Isolation, identification, and characterization of a strain resistant to cadmium, lead, and zinc from the rhizosphere of hyperaccumulator Celosia argentea Linn
WANG An-qi1, LIN Hua1,2,3, LIU Jie1,2, LIN Yi1, YANG Xue-meng1, LAI Cai-xing1, DONG Zi-han1, YU Guo1,4
1. Guangxi Key Laboratory of Environmental Pollution Control Theory and Technology, GuilinUniversity of Technology, Guilin 541006, China; 2. Guangxi Collaborative Innovation Center for Water Pollution Control and WaterSafety in Karst Area, Guilin University of Technology, Guilin 541006, China; 3. Guangxi Engineering Research Center ofComprehensive Treatment for Agricultural Non-Point Source Pollution, Guilin University of Technology, Guilin 541006, China; 4. School of Environment, Tsinghua University, Beijing 100084, China
Abstract：This study investigated the functional characteristics of plant growth-promoting rhizobacteria (PGPR) isolated from the rhizosphere soil of Celosia argentea Linn., a Cd-hyperaccumulator. A strain with high tolerance to Cd2-, Pb2-, and Zn2- was isolated. This strain was identified using physiological and biochemical characteristics analysis and sequence analysis of the 16S rDNA and nrdA functional genes. The effects of various culture conditions on the strain's growth and heavy metal removal capabilities, as well as its potential for promoting plant growth were examined. This strain was identified as Achromobacter sp., designated WL-37. The minimum inhibitory concentrations (MIC) of Cd2-, Pb2-, and Zn2- for strain WL-37 were determined to be 600, 1800, and 1000mg/L, respectively. Under optimized conditions (e.g., pH values and inoculation amounts), the strain achieved maximum removal rates of 69% for Cd2+, 95% for Pb2+, and 62% for Zn2+. Moreover, WL-37 exhibited multiple plant-promoting traits, including nitrogen fixation, ACC deaminase production, and siderophore production. In summary, the high-efficiency strain identified in this study represents a valuable resource for the remediation of multi-metal contaminated soils and supports the development of plant-microbe combined remediation technologies.
王桉祺, 林华, 刘杰, 林毅, 杨雪萌, 赖才星, 董梓涵, 俞果. 超富集植物青葙根际耐镉、铅、锌菌株的筛选鉴定与特性研究[J]. 中国环境科学, 2025, 45(5): 2631-2642.
WANG An-qi, LIN Hua, LIU Jie, LIN Yi, YANG Xue-meng, LAI Cai-xing, DONG Zi-han, YU Guo. Isolation, identification, and characterization of a strain resistant to cadmium, lead, and zinc from the rhizosphere of hyperaccumulator Celosia argentea Linn. CHINA ENVIRONMENTAL SCIENCECE, 2025, 45(5): 2631-2642.
[1] Zhang H Y, Zhou Q Q, Liu R Y, et al. Enhancing zinc biofortification and mitigating cadmium toxicity in soil–earthworm–spinach systems using different zinc sources [J]. Journal of Hazardous Materials, 2024,476:135243. [2] Liu N Q, Zhao J, Du J W, et al. Non-phytoremediation and phytoremediation technologies of integrated remediation for water and soil heavy metal pollution: A comprehensive review [J]. Science of the Total Environment, 2024,948:174237. [3] Sodhi K K, Kumar M, Agrawal P K, et al. Perspectives on arsenic toxicity, carcinogenicity and its systemic remediation strategies [J]. Environmental Technology and Innovation, 2019,16(1):100462. [4] Azhar U, Ahmad H, Shafqat H, et al. Remediation techniques for elimination of heavy metal pollutants from soil: A review [J]. Environmental Research, 2022,214:113918. [5] Li X H, Gao Y, Ning X L, et al. Research progress and hotspots on microbial remediation of heavy metal-contaminated soil: A systematic review and future perspectives [J]. Environmental Science and Pollution Research, 2023,30:118192–118212. [6] Mishra S, Lin Z, Pang S, et al. Biosurfactant is a powerful tool for the bioremediation of heavy metals from contaminated soils [J]. Journal of Hazardous Materials, 2021,418:126253. [7] Song L P, Zhou J H, Xu X Y, et al. Inoculation of cadmium-tolerant bacteria to regulate microbial activity and key bacterial population in cadmium-contaminated soils during bioremediation [J]. Ecotoxicology and Environmental Safety, 2024,271:115957. [8] Li P H, Chen J L, Ying S M, et al. Different responses of Sinorhizobium sp. upon Pb and Zn exposure: Mineralization versus complexation [J]. Environmental Pollution, 2024,343:123260. [9] Ru X, Liao J, Liang L, et al. Quantification of the relationship between multiple metal (loid) distribution and integrated effect of internalexternal factors in riverbed sediments across Xijiang River basin, South China [J]. Science of the Total Environment, 2018,643:527-538. [10] Luo Y, Pang J, Li C, et al. Long-term and high-bioavailable potentially toxic elements (PTEs) strongly influence the microbiota in electroplating sites [J]. Science of the Total Environment, 2022,814:151933. [11] Mathivanan K, Chandirika J U, Vinothkanna A, et al. Bacterial adaptive strategies to cope with metal toxicity in the contaminated environment-A review [J]. Ecotoxicology and Environmental Safety, 2021,226:112863. [12] Li J, Gurajala H K, Wu L, et al. Hyperaccumulator plants from China: A synthesis of the current state of knowledge [J]. Environmental Science and Technology, 2018,52:11980-11994. [13] Halim A, Rahman M M, Megharaj M, et al. Cadmium immobilization in the rhizosphere and plant cellular detoxification: role of plant-growthpromoting rhizobacteria as a Sustainable Solution [J]. Journal of Agricultural and Food Chemistry, 2020,68(47):13497-13529. [14] Cao M Y, Narayanan M, Shi X J, et al. Optimistic contributions of plant growth-promoting bacteria for sustainable agriculture and climate stress alleviation [J]. Environmental Research, 2023,217:114924. [15] Kumar M, Poonam, Ahmad S, et al. Plant growth promoting microbes: Diverse roles for sustainable and ecofriendly agriculture [J]. Energy Nexus, 2022,7:100133. [16] Teng Z D, Shao W, Zhang K Y, et al. Characterization of phosphate solubilizing bacteria isolated from heavy metal contaminated soils and their potential for lead immobilization [J]. Journal of Environmental Management, 2019,231:189-197. [17] Guo J, Muhammad H, Lu X, et al. Prospects and applications of plant growth promoting rhizobacteria to mitigate soil metal contamination: A review [J]. Chemosphere, 2020,246:125823. [18] Xie Y, Bu H S, Feng Q J, et al. Identification of Cd-resistant microorganisms from heavy metal-contaminated soil and its potential in promoting the growth and Cd accumulation of bermudagrass [J]. Environmental Research, 2021,200:111730. [19] Yu G, Jiang P P, Fu X F, et al. Phytoextraction of cadmiumcontaminated soil by Celosia argentea Linn.: A long-term field study [J]. Environmental Pollution, 2020,266(1):115408. [20] 游胜雄,王义安,林华,等.一株新型Kerstersia gyiorum菌株CY-2对Cr( ) Ⅵ 的还原性能[J]. 环境工程学报, 2023,17(12):4096-4106. You S X, Wang Y A, Lin H, et al. Reduction properties of Cr( ) by Ⅵ a novel Kerstersia gyiorum strain CY-2[J]. Chinese Journal of Environmental Engineering, 2023,17(12):4096-4106. [21] Narde G K, Kapley A, Purohit H J. Isolation and characterization of Citrobacter Strain HPC255 for broad-range substrate specificity for chlorophenols [J]. Current Microbiology, 2004,48(6):419-423. [22] Coward A, Kenna D T D, Perry C, et al. Use of nrdA gene sequence clustering to estimate the prevalence of different Achromobacter species among Cystic Fibrosis patients in the UK [J]. Journal of Cystic Fibrosis, 2016,15(4)479-485. [23] Biswas B B, Basu P S, Pal M K. Gram staining and its molecular mechanism [J]. International Review of Cytology, 1970,29:1-27. [24] Truu J, Talpsep E, Heinaru E, et al. Comparison of API 20NE and Biolog GN identification systems assessed by techniques of multivariate analyses [J]. Journal of Microbiological Methods, 1999,36(3):193-201. [25] Babu A G, Shea P J, Sudhakar D, et al. Potential use of Pseudomonas koreensis AGB-l in association with Miscanthus sinensis to remediate heavy metal (loid) -contaminated mining site soil [J]. Journal of Environmental Management, 2015,151:160-166. [26] Chen L, Luo S L, Xiao X, et al. Application of plant growthpromoting endophytes (PGPE) isolated from Solanum nigrum L. for phytoextraction of Cd-polluted soils [J]. Applied Soil Ecology, 2010,46(3):383-389. [27] Machuca A, Milagres A M F. Use of CAS-agar plate modified to study the effect of different variables on the siderophore production by Aspergillus [J]. Letters in Applied Microbiology, 2003,36(3):177–181. [28] Schwyn B, Neilands J B. Universal chemical assay for the detection and determination of siderophores [J]. Analytical Biochemistry, 1987, 160(1):47-56. [29] Shenker M, Oliver I, Helmann M, et al. Utilization by tomatoes of iron mediated by a siderophore produced by Rhizopus arrhizus [J]. Journal of Plant Nutrition, 1992,15(10):2173-2182. [30] Xu J Y, Han Y H, Chen Y S, et al. Arsenic transformation and plant growth promotion characteristics of As-resistant endophytic bacteria from As-hyperaccumulator Pteris vittata [J]. Chemosphere, 2016,144:1233-1240. [31] Yuan Y D, Zu M T, Sun L P, et al. Isolation and Screening of 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase producing PGPR from Paeonia lactiflora rhizosphere and enhancement of plant growth [J]. Scientia Horticulturae, 2022,297:110956. [32] Cai F F, Yang C D, Ma T, et al. An endophytic Paenibacillus polymyxa hg18and its biocontrol potential against Fusarium oxysporum f. sp. Cucumerinum [J]. Biological Control, 2024,188:105380. [33] Andy A K, Masih S A, Gour V S. Isolation, screening and characterization of plant growth promoting rhizobacteria from rhizospheric soils of selected pulses [J]. Biocatalysis and Agricultural Biotechnology, 2020,27:101685. [34] Emenike C U, Jayanthi B, Agamuthhu P, et al. Biotransformation and removal of heavy metals: A review of phytoremediation and microbial remediation assessment on contaminated soil [J]. Environment Reviews, 2018,26(2):156-168. [35] Nanda M, Kumar V, Sharma D K. Multimetal tolerance mechanisms in bacteria: The resistance strategies acquired by bacteria that can be exploited to ‘clean-up’ heavy metal contaminants from water [J]. Aquatic Toxicology, 2019,212:1-10. [36] Jeong S W, Kim H K, Yang J E, et al. Removal of Pb(Ⅱ) by pellicle-like biofilm-producing Methylobacterium hispanicum EM2strain from aqueous media [J]. Water, 2019,11(10):2081. [37] Kan H Y, Yang S X, Sun L L, et al. Isolation, identification and adsorption capacity of a strain resistant to lead, zinc and chromium [J]. Microbiology China, 2020,47(12):3974-3986. [38] Ma B, Song W L, Zhang X X, et al. Potential application of novel cadmium-tolerant bacteria in bioremediation of Cd-contaminated soil [J]. Ecotoxicology and Environmental Safety, 2023,255:114766. [39] Wang Z B, Tan R, Gong J, et al. Process parameters and biological mechanism of efficient removal of Cd(II) ion from wastewater by a novel Bacillus subtilis TR1[J]. Chemosphere, 2023,318:137958. [40] Xu X, Hao R, Xu H, et al. Removal mechanism of Pb(II) by Penicillium polonicum: immobilization, adsorption, and bioaccumulation [J]. Scientfic Reports, 2020,10:9079. [41] 张祖烨,何梦园,乔月,等.Agromyces sp. CS16去除水体中重金属镉、镍、铜、锌离子的研究[J]. 微生物学报, 2023,63(7):2791-2808. Zhang Z Y, He M Y, Qiao Y, et al. Removal of cadmium, nickel, copper, and zinc ions from water by Agromyces sp. CS16[J]. Acta Microbiologica Sinica, 2023,63(7):2791-2808. [42] Abo-Alkasem M I, Hassan N H, Elsoud M M A. Microbial bioremediation as a tool for the removal of heavy metals [J]. Bulletin of the National Research Centre, 2023,47(31). [43] Tyagi S, Kumar V, Singh J, et al. Bioremediation of pulp and paper mill effluent by dominant aboriginal microbes and their consortium [J]. International Journal of Environmental Research, 2014,8(3):561–568. [44] Mielniczki-Pereira A A, Hna A B B, Bonatto D, et al. New insights into the Ca2+-ATPases that contribute to cadmium tolerance in yeast [J]. Toxicology Letters, 2011,207(2):104-111. [45] Fashola M O, Ngole-Jeme V M, Babalola O O. Heavy metal pollution from gold mines: environmental effects and bacterial strategies for resistance [J]. International Journal of Environmental Research and Public Health, 2016,13(11):1047. [46] Satya A, Harimawan A, Haryani G S, et al. Batch study of cadmium biosorption by carbon dioxide enriched Aphanothece sp. dried biomass [J]. Water, 2020,12(1):264. [47] Tayebi B, Ahangar A G. The influence of heavy metals on the development and activity of soil microorganisms [J]. International Journal of Plant, Animal and Environmental Sciences, 2014,4:74-85. [48] Fathollahi A, Khasteganan N, Coupe S J, et al. A meta-analysis of metal biosorption by suspended bacteria from three phyla [J]. Chemosphere, 2021,268:129290. [49] Wang X, Zhang Y, Sun X, et al. Eficient removal of hexavalent chromiuom from water by Bacillus sp. Y2-7with production of extracltular polymeric substances [J]. Environmental Technology, 2023,45(14):2698-2708. [50] Yin K, Wang Q N, Lv M, et al. Microorganism remediation strategies towards heavy metals [J]. Chemical Engineering Journal, 2019,360: 1553-1563. [51] Asksonthong R, Siripongvutikorn S, Usawakesmanee W. Heavy metal removal ability of Halomonas elongata and Tetragenococcus halophilus in a media model system as affected by pH and incubation time [J]. International Food Research Journal, 2018,25(1):234-240. [52] Ma X Y. Heavy metals remediation through lactic acid bacteria: Current status and future prospects [J]. Science of the Total Environment, 2024,946:174455. [53] Hložková K, Šuman J, Strnad H, et al. Characterization of pbt genes conferring increased Pb2+ and Cd2+ tolerance upon Achromobacter xylosoxidans A8[J]. Research in Microbiology, 2013,164(10):1009-1018. [54] Suman J, Kotrba P, Macek T. Putative P1B-type ATPase from the bacterium Achromobacter xylosoxidans A8alters Pb2+/Zn2+/Cd2+- resistance and accumulation in Saccharomyces cerevisiae [J]. Biochimica et Biophysica Acta (BBA)-Biomembranes, 2014,1838(5):1338-1343. [55] Subudhi S, Batta N, Pathak M, et al. Bioflocculant production and biosorption of zinc and lead by a novel bacterial species, Achromobacter sp. TERI-IASST N, isolated from oil refinery waste [J]. Chemosphere, 2014,113:116-124. [56] Liu S M, Liu H M, Chen R, et al. Role of two plant growth-promoting bacteria in remediating cadmium-contaminated soil combined with Miscanthus floridulus (Lab.) [J]. Plants, 2021,10(5):912. [57] Liang D H, Hu Y Y. Application of a heavy metal-resistant Achromobacter sp. for the simultaneous immobilization of cadmium and degradation of sulfamethoxazole from wastewater [J]. Journal of Hazardous Materials, 2021,402:124032. [58] Yang Y J, PratapSingh R, Song D, et al. Synergistic effect of Pseudomonas putida II-2and Achromobacter sp. QC36 for the effective biodegradation of the herbicide quinclorac [J]. Ecotoxicology and Environmental Safety, 2020,188:109826. [59] Sun L N, Zhang X H, Ouyang W K, et al. Lowered Cd toxicity, uptake and expression of metal transporter genes in maize plant by ACC deaminase-producing bacteria Achromobacter sp. [J]. Journal of Hazardous Materials, 2022,423:127036. [60] 王慧,靳海洋,陈哲,等.稻田高效固氮蓝细菌的筛选鉴定及其促生潜力[J]. 微生物学通报, 2024,51(10):4043-4057. Wang H, Jin H Y, Chen Z, et al. Screening and identification of efficient nitrogen-fixing cyanobacteria with plant growth-promoting potential from paddy fields [J]. Microbiology China, 2024,51(10): 4043-4057. [61] 陈伟,舒健虹,陈莹,等.黑麦草根际铁载体产生菌WN-H3的分离鉴定及其产铁载体培养条件的优化[J]. 生物技术通报, 2016, 32(10):219-226. Chen W, Shu J H, Chen Y, et al. Screening, identification and fermentation condition optimun of a siderophore-producing bacteria WN-H3 from rhizosphere of ryegrass [J]. Biotechnology Bulletin, 2016,32(10):219-226. [62] Herpell J B, Alickovic A, Diallo B. et al. Phyllosphere symbiont promotes plant growth through ACC deaminase production [J]. The ISME Journal, 2023,17:1267-1277. [63] Xu P, Wang E. Diversity and regulation of symbiotic nitrogen fixation in plants [J]. Current Biology, 2023,33(11):543-559.
