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Quantitative assessments of the host range and strain specificity of endophytic colonization by Klebsiella pneumoniae 342

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Abstract

Enteric bacteria, particularly Klebsiella, are common endophytes of plants. Endophytic colonization is important as these bacteria may be beneficial, either by providing fixed N or growth hormones to the host plant. In this work, we assessed the host range and strain specificity for endophytic colonization with Klebsiella pneumoniae 342 (Kp342) on five host plants. This strain was inoculated onto seedlings of Medicago sativa, Medicago truncatula, Arabidopsis thaliana, Triticum aestivum, and Oryza sativa. The type strain of K. pneumoniae, ATCC13883, was also inoculated on all of these hosts except M. truncatula. Both strains were labeled with GFP. Eight inoculum levels were used from 1 CFU to 107 CFU per plant plus uninoculated controls. Six days after inoculation, the number of cells colonizing the rhizosphere and interior were determined. Inoculation with about one CFU of Kp342 was adequate to obtain high colonization levels on the rhizosphere and roots of all host plants. The type strain could colonize the interior of the host plant but the highest colonization levels were generally 100-fold lower than those obtained from Kp342 and those levels required at least 1000 cells in the inoculum. Thus, Kp342 was a more efficient colonizer of the plant apoplast. In addition, the monocots inoculated in this work were colonized endophytically in much higher numbers than were the dicots. Cells of Kp342 congregate at lateral root junctions suggesting the cells enter the plant through cracks created by lateral root extensions. The strain and host effects observed here suggest that endophytic colonization is an active process controlled by genetic determinants from both partners.

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References

  • Baldani J I, Baldani V L D, Seldin L and Döbereiner J 1986 Characterization of Herbaspirillum seropedicae gen. nov., sp. nov., a root associated nitrogen fixing bacterium. Int. J. Syst. Bacteriol. 36, 86–93.

    Google Scholar 

  • Bastian F, Rapparini F, Baraldi R, Piccoli P and Bottini R 1999 Inoculation with Acetobacter diazotrophicus increases glucose and fructose content in shoots of Sorghum bicolor (L.) Moench. Symbiosis 27, 147–156.

    Google Scholar 

  • Broek A V, Michiels J, Vangool A and Vanderleyden J 1993 Spatialtemporal colonization patterns of Azospirillum brasilense on the wheat root surface and expression of the bacterial NifH gene during association. Mol. Plant Microb. Interact. 6, 592–600.

    Google Scholar 

  • Chelius M K and Triplett E W 2000 Immunolocalization of dinitrogenase reductase produced by Klebsiella pneumoniae in association with Zea mays L. Appl. Environ. Microbiol. 66, 783–787.

    Google Scholar 

  • Chelius M K and Triplett E W 2001 The diversity of Archaea and Bacteria in the roots of Zea mays L. Microb.Ecol. 41, 252–263.

    Google Scholar 

  • Cody CW, Prasher D C, Westler WM, Prendergast F G and Ward W W 1993 Chemical structure of the hexapeptide chromophore of the Aequorea green fluorescent protein. Biochemistry 32, 1212–1218.

    Google Scholar 

  • Dong Z M, Canny M J, McCully M E, Roboredo M R, C abadilla C F, Ortega E and Rodes R 1994 A nitrogen-fixing endophyte of sugarcane stems – a new role for the apoplast. Plant Physiol. 105, 1139–1147.

    Google Scholar 

  • Dong Y, Glasner J D, Blattner F R and Triplett E W 2001 Genomic interspecies microarray hybridization: rapid discovery of three thousand genes in the maize endophyte, Klebsiella pneumoniae 342, by microarray hybridization with Escherichia coli K12 open reading frames. Appl. Environ. Microbiol. 67, 1911–1921.

    Google Scholar 

  • Dong,Y, Iniguez, A L, Ahmer B M M and Triplett E W 2003 Kinetics and strain specificity of rhizosphere and endophytic colonization by enteric bacteria on seedlings of Medicago sativa and Medicago truncatula. Appl. Environ. Microbiol. 69, 1783–1790.

    Google Scholar 

  • Dos Reis F B, Da Silva L G, Reis V M and Döbereiner, J 2000 Occurrence of diazotrophic bacteria in different sugar canegenotypes. Pesquisa Agropecuaria Brasileira 35, 985–994.

    Google Scholar 

  • Egener T, Hurek T and Reinhold-Hurek, B 1999 Endophytic expression of nif genes of Azoarcus sp. strain BH72 in rice roots. Mol. Plant-Microbe Interact. 12, 813–819.

    Google Scholar 

  • Elbeltagy A, Nishioka K, Sato T, Suzuki H, Ye B, Hamada T, Isawa T, Mitsui H and Minamisawa K 2001 Endophytic colonization and in planta nitrogen fixation by a Herbaspirillum sp. isolated from wild rice species. Appl. Environ. Microbiol. 67, 5285–5293.

    Google Scholar 

  • Estrada P, Mavingui P, Cournoyer B, Fontaine F, Balandreau J and Caballero-Mellado J 2002 A N2 fixing endophytic Burkholderia sp. associated with maize plants cultivated in Mexico. Can. J. Microbiol. 48, 285–294.

    Google Scholar 

  • Fuentes-Ramirez L E, Bustillos-Cristales R, Tapia-Hernandez A, Jimenez-Salgado T, Wang E T, Martinez-Romero E and Caballero-Mellado, J 2002 Novel nitrogen-fixing acetic acid bacteria, Gluconacetobacter johannae sp. nov. and Gluconacetobacter azotocaptans sp. nov., associated with coffee plants. Int. J. Syst. Evol. Microbiol. 51, 1305–1314.

    Google Scholar 

  • Gandhi M, Golding, S, Yaron S and Mattews K R 2001 Use of green fluorescent protein expressing Salmonella Stanley to investigate survival, spatial location, and control on alfalfa sprouts. J. Food Protect. 64, 1891–1898.

    Google Scholar 

  • Gillis M, Kersters K, Hoste B, Janssens D, Kroppenstedt R M, Stephan M P, Teixeira K R S, Döbereiner J and Deley J 1989 Acetobacter diazotrophicus sp. nov., a nitrogen-fixing acetic acid bacterium associated with sugarcane. Int. J. Syst. Bacteriol. 39, 361–364.

    Google Scholar 

  • Gutierrez-Zamora M L and Martinez-Romero E 2001 Natural endophytic association between Rhizobium etli and maize (Zea mays L.). J. Biotechnol. 91, 117–126.

    Google Scholar 

  • Gyaneshwar P, James, E K, Mathan N, Teddy P M, Reinhold-Hurek B and Ladha J K 2001 Endophytic colonization of rice by a diazotrophic strain of Serratia marcescens. J. Bacteriol. 183, 2634–2645.

    Google Scholar 

  • James E K and Olivares F L 1997 Infection and colonization of sugar cane and other graminaceous plants by endophytic diazotrophs. Crit. Rev. Plant Sci. 17, 77–119.

    Google Scholar 

  • James E K, Reis V M, Olivares F L, Baldani J I and Döobereiner J 1994 Infection of sugar cane by the nitrogen-fixing bacterium Acetobacter diazotrophicus. J. Exp. Bot. 45, 757–766.

    Google Scholar 

  • James E K, Olivares F L, de Oliveira A L M, dos Reis F B, da Silva L G and Reis V M 1997 Further observations on the interaction between sugar cane and Gluconacetobacter diazotrophicus under laboratory and greenhouse conditions. J. Exp. Bot. 52, 747–760.

    Google Scholar 

  • Jensen H L 1942 Nitrogen fixation in leguminous plants. II. Is symbiotic nitrogen fixation influenced by Azotobacter? Proc. Linn. Soc. NSW 67, 205–212.

    Google Scholar 

  • Jimenez-Salgado T, Fuentes-Ramirez L E, Tapia-Hernandez A, Mascarua-Esparza M A, Martinez-Romero E and Caballero-Mellado J 1997 Coffea arabica L., a new host plant for Acetobacter diazotrophicus, and isolation of other nitrogen-fixing acetobacteria. Appl. Environ. Microbiol. 63, 3676–3683.

    Google Scholar 

  • Kirchhof G, Eckert B, Stoffels M, Baldani J I, Reis V M and Hartmann A 2001 Herbaspirillum frisingense sp. nov.., a new nitrogen-fixing bacterial species that occurs in C4-fibre plants. Int. J. Syst. Evol. Microbiol. 51, 157–168.

    Google Scholar 

  • Kirchhof G, Reis VM, Baldani J I, Eckert B, Döbereiner J and Hartmann A 1997 Occurrence, physiological and molecular analysis of endophytic diazotrophic bacteria in gramineous energy plants. Plant Soil 194, 45–55.

    Google Scholar 

  • Kovtunovych G, Lar O, Kamalova S, Kordyum V, Kleiner D and Kozyrovska N 1999 Correlation between pectate lyase activity and ability of diazotrophic Klebsiella oxytoca VN 13 to penetrate into plant tissues. Plant Soil 215, 1–6.

    Google Scholar 

  • Loganathan P, Sunita R, Parida A K and Nair S 1999 Isolation and characterization of two genetically distant groups of Acetobacter diazotrophicus from a new host plant Eleusine coracana L. J. Appl. Microbiol. 87, 167–172.

    Google Scholar 

  • Omoregie S N, Asemota H N, Osagie A U, Mantell S and Ahmad M H 1999 Occurrence of free-living bacteria in tubers of Dioscorea yams. Trop. Agric. 76, 250–255.

    Google Scholar 

  • Palus J A, Borneman J, Ludden PW and Triplett EW 1996 Isolation and characterization of endophytic diazotrophs from Zea mays L. and Zea luxurians Iltis and Doebley. Plant Soil 186, 135–142.

    Google Scholar 

  • Reinhold-Hurek B, Hurek T, Gillis M, Hoste B, Vancanneyt M, Kersters K and Deley J1993 Azoarcus gen. nov., nitrogen fixing proteobacteria associated with roots of Kallar grass (Leptochloa fusca (L.) Kunth.), and description of 2 species, Azoarcus in digens sp. nov. and Azoarcus communis sp. nov. Int. J. Syst. Bacteriol. 43, 574–584.

    Google Scholar 

  • Reis Jnr F B, Olivares F L, Oliveira L M N, Reis V L M, Baldani J I and Döbereiner J 1995 Infection and colonization of Acetobacter diazotrophicus in sugar cane plantlets. In International Symposium on Sustainable Agriculture for the Tropics – the Role of Biological Nitrogen Fixation, Eds. R M Boddey and A S de Resende. pp. 225–226. EMBRAPA, Rio de Janeiro.

    Google Scholar 

  • Reis VM, Baldani J I, Baldani V L D and Döbereiner, J 2000 Biological dinitrogen fixation in gramineae and palm trees. Crit. Rev. Plant Sci. 19, 227–247.

    Google Scholar 

  • Riggs P J, Chelius M K, Iniguez A L, Kaeppler S M and Triplett E W 2001 Enhanced maize productivity by inoculation with diazotrophic bacteria. Austr. J. Plant Physiol. 28, 829–836.

    Google Scholar 

  • Riggs P J, Moritz R L, Chelius MK, Dong Y, Iniguez A L, Kaeppler S M, Casler M D and Triplett E W 2002 Isolation and characterization of diazotrophic endophytes from grasses and their effects on plant growth. In Nitrogen Fixation: Global Perspectives, Proceedings of the 13th International Congress on Nitrogen Fixation. Eds. T R Finan, M R O'Brian, D B Layzell, J K Vessey and W E Newton. pp. 263–267. CABI Publishing, New York.

    Google Scholar 

  • Robleto E A, Scupham A J and Triplett E W1997 Trifolitioxin production in Rhizobium etli strain CE3 increases competitiveness for rhizosphere colonization and root noculation of Phaseolus vulgaris in soil. Mol. Plant Microb. Interact. 10, 228–233.

    Google Scholar 

  • Stein T, Hayen-Schneg N and Fendrik I 1997 Contribution of BNF by Azoarcus sp. BH72 in Sorghum vulgare. Soil Biol. Biochem. 29, 969–971.

    Google Scholar 

  • Tan Z Y, Hurek T, Gyaneshwar P, Ladha J K and Reinhold-Hurek B 2001 Novel endophytes of rice form a taxonomically distinct subgroup of Serratia marcescens. Syst. Appl. Microbiol. 24, 245–251.

    Google Scholar 

  • Tapia-Hernandez A, Bustillos-Cristales M R, Jimenez-Salgado T, Caballero-Mellado J and Fuentes-Ramirez L E 2000 Natural endophytic occurrence of Acetobacter diazotrophicus in pineapple plants. Microb. Ecol. 39, 49–55.

    Google Scholar 

  • Verma S C, Ladha J K and Tripathi A K 2001 Evaluation of plant growth promoting and colonization ability of endophytic diazotrophs from deep water rice. J. Biotechnol. 91, 127–141

    Google Scholar 

  • Weber O B, Baldani V L D, Teixeira K R S, Kirchhof G, Baldani J I and Döbereiner J 1999 Isolation and characterization of diazotrophic bacteria from banana and pineapple plants. Plant Soil 12, 103–113.

    Google Scholar 

  • Yanni Y G, Rizk R Y, Corich V, Squartini A, Ninke K, PhilipHollingsworth S, Orgambide G, deBruijn F, Stoltzfus J, Buckley D, Schmidt T M, Mateos P F, Ladha J K and Dazzo F B 1997 Natural endophytic association between Rhizobium leguminosarum bv. trifolii and rice roots and assessment of its potential to promote rice growth. Plant Soil 194, 99–114.

    Google Scholar 

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  1. Department of Agronomy, University of Wisconsin-Madison, 1575 Linden Drive, Madison, WI, 53706, USA

    Yuemei Dong, A. Leonardo Iniguez & Eric W. Triplett

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  1. Yuemei Dong
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  2. A. Leonardo Iniguez
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  3. Eric W. Triplett
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Dong, Y., Iniguez, A.L. & Triplett, E.W. Quantitative assessments of the host range and strain specificity of endophytic colonization by Klebsiella pneumoniae 342. Plant and Soil 257, 49–59 (2003). https://doi.org/10.1023/A:1026242814060

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