LAPORAN HASIL PENELITIAN DISERTASI DOKTOR TAHUN ANGGARAN 2011
ISOLASI, KARAKTERISASI & UJI AKTIFITAS MIKROBA PELARUT FOSFAT DAN PENGIKAT NITROGEN DARI MOL (Mikroorganisme Lokal) BONGGOL & BATANG PISANG (Musa paradisiaca)
SUSLAM PRATAMANINGTYAS
Dibiayai Oleh Direktorat Jenderal Pendidikan Tinggi, Kementerian Pendidikan Nasional, melalui DIPA Universitas Brawijaya REV.1 Nomor: 0636/023-04.2.16/15/2011 R, tanggal 30 Maret 2011 dan berdasarkan Surat Keputusan Rektor Nomor: 214/SK/2011 tanggal 2 Mei 2011
UNIVERSITAS BRAWIJAYA NOVEMBER 2011
RINGKASAN DAN SUMMARY Budidaya suatu jenis tanaman secara terus menerus secara intensif membawa konsekuensi pengurasan hara tanah yang berakibat menurunnya produktifitas lahan yang bersangkutan. Salah satu indikator menurunnya produktifitas lahan ini adalah tidak efektifnya pemupukan. Penambahan dosis pupuk yang dimaksudkan untuk mendongkrak produksi tidak dapat memberikan hasil seperti yang diharapkan. Dilatarbelakangi oleh kondisi tersebut, sekelompok petani padi di Kecamatan Sukorejo, Kabupaten Pasuruan Jawa Timur, berinisiatif membuat pupuk cair dari bahan-bahan sisa panen yang tersedia di lingkungan setempat, diantaranya adalah pupuk MOL bonggol pisang dan MOL batang pisang, yang ‘dianggap’ dapat menggantikan pupuk sintetis Urea dan SP36. Hasil analisis laboratorium menunjukkan bahwa kandungan nitrogen dan fosfor dari pupuk MOL terlalu rendah untuk bisa disebut sebagai pengganti pupuk sintetis. Isolasi, identifikasi dan uji kemampuan mengikat nitrogen dan kemampuan melarutkan fosfat oleh mikroorganisme yang dikandung MOL bonggol dan batang pisang dilakukan di Laboratorium Biologi, Fakultas Pertanian Universitas Widyagama Malang. Beberapa isolat fungi dan bakteri telah berhasil diisolasi. Dari semua isolat fungi yang ditemukan, tidak teridentifikasi ada isolat yang mampu mengikat nirogen bebas maupun yang mempunyai kemampuan melarutkan fosfat. Semua isolat bakteri juga tidak teridentifikasi mempunyai kemampuan mengikat nitrogen bebas, namun ditemukan satu isolat yang mempunyai kemampuan kuat dan cepat dalam melarutkan fosfat. Pengujian kemampuan mikroba mengikat nitrogen bebas dilakukan dengan inokulasi pada media Burk’s N-free. Apabila isolat mampu tumbuh, berarti isolat yang bersangkutan mempunyai kemampuan mengikat nitrogen bebas untuk pertumbuhannya. Pengujian kemampuan mikroba melarutkan fosfat dilakukan dengan inokulasi pada media Pikovskaya. Isolat yang mempunyai kemampuan melarutkan fosfat ditandai oleh terbentuknya halozone disekitar koloni Pengujian biokmia untuk identifikasi adalah Uji Gram & KOH 3%, Uji Katalase, dan beberapa uji lain sesuai standar pengujian idetifikasi bakteri menurut Bergey’s Manual of Determinative Bacteriology. Hasil Uji untuk identifikasi menyimpulkan bahwa isolat – 1 adalah Serratia marcescens. Hasil Uji pathogenisitas menyimpulkan bahwa strain Serratia marcescens yang ditemukan tidak besifat pathogen untuk padi (Oryza sativa). Kemampuan isolat-1 melarutkan P secara kualitatif adalah rata-rata Ø halozone adalah 1,95 Cm, sedangkan kemampuan melarutkan P secara kuantitatif adalah 357 mg/L. Produksi asam yang dihasilkan isolat-1 Serratia marcescens mampu menurunkan pH media menjadi 5,05 dibandingkan pH pada media kontrol 6,25.
SUMMARY Intensive cultivation of one plant species continuously, have a consequences to soil nutrient depletion and resulting in declining productivity of land were concerned. the One of declining indicator of land productivity is uneffective fertilization. The addition of fertilizer dose that is intended to boost production can not deliver results as expected. Motivated by these conditions, a group of rice farmers in the Sukorejo sub-district, Pasuruan regency-East Java, took the initiative to make liquid fertilizer from the harvesting residual which available in the local environment, including the indigenous microorganism (IMO) fertilizer banana root and pseudo-stem, which are 'deemed' can replace the synthetic fertilizers Urea and SP-36 The results of laboratory analysis indicates that the content of nitrogen and phosphorus from IMO fertilizers are too low to be called as a substitute for synthetic fertilizers. the isolation, identification and the test of the nitrogen fixing ability and the phosphate solubilizing ability of the microorganisms were contained in IMO from banana root and pseudo-stem were performed at the Laboratory of Biology, Agricultural FacultyWidyagama.Malang University Some isolates of fungi and bacteria have been isolated. All of the fungal isolates were found, are not identified as free nitrogen fixing or have the ability to solubilize phosphate. All of bactery isolates were also unidentified having the ability to fixing the free nitrogen, but one isolate was found has a strong ability and fast to solubilize phosphate. To testing the ability of microbial to fixing the free-nitrogen is performed by inoculation it on Burk's N-free. When the isolates were able to grow, it means that the isolates have the ability to fixing the free-nitrogen for it's growth. To testing the phosphate solubilizing ability of microbes was carried out by inoculating to the Pikovskaya medium. Isolates that have the ability to solubilize phosphate is characterized by the formation of halozone around the colonies. Biochemist testing for identification of bacteria is Gram Test & KOH-3% test, Catalase Test, and several other tests according to bacteria identification testing standards according to Bergey's Manual of Determinative Bacteriology. Test results for the identification concluded that isolates - 1 is Serratia marcescens. Results for pathogenisity test concluded that the strain Serratia marcescens was found that its not tend to be pathogen for rice (Oryza sativa). The qualitative ability of isolate-1 to solubilize P were shown with Ø halozone average is 1.95 cm, while the quantitative ability to solubilize P is 357 mg / L. The acid production result of Serratia marcescens isolate-1 have capable to lowering the pH of the media to 5.05 compared to control medium pH at 6.25.
DAFTAR PUSTAKA Abdulrachman, S., Z. Susanti, dan Suhana. 2000. Dinamika unsur NPK pada lahan sawah dalam jangka panjang. Laporan akhir PAATP. Balitpa. Sukamandi. Abdulrachman, S. 2002. Pengembangan metode pengelolaan hara spesifik lokasi pada padi sawah Dalam Prosiding Pengelolaan Hara P dan K pada Padi Sawah. Puslittanak. Bogor. p 39-58 Abdulrachman, S.& H. Sembiring. 2006. Penentuan Takaran Pupuk Fosfat untuk Tanaman Padi Sawah. Iptek Tanaman Pangan no. 1 Alikhani, H.A. et al. 2006. Phosphate solubilization activity of rhizobia native to Iranian soils. Plant and Soil 287:35–41 Anonymous. 2009. Sifat Biologi Tanah – MOL. Materi Sekolah Lapang SRI. TIM SRI COMDEV. PT. HM. SAMPOERNA,Tbk. Sukorejo – Pasuruan. Bayona, S.J., B. Chua, L-LY. Tan, R. Isaac F. 2009. Unknown Bacteria Identification: The Serratia marcescens Project. MICROBIOLOGY. BIO 120 LAB. FEJ. ALL RIGHTS RESERVED. Beever, R.E. and D.J.W. Burns. 1980. Phosphorus Uptake, Storage and Utilization by Fungi. Adv. Bot. Res. *: 127 – 219. Chen, Y.P., P.D. Rekha, A.B. Arun, F.T. Shen, W.-A. Lai, C.C. Young. 2006. Phosphate Solubilizing Bacteria from Subtropical Soil and Their Tricalcium Phosphate Solubilizing Ability. App. Soil Ecol. 34: 33 – 41. Dobermann, A. and T. Fairhurst. 2000. Nutrient disorders and nutrient management. IRRI and Potash & PPI /PPIC. Manila, Philipina. Elfiati, D. 2005. Peranan Mikroba Pelarut Fosfat terhadap Pertumbuhan Tanaman. Jurusan Kehutanan Fakultas Pertanian Universita Sumatera Utara. e-USU Repository. 10pp Gyaneshwar.P., G.N.Kumar , L.J. Parekh and P.S. Poole. 2002. Role of soil microorganism in improving P nutrition of Plants. Plant soil 245: 83-93. Heiling, M. & G. Hardarson. 2010. Biological Nitroen (N) Fixation – The source of N nutrient to increase yield. www-naweb.iaea.org/nafa/swmn/soil-science Hindersah, R. & T. Simarmata. 2004. Potensi Rizobakteri Azotobacter dalam Meningkatkan Kesehatan Tanah. JurnalNatur Indonesia 5(2): 127 – 133. Universitas Padjadjaran, Bandung. Hubbell, D.H. & G. Kidder. 2009. The Importance of Nitrogen. http://edis.ifas.ufl.edu. Isherwood, K.F. 1998. Fertilizer use in environment. In: N.Ahmed and A.Hamid (eds), Proc. Symp. Plant Nutrition Management for Sustainable Agricultural Growth. NFDC, Islamabad. Pp. 57 – 76. Khan, A.A., G. Jilani, M.S. Akhtar, S.M.S. Naqvi, M. Rasheed 2009. Phosphorus Solubilizing Bacteria: Occurrence, Mechanisms and their Role in Crop Production. J. agric. biol. sci. 1 (1):48-58 Lal. L. 2002. Phosphate biofertilizers. Agrotech. Publ. Academy, Udaipur. India. 224p. Mearyard.B. 1999. Phosphate enzymes from plants. Journal of Biological Education 33(2): 109-112. Mekhael, R. and S.Y. Yousif. 2009. The Role of Red Pigment Produced by Serratia marcescens as Antibacterial and Plasmid Curing Agent. The 2nd Kurdistan Conference
on Biological Sciences J. Duhok Univ. Vol.12, No.1, pp 268 – 274. University of Duhok, Kurdistan Pang. P.C.K. and H. Kolenk. 1986. Phosphomonoesterase activity in forest soils. Soil Biol. Biochem. 18 (1): 35-40. Ponmurugan. P., and C. Gopi. 2006. In vitro production of growth regulators and phosphatase activity by phosphate solubilizing bacteria. African Journal of Biotechnology 5(4): 348-350. Pradhan, L. and L.B. Sukla. 2005. Solubilization of inorganic phosphates by fungi isolated from agriculture soil. African Journal of Biotechnology Vol. 5 (10), pp. 850-854 Purwasasmita, M & K. Kunia. 2009. Mikroorganisme Lokal sebagai Pemicu Siklus Kehidupan dalam Bioreaktor Tanaman. Prosiding Seminar Nasional Teknik Kimia Indonesia. Bandung. ISBN 978-97998300-1-2 Rengel, Z. & Marschner, P. 2005. Nutrient availability and management in the rhizosphere: exploiting genotypic differences. New Phytologist 168, 305-312. Rodriguez, H. & R. Fraga. 1999. Phosphate Solubilizing Bacteria and Their Role in Plant Growth Promotion. Biotech. Adv. 17: 319 – 339. Samrot, A.V., K. Chandana, P. Senthilkumar and N.G. Kumar. 2011. Optimization of prodigiosin production by Serratia marcescens SU-10 and evaluation of its bioactivty. International Research Journal of Biotechnology Vol. 2(5) pp. 128 – 133. Suliasih dan Rahmat. 2007. Aktivitas Fosfatase dan Pelarutan Kalsium Fosfat oleh beberapa Bakteri Pelarut Fosfat. BIODIVERSITAS 8 (1): 23-26 Uphoff, N. & E. Fernandes. 2003. Sistem Intensifikasi Padi Tersebar Pesat. CIIFAD, 31 Warren Hall,Cornell University, Ithaca, NY 14853. van Veen, J. A., L.S. van Overbeek and J.D. van Elsas. 1997. Fate and Activity of Microorganisms Introduced into Soil. Microbiology and Molecular Biology Reviews, p. 121 – 135. Vessey, J.K. 2003. Plant growth promoting rhizobacteria as biofertilizers. Plant & Soil 255, 571-586. Williams, R.P. 1973. Biosynthesis of Prodigiosin, a Secondary Metabolite of Serratia marcescens. Applied Microbiology. American Society for Microbiology. 25:3. William, R.P. and S.M. Quadri. 1980. The Pigment of Serratia. In the Genus Serratia. Pp. 31 – 75. Edited by A. Von Graevenitz & S.J. Rubin. Boca Raton, FL: CRC Press Inc. Willson, A. 2005. Types of Nitrogen Fixation. The World of Nitrogen Fixation SeriesPart-1. AUSTRALIAN ORGANIC JOURNAL – AUTUMN 2005. Zhao, X., Y-x. Xie, Z-q. Xiong, X-y. Yan, G-x. Xing, Z-l. Zhu, 2009. Nitrogen fate and environmental consequence in paddy soil under rice-wheat rotation in the Taihu lake region, China. REGULAR ARTICLE. Springer Science + Business Media B.V.
