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Elektron Mikroskopi dan Imunogenisitas Baculovirus oryctes Isolat Yogyakarta Y. B. Sumardiyono
Jurnal Perlindungan Tanaman Indonesia Vol 1, No 1 (1995)
Publisher : Universitas Gadjah Mada

Palm rhinoceros beetle (Oryctes rhinoceros) was infected per os with Yogyakarta isolate of Baculovirus oryctes in laboratory condition. Midguts of infected beetle obtained were then extracted for further nucleoprotein purification by centrifugation method. Electron microscopy studies on purified nucleoprotein revealed rod-shape viruses with rounded end measured 190×94 nm in average. One end of the particle showed tail-like structure. Antibodies against the virus were obtained by immunization to rabbit, and reacted against either purified virus or extract of infected beetle, but not against extract of healthy beetle.

Konsentrasi PEG 6000 dan Senyawa Aditif Buffer Fosfat yang Diperlukan dalam Pemurnian Soybean Mosaic Virus Y. B. Sumardiyono; Susamto Somowiyarjo; Wuye Ria Andayani
Jurnal Perlindungan Tanaman Indonesia Vol 1, No 1 (1995)
Publisher : Universitas Gadjah Mada

The objective of study was to determine the concentration of PEG 6000 for precipitation of virus particles, and additive substance added to the resuspension buffer, during the purification of Soybean Mosaic Virus isolated from Yogyakarta. The result showed that precipitation with PEG 6000 of 6 per cent at final concentration, followed by either two or three cycles differential centrifugation and the virus obatained from each centrifugation resuspended in Phosphate buffer 0.05 M contained NaCl 0.1 M added with 0,005 M Na-EDTA, and then subjected to 10-50 per cent sucrose density gradient centrifugation. The purified virus obtained was infective, with the ultraviolet absorption maximum at 260 nm and minimum at 247 nm, ratio A280/A260 was 0.7343. based on extinction coefficient at 260 nm = 2.4, the yield was 0.306 mg/100 g infected leaves. SDS-PAGE indicating that coat protein molecule weight was 29.71 kD.

Reaktivitas Antibodi Poliklonal SSV terhadap Antigen Homolog dan Heterolog Sri Sulandari; Y. B. Sumardiyono; M. Roechan
Jurnal Perlindungan Tanaman Indonesia Vol 4, No 1 (1998)
Publisher : Universitas Gadjah Mada

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).

Deteksi Keragaman Virus Tungro dari Beberapa Daerah Endemis di Indonesia dengan Teknik PCR-RFLP R. Heru Praptana; Y. B. Sumardiyono; Sedyo Hartono; I Nyoman Widiarta; Muhammad Muhsin
Jurnal Perlindungan Tanaman Indonesia Vol 15, No 1 (2009)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.11763

Tungro is one of rice disease caused by two different viruses (rice tungro virus=RTV) i.e. Rice tungro bacilliform virus (RTBV) and Rice tungro spherical virus (RTSV) that are transmitted only by green leafhopper. Tungro had become a serious problem in several rice productions centre in Indonesia. Various components of management effort have been applied but they were inefficient in preventing the tungro disease development. Resistance variety is the most efficient component to tungro disease management. Complexity interactions of tungro disease components are mayor constraint in tungro disease management. Detection of molecular variability in rice tungro virus from several endemic areas in Indonesia were conducted by using PCR-RFLP technique. Existence of RTBV and RTSV in the infected plants collected from several endemic areas were successfully detected by PCR. The RFLP analysis with restriction enzymes BstYI and HindIII showed that there were significant difference among the RTSV originated from Java, Bali and Sulawesi.

Seleksi, Karakterisasi, dan Reaktivitas Antibodi Monoklonal Virus Kerdil Kedelai Sri Sulandari; Y. B. Sumardiyono; Roechan M.
Jurnal Perlindungan Tanaman Indonesia Vol 9, No 2 (2003)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.12233

Soybean Stunt Virus (SSV) is a member of Cucumber mosaic virus group that caused soybean stunt disease. The disease is the most important viral disease of soybean in Indonesia. The objective of the study was to obtain SSV monoclonal antibody for detection SSV in both infected seed and plant. Six hybridoma clones were obtained from fusion between spleen cells of BALB/c immunized with Soybean stunt virus and mouse myelome cell lines (NS-1); they were IgG3 type antibodies, and its titres were varied between 1: 1O to 1: 1OO. Using non-recoated I-ELISA method, the homolog antigen (SSV) was well detected but not the heterolog antigens (CMV isolated from banana, tobacco).

Epidemi Penyakit Daun Keriting Kuning Cabai Y. B. Sumardiyono; Sedyo Hartono; Sri Sulandari
Jurnal Perlindungan Tanaman Indonesia Vol 9, No 1 (2003)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.12269

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Pemendekan Waktu ELISA dalam Deteksi TMV Susamto Somowiyarjo; Y. B. Sumardiyono; Purwati Widiyati
Jurnal Perlindungan Tanaman Indonesia Vol 2, No 2 (1996)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.12891

In order to support the program for the management of viral diseases on garlic, a rapid diagnosis procedure was developed by shortening the incubation period of the indirect ELISA. No significant differences of ELISA absorbance were observed when the antigen was incubated for 4 h, 2 h, and 30 min at 37ºC. The incubation period for the antibody and conjugate could he reduced from 4 h at 37ºC or 18 h at 4ºC to 30 min al 37ºC. The shortest period of incubation for the assay could be obtained when each incubation time for the antigen, antibody and conjugate was 30 min. The detection limit for the shortened ELISA was 10^-4 for the virus in crude extracts and 0,5 μg/ml for the purified preparation.

Isolasi, Pemurnian dan Karakterisasi Parsial Soybean Stunt Virus Sri Sulandari; Y. B. Sumardiyono; Roechan Roechan
Jurnal Perlindungan Tanaman Indonesia Vol 3, No 1 (1997)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.12902

The objective of this study was isolation and characterization of Soybean Stunt Virus, the causal agent of soybean stunt disease. The virus was isolated with single lesion method through Chenopodium amaranticolor and Vigna unguiculata, and then propagated on Nicotiana tabacum var. Xanthi. Differential centrifugation method was used for purification. Virus isolate obtained from Bogor was used for futher studies. The result showed that SSV could be isolated on C. amaranticolor, but not on V. unguiculata and then propagated on Niconana tabacum var. Xanthi without any symptom. Purified virus showed A260/A280 = 1.55, lower than that of CMV. The virion were small isometric particles, about 30 nm in diameter. Coat protein consists of a single type of subunit protein, with molecular weight about 29 kD.

Penularan Penyakit Mosaik Kacang Panjang oleh Aphis craccivora Y. B. Sumardiyono; Supratoyo Supratoyo; Samsuri Samsuri
Jurnal Perlindungan Tanaman Indonesia Vol 3, No 1 (1997)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.12905

Transmission of cowpea mosaic disease (Vigna unguiculata (L.) Walp.) caused by Cowpea Aphid-borne Mosaic Virus (CAMV) through insect vector Aphis craccivora Koch was studied tn the laboratory and the field. Serial inoculation test in varied inoculation access period (1, 5, 10, and 15 minutes) was done to determine the retention time of virus. Field studies were undertaken to evaluate the effect of the number of initial inoculum and plant age on the disease progress. The results showed that viruliferous insect retained their inoculativity untill third series inoculation with 5, 10, and 15 minutes inoculation access period. lt indicated that the longest retention time was 45 minutes, and virus - insect relationship is non-persistent. The disease-spread changed with the number of initial inoculum and the age of the plant. The highest disease intensity of 97.92 per cent was observed in the plot with 2 diseased plants as inittal inoculum and the insects vectors were infested at 10 days plant of age. While the lowest was observed in the plot with one diseased plant as initial inoculum and infestation of insects vector at 30 days after planting.

Pemanfaatan Membran Nitroselulosa untuk Pengiriman Antigen Uji dalam Deteksi TMV dengan DIBA Susamto Somowiyarjo; Y. B. Sumardiyono; Suharno Suharno
Jurnal Perlindungan Tanaman Indonesia Vol 3, No 1 (1997)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.12969

Simplification of dot immunobinding assay (DIBA) by using TMV-infected samples which were stored and mailed in nitrocellulose membrane (NCM) was described. The antigenicity of TMV in DIBA could be maintained in leaf extract-dotted NCM which had been stored at 29ºC al least for 42 days. The method was developed for sending samples using infected leaf extract-dotted NCM to replace fresh samples. By this method, the antigenicity of the virus could he detected after they have been sent to 18 places which took time from 7 to 26 days. It ts anticipated that the simplicity of DIBA using mailed samples will lead to DIBA's rapid adoption for development of central diagnosis facilities to support the viral diseases management. It may also have wider use in DIBA-based screening and survey programs for plant viruses and could overcome plant quarantine restriction.

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