The Effect of Biofertilizer on The Diversity of N2O Reducing Bacteria in Paddy Fields of Sukabumi, Indonesia

Alfan Cahyadi(1), Iman Rusmana(2), Nisa Rachmania Mubarik(3),


(1) Study Program of Microbiology, Graduate School, Institut Pertanian Bogor
(2) Department of Biology, Faculty of Mathematic and Natural Sciences, Institut Pertanian Bogor
(3) Department of Biology, Faculty of Mathematic and Natural Sciences, Institut Pertanian Bogor

Abstract

Some of the methanotrophic bacteria and N2O reducing bacteria have been proven to be able to support the plant growth and increase the productivity of paddy. However effect of the methanotrophic and N2O reducing bacteria application as a biofertilizer to indigenous N2O reducing bacteria is still not well known yet. The aim of this study was to analyze the diversity of N2O reducing bacteria in lowland paddy soil based on a nosZ gene. Soil samples were taken from lowland paddy soils in Pelabuhan Ratu Sukabumi, West Java, Indonesia. There were two treatments for the paddy field soil, ie. biofertilizer-treated field 20% fertilizer (50 kg/ha) with the addition of biofertilizer and 100% fertilizer. PCR amplification of nosZ gene was successfully conducted using nosZF and nosZR primer pair. Denaturing Gradient Gel Electrophoresis (DGGE) process was conducted at 150 V for 5.5h. There were three differences nosZ bands were sequenced. The phylogenetic analysis showed that they were close to uncultured bacteria. Microbial diversity in the biofertilizer-treated field was higher than that of in the 100% fertilizer-treated field. The biofertilizer treatment has higher in microbial diversity than that of applied non-biofertilizer paddy fields. This research might have impact in the application of biofertilizers due to the emission of N2O as a green house gas from paddy fields farming activity. The biofertilizer has great potential application in sustainable environmental friendly agriculture systems.

Keywords

Denitrification; DGGE; Metagenomic; nosZ; N2O emission; Paddy fields

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References

Astuti, D. D. (2009). Physiological characterization and molecular identification of methanotrophic isolates from Bogor and Sukabumi (in Indonesia). Undergraduate Thesis, Bogor Agricultural University. pp. 1-4.

Aulakh, M. S., Wassmann, R., Bueno, C., Kreuzwieser, J. & Rennenberg, H. (2001). Characterization of root exudates at different growth stages of ten rice (Oryza sativa L.) cultivars. Plant Biology, 3(2), 139-148.

Bai, Q., Gattinger, A. & Zelles, L. (2000). Characterization of microbial consortia in paddy rice soil by phospholipid analysis. Microbial Ecology, 39(4), 273-281.

Bintarti, A. F., Rusmana, I. & Wahyudi, A. T. (2014). Methanotrophic bacteria from the ricefield. Annales Bogoriense, 18(2), 13-25.

Canfield, D. E., Glazer, A. N, & Falkowski, P. G. (2010). The evolution and future of Earth’s nitrogen cycle. Science, 330(6001), 192-196.

Chen, Z., Hou, H., Zheng, Y., Qin, H., Zhu, Y., Wu, J. & Wei, W. (2012). Influence of fertilisation regimes on a nosZ-containing denitrifying community in rice paddy soil. Journal of the Science of Food and Agriculture, 92(5), 1064-1072.

Dobermann, A. & White, P. F. (1999). Strategies for nutrient management in irrigated and rainfed lowland rice systems. Nutrient Cycling in Agroecosystems, 53, 1-18.

Fakruddin, & Mannan, K. S. (2013). Methods for analyzing diversity of microbial communities in natural environments. Ceylon Journal of Science, 42(1), 19-33.

Forster, P. M. F. (2007). Changes in Atmospheric Constituents and in Radiative Forcing. Cambridge (GB): Cambridge Univ Press. pp 129-234.

Gelsomino, A., Keijzer, W. A. C., Cacco, G. & Van Elsas, J. D. (1999). Assessment of bacterial community structure in soil by polymerase chain reaction and denaturing gradient gel electrophoresis. JJournal of Microbiological Methods, 38(1), 1-15.

Hadianta, R., Rusmana, I. & Mubarik, N. R. (2014). Diversity of Nitrogen fixing bacteria based on nifH gene in rice fields. Advances in Environmental Biology, 8(14), 63-69.

Hapsari, W. (2008). Isolation and characterization of methanotrophic bacteria from Bogor and Sukabumi rice field (in Indonesia). Undergraduate Thesis, Bogor Agricultural University. pp 3-4.

Henry S., Bru D., Stres B., Hallet S. & Philippot L. (2006). Quantitative detection of the nosZ gene, encoding nitrous oxide reductase, and comparison of the abundances of 16S rRNA, narG, nirK, and nosZ genes in soils. Applied and environmental microbiology, 72(8), 5181-5189.

Jung, J., Choi, S., Jung, H., Scow, K. M. & Park, W. (2013). Primers for amplification of nitrous oxide reductase genes associated with Firmicutes and Bacteroidetes in organic-compound-rich soils. Microbiology, 159(2), 307-315.

Knowles, R. (1982). Denitrification. Microbiological reviews, 46(1), 43-70.

MacKenzie, A. F., Fan, M. X. & Cadrin, F. (1998). Nitrous oxide emission in three years as affected by tillage, corn-soybean-alfalfa rotations, and nitrogen fertilization. Journal of Environmental Quality, 27(3), 698-703.

Montzka, S. A., Dlugokencky, E. J. & Butler, J. H. (2011). Non-CO2 greenhouse gases and climate change. Nature, 476(7358), 43-50.

Muyzer, G. & Smalla, K. (1998). Aplication of denaturing gradient gel elektrophoresis (DGGE) and temperature gradient gel electrophoresis (TGGE) in microbial ecology. Antonie van Leuwenhoek Journal of General and Molecular Microbiology, 73(1), 127-141.

Orellana, L. H., Rodriguez-R, L. M., Higgins, S., Chee-Sanford, J. C., Sanford, R., Ritalahti, K., Löffler, F. E. & Konstantinidis, K. T. (2014). Detecting nitrous oxide reductase (nosZ) genes in soil metagenomes: method development and implications for the nitrogen cycle. mBio, 5(3), 1-8.

Palmer, T. & Berks, B. C. (2012). The twin-arginine translocation (Tat) protein export pathway. Nature Reviews Microbiology, 10(7), 483-496.

Pingak, G. M. F., Sutanto H., Akhdiya A. & Rusmana, I. (2013). Effectivity of methanotrophic bacteria and Ochrobactrum anthropi as biofertilizer and emission reducer of CH4 and N2O in inorganic paddy fields. Journal of Medical and Bioengineering, 3(3), 217-221.

PERMENTAN: Peraturan Menteri Pertanian. (2007). Reference adoption of recommendations of NPK fertilizer in paddy fields (in Indonesia). Jakarta. Permentan No: 140(04), 2007.

Perez, P. G., Ye, J., Wang, S., Wang, X. & Huang, D. (2014). Analysis of the occurrence and activity of diazotrophic communities in organic and conventional horticultural soils. Applied soil ecology, 79, 37-48.

Setyaningsih, R., Rusmana, I., Setyanto, P. & Suwanto, A. (2010). Physiological characterization and molecular identification of denitrifying bacteria possesing nitrous oxide high reduction activity isolated from rice soils. Journal of Microbiology Indonesia, 4(2), 75-78.

Smith, K. A., Crutzen, P. J., Mosier, A. R. & Winiwarter W. (2010). Nitrous oxide and climate change. London (GB): Earthscan. pp.4-35.

Sukmawati, Rusmana, I. & Mubarik, N. R. (2016). The effectiveness of methanotrophic bacteria and Ochrobactrum anthropi to reduce CH4 and N2O emissions and to promote paddy growth in lowland paddy fields. Malaysian Journal of Microbiology, 12(1), 50-55.

Sutanto, H., Rusmana, I. & Mubarik, N. R. (2014). Community succesion of methanothrophic bacteria based on pmoA gene in ricefields. Advances in Environmental Biology, 8(14), 50-56.

Sylvia, D. M. (2005). Principles and Applications of Soil Microbiology. Upper Saddle River, New Jersey (US): Pearson Prentice Hall.

Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. (2013). Mega 6: Molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution, 30(12), 2725-2729.

Throback, I., Enwall, N., Jarvis, K. A. & Hallin, S. (2004). Reassessing PCR primers targeting nirS, nirK and nosZ genes for community surveys of denitrifying bacteria with DGGE. FEMS Microbiology Ecology, 49(3), 401-417.

Zumft, W. G. & Kroneck, P. M. (2007). Respiratory transformation of nitrous oxide (N2O) to dinitrogen by bacteria and archaea. Advances in Microbial Physiology, 52, 107-227.

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