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 Faculty-

Widyagama.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 Series-

Part-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.