Environmental Science and Pollution Research

, Volume 24, Issue 20, pp 16735–16750 | Cite as

Impact of metal stress on the production of secondary metabolites in Pteris vittata L. and associated rhizosphere bacterial communities

  • Hoang Nam Pham
  • Serge Michalet
  • Josselin Bodillis
  • Tien Dat Nguyen
  • Thi Kieu Oanh Nguyen
  • Thi Phuong Quynh Le
  • Mohamed Haddad
  • Sylvie Nazaret
  • Marie-Geneviève Dijoux-Franca
Research Article
  • 182 Downloads

Abstract

Plants adapt to metal stress by modifying their metabolism including the production of secondary metabolites in plant tissues. Such changes may impact the diversity and functions of plant associated microbial communities. Our study aimed to evaluate the influence of metals on the secondary metabolism of plants and the indirect impact on rhizosphere bacterial communities. We then compared the secondary metabolites of the hyperaccumulator Pteris vittata L. collected from a contaminated mining site to a non-contaminated site in Vietnam and identified the discriminant metabolites. Our data showed a significant increase in chlorogenic acid derivatives and A-type procyanidin in plant roots at the contaminated site. We hypothesized that the intensive production of these compounds could be part of the antioxidant defense mechanism in response to metals. In parallel, the structure and diversity of bulk soil and rhizosphere communities was studied using high-throughput sequencing. The results showed strong differences in bacterial composition, characterized by the dominance of Proteobacteria and Nitrospira in the contaminated bulk soil, and the enrichment of some potential human pathogens, i.e., Acinetobacter, Mycobacterium, and Cupriavidus in P. vittata’s rhizosphere at the mining site. Overall, metal pollution modified the production of P. vittata secondary metabolites and altered the diversity and structure of bacterial communities. Further investigations are needed to understand whether the plant recruits specific bacteria to adapt to metal stress.

Keywords

Bacterial communities Metal stress Pteris vittata Rhizosphere Secondary metabolites 

Abbreviations

UHPLC-DAD-ESI/QTOF-MS

Ultrahigh-performance liquid chromatography with diode array detection coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry

UV

Ultraviolet

ESI

Electrospray ionization

MSMS

Tandem mass spectrometry

ESI/MS2

Electrospray ionization tandem mass spectrometry

HRMS

High-resolution mass spectrometry

1H-NMR

Proton nuclear magnetic resonance

RT

Retention time

SPE

Solid phase extraction

PVP

Soil under P. vittata polluted

BSP

Bulk soil polluted

DLP

Soil under Dicranopteris linearis polluted

PVC

Soil under P. vittata control

HSD

Honest significant difference

PCA

Principal component analysis

ANOVA

Analysis of variance

CFU

Colony forming unit

QPCR

Quantitative real-time polymerase chain reaction

OTU

Operational taxonomic unit

DGGE

Denaturing gradient gel electrophoresis

DPPH

2,2-diphenyl-1-picrylhydrazyl

Notes

Acknowledgments

Hoang Nam Pham wish to gratefully thank the Vietnam Ministry of Education and Training, the University of Sciences and Technologies of Hanoi, and team “Environmental resistance and bacterial efflux,” UMR 5557 CNRS Microbial Ecology, for the financial support. We thank the platforms CESN (Centre d’Etudes des Substances Naturelles) and PARMIC (Plateau d’Analyse du Risque MICrobiologique, UMR Ecologie Microbienne, Université Lyon1) for the equipment facilities, and Institut of Marine Biochemistry (Vietnam Academy of Science and Technology, Hanoi) is also acknowledged for providing valuable assessment during the development of this subject.

Supplementary material

11356_2017_9167_MOESM1_ESM.docx (157 kb)
Fig. S1Typical UHPLC/DAD chromatogram at 280 nm of P. vittata root extracts. PVP: From plants grown on contaminated soil (Ha Thuong); PVC: From plants grown on non-polluted soil (USTH) (DOCX 157 kb).
11356_2017_9167_MOESM2_ESM.docx (183 kb)
Fig. S2Typical UHPLC/DAD chromatogram at 280 nm of P. vittata stems extracts. PVP: From plants grown on contaminated soil (Ha Thuong); PVC: From plants grown on non-polluted soil (USTH) (DOCX 182 kb).
11356_2017_9167_MOESM3_ESM.docx (186 kb)
Fig. S3Typical UHPLC/DAD chromatogram at 280 nm of P. vittata leaves extracts. PVP: From plants grown on contaminated soil (Ha Thuong); PVC: From plants grown on non-polluted soil (USTH) (DOCX 185 kb).
11356_2017_9167_MOESM4_ESM.docx (12 kb)
Table S1(DOCX 11 kb).

References

  1. Ahmed SI, Hayat MQ, Tahir M et al (2016) Pharmacologically active flavonoids from the anticancer, antioxidant and antimicrobial extracts of Cassia angustifolia Vahl. BMC Complement Altern Med 16:460. doi:10.1186/s12906-016-1443-z CrossRefGoogle Scholar
  2. Ali MB, Singh N, Shohael AM et al (2006) Phenolics metabolism and lignin synthesis in root suspension cultures of Panax ginseng in response to copper stress. Plant Sci 171:147–154. doi:10.1016/j.plantsci.2006.03.005 CrossRefGoogle Scholar
  3. Amadou C, Pascal G, Mangenot S et al (2008) Genome sequence of the β-rhizobium Cupriavidus taiwanensis and comparative genomics of rhizobia. Genome Res 18:1472–1483. doi:10.1101/gr.076448.108 CrossRefGoogle Scholar
  4. Anh BTK, Kim DD, Tua TV et al (2011) Phytoremediation potential of indigenous plants from Thai Nguyen province, Vietnam. J Environ Biol 32:257–262Google Scholar
  5. An ZZ, Huang ZC, Lei M et al (2006) Zinc tolerance and accumulation in Pteris vittata L. and its potential for phytoremediation of Zn- and As-contaminated soil. Chemosphere 62:796–802. doi:10.1016/j.chemosphere.2005.04.084 CrossRefGoogle Scholar
  6. Azarbad H, Niklińska M, van Gestel CAM et al (2013) Microbial community structure and functioning along metal pollution gradients. Environ Toxicol Chem 32:1992–2002. doi:10.1002/etc.2269 CrossRefGoogle Scholar
  7. Bardon C, Piola F, Bellvert F et al (2014) Evidence for biological denitrification inhibition (BDI) by plant secondary metabolites. New Phytol 204:620–630. doi:10.1111/nph.12944 CrossRefGoogle Scholar
  8. Berg J, Brandt KK, Al-Soud WA et al (2012) Selection for Cu-tolerant bacterial communities with altered composition, but unaltered richness, via long-term Cu exposure. Appl Environ Microbiol 78:7438–7446. doi:10.1128/AEM.01071-12 CrossRefGoogle Scholar
  9. Clifford MN, Johnston KL, Knight S, Kuhnert N (2003) Hierarchical scheme for LC-MSn identification of chlorogenic acids. J Agric Food Chem 51:2900–2911. doi:10.1021/jf026187q CrossRefGoogle Scholar
  10. Courts FL, Williamson G (2015) The occurrence, fate and biological activities of C-glycosyl flavonoids in the human diet. Crit Rev Food Sci Nutr 55:1352–1367. doi:10.1080/10408398.2012.694497 CrossRefGoogle Scholar
  11. Crupi P, Genghi R, Antonacci D (2014) In-time and in-space tandem mass spectrometry to determine the metabolic profiling of flavonoids in a typical sweet cherry (Prunus avium L.) cultivar from Southern Italy. J Mass Spectrom 49:1025–1034. doi:10.1002/jms.3423 CrossRefGoogle Scholar
  12. Danh LT, Truong P, Mammucari R, Foster N (2014) A critical review of the arsenic uptake mechanisms and phytoremediation potential of Pteris vittata. Int J Phytoremediation 16:429–453. doi:10.1080/15226514.2013.798613 CrossRefGoogle Scholar
  13. Dixon RA, Paiva NL (1995) Stress-induced phenylpropanoid metabolism. Plant Cell 7:1085–1097. doi:10.1105/tpc.7.7.1085 CrossRefGoogle Scholar
  14. Edgar RC, Haas BJ, Clemente JC et al (2011) UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 27:2194–2200. doi:10.1093/bioinformatics/btr381 CrossRefGoogle Scholar
  15. Ekelund F, Olsson S, Johansen A (2003) Changes in the succession and diversity of protozoan and microbial populations in soil spiked with a range of copper concentrations. Soil Biol Biochem 35:1507–1516. doi:10.1016/S0038-0717(03)00249-9 CrossRefGoogle Scholar
  16. Epelde L, Martín-Sánchez I, González-Oreja JA et al (2012) Impact of sources of environmental degradation on microbial community dynamics in non-polluted and metal-polluted soils. Sci Total Environ 433:264–272. doi:10.1016/j.scitotenv.2012.06.049 CrossRefGoogle Scholar
  17. Gong X-L, Chen Z-H, Liang N-C (2007) Advances in study on chemical constituents and pharmacological activities of plants of genus Pteris. Chin J Chin Mater Medica 32:1382–1387Google Scholar
  18. Gracelin DHS, Britto AJD, Kumar PBJR (2012) Qualitative and quantitative analysis of phytochamicals in five Pteris species. Int J Pharm Pharm Sci 5:105–107Google Scholar
  19. Gu L, Kelm MA, Hammerstone JF et al (2003) Screening of foods containing proanthocyanidins and their structural characterization using LC-MS/MS and thiolytic degradation. J Agric Food Chem 51:7513–7521. doi:10.1021/jf034815d CrossRefGoogle Scholar
  20. Guo Z, Megharaj M, Beer M et al (2009) Heavy metal impact on bacterial biomass based on DNA analyses and uptake by wild plants in the abandoned copper mine soils. Bioresour Technol 100:3831–3836. doi:10.1016/j.biortech.2009.02.043 CrossRefGoogle Scholar
  21. Hartmann A, Schmid M, Tuinen D van, Berg G (2008) Plant-driven selection of microbes. Plant Soil 321:235–257. doi: 10.1007/s11104-008-9814-y
  22. Hong C, Si Y, Xing Y, Li Y (2015) Illumina MiSeq sequencing investigation on the contrasting soil bacterial community structures in different iron mining areas. Environ Sci Pollut Res Int 22:10788–10799. doi:10.1007/s11356-015-4186-3 CrossRefGoogle Scholar
  23. Imperato F (2000) Kaempferol and quercetin 3-O-(X″,X″-di-protocatechuoyl)-glucuronides from Pteris vittata. Am Fern J 90:141–144. doi:10.2307/1547491 CrossRefGoogle Scholar
  24. Jaishankar M, Tseten T, Anbalagan N et al (2014) Toxicity, mechanism and health effects of some heavy metals. Interdiscip Toxicol 7:60–72. doi:10.2478/intox-2014-0009 CrossRefGoogle Scholar
  25. Jaishee N, Chakraborty U (2015) Comparative assessment of phytochemicals and HPLC analyses of phenolics present in Dicranopteris linearis (N. Burm.) Underw and Pteris vittata L. Int J Pharm Pharm Sci Res 5:1–7Google Scholar
  26. Kachenko AG, Singh B, Bhatia NP (2007) Heavy metal tolerance in common fern species. Aust J Bot 55:63–73. doi:10.1071/BT06063 CrossRefGoogle Scholar
  27. Kajdžanoska M, Gjamovski V, Stefova M (2010) HPLC-DAD-ESI-MSn identification of phenolic compounds in cultivated strawberries from Macedonia. Maced J Chem Chem Eng 29:181–194Google Scholar
  28. Karafin M, Romagnoli M, Fink DL et al (2010) Fatal infection caused by Cupriavidus gilardii in a child with aplastic anemia. J Clin Microbiol 48:1005–1007. doi:10.1128/JCM.01482-09 CrossRefGoogle Scholar
  29. Karamać M (2009) Chelation of Cu(II), Zn(II), and Fe(II) by tannin constituents of selected edible nuts. Int J Mol Sci 10:5485–5497. doi:10.3390/ijms10125485 CrossRefGoogle Scholar
  30. Kelly JJ, Häggblom M, Tate RL III (1999) Changes in soil microbial communities over time resulting from one time application of zinc: a laboratory microcosm study. Soil Biol Biochem 31:1455–1465. doi:10.1016/S0038-0717(99)00059-0 CrossRefGoogle Scholar
  31. Konopka A, Zakharova T, Bischoff M et al (1999) Microbial biomass and activity in lead-contaminated soil. Appl Environ Microbiol 65:2256–2259Google Scholar
  32. Kozich JJ, Westcott SL, Baxter NT et al (2013) Development of a dual-index sequencing strategy and curation pipeline for analyzing amplicon sequence data on the MiSeq Illumina sequencing platform. Appl Environ Microbiol 79:5112–5120. doi:10.1128/AEM.01043-13 CrossRefGoogle Scholar
  33. Langevin S, Vincelette J, Bekal S, Gaudreau C (2011) First case of invasive human infection caused by Cupriavidus metallidurans. J Clin Microbiol 49:744–745. doi:10.1128/JCM.01947-10 CrossRefGoogle Scholar
  34. Lenart-Boroń A, Boroń P (2014) The effect of industrial heavy metal pollution on microbial abundance and diversity in soils—a review. In: Hernandez Soriano MC (ed) Environmental risk assessment of soil contamination. InTech. p759–783. doi: 10.5772/57406
  35. Leong CNA, Tako M, Hanashiro I, Tamaki H (2008) Antioxidant flavonoid glycosides from the leaves of Ficus pumila L. Food Chem 109:415–420. doi:10.1016/j.foodchem.2007.12.069 CrossRefGoogle Scholar
  36. Lin L-Z, Harnly JM (2010) Phenolic component profiles of mustard greens, yu choy, and 15 other brassica vegetables. J Agric Food Chem 58:6850–6857. doi:10.1021/jf1004786 CrossRefGoogle Scholar
  37. Lou Z, Wang H, Zhu S et al (2011) Antibacterial activity and mechanism of action of chlorogenic acid. J Food Sci 76:M398–M403. doi:10.1111/j.1750-3841.2011.02213.x CrossRefGoogle Scholar
  38. Ma LQ, Komar KM, Tu C et al (2001) A fern that hyperaccumulates arsenic. Nature 409:579. doi:10.1038/35054664 CrossRefGoogle Scholar
  39. Mabry TJ, Markham KR, Thomas MB (1970) The systematic identification of flavonoids. Springer Berlin Heidelberg, Berlin, Heidelberg doi:10.1007/978-3-642-88458-0
  40. Maldonado PD, Rivero-Cruz I, Mata R, Pedraza-Chaverrí J (2005) Antioxidant activity of A-type proanthocyanidins from Geranium niveum (Geraniaceae). J Agric Food Chem 53:1996–2001. doi:10.1021/jf0483725 CrossRefGoogle Scholar
  41. Marques V, Farah A (2009) Chlorogenic acids and related compounds in medicinal plants and infusions. Food Chem 113:1370–1376. doi:10.1016/j.foodchem.2008.08.086 CrossRefGoogle Scholar
  42. Martucci MEP, De Vos RCH, Carollo CA, Gobbo-Neto L (2014) Metabolomics as a potential chemotaxonomical tool: application in the genus Vernonia schreb. PLoS One 9:e93149. doi:10.1371/journal.pone.0093149 CrossRefGoogle Scholar
  43. Mendoza-Wilson AM, Castro-Arredondo SI, Espinosa-Plascencia A et al (2016) Chemical composition and antioxidant-prooxidant potential of a polyphenolic extract and a proanthocyanidin-rich fraction of apple skin. Heliyon 2:e00073. doi:10.1016/j.heliyon.2016.e00073 CrossRefGoogle Scholar
  44. Michalet S, Rohr J, Warshan D et al (2013) Phytochemical analysis of mature tree root exudates in situ and their role in shaping soil microbial communities in relation to tree N-acquisition strategy. Plant Physiol Biochem 72:169–177. doi:10.1016/j.plaphy.2013.05.003 CrossRefGoogle Scholar
  45. Nguyen THH, Sakakibara M, Sano S, Mai TN (2011) Uptake of metals and metalloids by plants growing in a lead-zinc mine area, Northern Vietnam. J Hazard Mater 186:1384–1391. doi:10.1016/j.jhazmat.2010.12.020 CrossRefGoogle Scholar
  46. Nunes I, Jacquiod S, Brejnrod A, et al (2016) Coping with copper: legacy effect of copper on potential activity of soil bacteria following a century of exposure. FEMS Microbiol Ecol 92:fiw175. doi: 10.1093/femsec/fiw175
  47. Paul T, Das B, Apte K, Suchitra (2012) Hypoglycemic activity of Pteris vittata L., a fern on alloxan induced diabetic rats. Inventi Impact Planta active 2:88–91.Google Scholar
  48. Plumb GW, Price KR, Williamson G (1999) Antioxidant properties of flavonol glycosides from green beans. Redox Rep Commun Free Radic Res 4:123–127. doi:10.1179/135100099101534800 CrossRefGoogle Scholar
  49. Poehlein A, Kusian B, Friedrich B et al (2011) Complete genome sequence of the type strain Cupriavidus necator N-1. J Bacteriol 193:5017. doi:10.1128/JB.05660-11 CrossRefGoogle Scholar
  50. Qiang L, Luo F, Zhao X et al (2015) Identification of proanthocyanidins from litchi ( Litchi chinensis Sonn .) pulp by LC-ESI-Q-TOF-MS and their antioxidant activity. PLoS One 10:e0120480. doi:10.1371/journal.pone.0120480 CrossRefGoogle Scholar
  51. Quast C, Pruesse E, Yilmaz P et al (2013) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res 41:590–596. doi:10.1093/nar/gks1219 CrossRefGoogle Scholar
  52. R Core Team (2014) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL http://www.R-project.org/
  53. Salatino MLF, Prado J (1998) Flavonoid glycosides of Pteridaceae from Brazil. Biochem Syst Ecol 26:761–769. doi:10.1016/S0305-1978(98)00032-5 CrossRefGoogle Scholar
  54. Sánchez-Andrea I, Rodríguez N, Amils R, Sanz JL (2011) Microbial diversity in anaerobic sediments at Río Tinto, a naturally acidic environment with a high heavy metal content. Appl Environ Microbiol 77:6085–6093. doi:10.1128/AEM.00654-11 CrossRefGoogle Scholar
  55. Scherer J, Nies DH (2009) CzcP is a novel efflux system contributing to transition metal resistance in Cupriavidus metallidurans CH34. Mol Microbiol 73:601–621. doi:10.1111/j.1365-2958.2009.06792.x CrossRefGoogle Scholar
  56. Schloss PD, Westcott SL, Ryabin T et al (2009) Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl Environ Microbiol 75:7537–7541. doi:10.1128/AEM.01541-09 CrossRefGoogle Scholar
  57. Sharma SS, Dietz K-J (2009) The relationship between metal toxicity and cellular redox imbalance. Trends Plant Sci 14:43–50. doi:10.1016/j.tplants.2008.10.007 CrossRefGoogle Scholar
  58. Sheik CS, Mitchell TW, Rizvi FZ et al (2012) Exposure of soil microbial communities to chromium and arsenic alters their diversity and structure. PLoS One 7:e40059. doi:10.1371/journal.pone.0040059 CrossRefGoogle Scholar
  59. Singh BK, Quince C, Macdonald CA et al (2014) Loss of microbial diversity in soils is coincident with reductions in some specialized functions. Environ Microbiol 16:2408–2420. doi:10.1111/1462-2920.12353 CrossRefGoogle Scholar
  60. Singh M, Govindarajan R, Rawat AKS, Khare PB (2008) Antimicrobial flavonoid rutin from Pteris vittata L. against pathogenic gastrointestinal microflora. Am Fern J 98:98–103CrossRefGoogle Scholar
  61. Singh S, Parihar P, Singh R et al (2016) Heavy metal tolerance in plants: role of transcriptomics, proteomics, metabolomics, and ionomics. Front Plant Sci 6:1143. doi:10.3389/fpls.2015.01143 Google Scholar
  62. Thijs S, Sillen W, Rineau F et al (2016) Towards an enhanced understanding of plant–microbiome interactions to improve phytoremediation: engineering the metaorganism. Front Microbiol doi. doi:10.3389/fmicb.2016.00341
  63. Tu C, Ma LQ (2002) Effects of arsenic concentrations and forms on arsenic uptake by the hyperaccumulator ladder brake. J Environ Qual 31:641–647CrossRefGoogle Scholar
  64. Tu C, Ma LQ, Bondada B (2002) Arsenic accumulation in the hyperaccumulator Chinese brake and its utilization potential for phytoremediation. J Environ Qual 31:1671–1675CrossRefGoogle Scholar
  65. Turpeinen R, Kairesalo T, Häggblom MM (2004) Microbial community structure and activity in arsenic-, chromium- and copper-contaminated soils. FEMS Microbiol Ecol 47:39–50. doi:10.1016/S0168-6496(03)00232-0 CrossRefGoogle Scholar
  66. von Rozycki T, Nies DH (2009) Cupriavidus metallidurans: evolution of a metal-resistant bacterium. Antonie Van Leeuwenhoek 96:115–139. doi:10.1007/s10482-008-9284-5 CrossRefGoogle Scholar
  67. Vukics V, Guttman A (2010) Structural characterization of flavonoid glycosides by multi-stage mass spectrometry. Mass Spectrom Rev 29:1–16. doi:10.1002/mas.20212 Google Scholar
  68. Wahid F, Khan T, Shehzad O et al (2016) Phytochemical analysis and effects of Pteris vittata extract on visual processes. J Nat Med 70:8–17. doi:10.1007/s11418-015-0930-8 CrossRefGoogle Scholar
  69. Wakelin SA, Chu G, Lardner R et al (2010) A single application of Cu to field soil has long-term effects on bacterial community structure, diversity, and soil processes. Pedobiologia 53:149–158. doi:10.1016/j.pedobi.2009.09.002 CrossRefGoogle Scholar
  70. Wang J, Zhao F-J, Meharg AA et al (2002) Mechanisms of arsenic hyperaccumulation in Pteris vittata. Uptake kinetics, interactions with phosphate, and arsenic speciation. Plant Physiol 130:1552–1561. doi:10.1104/pp.008185 CrossRefGoogle Scholar
  71. Wang Q, Garrity GM, Tiedje JM, Cole JR (2007) Naïve bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73:5261–5267. doi:10.1128/AEM.00062-07 CrossRefGoogle Scholar
  72. Wang X, Chen M, Xiao J et al (2015) Genome sequence analysis of the naphthenic acid degrading and metal resistant bacterium Cupriavidus gilardii CR3. PLoS One 10:e0132881. doi:10.1371/journal.pone.0132881 CrossRefGoogle Scholar
  73. Whitaker BD, Stommel JR (2003) Distribution of hydroxycinnamic acid conjugates in fruit of commercial eggplant (Solanum melongena L.) cultivars. J Agric Food Chem 51:3448–3454. doi:10.1021/jf026250b CrossRefGoogle Scholar
  74. Yang J, Guo J, Yuan J (2008) In vitro antioxidant properties of rutin. LWT - Food Sci Technol 41:1060–1066. doi:10.1016/j.lwt.2007.06.010 CrossRefGoogle Scholar
  75. Zhang X, Niu J, Liang Y et al (2016) Metagenome-scale analysis yields insights into the structure and function of microbial communities in a copper bioleaching heap. BMC Genet 17:21. doi:10.1186/s12863-016-0330-4 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Hoang Nam Pham
    • 1
    • 2
  • Serge Michalet
    • 1
  • Josselin Bodillis
    • 1
  • Tien Dat Nguyen
    • 3
  • Thi Kieu Oanh Nguyen
    • 2
  • Thi Phuong Quynh Le
    • 4
  • Mohamed Haddad
    • 5
  • Sylvie Nazaret
    • 1
  • Marie-Geneviève Dijoux-Franca
    • 1
  1. 1.UMR 5557, Ecologie Microbienne, CNRS, INRA, VetagroSup, UCBLUniversité de LyonVilleurbanneFrance
  2. 2.University of Science and Technology of Hanoi, Vietnam Academy of Science and TechnologyHanoiVietnam
  3. 3.Institute of Marine BiochemistryVietnam Academy of Science and TechnologyHanoiVietnam
  4. 4.Institute of Natural Products ChemistryVietnam Academy of Science and TechnologyHanoiVietnam
  5. 5.UMR 152 Pharma-DEVUniversité de Toulouse, IRD, UPSToulouseFrance

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