Current Microbiology

, Volume 52, Issue 3, pp 186–190 | Cite as

Colonization of Maize and Rice Plants by Strain Bacillus megaterium C4

Article

Abstract

Bacillus megaterium C4, a nitrogen-fixing bacterium, was marked with the gfp gene. Maize and rice seedlings were inoculated with the, GFP-labeled B. megaterium C4 and then grown in gnotobiotic condition. Observation by confocal laser scanning microscope showed that the GFP-labeled bacterial cells infected the maize roots through the cracks formed at the lateral root junctions and then penetrated into cortex, xylem, and pith, and that the bacteria migrated slowly from roots to stems and leaves. The bacteria were mainly located in the intercellular spaces, although a few bacterial cells were also present within the xylem vessels, root hair cells, epidermis, cortical parenchyma, and pith cells. In addition, microscopic observation also revealed clearly that the root tip in the zone of elongation and differentiation and the junction between the primary and the lateral roots were the two sites for the bacteria entry into rice root. Therefore, we conclude that this Gram-positive nitrogen-fixer has a colonization pattern similar to those of many Gram-negative diazotrophs, such as Azospirillun brasilense Yu62 and Azoarcus sp. As far as we know, this is the first detailed report of the colonization pattern for Gram-positive diazotrophic Bacillus.

Literature Cited

  1. 1.
    An QL, Yang XJ, Dong YM, Feng U, Kuang BJ, Li JD (2001) Using confocal laser scanning microscope to visualize the infection of rice by GFP-labelled Klebsiella oxytoca SA2, Acta Botanica Sinica 43:558–564Google Scholar
  2. 2.
    Chi F, -Sheii SH, Chen SF, Jing YX (2004) Migration of Azospirillum brasilense Yu62 from root to stem and leaves inside rice and tobacco plants. Acta Botanica Sinica 46:1065–1070Google Scholar
  3. 3.
    Ding YQ, Wang JP, Liu Y, Chen SF (2005) Isolation and identification of nitrogen-fixing bacilli from plant rhizosphere in Beijing region. J Appl Microbiol (in press)Google Scholar
  4. 4.
    Dunn AK, Handelsman J (1999) A vector for promoter trapping in Bacillus cereus. Gene 226:297–305CrossRefPubMedGoogle Scholar
  5. 5.
    Forester RC (1986) The infrastructure of the rhizoplane and rhizosphere, Annu Rew Phytopathol 24:211–234Google Scholar
  6. 6.
    Hurek T, Handley LL, Reinhold-Hurek B, Piche Y (2002) Azoarcus grass endophytes contribute fixed nitrogen to the plant in an unculturable state. Mol Plant-Microbe Interact 15:233–242PubMedGoogle Scholar
  7. 7.
    Iniguez AL, Dong Y, Triplet EW (2004) Nitrogen fixation in wheat provided by Klebsiella pnewnoniae 342. Mol Plant-Microbe Interact 17:1078–1085PubMedGoogle Scholar
  8. 8.
    James EK, (2000) Nitrogen fixation in endophytic and associative symbiosis. Field Crops Res 65:197–209CrossRefGoogle Scholar
  9. 9.
    Lancashire JF, Terry TD, Blackall PJ, Joinings MP (2005) Plasmid-encoded Tet B tetracycline resistance in Haemophilus parasuis. Antimicrob Agents Chemother 49:1927–1931CrossRefPubMedGoogle Scholar
  10. 10.
    Li L, Yang C, Liu ZD? Li FD, Yu ZN (2000) Screening of acrystalliferous mutants from Bacillus, thurimiensis. and their transformation properties. Acta Microbiol Sin 40:85–90 (in Chinese with English abstract)Google Scholar
  11. 11.
    Liu Y, Chen SF, Li JL (2003) Colonization pattern of Azospirillum brasilense Yu62 on maize roots. Acta Botan Sin 45:748–752Google Scholar
  12. 12.
    Nair JR, Singh G, Sekar V (2002) Isolation and characterization of a novel Bacillus strain from coffee phyllosphere showing antifungal activity. J Appl Microbiol 93:772–780CrossRefPubMedGoogle Scholar
  13. 13.
    Newman KL, Almeida RPP, Purcell AH, Lindow SE (2003) Use of a green fluoresent strain for analysis of Xylella fastidiosa colonization of Vitis vinifera. Appl Environ Microbiol 69:7319–7327CrossRefPubMedGoogle Scholar
  14. 14.
    Patriquin DG5 Dobereiner J, Jain DK (1983) Sites and progresses of association between diazotrophs and grasses. Can J Microbiol 29:900–915Google Scholar
  15. 15.
    Reinhold-Hurek B, Hurek T (1998) Life in grasses: diazotrophic endophytes. Trends Microbiol 61:131–144Google Scholar
  16. 16.
    Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual (3rd ed). Cold Spring Harbor Laboratory PressGoogle Scholar
  17. 17.
    Schloter M, Kirchhof G, Heinzmann J, Dobereiner J, Hartmann A (1994) Immunological studies of the wheat-root-colonization by the Azospirillum brasilense strains Sp7 and Sp245 using strain-specific monoclonal antibodies. In: Hegazi NA, Fayez M, Monib N (eds) Nitrogen fixation with nonn-legumes. Cairo: American University in Cairo Press, pp 291–297Google Scholar
  18. 18.
    Schloter M, Hartmann A (1998) Endophytic and surface colonization of wheat roots (Triticwn aestivum) by different Azospirillum brasilense strains studied with strain-specific monoclonal antibodies. Symbiosis 25:159–179Google Scholar
  19. 19.
    Sevilla M, Burris RH, Gunapala N, Kennedy C (2001) Comparison of benefit to sugarcane plant growth and 15N2 incorporation following inoculation of sterile plants with Acetobacter diazotrophicus wild-type and Nif mutant strains. Mol Plant Microbe Interact 4:358–366Google Scholar
  20. 20.
    Tian T, Qi XC, Wang Q, Mei RH (2004) Colonization study of GFP-tegged Bacillus strains on wheat surface. Acta Phytopathol Sin 34:346–351 (in Chinese with English abstract)Google Scholar
  21. 21.
    Vasse J, Frey P, Trigalet A (1995) Microscopic studies of intercellular infection and protoxylem invasion of tomato roots Pseudomonas solanacecarum. Mol Plant Microbe Interact 8:241–251Google Scholar
  22. 22.
    Yan Z, Reddy MS, Kloepper JW (2003) Survival and colonization of rhizobacteria in a tomato transplant system. Can J Microbiol 49:383–389CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  1. 1.State Key Laboratory for Agrobiotechnology and College of Biological SciencesChina Agricultural UniversityP. R. China
  2. 2.Laboratory for Agro-Microbial Resource and Application of Ministry of AgricultureP. R. China
  3. 3.Chemical and Biological SchoolShenYang Normal UniversityP. R. China

Personalised recommendations