Abstract
The apoplastic fluids of field-grown Zea mays and Zea luxurians plants were isolated from surface sterilized stem tissue by centrifugation and spread on agar plates containing a nitrogen-free, defined medium. The predominant bacterium isolated from these plates was characterized further. The ability of this bacterium to fix nitrogen was confirmed by its ability to grow on a semi-solid, nitrogen-free medium and reduce 15N2 to 15NH3 and acetylene to ethylene. Protions of the nifH and 16S rRNA genes from this organism were amplified by PCR and sequenced. The nifH gene, which codes for dinitrogenase reductase, from this organism is closely related to nifH from Klebsiella pneumoniae. Similarly, the 16S rRNA gene sequences and carbon utilization tests grouped it closely with K. pneumoniae. Based an these data, the isolates from Z. mays and Z. luxurians are tentatively classified as Klebsiella spp. (Zea). The ability of this bacterium to contribute to the nitrogen economy of the corn plant is unknown.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Angert E R, Clements K D and Pace N R 1993 The largest bacterium. Nature 362, 239–241.
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.
Baldani V L D, Baldani J I, Olivares F and Döbereiner J 1992 Identification and ecology of Herbaspirillum seropedicae and the closely related Pseudomonas rubrisubalbicans. Symbiosis 13, 65–73.
Bertolini M, Lorenzoni C, Marocco A and Maggiore T 1993 Soluble solids content in the stakl of maize (Zea mays L.) lines and hybrids. Maydica 8, 321–324.
Boddey R M, Urquiaga S, Ries V and Dobereiner J 1991 Biological nitrogen fixation associated with sugar cane. Plant and Soil 137, 111–117.
Burris R H 1972 Nitrogen fixation-assay methods and techniques. Meth. Enzymol. 24B, 415–431.
Cavalcante V A and Döbereiner J 1988 A new acid-tolerant nitrogenfixing bacterium associated with sugarcane. Plant and Soil 108, 23–31.
Dixon R, Vanderleyden J and Romero D 1993 Extension of nitrogen fixation to other crops. In New New Horizons in Nitrogen Fixation. Eds. R Palacios, J Mora and W E Newton, pp 765–768. Kluwer Academic Publishers, Dordrecht, the Netherlands.
Döbereiner J, Reis V and Lararini A C 1988 New N2 fixing bacteria in association with cereals and sugarcane. In Nitrogen Fixation: Hundred Years After. Eds. H Bothe, F J de Bruijn and W E Newton. pp 717–722. Gustav Fischer, Stuttgart, Germany.
Döbereiner J, Reis V M, Paula M A and Olivares F 1993 Endophytic diazotrophs in sugar cane, cereals and tuber plants. In New Horizons in Nitrogen Fixation. Eds. R Palacios, J Mora and W E Newton. pp 671–676. Kluwer Academic Publishers, Dordrecht, the Netherlands.
Dong Z, Canny M J, McCully M E, Roboredo M R Cabadilla C F, Ortega E and Rodés R 1994 nitrogen-fixing endophyte of sugarcane stems. A new role for the apoplast. Plant Physiol. 105, 1139–1147.
Ela S W, Anderson M A and Brill W J 1982 Screening and selection of maize to enhance associative bacterial nitrogen fixation. Plant Physiol. 70, 1564–1567.
Fisher P J, Petrini O and Scott H M L 1992 The distribution of some fungal and bacterial endophytes in maize (Zea mays L.). New Phytol. 122, 299–305.
Fuentes-Ramirez L E, Jimenez-Salgado T and Abarca-Ocampo I R 1993 Acetobacter diazotrophicus, an indoleacetic acid producing bacterium isolated from sugarcane cultivars of Mexico. Plant and Soil 154, 145–150.
Klingler J M, Stowe R P, Obenhuber D C, Groves T O, Mishra S K and Pierson D L 1992 Evaluation of the Biolog automated microbial identification system. Appl. Environ. Microbiol. 58, 2089–2092.
Kumar S, Tamura K and Nei M 1993 MEGA: molecular evolutionary genetics analysis, version 1.0. Pennsylvania State University, University Park, USA.
Li R P and MacRae I C 1991 Specific association of diazotrophic acetobacters with sugarcane. Soil Biol. Biochem. 23, 999–1002.
Li R P and MacRae I C 1991 Specific identification and numeration of Acetobacter diazotrophicus in sugarcane. Soil Biol. Biochem. 24, 413–419.
McInroy J A and Kloepper J W 1991 Analysis of population densities and identification of endophytic bacteria of maize and cotton in the field. In Plant Growth-Promoting Rhizobacteria-Progress and Prospects. Eds. C Keel, B Koller and G Défago. Bull. SROP 14, 328–331.
McInroy J A and Kloepper J W 1995 Population dynamics of endophytic bacteria in field-grown sweet corn and cotton. Can. J. Microbiol. 41, 895–901.
Paula M A, Reis V M and Döbereiner J 1991 Interactions of Glomus clarum with Acetobacter diazotrophicus in infection of sweet potato (Ipomoea batatas), sugarcane (Saccharum spp.) and sweet sorghum (Sorghum vulgare). Biol. Fert. Soils 11, 111–115.
Peterson G L 1977 A simplication of the protein assay method of Lowry et al. which is more generally applicable. Anal. Biochem. 83, 346–356.
Pimentel J P, Olivares F, Pitard R M, Urquiaga S, Akiba F and Döbereiner J 1991 Dinitrogen fixation and infection of grass leaves by Pseudomonas rubrisubalbicans and Herbaspirillum seropedicae. Plant and Soil 137, 61–65.
Reis V M, de Paula M A, Urquiaga S and Dobereiner J 1990 Infection of sugar cane by Acetobacter diazotrophicus is aided by VA mycorrhyzae. In Nitrogen Fixation: Achievements and Objectives. Eds. P M Gresshoff, L E Roth, G Stacey and W E Newton. p 688. Chapman and Hall, New York, USA.
Sambrook J, Fritsch E F and Maniatis T 1989 Molecular Cloning: a laboratory manual. Cold Spring Harbor Laboratory. Cold Spring Harbor, New York, USA.
Ueda T, Suga Y, Yahiro N and Matsuguchi T 1995 Remarkable N2-fixing bacterial diversity detected in rice roots by molecular evolutionary analysis of nifH gene sequences. J. Bacteriol. 177, 1414–1417.
Urquiaga S, Cruz K H S and Buddey R M 1992 Contribution of nitrogen fixation to sugar cane. N-15 and nitrogen balance estimates. Soil Sci. Soc. Am. J. 56, 105–114.
Wittwer Carl T 1992 Buffers and reaction components for rapid cycling. Rapid Cyclist 1, 1–6.
Young J P W 1992 Phylogenetic classification of nitrogen-fixing organisms. In Biological Nitrogen Fixation. Eds. G Stacey, R H Burris and H J Evans. pp 43–86. Chapman and Hall, New York, USA.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Palus, J.A., Borneman, J., Ludden, P.W. et al. A diazotrophic bacterial endophyte isolated from stems of Zea mays L. and Zea luxurians Iltis and Doebley. Plant Soil 186, 135–142 (1996). https://doi.org/10.1007/BF00035067
Issue Date:
DOI: https://doi.org/10.1007/BF00035067