1887

Abstract

In nitrogen-fixing rhizobia, emerging evidence shows significant roles for polyamines in growth and abiotic stress resistance. In this work we show that a polyamine-deficient ornithine decarboxylase null mutant () derived from Rm8530 had significant phenotypic differences from the wild-type, including greatly reduced production of exopolysaccharides (EPS; ostensibly both succinoglycan and galactoglucan), increased sensitivity to oxidative stress and decreased swimming motility. The introduction of the gene borne on a plasmid into the mutant restored wild-type phenotypes for EPS production, growth under oxidative stress and swimming. The production of calcofluor-binding EPS (succinoglycan) by the mutant was also completely or mostly restored in the presence of exogenous spermidine (Spd), norspermidine (NSpd) or spermine (Spm). The mutant formed about 25 % more biofilm than the wild-type, and its ability to form biofilm was significantly inhibited by exogenous Spd, NSpd or Spm. The mutant formed a less efficient symbiosis with alfalfa, resulting in plants with significantly less biomass and height, more nodules but less nodule biomass, and 25 % less nitrogen-fixing activity. Exogenously supplied Put was not able to revert these phenotypes and caused a similar increase in plant height and dry weight in uninoculated plants and in those inoculated with the wild-type or mutant. We discuss ways in which polyamines might affect the phenotypes of the mutant.

Funding
This study was supported by the:
  • DGAPA-PAPIIT (Award IN206317)
    • Principle Award Recipient: Michael F. Dunn
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000886
2020-01-14
2021-10-23
Loading full text...

Full text loading...

/deliver/fulltext/micro/166/3/278.html?itemId=/content/journal/micro/10.1099/mic.0.000886&mimeType=html&fmt=ahah

References

  1. Igarashi K, Kashiwagi K. Modulation of protein synthesis by polyamines. IUBMB Life 2015; 67:160–169 [View Article][PubMed]
    [Google Scholar]
  2. Michael AJ. Polyamine function in archaea and bacteria. J Biol Chem 2018; 293:18693–18701 [View Article][PubMed]
    [Google Scholar]
  3. Shah P, Swiatlo E. A multifaceted role for polyamines in bacterial pathogens. Mol Microbiol 2008; 68:4–16 [View Article][PubMed]
    [Google Scholar]
  4. Xie S-S, Wu H-J, Zang H-Y, Wu L-M, Zhu Q-Q et al. Plant growth promotion by spermidine-producing Bacillus subtilis OKB105. Mol Plant Microbe Interact 2014; 27:655–663 [View Article][PubMed]
    [Google Scholar]
  5. Michael AJ. Polyamines in eukaryotes, bacteria, and archaea. J Biol Chem 2016; 291:14896–14903 [View Article][PubMed]
    [Google Scholar]
  6. Altaf MM, Ahmad I. Biofilm formation on plant surfaces by rhizobacteria: Impact on plant growth and ecological significance. In Gupta VK. editor The Handbook of Microbial Bioresuroces Wallingford, Oxfordshire: CAB International; 2016 pp 81–95
    [Google Scholar]
  7. Becerra-Rivera VA, Dunn MF. Polyamine biosynthesis and biological roles in rhizobia. FEMS Microbiol Lett 2019; 366:7 [View Article]
    [Google Scholar]
  8. Geddes BA, Oresnik IJ. The mechanism of symbiotic nitrogen fixation. In Hurst CJ. editor The Mechanistic Benefits of Microbial Symbiosis Switzerland: Springer; 2016 pp 69–97
    [Google Scholar]
  9. Janczarek M. Exopolysaccharide production in rhizobia is regulated by environmental factors. In de Bruijn FJ. editor Biological Nitrogen Fixation 1 New Jersey, USA: John Wiley and Sons; 2015 pp 365–380
    [Google Scholar]
  10. Poole P, Ramachandran V, Terpolilli J. Rhizobia: from saprophytes to endosymbionts. Nat Rev Microbiol 2018; 16:291–303 [View Article][PubMed]
    [Google Scholar]
  11. Tambalo DD, Yost CK, Hynes MF. Motility and chemotaxis in the rhizobia. In de Bruijn FJ. editor Biological Nitrogen Fixation 1 New Jersey, USA: John Wiley and Sons; 2015 pp 337–348
    [Google Scholar]
  12. Dunn MF. Rhizobial amino acid metabolism: polyamine biosynthesis and functions. In D’Mello FJ. editor Handbook of Microbial Metabolism of Amino Acids Wallingford, Oxfordshire UK: CAB International; 2017 pp 352–370
    [Google Scholar]
  13. López-Gómez M, Cobos-Porras L, Prell J, Lluch C. Homospermidine synthase contributes to salt tolerance in free-living Rhizobium tropici and in symbiosis with Phaseolus vulgaris . Plant Soil 2016b; 404:413–425 [View Article]
    [Google Scholar]
  14. Braeken K, Daniels R, Vos K, Fauvart M, Bachaspatimayum D et al. Genetic determinants of swarming in Rhizobium etli . Microb Ecol 2008; 55:54–64 [View Article][PubMed]
    [Google Scholar]
  15. Kim SH, Wang Y, Khomutov M, Khomutov A, Fuqua C et al. The Essential Role of Spermidine in Growth of Agrobacterium tumefaciens Is Determined by the 1,3-Diaminopropane Moiety. ACS Chem Biol 2016; 11:491–499 [View Article]
    [Google Scholar]
  16. Wang Y, Kim SH, Natarajan R, Heindl JE, Bruger EL et al. Spermidine inversely influences surface interactions and planktonic growth in Agrobacterium tumefaciens . J Bacteriol 2016; 198:2682–2691 [View Article][PubMed]
    [Google Scholar]
  17. Becerra-Rivera VA, Bergström E, Thomas-Oates J, Dunn MF. Polyamines are required for normal growth in Sinorhizobium meliloti . Microbiology 2018; 164:600–613 [View Article][PubMed]
    [Google Scholar]
  18. Hernández VM, Girard L, Hernández-Lucas I, Vázquez A, Ortíz-Ortíz C et al. Genetic and biochemical characterization of arginine biosynthesis in Sinorhizobium meliloti 1021. Microbiology 2015; 161:1671–1682 [View Article][PubMed]
    [Google Scholar]
  19. Wang Y, Xu J, Chen A, Wang Y, Zhu J et al. GGDEF and EAL proteins play different roles in the control of Sinorhizobium meliloti growth, motility, exopolysaccharide production, and competitive nodulation on host alfalfa. Acta Biochim Biophys Sin 2010; 42:410–417 [View Article][PubMed]
    [Google Scholar]
  20. Dunn MF, Karr AL. Isolation of an extracellular polysaccharide (EPS) depolymerase produced by Bradyrhizobium japonicum . Curr Microbiol 1990; 20:359–363 [View Article]
    [Google Scholar]
  21. Bernabéu-Roda L, Calatrava-Morales N, Cuéllar V, Soto MJ. Characterization of surface motility in Sinorhizobium meliloti: regulation and role in symbiosis. Symbiosis 2015; 67:79–90 [View Article]
    [Google Scholar]
  22. O'Toole GA, Kolter R. Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis. Mol Microbiol 1998; 28:449–461 [View Article][PubMed]
    [Google Scholar]
  23. Hardy RW, Holsten RD, Jackson EK, Burns RC. The acetylene-ethylene assay for n(2) fixation: laboratory and field evaluation. Plant Physiol 1968; 43:1185–1207 [View Article][PubMed]
    [Google Scholar]
  24. López-Gómez M, Hidalgo-Castellanos J, Muñoz-Sánchez JR, Marín-Peña AJ, Lluch C et al. Polyamines contribute to salinity tolerance in the symbiosis Medicago truncatula-Sinorhizobium meliloti by preventing oxidative damage. Plant Physiol Biochem 2017; 116:9–17 [View Article][PubMed]
    [Google Scholar]
  25. Reuber TL, Walker GC. Biosynthesis of succinoglycan, a symbiotically important exopolysaccharide of Rhizobium meliloti . Cell 1993; 74:269–280 [View Article][PubMed]
    [Google Scholar]
  26. Hawkins JP, Geddes BA, Oresnik IJ. Common dyes used to determine bacterial polysaccharides on agar are affected by medium acidification. Can J Microbiol 2017; 63:559–562 [View Article][PubMed]
    [Google Scholar]
  27. Calatrava-Morales N, McIntosh M, Soto M. Regulation mediated by N-acyl homoserine lactone quorum sensing signals in the Rhizobium-legume symbiosis. Genes 2018; 9:9L263 [View Article]
    [Google Scholar]
  28. Amaya-Gómez CV, Hirsch AM, Soto MJ. Biofilm formation assessment in Sinorhizobium meliloti reveals interlinked control with surface motility. BMC Microbiol 2015; 15:58 [View Article][PubMed]
    [Google Scholar]
  29. Schäper S, Krol E, Skotnicka D, Kaever V, Hilker R et al. Cyclic di-GMP regulates multiple cellular functions in the symbiotic alphaproteobacterium Sinorhizobium meliloti . J Bacteriol 2016; 198:521–535 [View Article]
    [Google Scholar]
  30. Pérez-Mendoza D, Rodríguez-Carvajal Miguel Ángel, Romero-Jiménez L, Farias GdeA, Lloret J et al. Novel mixed-linkage β-glucan activated by c-di-GMP in Sinorhizobium meliloti . Proc Natl Acad Sci U S A 2015; 112:E757–E765 [View Article][PubMed]
    [Google Scholar]
  31. D'Argenio DA, Miller SI. Cyclic di-GMP as a bacterial second messenger. Microbiology 2004; 150:2497–2502 [View Article]
    [Google Scholar]
  32. Costerton JW, Lewandowski Z, Caldwell DE, Korber DR, Lappin-Scott HM. Microbial biofilms. Annu Rev Microbiol 1995; 49:711–745 [View Article]
    [Google Scholar]
  33. Rinaudi LV, González JE. The low-molecular-weight fraction of exopolysaccharide II from Sinorhizobium meliloti is a crucial determinant of biofilm formation. J Bacteriol 2009; 191:7216–7224 [View Article]
    [Google Scholar]
  34. Sorroche FG, Spesia MB, Zorreguieta A, Giordano W. A positive correlation between bacterial autoaggregation and biofilm formation in native Sinorhizobium meliloti isolates from Argentina. Appl Environ Microbiol 2012; 78:4092–4101 [View Article][PubMed]
    [Google Scholar]
  35. Flemming HC, Wingender J, Griegbe C, Mayer C. Physico-chemical properties of biofilms. In Evans LV. editor Biofilms: Recent Advances in their Study and Control Amsterdam: Harwood Academic Publishers; 2000 pp 19–34
    [Google Scholar]
  36. Fujishige NA, Kapadia NN, De Hoff PL, Hirsch AM. Investigations of Rhizobium biofilm formation. FEMS Microbiol Ecol 2006; 56:195–206 [View Article]
    [Google Scholar]
  37. Sturgill G, Rather PN. Evidence that putrescine acts as an extracellular signal required for swarming in Proteus mirabilis . Mol Microbiol 2004; 51:437–446 [View Article]
    [Google Scholar]
  38. Corral-Lugo A, De la Torre J, Matilla MA, Fernández M, Morel B et al. Assessment of the contribution of chemoreceptor-based signalling to biofilm formation. Environ Microbiol 2016; 18:3355–3372 [View Article][PubMed]
    [Google Scholar]
  39. Kurihara S, Suzuki H, Tsuboi Y, Benno Y. Dependence of swarming in Escherichia coli K-12 on spermidine and the spermidine importer. FEMS Microbiol Lett 2009; 294:97–101 [View Article]
    [Google Scholar]
  40. Skiebe E, de Berardinis V, Morczinek P, Kerrinnes T, Faber F et al. Surface-Associated motility, a common trait of clinical isolates of Acinetobacter baumannii, depends on 1,3-diaminopropane. Int J Med Microbiol 2012; 302:117–128 [View Article][PubMed]
    [Google Scholar]
  41. Karatan E, Michael AJ. A wider role for polyamines in biofilm formation. Biotechnol Lett 2013; 35:1715–1717 [View Article]
    [Google Scholar]
  42. Király Z, El-Zahaby HM, Klement Z. Role of extracellular polysaccharide (EPS) slime of plant pathogenic bacteria in protecting cells to reactive oxygen species. J Phytopathol 1997; 145:59–68 [View Article]
    [Google Scholar]
  43. Lehman AP, Long SR. Exopolysaccharides from Sinorhizobium meliloti can protect against H2O2-dependent damage. J Bacteriol 2013; 195:5362–5369 [View Article]
    [Google Scholar]
  44. Tkachenko AG, Nesterova LY. Polyamines as modulators of gene expression under oxidative stress in Escherichia coli . Biochemistry 2003; 68:850–856 [View Article][PubMed]
    [Google Scholar]
  45. Hérouart D, Sigaud S, Moreau S, Frendo P, Touati D et al. Cloning and characterization of the katA gene of Rhizobium meliloti encoding a hydrogen peroxide-inducible catalase. J Bacteriol 1996; 178:6802–6809 [View Article]
    [Google Scholar]
  46. Lehman AP, Long SR. OxyR-Dependent Transcription Response of Sinorhizobium meliloti to Oxidative Stress. J Bacteriol 2018; 200:e00622–17 [View Article][PubMed]
    [Google Scholar]
  47. Glazebrook J, Walker GC. A novel exopolysaccharide can function in place of the calcofluor-binding exopolysaccharide in nodulation of alfalfa by Rhizobium meliloti . Cell 1989; 56:661–672 [View Article][PubMed]
    [Google Scholar]
  48. Hardarson G, Heichel GH, Vance CP, Barnes DK. Evaluation of alfalfa and Rhizobium meliloti for compatibility in nodulation and nodule effectiveness 1 . Crop Sci 1981; 21:562–567 [View Article]
    [Google Scholar]
  49. Choi D, Lee Y, Cho H-T, Kende H. Regulation of expansin gene expression affects growth and development in transgenic rice plants. Plant Cell 2003; 15:1386–1398 [View Article]
    [Google Scholar]
  50. López-Gómez M, Hidalgo-Castellanos J, Lluch C, Herrera-Cervera JA. 24-Epibrassinolide ameliorates salt stress effects in the symbiosis Medicago truncatula-Sinorhizobium meliloti and regulates the nodulation in cross-talk with polyamines. Plant Physiol Biochem 2016a; 108:212–221 [View Article][PubMed]
    [Google Scholar]
  51. Palma F, López-Gómez M, Tejera NA, Lluch C. Salicylic acid improves the salinity tolerance of Medicago sativa in symbiosis with Sinorhizobium meliloti by preventing nitrogen fixation inhibition. Plant Sci 2013; 208:75–82 [View Article][PubMed]
    [Google Scholar]
  52. Palma F, López-Gómez M, Tejera NA, Lluch C. Involvement of abscisic acid in the response of Medicago sativa plants in symbiosis with Sinorhizobium meliloti to salinity. Plant Sci 2014; 223:16–24 [View Article][PubMed]
    [Google Scholar]
  53. Radhakrishnan R, Lee I-J. Ameliorative effects of spermine against osmotic stress through antioxidants and abscisic acid changes in soybean pods and seeds. Acta Physiol Plant 2013; 35:263–269 [View Article]
    [Google Scholar]
  54. Terakado-Tonooka J, Fujihara S. Involvement of polyamines in the root nodule regulation of soybeans (Glycine max). Plant Root 2008; 2:46–53 [View Article]
    [Google Scholar]
  55. Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA et al. Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene 1995; 166:175–176 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000886
Loading
/content/journal/micro/10.1099/mic.0.000886
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error