Influence of ammonium and moisture on survival and nifH transcription in the diazotrophic Pseudomonas mendocina S10

  • Sushma SHARMA University of Delhi, Department of Zoology
  • Dileep K. SINGH University of Delhi, Department of Zoology
Keywords: ERIC-PCR; nitrogenase; nifH; Pseudomonas mendocina S10; real time PCR

Abstract

Nitrogen is important for crop productivity and usually added in form of urea into the soil which negatively affects the environment. It is important to utilize nitrogen fixing bacteria for improving the nitrogen content of soil in India. Here, we have isolated nitrogen fixing-bacteria Pseudomonas mendocina S10 from rhizospheric soil and studied its nitrogenase activity along with its survival under sterile soil conditions. Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR and Real-time quantitative PCR were employed to investigate the population and nifH transcripts level respectively in presence of ammonium and moisture additives. Strain S10 was capable of growth and expressing nifH transcripts in the presence of 2.5 mM ammonium and 20 percent water availability. Population of isolated strain and it’s nifH mRNA was found at low levels when exposed to 5 mM ammonium for 60 days of incubation period. However, viable bacterial count and nifH transcript levels remained low in the presence of 1.25 mM ammonium and zero percent water content. These findings indicate that isolated strain could tolerate ammonium up to 5 mM for 60 days and can maintained their cell viability in low moisture conditions. Results revealed the advantage of using gene expression to evaluate the physiological state of microorganism’s population in soil.

Metrics

Metrics Loading ...

References

Aaronson S (1970). Experimental microbial ecology. Academic Press, New York pp 236.

Alexander M (1977). An introduction to soil microbiology. John Wiley Sons, New York pp 467.

Burgmann H, Widmer F, Sigler WV, Zeyer J (2003). mRNA extraction and reverse transcription-pcr protocol for detection of nifH gene expression by Azotobacter vinelandii in soil. Applied and Environmental Microbiology 69(4):1928-1935.

Cattaneo MV, Masson C, Greer CW (1997). The influence of moisture on microbial transport, survival and 2, 4-D biodegradation with a genetically marked Burkholderia cepacia in unsaturated soil columns. Biodegradation 8:87-96.

Cleveland CC, Townsend AR, Schimel DS (1999). Global patterns of terrestrial biological nitrogen (N2) fixation in natural ecosystems. Global Biogeochemical Cycles 13(2):623-645.

Cummings SP, Humphry DR, Santos SR, Andrews M, James EK (2006). The potential and pitfalls of exploiting nitrogen fixing bacteria in agricultural soils as a substitute for inorganic fertilizer. Environmental Biotechnology 2(1):1-10.

Fedorov AS, Troshina OY, Laurinavichene TV, Glazer VM, Babykin MM, Zinchenko VV, … Tsygankov AA (1998). Regulatory effect of ammonium on the nitrogenase activity of Rhodobacter sphaeroides and Rhodobacter capsulatus is not mediated by ADP-ribosylation of the Fe-protein of nitrogenase. Microbiology 67:610-615.

Fuhrmann J, Davey CB, Wollum AG (1986). Desiccation tolerance in clover rhizobia in sterile soils. Soil Science Society of America Journal 50:639-644.

Guerrero MG, Lara C (1987). Assimilation of inorganic nitrogen. In: The Cyanobacteria. Edited by Fay P and Van Baalen C (Ed). Elsevier Science, Amsterdam pp 163-185.

Hallenbeck PC, Meyer CM, Vignais PM (1982). Nitrogenase from the photosynthetic bacterium Rhodopseudomonas capsulata: purification and molecular properties. Journal of Bacteriology 149(2):708-717.

Hallenbeck PC (1992). Mutations affecting nitrogenase switch-off in Rhodobacter capsulatus. Biochimica et Biophysica Acta 1118:161-168.

Hsu S, Buckley DH (2009). Evidence for the functional significance of diazotroph community structure in soil. ISME Journal 3:124-136.

Hubner P, Masepohl B, Klipp W, Bickle TA (1993). nif gene expression studies in Rhodobacter capsulatus: ntrC-independent repression by high ammonium concentrations. Molecular Microbiology 10:123-132.

Indian Agricultural Government (2012). A report on Rajasthan.

Kloepper JW, Beauchamp CJ (1992). A review of issues related to measuring of plant roots by bacteria. Canadian Journal of Microbiology 38:1219-1232.

Lipman LJA, de Nijs A, Lam TJGM, Gastra W (1995). Identification of Escherichia coli strains from cows with clinical mastitis by serotyping and DNA polymorphism patterns with REP and ERIC primers. Veterinary Microbiology 43:13-19.

Livak KJ, Schmittgen TD (2001). Analysis of relative gene expression data using real time quantitative PCR and the 2−ΔΔCT method. Methods 25:402-408.

Ludden PW, Roberts GP (1995). The biochemistry and genetics of nitrogen fixation by photosynthetic bacteria. In: Blankenship RE (Eds). Anoxygenic photosynthetic bacteria. Dordrecht: Kluwer Academic Publishers pp 929-947.

Marshall J, Grobbelaar N, James S (1989). Seasonal changes in the nitrogenase activity and other metabolic parameters of cycad coralloid roots. Botanical Bulletin of Academia Sinica 30(4):285-289.

Ohmori M, Hattori A (1972). Effect of nitrate on nitrogen-fixation by the blue-green alga Anabaena cylindrica. Plant and Cell Physiology 13:589-599.

Okon Y, Labandera-Gonzalez CA (1994). Agronomic applications of Azospirillum: anevaluation of 20 years worldwide field inoculation. Soil Biology and Biochemistry 26:1591-1601.

Orchard VA, Cook FG (1983). Relation between soil respiration and soil moisture. Soil Biology and Biochemistry 15:447-453.

Orr CH, James A, Leifert C, Cooper JM, Cummings SP (2011). Diversity and activity of free-living nitrogen-fixing bacteria and total bacteria in organic and conventionally managed soils. Applied and Environmental Microbiology 77(3):911-919.

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. Research Microbiology 152:95-103.

Robertst GP, Brill WJ (1980). Gene-product relationships of the nif regulon of Klebsiella pneumoniae. Journal of Bacteriology 144(1):210-216.

Ronen Z, Vasiluk L, Abeliovich A, Nejidat A (2000). Activity and survival of tribromophenol-degrading bacteria in a contaminated desert soil. Soil Biology and Biochemistry 32:1643-1650.

Shoushtari NH, Pepper IL (1985). Mesquite rhizobia isolated from the Sonoran Desert: competitiveness and survival in soil. Soil Biology and Biochemistry 17:803-806.

VersavolicJ, Koeuth T, Lupski JR (1991). Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Research 19:6823-6831.

Victoria M, Alejandro N, Ricardo G (2017). Use of endophytic and rhizosphere bacteria to improve phytoremediation of arsenic-contaminated industrial soils by autochthonous Betula celtiberica. Applied Environmental Microbiology 15:83(8): e03411-16.

Welbaum G, Sturz AV, Dong Z, Nowak J (2004). Fertilizing soil microorganisms to improve productivity of agroecosystems. Critical Reviews in Plant Science 23:175-193.

Yakunin AF, Hallenbeck PC (1998). Short-term regulation of nitrogenase activity by NH4 in Rhodobacter capsulatus: multiple in vivo nitrogenase responses to NH4 addition. Journal of Bacteriology 180:6392-6395.

Published
2020-03-31
How to Cite
SHARMA, S., & SINGH, D. K. (2020). Influence of ammonium and moisture on survival and nifH transcription in the diazotrophic Pseudomonas mendocina S10. Notulae Scientia Biologicae, 12(1), 114-123. https://doi.org/10.15835/nsb12110545
Section
Research articles