Reaktivitas Antibodi Poliklonal SSV terhadap Antigen Homolog dan Heterolog
Sri Sulandari(1*), Y. B. Sumardiyono(2), M. Roechan(3)
(1) Fakultas Pertanian Universitas Gadjah Mada
(2) Fakultas Pertanian Universitas Gadjah Mada
(3) Pusat Penelitian Bioteknologi Tanaman Pangan
(*) Corresponding Author
Abstract
Polyclonal antibodies for Soybean Stunt Virus (SSV) were produced in white rabbit through the following procedures: approximately 100 mg of purified virions emulsified in Complete Freund’s Adjuvant (CFA) were injected intramuscularly first. In the second and third injection 150 mg of purified virions in Incomplete Freund’s Adjuvant (IFA) per injection were injected intramuscularly. Finally, about 300 mg of purified virions were injected intravenously as a booster. The injection were done at 2 weeks interval. Antiserum was collected 5 days after the final injection. Antisera was purified by precipitation in saturated ammonium sulfate. Purified antibody was tested for the titer and reactivity of antibodies against the homologous and heterologous antigen. The studies were conducted with non-precoated I-ELISA test. This research was able to obtain about 25 ml of crude antisera for SSV, the concentration of purified polyclonal antibodies was about 9 mg/ml. the titer of polyclonal antibodies was 10.000 in I-ELISA. Without absorbtion with sap of healthy plant, the antibodies could not be use to identify the infected and healthy plant samples. In the following test, the absorbed antibody was used. Using antibodies to SSV at a dilution of 1:1000 and 1:10.000 against sap extracts sample of healthy and infected plant at a dilution of 1:10 by non-precoated I-ELISA test, indicated that the antibody could be used to identify the healthy and infected samples. By the same test, the antibody could be reacted to both homologous antigen (SSV) and heterologous antigen (CMV isolated from banana).
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Artama, W.T. 1990. Antibodi Monokional, Teori, Produksi, Karakterisasi dan Penerapan. Pusat Antar Universitas-Bioteknologi, UGM, Yogyakarta.
Clarck, M.F. & Adams. 1977. Characteristics of the microplate method of Enzyme-linked immunosorbent assay for the detection of plant viruses, J. Gen. Virol.34:475-483.
C.M.I. 1972. Descriptions of Plant Viruses. No.1. [4] Habili, N. & I.B. Francki. 1975. Comparative Studies on Tomato Aspermy and Cucumber Mosaic Viruses. IV. Immunogenic and Serological Properties. Virology 64: 421 - 429.
Koenig, R. 1981. Indirect-Elisa methods for the broad specifity detection of plant viruses, J. Gen. Virol., 55:53-62.
Lange, L. 1985. Seed transmitted virus diseases. Biology, Detection and Control. Danish Government Institute of Seed Pathology for Developing Country.
Lankow, R.K., S.H. Woodhead, R.J. Patterson, R. Massey & G. Schohetman. 1984. Monoclonal antibody diagnostics in plant disease management. Plant Disease 68: 1100-1101.
Liddell, J.E. & A. Cryer. 1991. A practical guide to monoclonal antibodies. John Wiley & Sons Ltd., Baffins Lane, Chister-England, 188p.
Mahmood, T., Hein G.L., & French, R.C. 1997. Development of serological procedures for rapid and reliable detection of Wheat streak mosaic virus in single wheat curl mite. Plant Disease 81: 250-253.
Marco, S. & S. Cohen 1979. Rapid detection and titer evaluation of viruses in pepper by Enzyme-linked Immunosorbent Assay. Phytopathology 69: 1259 - 1262.
Matthews, REF. 1992. Fundamental of Plant Virology. Academic Press, New York.
Mernaugh, R.L G.R. Mernaugh & G.R. Kovacs. 1990. The immune response: antigen, antibodies, antigen-antibody interaction, Dalam Hampton, R., E. Ball, S. De Boer (Eds.). Serological methods for detection and identification of viral and bacterial plant pathogens. APS Press, St.Paul, Minnesota. USA. p:3-26.
Roechan, M., M. Iwaki & D.M. Tantera. 1975. Virus disease of legume plants in Indonesia. Soybean stunt virus. Contr. Centr. Res. Inst. Agric. Bogor 15:1-16.
Roechan, M. 1992. Penyakit-penyakit virus pada tanaman kedelai. Disertasi S3. Univ. Pajajaran, Bandung. (Tidak dipublikasikan).
Seifer, D.L., Harvey,. T.L., Martin, TJ., & Jensen, S.G. 1997. Identification of the wheat curl mite as the vector of the High Plains virus of corn and wheat. Plant Disease 81: 1161 - 1166.
Sinclair, J.B & P.A. Backman. 1993. Compendium of Soybean Diseases. 2nd Ed. APS Press. St. Paul, Minnesota. 106 p.
Suastika, G., K. Tomaru, J. Kurihara & K.T. Natsuaki. 1996. A cucumber mosaic Cucumovirus isolate from banana mosaic disease in Indonesia. Biological control in sustainable tropical agriculture. Nodai Center for International Programs, Tokyo University of Agriculture. JSPS- DGHE Program p: 60-71.
Sudjono, M. Amir & R. Martoatmodjo. 1985. Penyakit kedelai dan penanggulangannya. Dalam Somaatmadja, S. (Ed.). Kedelai. Balai Penelitian Tanainan Pangan. Bogor. 3.
Suseno, R. 1987. Virus terbawa benih tanaman pangan serta hubungannya dengan sertifikasi dan pengawasan mutu benih, Lokakarya Patogen Terbawa Benih, Pengembangan Pengawasan Mutu dan Sertifikasi Benih, Bogor, 8-9 Desember 1987.
Suseno, R., G. Suastika, & Y.M. Kusumah. 1992. Studi beberapa aspek pengendalian virus terbawa benih kedelai yang ditularkan kutu daun (SSV dan SMV), Seminar hasil penelitian Pendukung Pengendalian Hama Terpadu, Cisarua, 7-8 September 1992.
Sutula, C.L., J.M. Gillett, S.M. Morrissey & D.C. Ramsdell. 1986. Interpreting ELISA Data and Establishing the Positive-Negative Threshold. Plant Disease 70: 722 - 726.
Tamada, T., T. Goto., I. Chiba & T. Suwa. 1970. Soybean dwarf, a new virus diseases. Ann. Phytopath. Soc. Japan 35:282-285.
Torrance, L. 1992. Serological methods to detect plant viruses: production and use of monoclonal antibodies. Dalam Duncan, J.M. & L. Torance (Eds.). Technique for the rapid detection of plant pathogens. Blackkwell Scientific. Publ.
Van Regenmortel, M.H.V. 1981. Serology of Plant Viruses. Acad. Press., New York.
Wahyuni, W.S., R.G. Dietzgen, K. Hanada & R.I.B. Francki. 1992. Serological and biological variation between and within subgroup I and II strains of Cucumber mosaic virus. Plant Pathol. 41: 282-297.
DOI: https://doi.org/10.22146/jpti.9883
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