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Identification and characterization of plant growth–promoting endophyte RE02 from Trifolium repens L. in mining smelter

Abstract

Endophyte-assisted phytoremediation is considered to be an effective approach for bioremediation of heavy metal–contaminated soil; however, few information is available on Trifolium repens L. and its endophytes to remediate heavy metal–polluted soils. In this study, heavy metal–resistant endophytes were isolated from T. repens growing in mining smelter and identified by BIOLOG system. The isolate was also evaluated for promoting plant growth in heavy metal–contaminated soils in pot experiments. A total of eight Cd2+-resistant endophytes were isolated and these isolates preferred to grow on l-aspartic acid and α-d-glucose. All the isolates had at least two plant growth–promoting properties including siderophore production, phosphate solubilization activity, indole acetic acid (IAA) production, and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity. Strain RE02, identified as Pseudomonas putida by Biolog system, showed the highest Cd tolerance and could reduce Cd concentration from 20 to 1.84 mg L−1 in about 49 h in liquid medium, amounting to about 90.8%. Among the five endophytes which have positive effect on the growth of T. repens, RE02, whose IAA production ability was 7.06 mg L−1 and phosphate solubilization was 134.76 mg L−1, could improve T. repens root and shoot biomass by 25.9% and 37.7% in cadmium-contained soil, respectively. Our research may provide a new microbial-enhanced phytoremediation of heavy metal–polluted soils and improve the remediation efficiency.

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References

  1. Afzal M, Khan QM, Sessitsch A (2014) Endophytic bacteria: prospects and applications for the phytoremediation of organic pollutants. Chemosphere 117:232–242

  2. Ali H, Naseer M, Sajad MA (2012) Phytoremediation of heavy metals by Trifolium alexandrinum. Int J Environ Sci 2:1459–1469

  3. Ali S, Charles TC, Glick BR (2014) Amelioration of high salinity stress damage by plant growth-promoting bacterial endophytes that contain ACC deaminase. Plant Physiol Biochem 80:160–167

  4. Behera BC, Yadav H, Singh SK, Sethi BK, Mishra RR, Kumari S, Thatoi H (2017) Alkaline phosphatase activity of a phosphate solubilizing Alcaligenes faecalis, isolated from mangrove soil. Biotechnol Res Inno 1:101–111

  5. Bhattacharya A (2011) Siderophore mediated metal uptake by Pseudomonas fluorescens and its comparison to iron (iii) chelation. Cey J Sci (Bio Sci) 39:147–155

  6. Boeris PS, Agustín MDR, Acevedo DF, Lucchesi GI (2016) Biosorption of aluminum through the use of non-viable biomass of Pseudomonas putida. J Biotechnol 236:57–63

  7. Das S, Chou M, Jean J, Yang H, Kim PJ (2017) Arsenic-enrichment enhanced root exudates and altered rhizosphere microbial communities and activities in hyperaccumulator Pteris vittata. J Hazard Mater 325:279–287

  8. Dong XZ, Cai MY (2001) Manual for Systematic Identification of Common Bacteria. Science Press, Beijing

  9. Fässler E, Evangelou MW, Robinson BH, Schulin R (2010) Effects of indole-3-acetic acid (IAA) on sunflower growth and heavy metal uptake in combination with ethylene diamine disuccinic acid (EDDS). Chemosphere 80:901–907

  10. Guo H, Luo S, Chen L, Xiao X, Xi Q, Wei W, Zeng G, Liu C, Wan Y, Chen J, He Y (2010) Bioremediation of heavy metals by growing hyperaccumulaor endophytic bacterium Bacillus sp. L14. Bioresour Technol 101:8599–8605

  11. Hassan SE (2017) Plant growth-promoting activities for bacterial and fungal endophytes isolated from medicinal plant of Teucrium polium L. J Adv Res 8:687–695

  12. Ji SH, Gururani MA, Chun S (2014) Isolation and characterization of plant growth promoting endophytic diazotrophic bacteria from Korean rice cultivars. Microbiol Res 169:83–98

  13. Jiang C, Sheng X, Qian M, Wang Q (2008) Isolation and characterization of a heavy metal-resistant Burkholderia sp. from heavy metal-contaminated paddy field soil and its potential in promoting plant growth and heavy metal accumulation in metal-polluted soil. Chemosphere 72:157–164

  14. Jiang Y, Chao S, Liu J, Yang Y, Chen Y, Zhang A, Cao H (2017) Source apportionment and health risk assessment of heavy metals in soil for a township in Jiangsu Province, China. Chemosphere 168:1658–1668

  15. Johnston-Monje D, Raizada MN (2011) Conservation and diversity of seed associated endophytes in Zea across boundaries of evolution, ethnography and ecology. PLoS One 6:e20396

  16. Kaneez F, Muhammad A, Asma I, Khan QM (2015) Bacterial rhizosphere and endosphere populations associated with grasses and trees to be used for phytoremediation of crude oil contaminated soil. Bull Environ Contam Toxicol 94:314–320

  17. Kang C, So J (2016) Heavy metal and antibiotic resistance of ureolytic bacteria and their immobilization of heavy metals. Ecol Eng 97:304–312

  18. Kang C, Oh SJ, Shin Y, Han S, Nam I, So J (2015) Bioremediation of lead by ureolytic bacteria isolated from soil at abandoned metal mines in South Korea. Ecol Eng 74:402–407

  19. Kang C, Kwon Y, So J (2016) Bioremediation of heavy metals by using bacterial mixtures. Ecol Eng 89:64–69

  20. Khalid S, Shahid M, Niazi NK, Murtaza B, Bibi I, Dumat C (2017) A comparison of technologies for remediation of heavy metal contaminated soils. J Geochem Explor 182:247–268

  21. Khan MU, Sessitsch A, Harris M, Fatima K, Imran A, Arslan M, Shabir G, Khan QM, Afzal M (2015a) Cr-resistant rhizo- and endophytic bacteria associated with Prosopis juliflora and their potential as phytoremediation enhancing agents in metal-degraded soils. Front Plant Sci 5:755

  22. Khan Z, Nisar MA, Hussain SZ, Arshad MN, Rehman A (2015b) Cadmium resistance mechanism in Escherichia coli P4 and its potential use to bioremediate environmental cadmium. Appl Microbiol Biotechnol 24:10745–10757

  23. Koulman A, Lee TV, Fraser K, Johnson L, Arcus V, Lott JS, Rasmussen S, Lane G (2012) Identification of extracellular siderophores and a related peptide from the endophytic fungus Epichloë festucae in culture and endophyte-infected Lolium perenne. Phytochemistry 75:128–139

  24. Leedjarv A, Ivask A, Virta M (2008) Interplay of different transporters in the mediation of divalent heavy metal resistance in Pseudomonas putida KT2440. J Bacteriol 190:2680–2689

  25. Liu Y, Xie A (2011) Enrichment features of Trifolium pratense L. under cadmium tress. J Henan Agric Sci 1:82–84

  26. Liu Z, Li YC, Zhang S, Fu Y, Fan X, Patel JS, Zhang M (2015) Characterization of phosphate-solubilizing bacteria isolated from calcareous soils. Appl Soil Ecol 96:217–224

  27. Luo S, Chen L, Chen J, Xiao X, Xu T, Wan Y, Rao C, Liu C, Liu Y, Lai C, Zeng G (2011) Analysis and characterization of cultivable heavy metal-resistant bacterial endophytes isolated from Cd-hyperaccumulator Solanum nigrum L. and their potential use for phytoremediation. Chemosphere 85:1130–1138

  28. Ma Y, Rajkumar M, Freitas H (2009) Improvement of plant growth and nickel uptake by nickel resistant-plant-growth promoting bacteria. J Hazard Mater 166:1154–1161

  29. Ma Y, Prasad MNV, Rajkumar M, Freitas H (2011a) Plant growth promoting rhizobacteria and endophytes accelerate phytoremediation of metalliferous soils. Biotechnol Adv 29:248–258

  30. Ma Y, Rajkumar M, Luo Y, Freitas H (2011b) Inoculation of endophytic bacteria on host and non-host plants—effects on plant growth and Ni uptake. J Hazard Mater 195:230–237

  31. Ma Y, Oliveira RS, Nai F, Rajkumar M, Luo Y, Rocha I, Freitas H (2015) The hyperaccumulator Sedum plumbizincicola harbors metal-resistant endophytic bacteria that improve its phytoextraction capacity in multi-metal contaminated soil. J Environ Manag 156:62–69

  32. Ma Y, Rajkumar M, Zhang C, Freitas H (2016) Beneficial role of bacterial endophytes in heavy metal phytoremediation. J Environ Manag 174:14–25

  33. Marzan LW, Hossain M, Mina SA, Akter Y, Chowdhury AMMA (2017) Isolation and biochemical characterization of heavy-metal resistant bacteria from tannery effluent in Chittagong city, Bangladesh: bioremediation viewpoint. Egypt. J. Aquat. Res 43:65–74

  34. Mohanty SR, Dubey G, Kollah B (2017) Endophytes of Jatropha curcas promote growth of maize. Rhizosphere 3:20–28

  35. Nautiyal CS (1999) An efcient microbiological growth medium for screening phosphate solubilizing microorganisms. FEMS Microbiol Lett 170:265–270

  36. Nichols SN, Hofmann RW, Williams WM (2015) Physiological drought resistance and accumulation of leaf phenolics in white clover interspecific hybrids. Environ Exp Bot 119:40–47

  37. Nithya C, Gnanalakshmi B, Pandian SK (2011) Assessment and characterization of heavy metal resistance in Palk Bay sediment bacteria. Mar Environ Res 71:283–294

  38. None, 1958. Bergey's manual of determinate bacteriology. 7th ed. Robert S. Breed, E. G. D. Murray and Nathan R. Smith, et al. The Williams and Wilkins Company, Baltimore, 1957, xviii+1,094 pp. 15×23 cm. Price $15.

  39. Onofre-Lemus J, Hernández-Lucas I, Girard L, Caballero-Mellado J (2009) ACC (1-aminocyclopropane-1-carboxylate) deaminase activity, a widespread trait in Burkholderia species, and its growth-promoting effect on tomato plants. Appl Environ Microbiol 20:6581–6590

  40. Passari AK, Mishra VK, Leo VV, Gupta VK, Singh BP (2016) Phytohormone production endowed with antagonistic potential and plant growth promoting abilities of culturable endophytic bacteria isolated from Clerodendrum colebrookianum Walp. Microbiol Res 193:57–73

  41. Qin S, Miao Q, Feng W, Wang Y, Zhu X, Xing K, Jiang J (2015) Biodiversity and plant growth promoting traits of culturable endophytic actinobacteria associated with Jatropha curcas L. growing in Panxi dry-hot valley soil. Appl Soil Ecol 93:47–55

  42. Rajkumar M, Ae N, Prasad MNV, Freitas H (2010) Potential of siderophore-producing bacteria for improving heavy metal phytoextraction. Trends Biotechnol 28:142–149

  43. Rajkumar M, Ma Y, Freitas H (2013) Improvement of Ni phytostabilization by inoculation of Ni resistant Bacillus megaterium SR28C. J Environ Manag 128:973–980

  44. Rathi M, Nandabalan YK (2017) Copper-tolerant rhizosphere bacteria—characterization and assessment of plant growth promoting factors. Environ Sci Pollut Res 24:9723–9733

  45. Santos SGD, Silva PRAD, Garcia AC, Zilli JÉ, Berbara RLL (2017) Dark septate endophyte decreases stress on rice plants. Braz J Microbiol 48:333–341

  46. Schwyn B, Neilands JB (1987) Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160:47–56

  47. Sessitsch A, Kuffner M, Kidd P, Vangronsveld J, Wenzel WW, Fallmann K, Puschenreiter M (2013) The role of plant-associated bacteria in the mobilization and phytoextraction of trace elements in contaminated soils. Soil Biol Biochem 60:182–194

  48. Shao D, Zhan Y, Zhou W, Zhu L (2016) Current status and temporal trend of heavy metals in farmland soil of the Yangtze River Delta Region: Field survey and meta-analysis. Environ Pollut 219:329–336

  49. Shehzadi M, Fatima K, Imran A, Mirza MS, Khan QM, Afzal M (2015) Ecology of bacterial endophytes associated with wetland plants growing in textile effluent for pollutant-degradation and plant growth-promotion potentials. G Bot Ital 150:1261–1270

  50. Solon AG, Robert PW (1951) Colorimetric estimation of indoleacetic acid. Plant Physiol:192–195

  51. Tiwari S, Sarangi BK, Thul ST (2016) Identification of arsenic resistant endophytic bacteria from Pteris vittata roots and characterization for arsenic remediation application. J Environ Manag 180:359–365

  52. Tran TN, Kim D, Ko S (2018) Synergistic effects of biogenic manganese oxide and Mn(II)-oxidizing bacterium Pseudomonas putida strain MnB1 on the degradation of 17 α-ethinylestradiol. J Hazard Mater 344:350–359

  53. Wang G, Chai K, Wu J, Liu F (2016) Effect of Pseudomonas putida on the degradation of epoxy resin varnish coating in seawater. Int Biodeterior Biodegrad 115:156–163

  54. Wang L, Lin H, Dong Y, He Y, Liu C (2018) Isolation of vanadium-resistance endophytic bacterium PRE01 from Pteris vittata in stone coal smelting district and characterization for potential use in phytoremediation. J Hazard Mater 341:1–9

  55. Xiao X, Luo S, Zeng G, Wei W, Wan Y, Chen L, Guo H, Cao Z, Yang L, Chen J, Xi Q (2010) Biosorption of cadmium by endophytic fungus (EF) Microsphaeropsis sp. LSE10 isolated from cadmium hyperaccumulator Solanum nigrum L. Bioresour Technol 101:1668–1674

  56. Xu J, Han Y, Chen Y, Zhu L, Ma LQ (2016) Arsenic transformation and plant growth promotion characteristics of As-resistant endophytic bacteria from As-hyperaccumulator Pteris vittata. Chemosphere 144:1233–1240

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Funding

This work was financially supported by the Major Science and Technology Program for Water Pollution Control and Treatment (2015ZX07205-003).

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Correspondence to Hai Lin.

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Liu, C., Lin, H., Dong, Y. et al. Identification and characterization of plant growth–promoting endophyte RE02 from Trifolium repens L. in mining smelter. Environ Sci Pollut Res 26, 17236–17247 (2019). https://doi.org/10.1007/s11356-019-04904-w

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Keywords

  • Cadmium contamination
  • Endophyte
  • T. repens
  • Endophyte-assisted phytoremediation
  • Plant growth–promoting
  • Cadmium absorption