Abstract
Himalayas are considered as a reservoir of diversified and dynamic gene pool. This study describes the response of a Himalayan psychrophilic diazotroph to low temperature diazotrophy. Seven cold adaptive N2 fixing bacteria were isolated and identified as Bacillus sp., Arthrobacter sp., Rhodococcus sp., Pseudomonas sp., etc. In order to examine the physiological response to low temperature diazotrophy, differential proteomic analysis of Pseudomonas migulae S10724 strain was carried out using two dimensional electrophoresis and MALDI–TOF–MS. Functional assessment of 66 differentially expressed proteins revealed several mechanisms thought to be involved in low temperature adaptation and nitrogen fixation, including general stress adaptation, protein and nucleic acid synthesis, energy metabolism, cell growth/maintenance, etc. Major fraction of the upregulated proteins was stress proteins, while majority of the downregulated proteins were related to cell division. Furthermore, MALDI–TOF–MS-based identification of randomly selected peptides encountered two exclusively expressed proteins: NifU family SUF system FeS assembly protein and membrane protein, suppressor for copper-sensitivity B precursor which might have a crucial role at low temperature nitrogen fixation. To the best of our knowledge, this is the first report of the isolation and differential proteomic analysis of psychrophilic diazotroph from Himalayan high altitude rhizospheric soil.
Similar content being viewed by others
References
Antony CP, Kumaresan D, Ferrando L, Boden R, Moussard H, Scavino AF, Shouche YS, Murrell JC (2010) Active methylotrophs in the sediments of Lonar Lake, a saline and alkaline ecosystem formed by meteor impact. ISME J 4:1470–1480
Dos Santos PC, Smith AD, Frazzon J, Cash VL, Johnson MK, Dean DR (2004) Iron-sulfur cluster assembly: NifU-directed activation of the nitrogenase Fe protein. J Biol Chem 279:19705–19711
Ekman M, Tollback P, Bergman B (2008) Proteomic analysis of the cyanobacterium of the Azolla symbiosis: identity, adaptation, and NifH modification. J Exp Bot 59(5):1023–1034
Franche C, Lindström K, Elmerich C (2009) Nitrogen-fixing bacteria associated with leguminous and non-leguminous plants. Plant Soil 321:35–59
Jacobson MR, Cash VL, Weiss MC, Laird NF, Newton WE, Dean DR (1989) Biochemical and genetic analysis of the nifUSVWZM cluster from Azotobacter vinelandii. Mol Gen Genet 219:49–57
Jain S, Rani A, Marla SS, Goel R (2010) Differential proteomic analysis of psychrotolerant Pseudomonas putida 710A and alkaliphilic Pseudomonas monteilli 97AN for cadmium stress. Int J Biol Med Res 1(4):234–241
Mierzejewska J, Jagura-Burdzy G (2012) Prokaryotic ParA–ParB–parS system links bacterial chromosome segregation with the cell cycle. Plasmid 67:1–14
Morita RY (1975) Psychrophilic bacteria. Bacteriol Rev 39(2):144
Nikodinovic-Runic J, Flanagan M, Hume AR, Cagney G, O’Conor KE (2009) Analysis of the Pseudomonas putida CA-3 proteome during growth on styrene under nitrogen-limiting and non limiting conditions. Microbiology 155:3348–3361
Poly F, Monrozier LJ, Bally R (2001) Improvement in the RFLP procedure for studying the diversity of nifH genes in communities of nitrogen fixers in soil. Res Microbiol 152:95–103
Prema Latha K, Soni R, Khan M, Marla SS, Goel R (2009) Exploration of Csp genes from temperate and glacier soils of the Indian Himalayas and in silico analysis of encoding proteins. Curr Microbiol 58:343–348
Rennie RJ (1981) A single medium for the isolation of acetylene reducing (dinitrogen-fixing) bacteria from soils. Can J Microbiol 27:8–14
Shivaji S, Pratibha MS, Sailaja B, Kishore KH, Singh AK, Begum Z, Anarasi U, Prabagaran SR, Reddy GSN, Srinivas TNR (2011) Bacterial diversity of soil in the vicinity of Pindari glacier, Himalayan mountain ranges, India, using culturable bacteria and soil 16S rRNA gene clones. Extremophiles 15:1–22
Singh C, Soni R, Jain S, Roy S, Goel R (2010) Diversification of nitrogen fixing bacterial community using nifH gene as a biomarker in different geographical soils of Western Indian Himalayas. J Environ Biol 31:553–556
Soni R, Goel R (2011) nifH homologous from soil metagenome. Ekologija 57(3):87–95
Soni R, Saluja B, Goel R (2010) Bacterial community analysis using temporal temperature gradient gel electrophoresis (TTGE) of 16S rDNA PCR products of soil metagenome. Ekologija 56(3–4):94–98
Wang W, Vinocur B, Shoseyov O, Altman A (2004) Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response. Trends Plant Sci 9:244–252
Xu XM, Moller SG (2011) Iron-sulfur clusters: biogenesis, molecular mechanisms, and their functional significance. Antioxid Redox Signal 15(1):271–307
Zakeri F, Sadeghizadeh M, Kardan MR, Zahiri HS, Ahmadian G, Masoumi F, Sharafi H, Rigi G, Vali H, Noghabi KA (2012) Differential proteome analysis of a selected bacterial strain isolated from a high background radiation area in response to radium stress. J Proteomics 75(15):4820–4832
Acknowledgments
This work is funded by the National Bureau of Agriculturally Important Microorganisms (NBAIM)/Indian Council of Agricultural Research (ICAR), Grant to RG. First author (DCS) acknowledge ICAR, Research Fellowship during the course of this study.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Suyal, D.C., Yadav, A., Shouche, Y. et al. Differential Proteomics in Response to Low Temperature Diazotrophy of Himalayan Psychrophilic Nitrogen Fixing Pseudomonas migulae S10724 Strain. Curr Microbiol 68, 543–550 (2014). https://doi.org/10.1007/s00284-013-0508-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00284-013-0508-1