Removal of bisphenols from contaminated waters using Phragmites australis and bacteria (2021-2025)

Experimental data were generated between 2021–2025 under controlled laboratory conditions. Methodology: Bacterial Isolation and Identification Bacterial strains were isolated from the rhizosphere of Phragmites australis collected in 2021 from a phenol-contaminated pond (Staw Kalina, southern Poland). Root suspensions were enriched in basal salt medium (BSM) supplemented with bisphenols (BPA, BPS, BPF; 1–10 mg L⁻¹). Morphologically distinct strains were purified and screened for plant growth-promoting (PGP) traits, including phosphate solubilization, siderophore and IAA production, ACC deaminase activity, cellulolytic activity, motility, and biosurfactant production. Two strains (BP1 and BP2) were selected for further experiments. Identification was performed using whole-cell fatty acid profiling and multilocus sequence analysis (16S rRNA, gyrB, rpoD), followed by ANI comparison with type strains. Floating Treatment Wetland (FTW) Experiment FTW microcosms (20 L tanks) were constructed using P. australis and pond water artificially contaminated with BPA and BPS (5 mg L⁻¹ each) and BPF (2.5 mg L⁻¹). Five treatments were established: plant control, natural attenuation, bioaugmentation, phytoremediation, and bacterial-assisted phytoremediation. Microcosms were maintained at 22°C (16/8 h light/dark cycle) for 21 days with three biological replicates per treatment. Bacterial strains were immobilized in alginate beads prior to inoculation. Bisphenol Quantification and Removal Analysis Bisphenol concentrations were determined using HPLC-UV following solid-phase extraction (SPE). Removal efficiency (RE%) and biodegradation percentage (Bp%) were calculated considering adsorption, bioaccumulation, and abiotic removal. Kinetic modeling followed a pseudo-first-order model. Computational analyses were performed in Python (pandas, NumPy). Plant and Water Analyses Water quality parameters (pH, DO, TDS, TSS, COD, TOC, BOD₇) were measured using standardized analytical procedures. Plant physiological status was assessed through biomass determination, oxidative stress markers (H₂O₂, MDA, catalase activity), chlorophyll fluorescence, and RT-qPCR analysis of photosynthesis-related genes. Phytotoxicity of treated water was evaluated using Pisum sativum germination assays. Statistical Analysis Data were analyzed using two-way ANOVA/MANOVA with post hoc tests (p < 0.05). Data are provided in Excel (.xlsx), PDF, PNG, and IPBYN formats. The dataset includes: Plant growth-promoting (PGP) bacterial traits: – Indole-3-acetic acid (IAA) production – ACC deaminase activity – Phosphate solubilization – Biosurfactant production Water quality parameters (COD, BOD, pH, DO, TSS, VDS, TOC) Plant biometric parameters (fresh weight, root length, hypocotyl length) Toxicity bioassay data (seed germination and growth measurements) Gene expression data (qPCR results including Ct, ΔCt, ΔΔCt, and relative expression 2^-ΔΔCt) Chlorophyll fluorescence parameters (OJIP test measurements) Kinetic modelling The dataset contains raw and processed experimental data generated during the study. File Structure: Each Excel file corresponds to a specific experimental component or figure. Files contain labeled worksheets. Most of variables and measurement units are described here: Polish name English name Description Unit Szczep Strain Bacterial strain identifier - Układ Treatment Experimental treatment group - Powtórzenie Replicate Biological replicate number - Korzeń_długość_cm Root_length_cm Root length measured in centimeters cm Hypokotyl_długość_cm Hypocotyl_length_cm Hypocotyl length measured in centimeters cm Mokra masa_g Fresh_weight_g Fresh plant biomass in grams g Sucha masa_g Dry_weight_g Dry plant biomass in grams g IAA_koncentracja_ug_mL IAA_concentration_ug_mL Indole-3-acetic acid concentration (µg mL⁻¹) µg mL⁻¹ ACC_koncentracja_umol_mL ACC_concentration_umol_mL ACC concentration (µmol mL⁻¹) µmol mL⁻¹ Przejaśnienie Halo_zone_mm Halo zone diameter indicating phosphate solubilization (mm) mm Kolonia Colony_diameter_cm Bacterial colony diameter (cm) cm Absorbancja Absorbance_λ Spectrophotometric absorbance at specified wavelength unitless Ct Ct_value Cycle threshold value in qPCR cycles ΔCt ΔCt Delta Ct (Ct_target − Ct_reference) cycles ΔΔCt ΔΔCt Delta Delta Ct (ΔCt_sample − ΔCt_control) cycles Relatywna ekspresja_2^-ΔΔCt Relative_expression_2^-ΔΔCt Relative gene expression calculated using 2^-ΔΔCt method fold change本实验数据于2021–2025年在可控实验室条件下生成。 研究方法： ## 细菌分离与鉴定 实验菌株分离自2021年采自波兰南部卡利纳池塘（Staw Kalina）苯酚污染水域的芦苇（*Phragmites australis*）根际。将根际悬浮液接种于添加双酚类化合物的基础盐培养基（BSM, Basal Salt Medium）中进行富集培养，添加物为双酚A（BPA, Bisphenol A）、双酚S（BPS, Bisphenol S）、双酚F（BPF, Bisphenol F），浓度范围为1~10 mg·L⁻¹。筛选形态各异的菌株并纯化，随后检测其植物促生（PGP, Plant Growth-Promoting）性状，包括磷酸盐溶解能力、嗜铁素与吲哚-3-乙酸（IAA）合成能力、ACC脱氨酶活性、纤维素分解活性、运动性以及生物表面活性剂合成能力。 最终选取BP1与BP2两株菌株开展后续实验。鉴定采用全细胞脂肪酸谱分析与多位点序列分析（16S rRNA、gyrB、rpoD），并与模式菌株进行平均核苷酸同源性（ANI, Average Nucleotide Identity）比对。 ## 浮式处理湿地（FTW, Floating Treatment Wetland）实验 构建20 L体积的浮式处理湿地微宇宙体系，以芦苇与人工添加双酚A（5 mg·L⁻¹）、双酚S（5 mg·L⁻¹）与双酚F（2.5 mg·L⁻¹）的池塘水为实验材料。设置5组处理：空白植物对照组、自然衰减组、生物强化组、植物修复组以及细菌辅助植物修复组。 微宇宙体系于22℃、16 h光照/8 h黑暗周期下培养21天，每组设置3个生物学重复。接种前将目标菌株固定于海藻酸钙微球中。 ## 双酚类化合物定量与去除率分析 采用固相萃取（SPE, Solid Phase Extraction）结合高效液相色谱-紫外检测法（HPLC-UV）测定双酚类化合物浓度。计算去除效率（RE%）与生物降解率（Bp%）时，需考虑吸附作用、生物富集作用与非生物降解的影响。 动力学建模采用准一级动力学模型。数据分析采用Python（pandas、NumPy库）完成。 ## 植物与水体分析 采用标准分析方法测定水体理化参数：pH值、溶解氧（DO）、总溶解固体（TDS）、总悬浮固体（TSS）、化学需氧量（COD）、总有机碳（TOC）与五日生化需氧量（BOD₇）。 通过生物量测定、氧化应激标志物（过氧化氢H₂O₂、丙二醛MDA、过氧化氢酶活性）、叶绿素荧光以及光合作用相关基因的实时定量PCR（RT-qPCR）分析评估植物生理状态。 采用豌豆（*Pisum sativum*）发芽实验评估处理后水体的植物毒性。 ## 统计学分析 采用双因素方差分析/多元方差分析（two-way ANOVA/MANOVA）结合事后检验进行数据分析，显著性阈值设为p < 0.05。 本数据集以Excel（.xlsx）、PDF、PNG以及IPBYN格式提供，包含以下内容： 1. 植物促生（PGP）细菌性状： - 吲哚-3-乙酸（IAA）合成能力 - ACC脱氨酶活性 - 磷酸盐溶解能力 - 生物表面活性剂合成能力 2. 水体理化参数（COD、BOD、pH、DO、TSS、VDS、TOC） 3. 植物生物统计参数：鲜重、根长、下胚轴长度 4. 毒性生物测定数据：种子发芽与生长测量值 5. 基因表达数据：实时定量PCR结果，包含Ct值、ΔCt值、ΔΔCt值以及2^-ΔΔCt相对表达量 6. 叶绿素荧光参数：OJIP测试测量值 7. 动力学建模数据 本数据集包含本研究中产生的原始与处理后实验数据。 ## 文件结构 每个Excel文件对应一个特定的实验组分或图表，文件内包含命名清晰的工作表。以下为多数变量与测量单位的说明： | 波兰语名称 | 英文名称 | 描述 | 单位 | | --- | --- | --- | --- | | Szczep | Strain | 细菌菌株标识符 | 无 | | Układ | Treatment | 实验处理组 | 无 | | Powtórzenie | Replicate | 生物学重复编号 | 无 | | Korzeń_długość_cm | Root_length_cm | 以厘米为单位的根长 | cm | | Hypokotyl_długość_cm | Hypocotyl_length_cm | 以厘米为单位的下胚轴长度 | cm | | Mokra masa_g | Fresh_weight_g | 以克为单位的植物鲜生物量 | g | | Sucha masa_g | Dry_weight_g | 以克为单位的植物干生物量 | g | | IAA_koncentracja_ug_mL | IAA_concentration_ug_mL | 吲哚-3-乙酸浓度（µg·mL⁻¹） | µg·mL⁻¹ | | ACC_koncentracja_umol_mL | ACC_concentration_umol_mL | ACC浓度（µmol·mL⁻¹） | µmol·mL⁻¹ | | Przejaśnienie | Halo_zone_mm | 表征磷酸盐溶解能力的溶磷圈直径 | mm | | Kolonia | Colony_diameter_cm | 细菌菌落直径 | cm | | Absorbancja | Absorbance_λ | 特定波长下的分光光度吸光度 | 无量纲 | | Ct | Ct_value | 实时定量PCR的循环阈值 | 循环数 | | ΔCt | ΔCt | 相对循环阈值（靶基因Ct值 − 内参基因Ct值） | 循环数 | | ΔΔCt | ΔΔCt | 相对循环阈值差值（样品ΔCt值 − 对照组ΔCt值） | 循环数 | | Relatywna ekspresja_2^-ΔΔCt | Relative_expression_2^-ΔΔCt | 采用2^-ΔΔCt法计算的基因相对表达量 | 倍数变化