AiM  Vol.6 No.3 , March 2016
Transcriptional Analysis of Pseudomonas stutzeri A1501 Associated with Host Rice
Abstract: Pseudomonas stutzeriA1501, associative and endophytic nitrogen-fixing bacterium showed the capacity of colonization in the rice roots and considered as the good colonizer in the rice plant. The experiment was conducted to study the expression of genes potentiality relevant to the association of nitrogen fixing Pseudomonas stutzeri with host rice and reveal the molecular mechanism by which underlying interaction between bacteria and host rice. The bacteria were shown to be uniformly distributed on the rhizoplane of the root and the density of bacteria was found at the intercellular junction and micro colony developed on the surface of the epidermal cells and on the cellular junctions. Root exudates of rice were the major components of carbon and energy sources for bacteria. RT-PCR analyses of pilK, metE, rpoN and fdhE genes expression of P. stutzeri A1501 were performed at positive and negative (control) conditions. After 1 h, it was found that pilK, metE and rpoN transcription were increased 5.7, 6.4 and 3.4-fold, respectively, whereas in the fdhE gene has no expression. Consequently, after 4 h pilk, fdhE, metE and rpoN were decreased -1.9, -4.4, -0.2 and -0.8-fold, respectively. The gene pilK, expression was up-regulation after 1 h and down-regulation after 4 h that has twitching motility to convey the bacterial cell to point of attachment in to host plant. The gene expressions of the bacteria, pilK, metE, rpoN and fdhE were up- and down-regulated during the influence of root exudates which regulated the colonization of bacteria during plant-microbe interaction.
Cite this paper: Alam, K. , Zhang, T. , Yan, Y. , Zhang, W. , Lin, M. and Lu, W. (2016) Transcriptional Analysis of Pseudomonas stutzeri A1501 Associated with Host Rice. Advances in Microbiology, 6, 210-221. doi: 10.4236/aim.2016.63021.

[1]   Mark, G.L., Dow, J.M., Kiely, P.D., Higgins, H., Haynes, J., Baysse, C., Abbas, A., Foley, T., Franks, A., Morrissey, J. and O’Gara, F. (2005) Transcriptome Profiling of Bacterial Responses to Root Exudates Identifies Genes Involved in Microbe-Plant Interactions. Proceedings of the National. Academy of Sciences of the United States of America, 102, 17454-17459.

[2]   Macario, B.J., Sara, A.F., Elsa, V.Z., Eduardo, P.C., Stephane, B. and Edgar, Z. (2003) Chemical Characterization of Root Exudates from Rice (Oryza sativa) and Their Effects on Chemotactic Response of Endophytic Bacteria. Plant and Soil, 249, 271-277.

[3]   Hawes, M.C. and Pueppke, S.G. (1986) Isolated Peripheral Root Cap Cells: Yield from Different Plants, and Callus Formation from Single Cells. American Journal of Botany, 73, 1466-1473.

[4]   Lynch, J.M. and Whipps, J.M. (1990) Substrate Flow in the Rhizosphere. Plant and Soil, 129, 1-10.

[5]   Song, S.C. and Lin, L.P. (1999) The Transition of Rhizobium freedii Lipopolysaccarides Induced by Soybean Root Exudation. Botanical Bulletin of Academia Sinica, 40, 73-78.

[6]   Morrissey, J.P., Dow, J.M., Mark, G.L. and O’Gara, F. (2004) Are Microbes at the Root of a Solution to World Food Production? Rational Exploitation of Interactions between Microbes and Plants Can Help to Transform Agriculture. EMBO Reports, 5, 922-926.

[7]   Reinhold-Hurek, B. and Hurek, T. (2011) Living inside Plants: Bacterial Endophytes. Current Opinion in Plant Biology, 14, 435-443.

[8]   Krause, A., Ramakumar, A., Bartels, D., et al. (2006) Complete Genome of the Mutualistic, N2-Fixing Grass Endophyte Azoarcus sp. Strain BH72. Nature Biotechnology, 24, 1385-1391.

[9]   Fouts, D.E., Tyler, H.L., DeBoy, R.T., Daugherty, S., Ren, Q., et al. (2008) Complete Genome Sequence of the N2-Fixing Broad Host Range Endophyte Klebsiella pneumoniae 342 and Virulence Predictions Verified in Mice. PLoS Genet, 4, e1000141.

[10]   Yan, Y., Yang, J., Dou, Y., et al. (2008) Nitrogen Fixation Island and Rhizosphere Competence Traits in the Genome of Root-Associated Pseudomonas stutzeri A1501.Proceedings of the National Academy of Sciences of the USA, 21, 7564-7569.

[11]   Bertalan, M., Albano, R., de Pa′dua, V., et al. (2009) Complete Genome Sequence of the Sugarcane Nitrogen-Fixing Endophyte Gluconacetobacter diazotrophicus PAl5. BMC Genomics, 10, 450.

[12]   Kaneko, T., Minamisawa, K., Isawa, T., et al. (2010) Complete Genomic Structure of the Cultivated Rice Endophyte Azospirillum sp. B510. DNA Research, 17, 37-50.

[13]   Pedrosa, F.O., Monteiro, R.A., Wassem, R., et al. (2011) Genome of Herbaspirillum seropedicae Strain SmR1, a Specialized Diazotrophic Endophyte of Tropical Grasses. PLoS Genetics, 7, e1002064.

[14]   Cartieaux, F., Thibaud, M.C., Zimmerli, L., et al. (2003) Transcriptome Analysis of Arabidopsis Colonized by a Plant-Growth Promoting Rhizobacterium Reveals a General Effect on Disease Resistance. The Plant Journal, 36, 177-188.

[15]   Verhagen, B.W.M., Glazebrook, J., Zhu, T., Chang, H.S., van Loon, L.C. and Pieterse, C.M.J. (2004) The Transcriptome of Rhizobacteria-Induced Systemic Resistance in Arabidopsis. Molecular Plant-Microbe Interactions, 17, 895-908.

[16]   Cheng, Z., McConkey, B.J. and Glick, B.R. (2010) Proteomic Studies of Plant Bacterial Interactions. Soil Biology and Biochemistry, 42, 1673-1684.

[17]   Walker, V., Bertrand, C., Bellvert, F., Moenne-Loccoz, Y. and Bally, R. (2011) Host Plant Secondary Metabolite Profiling Shows a Complex, Strain-Dependent Response of Maize to Plant Growth-Promoting Rhizobacteria of the Genus Azospirillum. New Phytologist, 189, 494-506.

[18]   Hoagland, D.R. (1975) Mineral Nutrition. In: De Kaufman, P.B., Labavitch, J., Anderson-Prouty, A. and Ghosheh, N.S., Eds., Laboratory Experiments and Plant Physiology, Macmillan Publishing Co. Inc., New York, 129-134.

[19]   Rediers, H., Bonnecarrère, V., Rainey, P.B., Hamonts, K., Vanderleyden, J. and De Mot, R. (2003) Development and Application of a dapB-Based in Vivo Expression Technology System to Study Colonization of Rice by the Endophytic Nitrogen-Fixing Bacterium Pseudomonas stutzeri A15. Applied and Environmental Microbiology, 69, 6864-6874.

[20]   Keitaro, T., Ryota, H., Takuro, S., Tadao, W., Kazuki, S. and Akira, O. (2009) Metabolite Profiling of Rice Root Exudate under Phosphorus Defficiency. Proceedings of the International Plant Nutrition Colloquium XVI, Sacramento, 26-30 August 2009.

[21]   Bao, T., Sun, T. and Sun, L. (2011) Low Molecular Weight Organic Acids in Root Exudates and Cadmium Accumulation in Cadmium Hyper Accumulator Solanum nigrum L. and Non-Hyper Accumulator Solanum lycopersicum L. African Journal of Biotechnology, 10, 17180-17185.

[22]   Yan, Y., Ping, S., Peng, J., Han, Y., Li, L., Yang, J., et al. (2010) Global Transcriptional Analysis of Nitrogen Fixation and Ammonium Repression in Root-Associated Pseudomonas stutzeri A1501. BMC Genomics, 11, 11.

[23]   Bowen, G.D. (1979) Integrated and Experimental Approaches to the Study of Growth of Organisms around Roots. In: Schippers, B. and Gams, W., Eds., Soil-Borne Plant Pathogens, Academic Press, London, 207-227.

[24]   Bennet, R.A. and Lynch, J.M. (1981) Bacterial Growth and Development in the Rhizosphere of Gnotobiotic Cereal Plants. Journal of General Microbiology, 125, 95-102.

[25]   Bacilio-Jiménez, M., Aguilar-Flores, S., Del Valle, M.V., Pérez, A., Zepeda, A. and Zenteno, E. (2001) Endophytic Bacteria in Rice Seeds Inhibit Early Colonization of Roots by Azospirillum brasilense. Soil Biology and Biochemistry, 33, 167-172.

[26]   Ramey, B.E., Koutsoudis, M., Von Bodman, S.B. and Fuqua, C. (2004) Biofilm Formation in Plant Microbe Associations. Current Opinion in Microbiology, 7, 602-609.

[27]   Roncato-Maccari, L.D., Ramos, H.J., Pedrosa, F.O., Alquini, Y., Chubatsu, L.S., Yates, M.G., Rigo, L.U., Steffens, M.B. and Souza, E.M. (2003) Endophytic Herbaspirillum seropedicae Expresses nif Genes in Graminous Plants. FEMS Microbiology Ecology, 45, 39-47.

[28]   Gough, C. and Cullimore, J. (2011) Lipo-Chitooligosaccharide Signaling in Endosymbiotic Plant-Microbe Interactions. Molecular Plant-Microbe Interactions, 24, 867-878.

[29]   Jofre, E., Lagares, A. and Mori, G. (2004) Disruption of dTDP-Rhamnose Biosynthesis Modifies Lipopolysaccharide Core, Exopolysaccharide Production and Root Colonization in Azospirillum brasilense. FEMS Microbiology Letters, 231, 267-275.

[30]   Balsanelli, E., Serrato, R.V., de Baura, V.A., Sassaki, G., Yates, M.G., Rigo, L.U., et al. (2010) Herbaspirillum seropedicae rfbB and rfbC Genes Are Required for Maize Colonization. Environmental Microbiology, 12, 2233-2244.

[31]   Burdman, S., Dulguerova, G., Okon, Y. and Jurkevitch, E. (2001) Purification of the Major Outer Membrane Protein of Azospirillum brasilense, Its Affinity to Plant Roots, and Its Involvement in Cell Aggregation. Molecular Plant-Microbe Interactions, 14, 555-561.

[32]   Gantar, M.P., Rowell, N.W., Kerby, N. and Sutherland, W. (1995) Role of Extracellular Polysaccharide in the Colonization of Wheat Triticum vulgare L. Roots by N2-Fixing Cyanobacteria. Biology and Fertility of Soils, 19, 41-48.

[33]   Agarwhal, S. and Shende, T.S. (1987) Tetrazolium Reducing Microorganisms Inside the Root of Brassica Species. Current Science, 56, 187-188.

[34]   Reinhold, B., Hurek, T., Niemann, E.G. and Fendrik, I. (1986) Close Association of Azospirillum and Diazotrophic Rods with Different Root Zones of Kallar Grass. Applied and Environmental Microbiology, 52, 520-526.

[35]   Alexandre, G. and Zhulin, I.B. (2007) Chemotaxis in Soil Diazotrophs: Survival and Adaptative Response. In: Elmerich, C. and Newton, W.E., Eds., Associative and Endophytic Nitrogen-Fixing Bacteria and Cyanobacterial Associations, Kluwer Academic Publishers, Dordrecht, 73-84.

[36]   Vanbleu, E. and Vanderleyden, J. (2007) Molecular Genetics of Rhizosphere and Plant-Root Colonization. In: Elmerich, C. and Newton, W.E., Eds., Associative and Endophytic Nitrogen-Fixing Bacteria and Cyanobacterial Associations, Kluwer Academic Publishers, Dordrecht, 85-112.

[37]   Compant, S., Clément, C. and Sessitsch, A. (2010) Plant Growth-Promoting Bacteria in the Rhizo- and Endosphere of Plants: Their Role, Colonization, Mechanisms Involved and Prospects for Utilization. Soil Biology and Biochemistry, 42, 669-678.

[38]   Lugtenberg, B.J., Dekkers, L. and Bloemberg, G.V. (2001) Molecular Determinants of Rhizosphere Colonization by Pseudomonas. Annual Review of Phytopathology, 39, 461-490.

[39]   Sarkar, A. and Reinhold, H.B. (2014) Transcriptional Profiling of Nitrogen Fixation and the Role of NifA in the Diazotrophic Endophyte Azoarcus sp. Strain BH72. PLoS ONE, 9, e86527.

[40]   Darzins, A. and Russell, M.A. (1997) Molecular Genetic Analysis of Type-4 Pilus Biogenesis and Twitching Motility Using Pseudomonas aeruginosa as a Model System—A Review. Gene, 192, 109-115.

[41]   Dorr, J., Hurek, T. and Reinhold, H.B. (1998) Type IV Pili Are Involved in Plant-Microbe and Fungus-Microbe Interactions. Molecular Microbiology, 30, 7-17.

[42]   Bohm, M., Hurek, T. and Reinhold-Hurek, B. (2007) Twitching Motility Is Essential for Endophytic Rice Colonization by the N2-Fixing Endophyte Azoarcus sp. Strain BH72. Molecular Plant-Microbe Interactions, 20, 526-533.