Some Characteristics of a Plant Growth Promoting Enterobacter sp. Isolated from the Roots of Maize

Frank Ogbo, Julius Okonkwo

Department of Applied Microbiology & Brewing, Nnamdi Azikiwe University, Awka, Nigeria.

DOI: 10.4236/aim.2012.23046                

Abstract

Some properties of an Enterobacter sp. isolated from the roots of maize are described. Isolation was carried out using the semisolid enrichment culture technique and subsequent plating, both on nitrogen free biotin medium. Morphological, biochemical and phylogenetic characterization using the MicroSeq^TM 16S rDNA technique were employed in identification of isolate, which was revealed to be closest matched at 99.4% with Enterobacter asburiae. The isolate possessed properties of plant growth promoting bacteria. Thus, it produced indole-3-acetic, plant polymer hydrolyzing enzymes, pectinase and cellulase as well as ammonia in vitro. The isolate grew well in the presence of both 3% NaCl and 10 μg of streptomycin. In plate bioassays, isolate promoted the germination of both maize and rice seeds as well as root and lateral root growth resulting in weight increases of seedlings over their controls. Experiments to quantify ability of isolate to promote plant growth was performed using hydroponics solutions and as appropriate, an inoculum of the isolate. Pot experiments were also employed. Results from these studies showed that isolate enhanced nitrogen accumulation and significantly (p < 0.05), improved the growth of maze seedlings over controls. Isolate has potential for utilizetion as inocula for sustainable production of cereals.

Keywords

Cereals; Plant Growth Promoting Enterobacter

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	B. Lugtenberg and F. Kamilova, “Plant-Growth-Promoting Rhizobacteria,” Annual Review of Microbiology, Vol. 63, 2009, pp. 541-556. doi:10.1146/annurev.micro.62.081307.162918
[2]	Food and Agriculture Organization, “Towards 2015/2030,” World Agriculture: Summary Report, 2002. ftp://ftp.fao.org/docrep/fao/004/y3557e/y3557e.pdf
[3]	M. Charpentier and G. Oldroyd, “How Close Are We to Nitrogen-Fixing Cereals?” Current Opinion in Plant Biology, Vol. 13, No. 5, 2010, pp. 556-564. doi:10.1016/j.pbi.2010.08.003
[4]	Y. Bashan, G. Holguin and R. Lifshitz, “Isolation and Characterization of Plant Growth-Promoting Rhizobacteria,” In: B. R. Glick and J. E. Thompson, Eds., Methods in Plant Molecular Biology and Biotechnology, CRC Press, Boca Raton, 1993, pp. 331-350.
[5]	C. E. A. Rodríguez, “Improved Medium for Isolation of Azospirillum sp.,” Applied and Environmental Microbiology, Vol. 44, No. 4, 1982, pp. 990-991.
[6]	D. Egamberdiyeva, “The Effect of Plant Growth Promoting Bacteria on Growth and Nutrient Uptake of Maize in Two Different Soils,” Applied Soil Ecology, Vol. 36, No. 2-3, 2007, pp. 184-189.
[7]	R. M. Teather and P. J. Wood, “Use of Congo red-Polysaccharide Interactions in Enumeration and Characterization of Cellulolytic bacteria from the Bovine Rumen,” Applied and Environmental Microbiology, Vol. 43, No. 4, 1982, pp. 777-780.
[8]	T. M. Fernandes-Salom?o, A. C. R. Amorim, V. M. Cha- ves-Alves, J. L. C. Coelho, D. O. Silva and E. F. Araújo, “Isolation of Pectinase Hyper Producing Mutants of Penicillium expansum,” Revista de Microbiologia, Vol. 27, No. 1, 1996, pp. 15-18.
[9]	J. E. Loper and M. N. Schroth, “Influence of Bacterial Sources on Indole-3 Acetic Acid on Root Elongation of Sugar Beet,” Phytopathology, Vol. 76, No. 4, 1986, pp. 386-389. doi:10.1094/Phyto-76-386
[10]	J. C. Cappucino and N. Sherman, “Microbiology: A Laboratory Manual,” 3rd Edition, Benjamin/Cumming Pub. Co., New York, 1992.
[11]	H. Lock, “Production of Hydrocyanic Acid by Bacteria,” Physiologia Plantarum, Vol. 1, No. 2, 1948, pp. 142-146. doi:10.1111/j.1399-3054.1948.tb07118.x
[12]	D. W. Nelson and L. E. Sommers, “Total Nitrogen Analysis of Soil and Plant Tissues,” Journal of the Association of Official Analytical Chemists, Vol. 63, 1980, 1980, pp. 770-779.
[13]	F. Grimont and P. A. D. Grimont, “The Genus Enterobacter,” In: M. Dworkin, S. Falkow, E. Rosenberg, K. H. Schleifer and E. Stackebrandt, Eds., The Prokaryotes, Vol. 6, Proteobacteria: Gamma Subclass, Springer, Berlin, 2006, pp. 197-214. doi:10.1007/0-387-30746-X_9
[14]	H. Hoffmann, S. Stindl, W. Ludwig, A. Stumpf, A. Mehlen, J. Heesemann, D. Monget, K. H. Schleifer and A. Roggenkamp, “Reassignment of Enterobacter dissolvens to Enterobacter cloacae as E. cloacae Subspecies dissolvens comb. nov and Emended Description of Enterobacter asburiae and Enterobacter kobei,” Systematic and Applied Microbiology, Vol. 28, No. 3, 2005, pp. 196-205. doi:10.1016/j.syapm.2004.12.010
[15]	C. Bi, X. Zhang, L. O. Ingram and J. F. Preston, “Genetic Engineering of Enterobacter asburiae Strain JDR-1 for Efficient Production of Ethanol from Hemicellulose Hydrolysates,” Applied and Environmental Microbiology, Vol. 75, No. 18, 2009, pp. 5743-5749. doi:10.1128/AEM.01180-09
[16]	J. Czaban, A. Gajda and B. Wroblewska, “The Motility of Bacteria from Rhizosphere and Different Zones of Winter Wheat Roots,” Polish Journal of Environmental Studies, Vol. 16, No. 2, 2007, pp. 301-308.
[17]	A. Quadt-Hallmann and J. W. Kloepper, “Immunological Detection and Localization of the Cotton Endophyte Enterobacter asburiae JM22 in Different Plant Species,” Canadian Journal of Microbiology, Vol. 42, No. 11, 1996, pp. 1144-1154. doi:10.1139/m96-146
[18]	V. Van?ura and N. Hanzlikova, “Root Exudates of Plants IV. Differences in Chemical Composition of Seed and Seedlings Exudates,” Plant and Soil, Vol. 36, No. 1-3, 1972, pp. 271-282. doi:10.1007/BF01373482
[19]	J. A. McInroy and J. W. Kloepper, “Survey of Indigenous Bacterial Endophytes from Cotton and Sweet Corn,” Plant and Soil, Vol. 173, No. 2, 1995 pp. 337-342. doi:10.1007/BF00011472
[20]	C. A. Asis and K. Adachi, “Isolation of Endophytic Di- azotroph Pantoea agglomerans and Nondiazotroph Enterobacter asburiae from Sweet Potato Stem in Japan,” Letters in Applied Microbiology, Vol. 38, No. 19-23, 2003, pp. 19-23. doi:10.1046/j.1472-765X.2003.01434.x
[21]	M. Zakria, A. Ohsako, Y. Saeki, A. Yamamoto and S. Akao, “Colonization and Growth Promotion Characteristics of Enterobacter sp. and Herbaspirillum sp. on Brassica oleracea,” Soil Science and Plant Nutrition, Vol. 54, No. 4, 2008 pp. 507-516. doi:10.1111/j.1747-0765.2008.00265.x
[22]	Y. E. Morales-García, D. A. Juárez-Hernández, C. Aragón-Hernández, M. A. Mascarua-Esparza, M. R. Bustillos-Cristales, L. E. Fuentes-Ramírez, R. D. Martínez-Contreras and J. Mu?oz-Rojas, “Growth Response of Maize Plantlets Inoculated with Enterobacter sp., as a Model for Alternative Agriculture,” Revista Argentina de Microbiología, Vol. 43, No. 4, 2011 pp. 287-293.
[23]	K. I. Al-Mughrabi, “Biological Control of Fusarium Dry Rot and Other Potato Tuber Diseases Using Pseudomonas fluorescens and Enterobacter cloacae,” Biological Control, Vol. 53, No. 3, 2010 pp. 280-284. doi:10.1016/j.biocontrol.2010.01.010
[24]	D. J. Brenner, A. C. Mcwhorter, A. Kai, A. G. Steigerwalt and J. J. Farmer, “Enterobacter asburiae Sp-Nov, a New Species Found in Clinical Specimens, and Reassignment of Erwinia dissolvens and Erwinia nimipressuralis to the Genus Enterobacter as Enterobacter dissolvens Comb-Nov and Enterobacter nimipressuralis Comb-Nov.,” Journal of Clinical Microbiology, Vol. 23, No. 6, 1986, pp. 1114-1120.
[25]	M. Ahemad and M. S. Khan, “Plant Growth Promoting Activities of Phosphate Solubilizing Enterobacter asburiae as Influenced by Fungicides,” EurAsian Journal of BioSciences, Vol. 4, No. 11, 2010, pp. 88-95. doi:10.5053/ejobios.2010.4.0.11
[26]	A. Rogers, K. McDonald, M. F. Muehlbauer, A. Hoffman, K. Koenig, L. Newman, S. Taghavi and D. Lelie, “Inoculation of hybrid Poplar with the Endophytic Bacterium Enterobacter sp. 638 Increases Biomass but Does Not Impact Leaf Level Physiology,” GCB Bioenergy, Vol. 4, No. 3, 2012, pp. 364-370. doi:10.1111/j.1757-1707.2011.01119.x
[27]	Y. Zhao, “Auxin Biosynthesis and Its Role in Plant De- velopment,” Annual Review of Plant Biology, Vol. 61, pp. 49-64. doi:10.1146/annurev-arplant-042809-112308
[28]	J. Zhu, Z. Li, L. Wang and S. Shen, “Temperature Sensitivity of a nifA-Like Gene in Enterobacter cloacae,” Journal of Bacteriology, Vol. 166, No. 1, pp. 357-359.
[29]	J. Gu, G. Yu, J. Zhu and S. Shen, “The N-Terminal Domain of NifA Determines the Temperature Sensitivity of NifA in Klebsiella pneumoniae and Enterobacter cloacae,” Science in China Series C: Life Sciences, Vol. 43, No. 1, 2000, pp. 8-15. doi:10.1007/BF02881712