONPs solutions of different concentrations on mitigation effects on 12 traits of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity during the 7-day growth experiment. ZnONPS ))

[0104]

[0105] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. The following treatments were used in a 7-day seedling growth experiment: 0.50% PS + 25 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate); 0.50% PS + 50 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate); and 0.50% PS + 75 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 75 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate). Treatments for 7-day seedling growth were as follows: 0.50% PS + 100 μmol / L ZnONPs (soil-based culture), 0.50% PS + 125 μmol / L ZnONPs (soil-based culture), and 0.50% PS + 150 μmol / L ZnONPs (soil-based culture). ZnONPs treatment (soaking in a mixture of 0.50% PS and 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate).

[0106] Table 6. Membership values ​​(U(RMEI)) of foliar spraying with different concentrations of ZnONPs solution on the mitigation effect of 0.50% PS microplastic toxicity on 12 traits of TS163 maize genotype seedlings during the 21-day growth experiment. ZnONPS ))

[0107]

[0108]

[0109] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. Treatments for 21-day seedling growth were as follows: 0.50% PS + 25 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 25 μmol / L ZnONPs solution); 0.50% PS + 50 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 50 μmol / L ZnONPs solution); and 0.50% PS + 75 μmol / L ZnONPs (seed treatment for 21-day seedling growth). ZnONPs treatments (seed soaking in a mixture of 0.50% PS and 75 μmol / L ZnONPs + culturing in a microplastic soil substrate with 0.50% PS + foliar spraying of seedlings with 75 μmol / L ZnONPs solution), 0.50% PS + 100 μmol / L ZnONPs treatments for a 21-day seedling growth experiment, 0.50% PS + 125 μmol / L ZnONPs treatments for a 21-day seedling growth experiment, 0.50% PS + 125 μmol / L ZnONPs treatments for a 21-day seedling growth experiment. The treatment consisted of: seed soaking in a mixture of ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 125 μmol / L ZnONPs solution; and a 0.50% PS + 150 μmol / L ZnONPs treatment for a 21-day seedling growth test (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 150 μmol / L ZnONPs solution).

[0110] 10. Comprehensive evaluation of the mitigating effect of different concentrations of GO solution soaking or foliar spraying on the toxicity of 0.50% PS microplastics in maize seedlings: Similarly, we evaluated the RMEI of 12 traits in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or foliar spraying with different concentrations of GO solution. GO The values ​​(Tables 7 and 8) were used as evaluation indicators for the mitigating effect of different concentrations of GO solution soaking on seeds or foliar spraying on TS163 maize genotype seedlings under 0.50% PS microplastic toxicity. The membership function method was employed, based on the attached values... Figure 1 Formula (8) was used to calculate the mitigation effect of different concentrations of GO solution soaking or foliar spraying on 12 traits of TS163 maize genotype seedlings cultured on day 7 or day 21 of 0.50% PS microplastic toxicity. GO (Tables 9 and 10), then according to the appendix Figure 1 Formula (9) was used to evaluate the overall mitigation effect of different concentrations of GO solution soaking in seeds or foliar spraying on TS163 maize seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity (CME). GO The results are attached. Figure 17 The results showed that soaking seeds in six different concentrations of GO solution had a negative effect on the CME of TS163 maize genotype seedlings cultured on day 7 under 0.50% PS microplastic toxicity. GO The values ​​ranged from 0.131 (0.50% PS + 2.5 mg / L GO treatment) to 0.900 (0.50% PS + 12.5 mg / L GO treatment); the difference was that foliar spraying with six GO solutions had a different effect on the CME of TS163 maize genotype seedlings cultured for 21 days under 0.50% PS microplastic toxicity. GO The values ​​ranged from 0.088 (0.50% PS + 10.0 mg / L GO treatment) to 0.847 (0.50% PS + 5.0 mg / L GO treatment). It is speculated that soaking seeds in a 12.5 mg / L GO solution provided the best overall mitigation effect on maize genotype seedlings cultured for 7 days under 0.50% PS microplastic toxicity, while foliar spraying with a 5.0 mg / L GO solution provided the best overall mitigation effect on maize genotype seedlings cultured for 21 days under 0.50% PS microplastic toxicity.

[0111] Table 7. Relative mitigation index (RMEI) of seeds soaked in different concentrations of GO solution on 12 traits of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity during a 7-day growth experiment. GO )value

[0112]

[0113]

[0114] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. The following treatments were used in a 7-day seedling growth experiment: 0.50% PS + 2.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 2.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 5.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 5.0 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 7.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 7.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); and 0.50% PS + 10.0 mg / L GO. GO refers to the following treatments used in the 7-day seedling growth experiment: 0.50% PS + 10.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 10.0 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 12.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 12.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); and 0.50% PS + 15.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in 0.50% PS microplastic soil substrate).

[0115] Table 8. Relative mitigation index (RMEI) of foliar spraying with different concentrations of GO solution on 12 traits of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity during the 21-day growth experiment. GO )value

[0116]

[0117] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. The following treatments were used in a 21-day seedling growth experiment: 0.50% PS + 2.5 mg / L GO (seed soaking in a mixture of 0.50% PS and 2.5 mg / L GO + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 2.5 mg / L GO solution); 0.50% PS + 5.0 mg / L GO (seed soaking in a mixture of 0.50% PS and 5.0 mg / L GO + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 5.0 mg / L GO solution); and 0.50% PS + 7.5 mg / L GO (seed treatment of 0.50% PS + 7.5 mg / L GO solution). Treatments included: 1) Seedling soaking in a GO mixture + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 7.5 mg / L GO solution; 2) Treatments with 0.50% PS + 10.0 mg / L GO for 21-day seedling growth: 0.50% PS + 10.0 mg / L GO mixture (seed soaking in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 10.0 mg / L GO solution); 3) Treatments with 0.50% PS + 12.5 mg / L GO for 21-day seedling growth: 0.50% PS + 12.5 mg / L GO mixture (seed soaking in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 12.5 mg / L GO solution); 4) Treatments with 0.50% PS + 15.0 mg / L GO. GO was used in a 21-day seedling growth experiment, consisting of 0.50% PS + 15.0 mg / L GO treatment (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 15.0 mg / L GO solution).

[0118] Table 9. Membership values ​​(U(RMEI)) of seed soaking in different concentrations of GO solution on mitigation effects of 0.50% PS microplastic toxicity on 12 traits of TS163 maize genotype seedlings during the 7-day growth experiment. GO ))

[0119]

[0120] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. The following treatments were used in a 7-day seedling growth experiment: 0.50% PS + 2.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 2.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 5.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 5.0 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 7.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 7.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); and 0.50% PS + 10.0 mg / L GO. GO refers to the following treatments used in the 7-day seedling growth experiment: 0.50% PS + 10.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 10.0 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 12.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 12.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); and 0.50% PS + 15.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in 0.50% PS microplastic soil substrate).

[0121] Table 10. Membership values ​​(U(RMEI)) of foliar spraying with different concentrations of GO solution on the mitigation effect of 0.50% PS microplastic toxicity on 12 traits of TS163 maize genotype seedlings during the 21-day growth experiment. GO ))

[0122]

[0123]

[0124] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. The following treatments were used in a 21-day seedling growth experiment: 0.50% PS + 2.5 mg / L GO (seed soaking in a mixture of 0.50% PS and 2.5 mg / L GO + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 2.5 mg / L GO solution); 0.50% PS + 5.0 mg / L GO (seed soaking in a mixture of 0.50% PS and 5.0 mg / L GO + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 5.0 mg / L GO solution); and 0.50% PS + 7.5 mg / L GO (seed treatment of 0.50% PS + 7.5 mg / L GO solution). Treatments included: 1) Seedling soaking in a GO mixture + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 7.5 mg / L GO solution; 2) Treatments with 0.50% PS + 10.0 mg / L GO for 21-day seedling growth: 0.50% PS + 10.0 mg / L GO mixture (seed soaking in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 10.0 mg / L GO solution); 3) Treatments with 0.50% PS + 12.5 mg / L GO for 21-day seedling growth: 0.50% PS + 12.5 mg / L GO mixture (seed soaking in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 12.5 mg / L GO solution); 4) Treatments with 0.50% PS + 15.0 mg / L GO. GO was used in a 21-day seedling growth experiment, consisting of 0.50% PS + 15.0 mg / L GO treatment (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 15.0 mg / L GO solution).

[0125] 11. Comprehensive evaluation of the mitigation strength of different concentrations of ZnONPs solution soaking or foliar spraying on maize seedlings under 0.50% PS microplastic toxicity: We also evaluated the AMIC of 12 traits in TS163 maize genotype seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity by different concentrations of ZnONPs solution soaking or foliar spraying. ZnONPSThe values ​​(Tables 11 and 12) were used as evaluation indicators for the mitigation intensity of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity by soaking seeds or foliar spraying with different concentrations of ZnONPs solution. The membership function method was used, based on the attached values. Figure 1 Formula (10) was used to calculate the mitigation strength membership value (U(AMIC)) of 12 traits in TS163 maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with different concentrations of ZnONPs solution. ZnONPs (Tables 13 and 14), then according to the appendix Figure 1 Formula (11) measures the overall mitigation intensity (CMI) of different concentrations of ZnONPs solution soaking or foliar spraying on TS163 maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity. ZnONPs The results are attached. Figure 18 The results showed that soaking seeds in six concentrations of ZnONPs solutions affected the CMI of TS163 maize genotype seedlings cultured on day 7 under 0.50% PS microplastic toxicity. ZnONPs The CMI ranged from 0.231 (0.50% PS + 125 μmol / L ZnONPs treatment) to 0.720 (0.50% PS + 75 μmol / L ZnONPs treatment); similarly, the CMI of foliar spraying with six concentrations of ZnONPs solutions on 21-day-old TS163 maize genotype seedlings cultured under 0.50% PS microplastic toxicity was also observed. ZnONPs The values ​​ranged from 0.170 (0.50% PS + 100 μmol / L ZnONPs treatment) to 0.780 (0.50% PS + 75 μmol / L ZnONPs treatment). It was predicted that soaking seeds in a 75 μmol / L ZnONPs solution and foliar spraying would provide the greatest overall mitigation effect on maize genotype seedlings cultured for 7 days and 21 days under 0.50% PS microplastic toxicity.

[0126] Table 11. Absolute mitigation strength coefficients (AMICs) of 12 traits in TS163 maize genotype seedlings under 0.50% PS microplastic toxicity after soaking seeds in ZnONPs solutions of different concentrations during a 7-day growth experiment. ZnONPS )value

[0127]

[0128] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. The following treatments were used in a 7-day seedling growth experiment: 0.50% PS + 25 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate); 0.50% PS + 50 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate); and 0.50% PS + 75 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 75 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate). Treatments for 7-day seedling growth were as follows: 0.50% PS + 100 μmol / L ZnONPs (soil-based culture), 0.50% PS + 125 μmol / L ZnONPs (soil-based culture), and 0.50% PS + 150 μmol / L ZnONPs (soil-based culture). ZnONPs treatment (soaking in a mixture of 0.50% PS and 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate).

[0129] Table 12. Absolute mitigation intensity coefficients (AMICs) of foliar spraying with different concentrations of ZnONPs solution on 12 traits of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity during a 21-day growth experiment. ZnONPS )value

[0130]

[0131] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. Treatments for 21-day seedling growth were as follows: 0.50% PS + 25 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 25 μmol / L ZnONPs solution); 0.50% PS + 50 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 50 μmol / L ZnONPs solution); and 0.50% PS + 75 μmol / L ZnONPs (seed treatment for 21-day seedling growth). ZnONPs treatments (seed soaking in a mixture of 0.50% PS and 75 μmol / L ZnONPs + culturing in a microplastic soil substrate with 0.50% PS + foliar spraying of seedlings with 75 μmol / L ZnONPs solution), 0.50% PS + 100 μmol / L ZnONPs treatments for a 21-day seedling growth experiment, 0.50% PS + 125 μmol / L ZnONPs treatments for a 21-day seedling growth experiment, 0.50% PS + 125 μmol / L ZnONPs treatments for a 21-day seedling growth experiment. The treatment consisted of: seed soaking in a mixture of ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 125 μmol / L ZnONPs solution; and a 0.50% PS + 150 μmol / L ZnONPs treatment for a 21-day seedling growth test (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 150 μmol / L ZnONPs solution).

[0132] Table 13. Membership values ​​(U(AMIC)) of the mitigation strength of seed soaking in ZnONPs solutions of different concentrations to 12 traits of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity during the 7-day growth experiment. ZnONPS ))

[0133]

[0134] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. The following treatments were used in a 7-day seedling growth experiment: 0.50% PS + 25 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate); 0.50% PS + 50 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate); and 0.50% PS + 75 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 75 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate). Treatments for 7-day seedling growth were as follows: 0.50% PS + 100 μmol / L ZnONPs (soil-based culture), 0.50% PS + 125 μmol / L ZnONPs (soil-based culture), and 0.50% PS + 150 μmol / L ZnONPs (soil-based culture). ZnONPs treatment (soaking in a mixture of 0.50% PS and 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate).

[0135] Table 14. Membership values ​​(U(AMIC)) of the mitigation strength of foliar spraying with different concentrations of ZnONPs solution on 12 traits of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity during a 21-day growth experiment. ZnONPS ))

[0136]

[0137] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. Treatments for 21-day seedling growth were as follows: 0.50% PS + 25 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 25 μmol / L ZnONPs solution); 0.50% PS + 50 μmol / L ZnONPs (seed soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 50 μmol / L ZnONPs solution); and 0.50% PS + 75 μmol / L ZnONPs (seed treatment for 21-day seedling growth). ZnONPs treatments (seed soaking in a mixture of 0.50% PS and 75 μmol / L ZnONPs + culturing in a microplastic soil substrate with 0.50% PS + foliar spraying of seedlings with 75 μmol / L ZnONPs solution), 0.50% PS + 100 μmol / L ZnONPs treatments for a 21-day seedling growth experiment, 0.50% PS + 125 μmol / L ZnONPs treatments for a 21-day seedling growth experiment, 0.50% PS + 125 μmol / L ZnONPs treatments for a 21-day seedling growth experiment. The treatment consisted of: seed soaking in a mixture of ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 125 μmol / L ZnONPs solution; and a 0.50% PS + 150 μmol / L ZnONPs treatment for a 21-day seedling growth test (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 150 μmol / L ZnONPs solution).

[0138] 12. Comprehensive evaluation of the mitigation strength of different concentrations of GO solution soaking or foliar spraying on maize seedlings under 0.50% PS microplastic toxicity: We further evaluated the AMIC of 12 traits in TS163 maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or foliar spraying with different concentrations of GO solution. GO The values ​​(Tables 15 and 16) were used as evaluation indicators for the mitigation intensity of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity by soaking seeds or foliar spraying with different concentrations of GO solution. The membership function method was employed, based on the attached values... Figure 1 Formula (12) was used to calculate the mitigation strength membership value (U(AMIC)) of 12 traits in TS163 maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with different concentrations of GO solution. GO (Tables 17 and 18), then according to the appendix Figure 1 Formula (13) measures the overall mitigation strength (CMI) of different concentrations of GO solution applied to seeds or leaves after soaking or foliar spraying under 0.50% PS microplastic toxicity for TS163 maize genotype seedlings cultured for 7 days or 21 days. GO The results are attached. Figure 19 The results showed that soaking seeds in six different concentrations of GO solution affected the CMI of TS163 maize genotype seedlings cultured on day 7 under 0.50% PS microplastic toxicity. GO The CMI ranged from 0.141 (0.50% PS + 2.5 mg / L GO treatment) to 0.908 (0.50% PS + 12.5 mg / L GO treatment); while the foliar spraying of six GO concentrations had an effect on the CMI of TS163 maize genotype seedlings cultured on day 21 under 0.50% PS microplastic toxicity. GO The values ​​ranged from 0.095 (0.50% PS + 10.0 mg / L GO treatment) to 0.860 (0.50% PS + 5.0 mg / L GO treatment). It was predicted that soaking seeds in a 75 μmol / L ZnONPs solution and foliar spraying would provide the greatest overall mitigation for maize genotype seedlings cultured on day 7 and day 21 of exposure to 0.50% PS microplastic toxicity.

[0139] Table 15. Absolute mitigation strength coefficients (AMICs) of 12 traits in TS163 maize genotype seedlings under 0.50% PS microplastic toxicity after soaking seeds in different concentrations of GO solution during a 7-day growth experiment. GO )value

[0140]

[0141] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. The following treatments were used in a 7-day seedling growth experiment: 0.50% PS + 2.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 2.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 5.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 5.0 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 7.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 7.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); and 0.50% PS + 10.0 mg / L GO. GO refers to the following treatments used in the 7-day seedling growth experiment: 0.50% PS + 10.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 10.0 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 12.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 12.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); and 0.50% PS + 15.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in 0.50% PS microplastic soil substrate).

[0142] Table 16. Absolute mitigation intensity coefficients (AMICs) of foliar spraying with different concentrations of GO solution on 12 traits of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity during a 21-day growth experiment. GO )value

[0143]

[0144] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. The following treatments were used in a 21-day seedling growth experiment: 0.50% PS + 2.5 mg / L GO (seed soaking in a mixture of 0.50% PS and 2.5 mg / L GO + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 2.5 mg / L GO solution); 0.50% PS + 5.0 mg / L GO (seed soaking in a mixture of 0.50% PS and 5.0 mg / L GO + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 5.0 mg / L GO solution); and 0.50% PS + 7.5 mg / L GO (seed treatment of 0.50% PS + 7.5 mg / L GO solution). Treatments included: 1) Seedling soaking in a GO mixture + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 7.5 mg / L GO solution; 2) Treatments with 0.50% PS + 10.0 mg / L GO for 21-day seedling growth: 0.50% PS + 10.0 mg / L GO mixture (seed soaking in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 10.0 mg / L GO solution); 3) Treatments with 0.50% PS + 12.5 mg / L GO for 21-day seedling growth: 0.50% PS + 12.5 mg / L GO mixture (seed soaking in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 12.5 mg / L GO solution); 4) Treatments with 0.50% PS + 15.0 mg / L GO. GO was used in a 21-day seedling growth experiment, consisting of 0.50% PS + 15.0 mg / L GO treatment (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 15.0 mg / L GO solution).

[0145] Table 17. Membership values ​​(U(AMIC)) of the mitigation strength of seed soaking in different concentrations of GO solution on 12 traits of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity during the 7-day growth experiment. GO ))

[0146]

[0147] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. The following treatments were used in a 7-day seedling growth experiment: 0.50% PS + 2.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 2.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 5.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 5.0 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 7.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 7.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); and 0.50% PS + 10.0 mg / L GO. GO refers to the following treatments used in the 7-day seedling growth experiment: 0.50% PS + 10.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 10.0 mg / L GO + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 12.5 mg / L GO (seed soaking in a mixture of 0.50% PS + 12.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); and 0.50% PS + 15.0 mg / L GO (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in 0.50% PS microplastic soil substrate).

[0148] Table 18. Membership values ​​(U(AMIC)) of the mitigation strength of foliar spraying with different concentrations of GO solution on 12 traits of TS163 maize genotype seedlings under 0.50% PS microplastic toxicity during a 21-day growth experiment. GO ))

[0149]

[0150] Note: SL is seedling length, SFW is seedling fresh weight, SDW is seedling dry weight, RL is root length, RFW is root fresh weight, RDW is root dry weight, SD is stem diameter, RD is root diameter, PDW is total plant dry biomass, SPAD is leaf chlorophyll SPAD value, SSI is seedling vigor index, and RV is root activity. The following treatments were used in a 21-day seedling growth experiment: 0.50% PS + 2.5 mg / L GO (seed soaking in a mixture of 0.50% PS and 2.5 mg / L GO + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 2.5 mg / L GO solution); 0.50% PS + 5.0 mg / L GO (seed soaking in a mixture of 0.50% PS and 5.0 mg / L GO + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 5.0 mg / L GO solution); and 0.50% PS + 7.5 mg / L GO (seed treatment of 0.50% PS + 7.5 mg / L GO solution). Treatments included: 1) Seedling soaking in a GO mixture + culturing in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 7.5 mg / L GO solution; 2) Treatments with 0.50% PS + 10.0 mg / L GO for 21-day seedling growth: 0.50% PS + 10.0 mg / L GO mixture (seed soaking in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 10.0 mg / L GO solution); 3) Treatments with 0.50% PS + 12.5 mg / L GO for 21-day seedling growth: 0.50% PS + 12.5 mg / L GO mixture (seed soaking in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 12.5 mg / L GO solution); 4) Treatments with 0.50% PS + 15.0 mg / L GO. GO was used in a 21-day seedling growth experiment, consisting of 0.50% PS + 15.0 mg / L GO treatment (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with 15.0 mg / L GO solution).

[0151] 13. Optimal ZnONPs / GO solution concentration for determining the best mitigation effect / maximum mitigation intensity of 0.50% PS microplastic toxicity in maize seedlings: Further evaluation was conducted by comprehensively comparing the intergroup clustering results of the growth status of all treatments in 7-day and 21-day growth trials of maize genotype seedlings treated with different concentrations of ZnONPs solution for seed soaking and foliar spraying to mitigate 0.50% PS microplastic toxicity (see appendix). Figure 12 and 13The comprehensive evaluation results of the mitigation effects of different concentrations of ZnONPs solution soaking on seeds or foliar spraying on TS163 maize genotype seedlings at 0.50% PS microplastic toxicity (see attached). Figure 16 The comprehensive evaluation results of the mitigation strength of different concentrations of ZnONPs solution soaking in seeds or foliar spraying on TS163 maize genotype seedlings at 0.50% PS microplastic toxicity (attached) and the results of the above-mentioned comprehensive evaluation of the mitigation strength of different concentrations of ZnONPs solution soaking in seeds or foliar spraying on foliar spraying (attached) Figure 18 We ultimately determined that the 0.50% PS + 75 μmol / L ZnONPs treatment (seed soaking in a mixture of 0.50% PS + 75 μmol / L ZnONPs + cultivation in a soil substrate with 0.50% PS microplastics) in the 7-day seedling growth test and the 0.50% PS + 75 μmol / L ZnONPs treatment (seed soaking in a mixture of 0.50% PS + 75 μmol / L ZnONPs + cultivation in a soil substrate with 0.50% PS microplastics + foliar spraying of seedlings with 75 μmol / L ZnONPs solution) in the 21-day seedling growth test were the optimal mitigation effects and the optimal mitigation strength of 0.50% PS microplastics concentration in TS163 maize genotype seedlings. Therefore, we can soak maize seeds in a 75 μmol / L ZnONPs solution and then cultivate them in a 0.50% PS microplastic environment for 7 days to achieve normal seedling emergence. Then, we continue to spray the leaves of each maize seedling in each pot with 1 mL of the 75 μmol / L ZnONPs solution every two days in the morning, for a total of 7 foliar sprays. This treatment showed the best overall mitigation effect and strongest overall mitigation intensity for maize seedlings cultivated for 7 and 21 days under 0.50% PS microplastic toxicity. Furthermore, we further evaluated the intergroup clustering results of the growth status of TS163 maize genotype seedlings in all treatments of the above-mentioned 7-day and 21-day growth experiments of seed soaking and foliar spraying with different concentrations of GO solution to mitigate 0.50% PS microplastic toxicity (see attached). Figure 14 and 15 The comprehensive evaluation results of the mitigation effects of different concentrations of GO solution soaking on seeds or foliar spraying on 0.50% PS microplastic toxicity in TS163 maize genotype seedlings (see attached). Figure 17The comprehensive evaluation results of the mitigation intensity of 0.50% PS microplastic toxicity in TS163 maize genotype seedlings by soaking seeds or spraying leaves with different concentrations of GO solution (19) finally found that the 0.50% PS + 12.5 mg / L GO treatment (soaking seeds in a mixture of 0.50% PS + 12.5 mg / L GO + culture in soil substrate with 0.50% PS microplastic) in the 7-day seedling growth test was the best mitigation effect and the optimal mitigation intensity of 0.50% PS microplastic toxicity in TS163 maize genotype seedlings. Therefore, we can soak corn seeds in a 12.5 mg / L GO solution and then cultivate them in a 0.50% PS microplastic environment for 7 days to achieve normal seedling emergence. Then, we can continue to spray the leaves of each corn seedling in each pot with a 5.0 mg / L GO solution every 2 days in the morning, for a total of 7 foliar sprays. This treatment has the best overall mitigation effect and the strongest overall mitigation intensity for corn seedlings cultivated under 0.50% PS microplastic toxicity for 7 days and 21 days.

[0152] As can be seen from the above embodiments, this invention starts from the practical application of mitigating PS microplastic toxicity in corn production. It fully considers the most sensitive growth period of corn seedlings to PS microplastic toxicity (i.e., from seed germination to early seedling development) as the starting point, and adopts the two most typical treatment methods, namely seed soaking and foliar spraying. It not only comprehensively studies the effects of soaking corn seeds in different concentrations of ZnONPs / GO solution or foliar spraying of corn seedlings on the growth, development, chlorophyll accumulation, and root characteristics of corn seedlings grown under 0.50% PS microplastic stress for 7 days and 21 days, but also systematically elucidates the mechanism of action of soaking corn seeds in different concentrations of ZnONPs / GO solution or foliar spraying in mitigating 0.50% PS microplastic toxicity in corn seedlings, laying a theoretical basis for the application of ZnONPs / GO solution to alleviate PS microplastic toxicity in corn seedlings in production. Furthermore, to more scientifically, simply, and accurately evaluate the comprehensive mitigation effect / intensity of different concentrations of ZnONPs / GO solution soaking in maize seeds or foliar spraying on maize seedlings 7 or 21 days old affected by 0.50% PS microplastic toxicity, this invention also innovatively proposes a relative mitigation effect index (RMEI) for individual traits of maize seedlings under 0.50% PS microplastic toxicity, based on the effects of different concentrations of ZnONPs / GO solution soaking in seeds or foliar spraying. ZnONPS and RMEI GO ) and Absolute Mitigation Intensity Coefficient (AMIC) ZnONPs and AMIC GOThe biological concepts and calculation formulas of ZnONPs / GO solution were used as evaluation indicators to assess the mitigating effects and intensity of different concentrations of ZnONPs / GO solution on maize seedlings exposed to 0.50% PS microplastic toxicity. The membership function method was employed to scientifically, objectively, comprehensively, and quantitatively evaluate the comprehensive mitigating effects of different concentrations of ZnONPs / GO solution on maize seedlings cultured for 7 or 21 days under 0.50% PS microplastic toxicity (CME). ZnONPs and CME GO ) and Comprehensive Mitigation Intensity (CMI) ZnONPs and CMI GO Furthermore, these comprehensive evaluation results were also compared with the inter-group cluster evaluation results of maize seedling growth status among all treatments in the 7-day and 21-day growth trials of maize genotype seedlings treated with different concentrations of ZnONPs / GO solutions for seed soaking and foliar spraying to alleviate the toxicity of 0.50% PS microplastics. This further validated our proposed use of RMEI. ZnONPS ,RMEI GO AMIC ZnONPs and AMIC GO It is feasible to scientifically, objectively, and quantitatively evaluate the comprehensive mitigation effect and intensity of different concentrations of ZnONPs / GO solution soaking in maize seeds or foliar spraying on maize seedlings cultured at 0.50% PS microplastic toxicity on days 7 or 21. This provides a scientific evaluation method for future evaluation of the comprehensive mitigation effect of different types of microplastic toxicity on maize seedlings. Furthermore, this invention scientifically proposes using 75 μmol / L ZnONPs solution to soak maize seeds or foliar spraying to alleviate the toxicity of 0.50% PS microplastic toxicity in maize seedlings. Application methods for mitigating PS microplastic toxicity in corn seedlings include soaking corn seeds in a 12.5 mg / L GO solution or foliar spraying corn seedlings with a 5.0 mg / L GO solution. This method is simple to implement, has good experimental repeatability, utilizes readily available and economical nano-ZnONPs / GO materials, requires small dosages, and significantly alleviates PS microplastic toxicity in corn seedlings. It provides a solid technical reference for the safe, efficient, and sustainable production of corn under PS microplastic exposure conditions, and has enormous application potential.

[0153] Example 3

[0154] This invention provides the application of ZnONPs in mitigating microplastic toxicity in maize seedlings, specifically:

[0155] Forty high-quality new maize seeds, sterilized with 70% ethanol (v / v), were soaked in 100 mL of a mixed solution of 0.50% PS + 75 μmol / L ZnONPs on a horizontal shaker for 24 hours. Ten of these soaked seeds of the corresponding maize genotype were then sown in pots containing 0.50% PS microplastic substrate and placed in an artificial climate chamber for cultivation. On day 7 of seedling cultivation, every two days, the leaves of each maize genotype seedling in each pot were sprayed with 10 mL of a 75 μmol / L ZnONPs solution in the morning, i.e., 1 mL of 75 μmol / L ZnONPs solution was sprayed on each seedling leaf each time. This process was continued for 14 days, for a total of 7 foliar sprays. During the 21-day cultivation period, the relative humidity was set at 65%, the temperature was set at alternating cycles of 25±0.5℃ and 20±0.5℃ for 12 hours each, the photoperiod was set at 16 / 8 hours of light / dark, and the light intensity was set at 300 μM m². -2 s -1 The CO2 concentration was 450 PPM. Simultaneously, 50 mL of ddH2O water was evenly applied to each pot every 3 days to replenish the seedlings' water supply. Then, on days 7 and 21 of cultivation, maize seedlings were tested for 12 traits: seedling length (SL), seedling fresh weight (SFW), seedling dry weight (SDW), root length (RL), root fresh weight (RFW), root dry weight (RDW), stem diameter (SD), root diameter (RD), total plant dry biomass (PDW), leaf chlorophyll SPAD value (SPAD), root activity (RV), and seedling vigor index (SSI). The experiment was performed in four biological replicates.

[0156] Example 4

[0157] This invention provides the application of GO in mitigating microplastic toxicity in maize seedlings, specifically:

[0158] Forty high-quality new maize seeds, sterilized with 70% ethanol (v / v), were soaked in 100 mL of a mixed solution of 0.50% PS and 12.5 mg / L GO on a horizontal shaker for 24 hours. Ten of these soaked seeds of the corresponding maize genotype were then sown in pots containing 0.50% PS microplastic substrate and cultured in an artificial climate chamber. On day 7 of seedling culture, every two days, the leaves of each maize genotype seedling in each pot were sprayed with 10 mL of a 5.0 mg / L GO solution in the morning, i.e., 1 mL of 5.0 mg / L GO solution was sprayed on each seedling leaf each time. This process was continued for 14 days, for a total of 7 foliar sprays. During the 21-day culture period, the relative humidity was set at 65%, the temperature was set at alternating cycles of 25±0.5℃ and 20±0.5℃ for 12 hours each, the photoperiod was set at 16 / 8 hours of light / dark, and the light intensity was set at 300 μM m². -2 s -1The CO2 concentration was 450 PPM. Simultaneously, 50 mL of ddH2O water was evenly applied to each pot every 3 days to replenish the seedlings' water supply. Then, on days 7 and 21 of cultivation, maize seedlings were tested for 12 traits: seedling length (SL), seedling fresh weight (SFW), seedling dry weight (SDW), root length (RL), root fresh weight (RFW), root dry weight (RDW), stem diameter (SD), root diameter (RD), total plant dry biomass (PDW), leaf chlorophyll SPAD value (SPAD), root activity (RV), and seedling vigor index (SSI). The experiment was performed in four biological replicates.

Claims

1. A comprehensive evaluation method for mitigating the toxicity of polystyrene microplastics in corn seedlings, the specific steps of which are as follows: (1) Preparation of high-quality corn seeds: Prepare high-quality new corn genotype seeds that are uniform in size, plump, vigorous and pure, harvested from the field in the current year, for later use; (2) Preparation of different types of solutions: 0.50 g of polystyrene microplastics were accurately weighed using an electronic balance and dissolved in 100 mL of double-distilled water to prepare a 0.50% PS microplastic solution, which was prepared fresh for each use; 12.0 mg of nano-sized zinc oxide (ZnONPs) was accurately weighed using an electronic balance and dissolved in 1 L of ddH2O water. The ZnONPs were dispersed by stirring with a magnetic stirrer for 30 min at room temperature, and then ultrasonically vibrated for 30 min using a 40 Hz ultrasonic cleaner to ensure that the ZnONPs were fully and uniformly dispersed in the ddH2O water, resulting in a 150 μmol / L ZnONPs stock solution. This stock solution was then diluted to prepare six concentrations of ZnONPs solutions: 25 μmol / L, 50 μmol / L, 75 μmol / L, 100 μmol / L, 125 μmol / L, and 150 μmol / L. Simultaneously, 100 mL of each of the six concentrations of ZnONPs solutions was taken, and 0.50 mg of each solution was added to the stock solution. Dissolving g of PS microplastics yielded six PS-ZnONPs mixtures with concentrations of 0.50% PS + 25 μmol / L ZnONPs, 0.50% PS + 50 μmol / L ZnONPs, 0.50% PS + 75 μmol / L ZnONPs, 0.50% PS + 100 μmol / L ZnONPs, 0.50% PS + 125 μmol / L ZnONPs, and 0.50% PS + 150 μmol / L ZnONPs, prepared fresh for each use. Additionally, 25 μL, 50 μL, 75 μL, 100 μL, 125 μL, and 150 μL of 1% monolayer graphene oxide (GO) were pipetted into volumetric flasks, respectively, and then diluted to 100 mL with ddH2O water to obtain concentrations of 2.5 mg / L, 5.0 mg / L, 7.5 mg / L, and 10.0 mg / L. Six GO solutions with concentrations of 12.5 mg / L, 12.5 mg / L, and 15.0 mg / L were prepared fresh for each use. Simultaneously, 100 mL of each of these six GO solutions was taken, and 0.50 g of PS microplastics was added to each solution to dissolve them, resulting in six PS / GO mixtures with concentrations of 0.50% PS + 2.5 mg / L GO, 0.50% PS + 5.0 mg / L GO, 0.50% PS + 7.5 mg / L GO, 0.50% PS + 10.0 mg / L GO, 0.50% PS + 12.5 mg / L GO, and 0.50% PS + 15.0 mg / L GO, prepared fresh for each use. In total, 26 types of solutions were prepared, including ddH2O water and a 0.50% PS microplastic solution. Six ZnONPs solutions with concentrations of 25 μmol / L, 50 μmol / L, 75 μmol / L, 100 μmol / L, 125 μmol / L, and 150 μmol / L; Six mixed solutions of PS and ZnONPs with concentrations of 0.50% PS + 25 μmol / L ZnONPs, 0.50% PS + 50 μmol / L ZnONPs, 0.50% PS + 75 μmol / L ZnONPs, 0.50% PS + 100 μmol / L ZnONPs, 0.50% PS + 125 μmol / L ZnONPs, and 0.50% PS + 150 μmol / L ZnONPs; and six GO solutions with concentrations of 2.5 mg / L, 5.0 mg / L, 7.5 mg / L, 10.0 mg / L, 12.5 mg / L, and 15.0 mg / L. Six PS and GO mixtures with concentrations of 0.50% PS + 2.5 mg / L GO, 0.50% PS + 5.0 mg / L GO, 0.50% PS + 7.5 mg / L GO, 0.50% PS + 10.0 mg / L GO, 0.50% PS + 12.5 mg / L GO, and 0.50% PS + 15.0 mg / L GO were prepared fresh for each use. (3) Different types of ZnONPs / GO solutions were used to soak corn seeds to alleviate the toxicity of polystyrene (PS) microplastics. The corn seedlings grew for 7 days. The seeds were disinfected with 70% ethanol for 10 min, rinsed with ddH2O water 5 times, and the water on the seed surface was dried with sterile filter paper to obtain the corresponding disinfected corn genotype seeds. The disinfected corn genotype seeds were then soaked in ddH2O water, 0.50% PS microplastic solution, 6 kinds of PS and ZnONPs mixtures, and 6 kinds of PS and GO mixtures, for a total of 14 types of solutions for 24 h. At the same time, two kinds of soil substrates with PS microplastics were prepared, namely 0.00% PS microplastic soil substrate (i.e., 0.00 g PS microplastics and 300.00 g nutrient soil were thoroughly mixed and then 250 mL of ddH2O water was added and stirred evenly) and 0.50% PS microplastic soil substrate (i.e. 1.50 g PS microplastics and 300.00 g nutrient soil were thoroughly mixed and then 250 mL of ddH2O water was added and stirred evenly) and 0.50% PS microplastic soil substrate (i.e. 1.50 g PS microplastics and 300.00 g nutrient soil were thoroughly mixed and then 250 mL of ddH2O water was added and stirred evenly). (The PS microplastics were thoroughly mixed with 300.00 g of nutrient soil, and then 250 mL of ddH2O water was added and stirred until a uniform soil substrate was formed.) Ten corn genotype seeds soaked in each of the 14 different solutions were then sown into pots containing the corresponding concentration of PS microplastic soil substrate and placed in an artificial climate chamber for cultivation. During cultivation, the relative humidity was set at 65%, the temperature was set at 25±0.5℃ / 20±0.5℃ alternating for 12 h, the photoperiod was set at 16 / 8 h light / dark, and the light intensity was set at 300 μM m². -2 s -1 The CO2 concentration was 450 PPM. In total, there were 14 treatments in the experiment: CK control (soaking in ddH2O water + culture in 0.00% PS microplastic soil substrate), 0.50% PS treatment (soaking in 0.50% PS microplastic solution + culture in 0.50% PS microplastic soil substrate), 0.50% PS + 25 μmol / L ZnONPs treatment (soaking in a mixture of 0.50% PS + 25 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate), 0.50% PS + 50 μmol / L ZnONPs treatment (soaking in a mixture of 0.50% PS + 50 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate), and 0.50% PS + 75 μmol / L ZnONPs treatment (soaking in a mixture of 0.50% PS + 75 μmol / L ZnONPs + 0.50% PS microplastic soil substrate). Treatments included: 0.50% PS + 100 μmol / L ZnONPs (soaking in a mixture of 0.50% PS + 100 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 125 μmol / L ZnONPs (soaking in a mixture of 0.50% PS + 125 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 150 μmol / L ZnONPs (soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + culture in 0.50% PS microplastic soil substrate); 0.50% PS + 2.5 mg / L GO (soaking in a mixture of 0.50% PS + 2.5 mg / L GO + culture in 0.50% PS microplastic soil substrate); and 0.50% PS + 5.0 mg / L GO. Treatments included: 0.50% PS + 5.0 mg / L GO (soaking in a mixture of 0.50% PS and 5.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate), 0.50% PS + 7.5 mg / L GO (soaking in a mixture of 0.50% PS and 7.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate), 0.50% PS + 10.0 mg / L GO (soaking in a mixture of 0.50% PS and 10.0 mg / L GO + culture in a 0.50% PS microplastic soil substrate), 0.50% PS + 12.5 mg / L GO (soaking in a mixture of 0.50% PS and 12.5 mg / L GO + culture in a 0.50% PS microplastic soil substrate), and 0.50% PS + 15.0 mg / L GO (soaking in a mixture of 0.50% PS and 15.0 mg / L GO + 0.50% PS microplastic soil substrate).The soil substrate was 50% PS microplastic soil culture medium), and each treatment was set up with 4 biological replicates; during the culture period, 50 mL of ddH2O water was evenly poured into each pot every 3 days. On the 7th day of culture, the seedling length (SL), seedling fresh weight (SFW), seedling dry weight (SDW), root length (RL), root fresh weight (RFW), root dry weight (RDW), stem diameter (SD), root diameter (RD), total plant dry biomass (PDW), leaf chlorophyll SPAD value (SPAD), root activity (RV) and seedling vigor index (SSI) of maize seedlings under these 14 treatments were measured respectively; SSI was calculated according to formula (1): (1); where SSI is the seedling strength index, SD is the stem diameter, SL is the seedling length, and PDW is the total dry biomass of the plant; (4) 21-day experiment on mitigating polystyrene (PS) microplastic toxicity in maize seedlings by foliar spraying with different types of ZnONPs / GO solutions: Maize genotype seedlings cultured under the corresponding 14 treatments in step (3) for 7 days were further cultured in an artificial climate chamber with the same environment for 14 days. Every 2 days, the leaves of the maize genotype seedlings under the corresponding treatments were sprayed with 10 mL of each of the 14 types of solutions of the corresponding concentration in the morning, for a total of 7 foliar sprays. In total, there were 14 treatments in the experiment, namely CK control treatment (dddH2O water soaking + 0.00% PS microplastic soil substrate culture + ddH2O water foliar spraying of seedlings), 0.50% PS treatment (0.50% concentration PS microplastic solution soaking + 0.50% PS microplastic soil substrate culture + ddH2O water foliar spraying of seedlings), 0.50% PS + 25 μmol / L ZnONPs treatment (0.50% PS + 25 μmol / L ZnONPs). Treatments included: seed soaking in a ZnONPs mixture + foliar spraying of seedlings with a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 25 μmol / L ZnONPs solution; treatments with 0.50% PS + 50 μmol / L ZnONPs mixture (seed soaking in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 50 μmol / L ZnONPs solution); treatments with 0.50% PS + 75 μmol / L ZnONPs mixture (seed soaking in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 75 μmol / L ZnONPs solution); and treatments with 0.50% PS + 100 μmol / L ZnONPs mixture (seed soaking in a 0.50% PS microplastic soil substrate + foliar spraying of seedlings with a 25 μmol / L ZnONPs solution). Treatments included: 1) Seedlings cultured in a PS microplastic soil substrate with foliar spraying of a 100 μmol / L ZnONPs solution; 2) Seedlings cultured in a 0.50% PS + 125 μmol / L ZnONPs solution (seed soaking in a mixture of 0.50% PS + 125 μmol / L ZnONPs + foliar spraying of a 0.50% PS microplastic soil substrate); 3) Seedlings cultured in a 0.50% PS + 150 μmol / L ZnONPs solution (seed soaking in a mixture of 0.50% PS + 150 μmol / L ZnONPs + foliar spraying of a 0.50% PS microplastic soil substrate); and 4) Seedlings cultured in a 0.50% PS + 2.5 mg / L GO solution (seed soaking in a mixture of 0.50% PS + 2.5 mg / L GO + 0.50% PS microplastic soil substrate). (GO solution foliar spraying of seedlings), 0.Treatments included: 50% PS + 5.0 mg / L GO (seed soaking in a mixture of 0.50% PS and 5.0 mg / L GO + culturing in a microplastic soil substrate + foliar spraying of seedlings with a 5.0 mg / L GO solution); 0.50% PS + 7.5 mg / L GO (seed soaking in a mixture of 0.50% PS and 7.5 mg / L GO + culturing in a microplastic soil substrate + foliar spraying of seedlings with a 7.5 mg / L GO solution); 0.50% PS + 10.0 mg / L GO (seed soaking in a mixture of 0.50% PS and 10.0 mg / L GO + culturing in a microplastic soil substrate + foliar spraying of seedlings with a 10.0 mg / L GO solution); and 0.50% PS + 12.5 mg / L GO (seed soaking in a mixture of 0.50% PS and 12.5 mg / L GO + 0.50% PS + 12.5 mg / L GO solution). The treatment included: PS microplastic soil substrate culture + foliar spraying of seedlings with 12.5 mg / L GO solution; 0.50% PS + 15.0 mg / L GO treatment (seed soaking in a mixture of 0.50% PS + 15.0 mg / L GO + foliar spraying of seedlings with 0.50% PS microplastic soil substrate culture + foliar spraying of seedlings with 15.0 mg / L GO solution), with 4 biological replicates for each treatment; relative humidity was set at 65% during culture; temperature was set at alternating cycles of 25±0.5℃ and 20±0.5℃ for 12 h each; photoperiod was set at 16 / 8 h light / dark; and light intensity was set at 300 μM m. -2 s -1 The CO2 concentration was 450 PPM. During the cultivation period, 50 mL of ddH2O water was evenly poured into each pot every 3 days. On the 21st day of seedling cultivation, 12 traits of maize genotype seedlings under these 14 treatments were measured. (5) Statistical analysis of different experimental data: Using ExCel software, the mean and standard deviation of 12 traits of corn seedlings in step (3) soaking corn seeds in different types of ZnONPs / GO solution to alleviate PS microplastic toxicity and step (4) foliar spraying of different types of ZnONPs / GO solution to alleviate PS microplastic toxicity and 21-day growth of corn seedlings were determined; IBM-SPSS Statistics 19 data statistical analysis software was used to perform joint variance analysis on all traits of corn seedlings in different types of ZnONPs / GO solution soaking seeds and foliar spraying to alleviate PS microplastic toxicity for 7 days and 21 days; online GENESCLOUD software was used to plot the growth of corn seedlings in different types of ZnONPs / GO solution soaking seeds and foliar spraying to alleviate PS microplastic toxicity for 7 days and 21 days respectively. The correlation coefficients of the 12 traits corresponding to the d-test were plotted. The IMLOG2 function was used to further standardize the mean values ​​of the 12 traits corresponding to the 7-day and 21-day growth tests of maize genotype seedlings treated with different types of ZnONPs / GO solutions for mitigating PS microplastic toxicity. IBM-SPSS Statistics 16 data analysis software was used to perform inter-group clustering among all treatments in the 7-day and 21-day growth tests of maize genotype seedlings treated with different types of ZnONPs / GO solutions for mitigating PS microplastic toxicity, evaluating the overall growth status of maize genotype seedlings at different cultivation stages and treatments. This allowed for the scientific and qualitative identification of the mitigation effects of different concentrations of ZnONPs / GO solutions on maize genotype seedlings at different cultivation stages under PS microplastic toxicity. (6) Relative mitigation effect index of different concentrations of ZnONPs / GO solution soaking or foliar spraying on individual traits of maize seedlings under 0.50% polystyrene (PS) microplastic toxicity: In order to more scientifically, objectively and accurately measure the mitigation effect of different concentrations of ZnONPs / GO solution soaking or foliar spraying on individual traits of maize genotype seedlings under 0.50% polystyrene (PS) microplastic toxicity, this invention newly defines the relative mitigation effect index (RMEI) of different concentrations of ZnONPs / GO solution soaking or foliar spraying on individual traits of maize seedlings under 0.50% polystyrene (PS) microplastic toxicity. ZnONPS and RMEI GO ); Formula (2): (2); where The relative mitigation index of the j-th trait in maize genotype seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. The values ​​for the j-th trait of maize seedlings treated with a 0.50% PS microplastic toxicity solution (i-ZnONPs) at concentration i-ZnONPs concentration for soaking seeds or foliar spraying on day 7 or day 21 of cultivation were measured. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity treatment is the result of the determination of the jth genotype. The average value of the j-th trait of maize genotype seedlings cultured on day 7 or day 21 after treatment with m types of solution soaking or foliar spraying; similarly, formula (3): and (3); In the formula The relative mitigation index of the j-th trait in maize seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. The values ​​for the j-th trait of maize seedlings treated with 0.50% PS microplastic toxicity solution (i-GO concentration) on day 7 or day 21 of culture were measured. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity treatment is the result of the determination of the jth genotype. The average value of the jth trait of maize seedlings cultured on day 7 or day 21 after treatment with n (n = 8) types of solution soaking or foliar spraying; The higher the value, the stronger the mitigating effect of soaking seeds or spraying leaves with the i-ZnONPs concentration solution on the j-th trait of maize genotype seedlings cultured on the 7th or 21st day under 0.50% PS microplastic toxicity. The higher the value, the stronger the mitigating effect of soaking seeds or spraying leaves with the i-GO concentration solution on the j-th trait of maize genotype seedlings cultured on the 7th or 21st day of 0.50% PS microplastic toxicity. (7) Absolute mitigation strength coefficient of different concentrations of ZnONPs / GO solution for seed soaking or foliar spraying on individual traits of maize seedlings under 0.50% polystyrene (PS) microplastic toxicity: In order to facilitate a more scientific, simple and objective evaluation of the mitigation strength of different concentrations of ZnONPs / GO solution for seed soaking or foliar spraying on individual traits of maize genotype seedlings under 0.50% polystyrene (PS) microplastic toxicity, this invention also newly defines the absolute mitigation strength coefficient (AMIC) of different concentrations of ZnONPs / GO solution for seed soaking or foliar spraying on individual traits of maize seedlings under 0.50% polystyrene (PS) microplastic toxicity. ZnONPs and AMIC GO ); Formula (4): (4); where The absolute mitigation strength coefficient of the j-th trait in maize genotype seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. The values ​​for the j-th trait of maize seedlings treated with a 0.50% PS microplastic toxicity solution (i-ZnONPs) at concentration i-ZnONPs concentration for soaking seeds or foliar spraying on day 7 or day 21 of cultivation were measured. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity treatment is the result of the determination of the jth genotype. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under CK treatment; similarly, formula (5): (5); In the formula The absolute mitigation strength coefficient of the j-th trait in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. The values ​​for the j-th trait of maize seedlings treated with 0.50% PS microplastic toxicity solution (i-GO concentration) on day 7 or day 21 of culture were measured. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity treatment is the result of the determination of the jth genotype. The value of the jth genotype in maize seedlings cultured on day 7 or day 21 under CK treatment; The higher the value, the greater the mitigation effect of soaking seeds or spraying leaves with the i-ZnONPs concentration solution on the j-th trait of maize genotype seedlings cultured on the 7th or 21st day of 0.50% PS microplastic toxicity. The higher the value, the greater the mitigation effect of soaking seeds in i-GO concentration solution or spraying leaves on the j-th trait of maize genotype seedlings cultured on the 7th or 21st day under 0.50% PS microplastic toxicity. (8) Comprehensive evaluation of the mitigation effect of different concentrations of ZnONPs / GO solution soaking seeds or foliar spraying on the toxicity of 0.50% polystyrene (PS) microplastics in maize seedlings: Further, based on the RMEI of different concentrations of ZnONPs / GO solution soaking seeds or foliar spraying on the toxicity of 0.50% PS microplastics in maize seedlings cultured for 7 days or 21 days, the results were obtained in step (6). ZnONPS and RMEI GO The values ​​were used as evaluation indicators for the mitigating effects of different concentrations of ZnONPs / GO solution on seed soaking or foliar spraying on maize genotype seedlings under 0.50% PS microplastic toxicity. The membership function method was used to scientifically, objectively, comprehensively, and quantitatively evaluate the comprehensive mitigating effects of different concentrations of ZnONPs / GO solution on maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity (CME). ZnONPs and CME GO ); Formula (6): (6); where: The membership value of the mitigation effect of soaking seeds or spraying leaves with i-ZnONPs concentration solution on the j-th trait of maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined. The relative mitigation index of the j-th trait in maize genotype seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. The index representing the maximum relative mitigation effect of ZnONPs solution applied to seeds or leaves during treatment with 0.50% PS microplastics at day 7 or 21 of culture was used to assess the j-th trait in maize seedlings of all ZnONPs concentrations. The minimum relative mitigation index for the j-th trait of maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or foliar spraying with ZnONPs solution at all concentrations; Formula (7): (7); In the formula The overall mitigation effect of soaking seeds or spraying leaves with i-ZnONPs concentration solution on maize seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity is shown, where k (k = 12) represents the number of traits measured. The membership value of the mitigation effect of soaking seeds or spraying leaves with i-ZnONPs concentration solution on the j-th trait of maize seedlings cultured on the 7th or 21st day of culture under 0.50% PS microplastic toxicity; Formula (8): (8); where: The membership value of the mitigation effect of soaking seeds or spraying leaves with i-GO concentration solution on the j-th trait of maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity is given. The relative mitigation index of the j-th trait in maize seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. The maximum relative mitigation index for the j-th trait in maize seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or foliar spraying with GO solution at all concentrations. The minimum relative mitigation index for the j-th trait of maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity, obtained by soaking seeds or foliar spraying with GO solution at all concentrations; Formula (9): (9); In the formula The overall mitigation strength of i-GO concentration solution applied to seeds or leaves during foliar spraying against maize seedlings cultured at 0.50% PS microplastic toxicity on day 7 or day 21 is given, where k (k = 12) represents the number of traits measured. Membership value of mitigation effect of soaking seeds or spraying leaves with i-GO concentration solution on the j-th trait of maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity; The higher the value, the stronger the overall mitigation effect of soaking seeds or spraying leaves with the i-ZnONPs concentration solution on maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. (9) Comprehensive evaluation of the mitigation strength of different concentrations of ZnONPs / GO solution soaking seeds or foliar spraying on maize seedlings under 0.50% polystyrene (PS) microplastic toxicity: Further, based on the AMIC of different concentrations of ZnONPs / GO solution soaking seeds or foliar spraying on maize seedlings cultured under 0.50% PS microplastic toxicity for 7 days or 21 days, the AMIC of 12 traits of maize genotype seedlings were calculated in step (7). ZnONPS and AMIC GO The values ​​were used as evaluation indicators for the mitigation intensity of different concentrations of ZnONPs / GO solution applied to seeds or leaves to maize genotype seedlings under 0.50% PS microplastic toxicity. The membership function method was employed to comprehensively, systematically, objectively, and quantitatively evaluate the overall mitigation intensity (CMI) of different concentrations of ZnONPs / GO solution applied to seeds or leaves to maize genotype seedlings cultured for 7 days or 21 days under 0.50% PS microplastic toxicity. ZnONPs and CMI GO ); Formula (10): (10); Where: The membership value of the mitigation strength of the j-th trait in maize seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. The absolute mitigation strength coefficient of the j-th trait in maize genotype seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution. The maximum absolute mitigation coefficient of the j-th trait in maize seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with ZnONPs solutions at all concentrations. The minimum absolute mitigation strength coefficient for the j-th trait in maize seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with ZnONPs solution at all concentrations; Formula (11): (11); where The overall mitigation strength of i-ZnONPs concentration solution applied to seeds or leaves during foliar spraying against maize seedlings cultured at 0.50% PS microplastic toxicity on day 7 or 21 is given, where k (k = 12) represents the number of traits measured. The membership value of the mitigation strength of the jth trait in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-ZnONPs concentration solution; Formula (12): (12); where: The membership value of the mitigation strength of the j-th trait in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. The absolute mitigation strength coefficient of the j-th trait in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. The maximum absolute mitigation coefficient of the j-th trait in maize seedlings cultured on day 7 or 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with GO solution at all concentrations. The minimum absolute mitigation strength coefficient for the j-th trait in maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity by soaking seeds or spraying leaves with GO solution at all concentrations; Formula (13): (13); where The overall mitigation strength of i-GO concentration solution applied to seeds or leaves during foliar spraying against 0.50% PS microplastic toxicity at day 7 or 21 is given, where k (k = 12) represents the number of traits measured. The membership value of the mitigation strength of the jth trait in maize seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity was determined by soaking seeds or spraying leaves with i-GO concentration solution. The higher the value, the greater the overall mitigation effect of soaking seeds or spraying leaves with i-ZnONPs concentration solution on maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. The higher the value, the greater the overall mitigation effect of soaking seeds or spraying leaves with the i-GO concentration solution on maize genotype seedlings cultured on day 7 or day 21 under 0.50% PS microplastic toxicity. (10) Optimal ZnONPs / GO solution concentration for best mitigation effect / maximum mitigation intensity of 0.50% polystyrene (PS) microplastic toxicity in maize seedlings: Further comparison was made by soaking seeds and foliar spraying with different types of ZnONPs / GO solutions in step (5) to mitigate PS microplastic toxicity in maize genotype seedlings for 7 days and 21 days. The results of intergroup cluster evaluation of the growth status of maize genotype seedlings in all treatments in experiment d, the comprehensive evaluation results of the mitigation effect of different concentrations of ZnONPs / GO solution soaking seeds or foliar spraying on maize genotype seedlings with 0.50% concentration of PS microplastic toxicity in step (8), and the comprehensive evaluation results of the mitigation intensity of different concentrations of ZnONPs / GO solution soaking seeds or foliar spraying on maize genotype seedlings with 0.50% concentration of PS microplastic toxicity in step (9) were comprehensively compared and analyzed. The mitigation effect of different concentrations of ZnONPs / GO solution soaking seeds or foliar spraying on maize genotype seedlings with 0.50% concentration of PS microplastic toxicity in different cultivation periods was comprehensively compared and analyzed. Finally, the optimal concentration of ZnONPs / GO solution for seed soaking or foliar spraying was determined as the best mitigation effect / maximum mitigation intensity of 0.50% concentration of PS microplastic toxicity in maize genotype seedlings at different cultivation periods.

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