Population and Biochemical Characteristics of Halotolerant N2 Fixing Rhizobacteria Isolated from Saline Soil Ecosystem

Fiqriah Hanum Khumairah, Triana Ulfah, Mieke Rochimi Setiawati, Betty Natalie Fitriatin, Tualar Simarmata

Abstract


Utilization of halotolerant N2 fixing rhizobacteria (HNFR) in saline soil ecosystem has the potential to reduce the negative impact of soil salinity. The aim of this study was to obtain isolates of HNFR in saline soil ecosystem and identify their biochemical characteristics. The research was conducted at Sukajaya Village (Cilamaya Kulon District, Karawang Regency) and Biology Laboratory (Department of Soil and Land Resources, Faculty of Agriculture, Padjadjaran University) from September 2019 to November 2020. Soil samples were obtained from rhizosphere of rice, mangrove, and grass vegetations located around the shoreline affected by sea water intrusion. Isolation and characterization activities were done by using salinized Asbhy selective media. The population of HNFR in saline soil ecosystem was analyzed by using total plate count method. Biochemical characteristics were determined by analyzing the IAA hormone production using spectrophotometer, nitrogenase activity using gas chromatography, and DNA analysis using 16S rRNA gene sequence. The results showed that the number of HNFR obtained was 15 isolates with the highest population shown by Ab-P4, Ab-P5, and Ab-R5 isolates. Ab-P5 and Ab-R5 isolates which were obtained from rhizosphere of rice and grass, respectively, were able to produce IAA phytohormones reaching 0.89 and 0.61 μg/l and nitrogenase activity reaching 1.982 and 0.331 μM C2H4/ml/h, respectively. DNA analysis showed that the Ab-P5 and Ab-R5 isolates were Bacillus cereus and Delftia tsuruhatensis, respectively, and these isolates potentially can be used as active ingredients for biofertilizers that could be applied in saline soil ecosystem.


Keywords


Halotolerant N2 fixing rhizobacteria; saline; biochemicals; biofertilizers

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References


Arfarita, N., Muhibuddin, A. & Imai, T. (2019) Exploration of indigenous free nitrogen-fixing bacteria from rhizosphere of Vigna radiata for agricultural land treatment. Journal of Degraded and Mining Lands Management. [Online] 6 (2), 1617–1623. Tersedia pada: https://doi.org/10.15243/jdmlm.2019.062.1617 [Diakses 7 Januari 2021].

Bric, J.M., Bostock, R.M. & Silverstonet, S.E. (1991) Rapid in situ assay for indoleacetic acid production by bacteria immobilized on a nitrocellulose membrane. Applied and Environmental Microbiology, 57 (2), 535–538.

Chaiharn, M. & Lumyong, S. (2011) Screening and optimization of indole-3-acetic acid production and phosphate solubilization from rhizobacteria aimed at improving plant growth. Current Microbiology. [Online] 62 (1), 173–181. Tersedia pada: https://doi.org/ 10.1007/s00284-010-9674-6 [Diakses 5 Maret 2021].

Cole, J.R., Wang, Q., Cardenas, E., Fish, J., Chai, B., Farris, R.J., Kulam-Syed-Mohideen, A.S., McGarrell, D.M., Marsh, T., Garrity, G.M. & Tiedje, J.M. (2009) The ribosomal database project: improved alignments and new tools for rRNA analysis. Nucleic Acids Research. [Online] 37, Issue suppl_1, D141–D145. Tersedia pada: https://doi.org/10.1093/nar/gkn879 [Diakses 17 Februari 2021].

Danapriatna, N., Hindersah, R. & Sastro, Y. (2010) Pengembangan pupuk hayati Azotobacter dan Azospirillum untuk meningkatkan produktivitas dan efisiensi penggunaan pupuk N di atas 15% pada tanaman padi. [Online] Badan Penelitian dan Pengembangan Pertanian, Kementerian Pertanian. Tersedia pada: http://old.litbang.pertanian.go.id/ks/one/ 644/file/315-316-PENGEMBANGAN-PUPUK.pdf [Di-akses 21 Januari 2021].

Dewi, T.K., Arum, E.S., Imamuddin, H. & Antonius, S. (2015) Karakterisasi mikroba perakaran (PGPR) agen penting pendukung pupuk organik hayati. Prosiding Seminar Nasional Masyarakat Biodiversitas Indonesia. [Online] 289–295. Tersedia pada: https://doi.org/ 10.13057/psnmbi/m010220 [Diakses 30 Maret 2021].

Dewi, A.K., Meylina, L. & Rusli, R. (2017) Isolasi bakteri dari tanah mangrove Rhizopora sp. di kota Bontang. Proceeding of Mulawarman Pharmaceuticals Conferences, 5, 59–68.

El-Swaify, S.A. (2000) Soil and water salinity. Dalam: Silva, J.A. & Uchida, R.S. (editor) Plant nutrient management in Hawaii’s soils-approaches for tropical and subtropical agriculture. Honolulu, University of Hawai'i Press, hlm. 151–158.

Erfandi, D. & Rachman, A. (2011) Identification of soil salinity due to seawater intrusion on rice field in the northern coast of Indramayu, West Java. Jurnal Tanah Tropika. [Online] 16 (2), 115–121. Tersedia pada: https://doi.org/10.5400/jts.2011.16.2.115 [Diakses 1 April 2021].

Gomez, K.A. & Gomez, A.A. (1984) Statistical procedures for agricultural research. 2nd edition. Toronto, John Wiley & Sons. Han, J., Sun, L., Dong, X., Cai, Z., Sun, X., Yang, H., Wang, Y. & Song, W. (2005) Characterization of a novel plant growth-promoting bacteria strain Delftia tsuruhatensis HR4 both as a diazotroph and a potential biocontrol agent against various plant pathogens. Systematic and Applied Microbiology. [Online] 28 (1), 66–76. Tersedia pada: https://doi.org/10.1016/j.syapm.2004.09.003 [Diakses 25 Februari 2021].

Hardy, R.W.F., Holsten, R.D., Jackson, E.K. & Burns, R.C. (1968) The acetylene etilene assay for N fixation: laboratory and field evaluation. Plant Physiology. [Online] 43 (8), 1185–1207. Tersedia pada: https:// academic.oup.com/plphys/article/43/8/1185/6093758 [Diakses 13 Februari 2021].

Harris, R.F. & Sommers, L.E. (1968) Plate-dilution frequency technique for assay of microbial ecology. Applied Microbiology. [Online] 16 (2), 330–334. Tersedia pada: https://www.ncbi.nlm.nih.gov/pmc/ articles/PMC547406 [Diakses 5 April 2021].

Hassan, T.U., Bano, A., Naz, I. & Hussain, M. (2018) Bacillus cereus: a competent plant growth promoting bacterium of saline sodic field. Pakistan Journal of Botany, 50 (3), 1029–1037.

Hawkes, C. (2001) Acetylene reduction method for measuring nitrogenase activity tested by Christine Hawkes. [Online] Tersedia pada: http:// www.biosci.utexas.edu/IB/faculty/hawkes/lab/protocols/acetylene_reduction [Diakses 18 Desember 2020].

Hucker, G.J. & Conn, H.J. (1923) Methods of gram staining. [Online] Tersedia pada: https://ecommons.cornell.edu/ bitstream/handle/1813/30787/1923%20Tech%20Bull%2093.pdf?sequence=2&isAllowed=y [Diakses 15 Januari 2021].

Jumadi, O., Liawati & Hartono (2015) Produksi zat pengatur tumbuh IAA (indole acetic acid) dan kemampuan pelarutan posfat pada isolat bakteri penambat nitrogen asal kabupaten Takalar. Jurnal Bionature, 16 (1), 43–48.

Kaburuan, R., Hapsoh, H. & Gusmawartati, G. (2014) Isolasi dan karakterisasi bakteri penambat nitrogen non-simbiotik tanah gambut cagar biosfer Giam Siak Kecil-Bukit Batu. Jurnal Agroteknologi, 5 (1), 35–39.

Kumawat, N., Kumar, R., Kumar, S. & Meena, V.S. (2017) Nutrient solubilizing microbes (NSMs): its role in sustainable crop production. Dalam: Meena, V., Mishra, P., Bisht, J. & Pattanayak, A. (editor) Agriculturally important microbes for sustainable agriculture. [e-book] Singapore, Springer, hlm. 25–61. Tersedia pada: https://doi.org/10.1007/978-981-10-5343-6_2 [Diakses 17 Februari 2021].

Kusmiyati, F., Sumarsono & Karno (2014) Pengaruh perbaikan tanah salin terhadap karakter fisiologis Calopogonium mucunoides. Jurnal Pastura, 4 (1), 1–6.

Lutfi, I. (2012) Ketersediaan unsur hara pada tanah garaman. [Online] Tersedia pada: https:// www.academia.edu/7911945/Unsur_Hara_pada_Tanah_Salin_Garaman_ [Diakses 5 Maret 2021].

Maulina, N.M.I., Khalimi, K., Wirya, G.N.A. & Suprapta, D.N. (2015) Potensi rizobakteri yang diisolasi dari rizosfir tanaman Graminae non-padi untuk memacu pertumbuhan bibit padi. Journal of Agricultural Science and Biotechnology. [Online] 4 (1), 23020–23113. Tersedia pada: http://ojs.unud.ac.id/index.php/JASB [Diakses 21 Januari 2021].

Morel, M.A., Ubalde, M.C., Braña, V. & Castro-Sowinski, S. (2011) Delftia sp. JD2: a potential Cr(VI)-reducing agent with plant growth-promoting activity. Archives of Microbiology. [Online] 193 (1), 63–68. Tersedia pada: https://doi.org/10.1007/s00203-010-0632-2 [Diakses 30 Maret 2021].

Noonan, L. & Freeman, J. (2020) Bacillus cereus and other non-anthracis Bacillus species. [Online] Tersedia pada: https://www.uptodate.com/contents/bacillus-cereus-and-other-non-anthracis-bacillus-species#H453848234 [Diakses 5 April 2021].

Numan, M., Bashir, S., Khan, Y., Mumtaz, R., Shinwari, Z.K., Khan, A.L., Khan, A. & Al-Harrasi, A. (2018) Plant growth promoting bacteria as an alternative strategy for salt tolerance in plants: a review. Microbiological Research. [Online] 209, 21–32. Tersedia pada: https://doi.org/10.1016/j.micres.2018.02.003 [Diakses 11 Januari 2021].

Nurhidayati, S., Faturrahman & Ghazali, M. (2015) Deteksi bakteri patogen yang berasosiasi dengan Kappaphycus alvarezii (Doty) bergejala penyakit ice-ice. Jurnal Sains Teknologi & Lingkungan, 1 (2), 24–30.

Nurmas, A., Rahman, A. & Andi Khaeruni, D. (2014) Exploration and characterization of indigenous Azotobacter for biofertilizer development of local upland rice of marginal land. Jurnal Agroteknos, 4 (2), 128–134.

Paul, S., Bandeppa, Aggarwal, C., Thakur, J.K., Rathi, M. & Khan, M.A. (2014) Effect of salt on growth and plant growth promoting activities of Azotobacter chroococcum isolated from saline soils. Journal of Environment and Ecology. [Online] 32 (4), 1255–1259. Tersedia pada: https://www.researchgate.net/ publication/272482828 [Diakses 7 Maret 2021].

Petti, C.A. (2007) Detection and identification of microorganisms by gene amplification and sequencing. Clinical Infectious Diseases. [Online] 44, (8), 1108–1114. Tersedia pada: https://doi.org/10.1086/512818 [Diakses 17 Februari 2021].

Rachman, A., Dariah, A. & Sutono, S. (2018) Pengelolaan sawah salin berkadar garam tinggi. [Online] Tersedia pada: https://balittanah.litbang.pertanian.go.id/ind/ dokumentasi/lainnya/lahan%20salin%20final.pdf [Diakses 15 April 2021].

Reetha, S., Bhuvaneswari, G., Thamizhiniyan, P. & Ravi, T. (2014) Isolation of indole acetic acid (IAA) producing rhizobacteria of Pseudomonas fluorescens and Bacillus subtilis and enhance growth of onion (Allium cepa L.). International Journal of Current Microbiology and Applied Sciences, 3 (2), 568–574.

Sabaté, D.C. & Audisio, M.C. (2013) Inhibitory activity of surfactin, produced by different Bacillus subtilis subsp. subtilis strains, against Listeria monocytogenes sensitive and bacteriocin-resistant strains. Microbiological Research. [Online] 168 (3), 125–129. Tersedia pada: https://doi.org/10.1016/ j.micres.2012.11.004 [Diakses 25 Maret 2021].

Schöttelndreier, M. & Falkengren-Grerup, U. (1999) Plant induced alteration in the rhizosphere and the utilisation of soil heterogeneity. Plant and Soil, 209, 297–309.

Sen, S. & Chandrasekhar, C.N. (2014) Effect of PGPR on growth promotion of rice (Oryza sativa L.) under salt stress. Asian Journal of Plant Science and Research. [Online] 4 (5), 62–67. Tersedia pada: https://www. imedpub.com/articles/effect-of-pgpr-on-growth-promotion-of-rice-oryza-sativa-l-under-salt-stress.pdf [Diakses 20 Maret 2021].

Setiawati, M.R., Suryatmana, P. & Hudaya, R. (2007) Peningkatan kandungan tanaman dan hasil padi gogo akibat aplikasi bakteri endofitik penambat N2 dan pupuk N anorganik pada tanah salin. Jurnal Himpunan Mahasiswa Pascasarjana Maluku (HMPM) Bandung. [Online] 3 (1), 1–9. Tersedia pada: https://www. researchgate.net/publication/265243612 [Diakses 2 Januari 2021].

Shaaban, M., Abid, M. & Abou-Shanab, R.A.I. (2013) Amelioration of salt affected soils in rice paddy system by application of organic and inorganic amendments. Plant, Soil and Environment, 59 (5), 227–233.

Shrivastava, P. & Kumar, R. (2015) Soil salinity: a serious environmental issue and plant growth promoting bacteria as one of the tools for its alleviation. Saudi Journal of Biological Sciences. [Online] 22 (2), 123–131. Tersedia pada: https://doi.org/10.1016/ j.sjbs.2014.12.001 [Diakses 11 Januari 2021].

Simarmata, T. (2013) Tropical bioresources to support biofertilizer industry and sustainable agriculture. [Online] International Seminar on Tropical Bio-Resources for Sustainable Bioindustry 2013; from Basic Research to Industry in Indonesia. Bandung, 30–31 Oktober. Tersedia pada: https://www.academia.edu/ 26245164/Tropical_Bioresources_to_Support_Biofertilizer_Industry_and_Sustainable_Agriculture_in_Indonesia_1 [Diakses 30 Januari 2021].

Sondang, Y., Anty, K. & Siregar, R. (2020) Potensi konsorsium bakteri pemacu pertumbuhan sebagai bahan aktif pupuk organik hayati pada tanaman jagung. In Agritech, 22 (2), 110–118.

Suganda, H., Setyorini, D., Kusnadi, H., Saripin, I. & Kurnia, U. (2003) Evaluasi pencemaran limbah industri tekstil untuk kelestarian lahan sawah. [Online] Seminar Nasional Multifungsi dan Konversi Lahan Pertanian. Bogor dan Jakarta, 2 dan 25 Oktober. Tersedia pada: https://balittanah.litbang.pertanian.go.id/ind/dokumentasi/prosiding/mflp2003/husein%20suganda16.pdf [Di-akses 2 Feburari 2021].

Susilowati, D.N. & Setyowati, M. (2016) Analisis aktivitas nitrogenase dan gen NIFH isolat bakteri rhizosfer tanaman padi dari lahan sawah pesisir Jawa Barat. Al-Kauniyah: Jurnal Biologi. [Online] 9 (2), 125–138. Tersedia pada: https://doi.org/10.15408/ kauniyah.v9i2.4036 [Diakses 11 Januari 2021].

Wang, Q., Garrity, G.M., Tiedje, J.M. & Cole, J.R. (2007) Naïve Bayesian Classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Applied and Environmental Microbiology, 73 (16), 5261–5267.

Wang, C., Yang, A., Yin, H. & Zhang, J. (2008) Influence of water stress on endogenous hormone contents and cell damage of maize seedlings. Journal of Integrative Plant Biology. [Online] 50 (4), 427–434. Tersedia pada: https://doi.org/10.1111/j.1774-7909.2008.00638.x [Di-akses 2 April 2021].

Widawati, S. & Muharam, A. (2012) Uji laboratorium Azospirillum sp. yang diisolasi dari beberapa ekosistem. Jurnal Hortikultura, 22 (3), 258–267.

Yumoto, I., Yamazaki, K., Kawasaki, K., Ezura, Y. & Shinano, H. (1998) Bacillus horti sp. nov., a new Gram-negative alkaliphilic bacillus. International Journal of Systematic Bacteriology, 48 (2), 565–571.

Yurnaliza, M., Siregar, W. & Priyani, N. (2011) Peran bakteri endofit penghasil IAA (indol acetic acid) terseleksi terhadap pertumbuhan tanaman padi (Oryza sativa L.). [Online] Seminar Nasional Biologi. Medan, 22 Januari. Tersedia pada: https://www.researchgate.net/ publication/323390414 [Diakses 30 Maret 2021].

Zelensky, G. (1999) Rice on saline soils of Russia. Dalam: Chataigner, J. (editor) Future of Water Management for Rice in Mediterranean Climate Areas: Proceedings of the Workshops. Montpellier, CIHEAM, hlm. 109–113.




DOI: http://dx.doi.org/10.21082/jbio.v18n1.2022.p1-10

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