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A nopA Deletion Mutant of Sinorhizobium fredii USDA257, a Soybean Symbiont, is Impaired in Nodulation

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Abstract

Sinorhizobium fredii USDA257 employs type III secretion system (T3SS) to deliver effector proteins into the host cells through pili. The nopA protein is the major component of USDA257 pili. The promoter region of USDA257 nopA possesses a well conserved tts box. Serial deletion analysis revealed that the tts box is absolutely essential for flavonoid induction of nopA. Deletion of nopA drastically lowered the number of nodules formed by USDA257 on cowpea and soybean cultivar Peking. In contrast to the parental strain, the USDA257 nopA mutant was able to form few nodules on soybean cultivars McCall and Williams 82. Light and transmission electron microscopy examination of these nodules revealed numerous starch grains both in the infected and uninfected cells.

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References

  1. Alfano JR, Bauer DW, Milos TM, Collmer A (1996) Analysis of the role of the Pseudomonas syringae pv. syringae hrpZ harpin in elicitation of the hypersensitive response in tobacco using functionally non-polar hrpZ deletion mutations, truncated hrpz fragments, and hrmA mutations. Mol Microbiol 19:715–728

    Article  CAS  PubMed  Google Scholar 

  2. Amadou C, Pascal G, Mangenot S, Glew M, Bontemps C, Capela D, Carrere S, Cruveiller S, Dossat C, Lajus A, Marchetti M, Poinsot V, Rouy B, Servin Z, Saad M, Schenowitz C, Barbe V, Batut J, Medigue C, Masson-Boivin C (2008) Genome sequence of the beta-rhizobium Cupriavidus taiwanensis and comparative genomics of rhizobia. Genome Res 18:1472–1483

    Article  CAS  PubMed  Google Scholar 

  3. Annapurna K, Krishnan HB (2003) Molecular aspects of soybean cultivar-specific nodulation by Sinorhizobium fredii USDA257. Indian J Exp Biol 41:1114–1123

    CAS  PubMed  Google Scholar 

  4. Bartsev AV, Deakin WJ, Boukli NM, McAlvin CB, Stacey G, Malnoë P, Broughton WJ, Staehelin C (2004) nopL, an effector protein of Rhizobium sp. NGR234, thwarts activation of plant defense reactions. Plant Physiol 134:871–879

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Deakin WJ, Broughton WJ (2009) Symbiotic use of pathogenic strategies: rhizobial protein secretion systems. Nat Rev Microbiol 7:312–320

    CAS  PubMed  Google Scholar 

  6. Deakin WJ, Marie C, Saad MM, Krishnan HB, Brougthon WJ (2005) nopA is associated with cell surface appendages produced by the type III secretion system of Rhizobium sp. strain NGR234. Mol Plant Microbe Interact 18:499–507

    Article  CAS  PubMed  Google Scholar 

  7. de Lyra MCCP, López-Baena FJ, Madinabeitia N, Vinardell JM, Espuny MR, Cubo MT, Bellogín RA, Ruiz-Sainz JE, Ollero FJ (2006) Inactivation of the Sinorhizobium fredii HH103 rhcJ gene abolishes nodulation outer proteins (Nops) secretion and decreases the symbiotic capacity with soybean. Int Microbiol 9:125–133

    Google Scholar 

  8. Finan TM, Wood JM, Jordan DC (1983) Symbiotic properties of C4-dicarboxylic acid transport mutants of Rhizobium leguminosarum. J Bacteriol 154:1403–1413

    CAS  PubMed Central  PubMed  Google Scholar 

  9. Galan JE, Collmer A (1999) Type III secretion machines: bacterial devices for protein delivery into host cells. Science 284:1322–1328

    Article  CAS  PubMed  Google Scholar 

  10. Harrison J, Jamet A, Muglia CI, Van de Sype G, Aguilar OM, Puppo A, Frendo P (2005) Glutathione plays a fundamental role in growth and symbiotic capacity of Sinorhizobium meliloti. J Bacteriol 187:168–174

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  11. He SY (1998) Type III protein secretion systems in plant and animal pathogenic bacteria. Annu Rev Phytopathol 36:363–392

    Article  CAS  PubMed  Google Scholar 

  12. Heron DS, Érsek T, Krishnan HB, Pueppke SG (1989) Nodulation mutants of Rhizobium fredii USDA257. Mol Plant Microbe Interact 2:4–10

    Article  Google Scholar 

  13. Kovacs LG, Balatti PA, Krishnan HB, Pueppke SG (1995) Transcriptional organization and expression of nolXWBTUV, a locus that regulates cultivar-specific nodulation of soybean by Rhizobium fredii USDA257. Mol Microbiol 17:923–933

    Article  CAS  PubMed  Google Scholar 

  14. Krause A, Doerfel A, Göttfert M (2002) Mutational and transcriptional analysis of the type III secretion system of Bradyrhizobium japonicum. Mol Plant Microbe Interact 15:1228–1235

    Article  CAS  PubMed  Google Scholar 

  15. Krishnan HB, Pueppke SG (1993) Flavonoid inducers of nodulation genes stimulate Rhizobium fredii USDA257 to export proteins into the environment. Mol Plant Microbe Interact 6:107–113

    Article  CAS  PubMed  Google Scholar 

  16. Krishnan HB, Lorio J, Kim WS, Jiang G, Kim KY, DeBoer M, Pueppke SG (2003) Extracellular proteins involved in soybean cultivar-specific nodulation are associated with pilus-like surface appendages and exported by a type III protein secretion system in Sinorhizobium fredii USDA257. Mol Plant Microbe Interact 16:617–625

    Article  CAS  PubMed  Google Scholar 

  17. Krishnan HB, Natarajan SS, Kim WS (2011) Distinct cell surface appendages produced by Sinorhizobium fredii USDA257 and S. fredii USDA191, cultivar-specific and nonspecific symbionts of soybean. Appl Environ Microbiol 77:6240–6248

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Lodwig EM, Hosie AH, Bourdès A, Findlay K, Allaway D, Karunakaran R, Downie JA, Poole PS (2003) Amino-acid cycling drives nitrogen fixation in the legume–Rhizobium symbiosis. Nature 422:722–726

    Article  CAS  PubMed  Google Scholar 

  19. Lo′pez-Baena FJ, Monreal JA, Pérez-Montaño F, Guasch-Vidal B, Bellogín RA, Vinardell JM, Ollero FJ (2009) The absence of Nops secretion in Sinorhizobium fredii HH103 increases GmPR1 expression in Williams soybean. Mol Plant Microbe Interact 22:1445–1454

    Article  Google Scholar 

  20. Lorio JC, Kim WS, Krishnan HB (2004) nopB, a soybean cultivar specificity protein from Sinorhizobium fredii USDA257, is a type III secreted protein. Mol Plant Microbe Interact 17:1259–1268

    Article  CAS  PubMed  Google Scholar 

  21. Meinhardt LW, Krishnan HB, Balatti PA, Pueppke SG (1993) Molecular cloning and characterization of a sym plasmid locus that regulates cultivar-specific nodulation of soybean by Rhizobium fredii USDA257. Mol Microbiol 9:17–29

    Article  CAS  PubMed  Google Scholar 

  22. Okazaki S, Zehner S, Hempel J, Lang K, Gottfert M (2009) Genetic organization and functional analysis of the type III secretion system of Bradyrhizobium elkanii. FEMS Microbiol Lett 295:88–95

    Article  CAS  PubMed  Google Scholar 

  23. Okazaki S et al (2010) Identification and functional analysis of type III effector proteins in Mesorhizobium loti. Mol Plant Microbe Interact 23:223–234

    Article  CAS  PubMed  Google Scholar 

  24. Pueppke SG, Broughton WJ (1999) Rhizobium sp. strain NGR234 and R. fredii USDA257 share exceptionally broad, nested host ranges. Mol Plant Microbe Interact 12:293–318

    Article  CAS  PubMed  Google Scholar 

  25. Quandt J, Hynes MF (1993) Versatile suicide vectors which allow direct selection for gene replacement in gram-negative bacteria. Gene 127:15–21

    Article  CAS  PubMed  Google Scholar 

  26. Schuldes J, Rodriguez Orbegoso M, Schmeisser C, Krishnan HB, Daniel R, Streit WR (2012) Complete genome sequence of the broad-host-range strain Sinorhizobium fredii USDA257. J Bacteriol 194:4483

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  27. Viprey V, Del Greco A, Golinowski W, Broughton WJ, Perret X (1998) Symbiotic implications of type III protein secretion machinery in Rhizobium. Mol Microbiol 28:1381–1389

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We thank Julio Lorio and Jeong Sun-Hyung for their technical assistance and Nathan Oehrle for reviewing this manuscript. Microscopy was carried out at the Electron Microscopy Facility of the University of Missouri, School of Veterinary Medicine. Names are necessary to report factually on available data; however, either the University of Missouri or USDA neither guarantees nor warrants the standard of product and the use of the name by the University of Missouri or USDA implies no approval of the product to the exclusion of others that may be suitable.

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  1. Plant Genetics Research Unit, Agricultural Research Service, U.S. Department of Agriculture, University of Missouri, Columbia, MO, 65211, USA

    Won-Seok Kim & Hari B. Krishnan

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  1. Won-Seok Kim
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  2. Hari B. Krishnan
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Correspondence to Hari B. Krishnan.

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Kim, WS., Krishnan, H.B. A nopA Deletion Mutant of Sinorhizobium fredii USDA257, a Soybean Symbiont, is Impaired in Nodulation. Curr Microbiol 68, 239–246 (2014). https://doi.org/10.1007/s00284-013-0469-4

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  • DOI: https://doi.org/10.1007/s00284-013-0469-4

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