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Susamto Somowiyarjo
Fakultas Pertanian Universitas Gadjah Mada
Agriculture, Biological Sciences & Forestry

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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

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (7177.294 KB) | DOI: 10.22146/jpti.9352

Abstract

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.
Deteksi dan Diferensiasi Virus Kerdil Pisang dengan Teknik PCR-RFLP Rahma Ayu Priani; Susamto Somowiyarjo; Sedyo Hartono; Siti Subandiyah
Jurnal Perlindungan Tanaman Indonesia Vol 16, No 1 (2010)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.11736

Abstract

Banana bunchy top disease (BBTD) can be caused by the infection of two different viruses, Banana bunchy top virus (BBTV) or Abaca bunchy top virus (ABTV). Both viruses can be transmitted persistently by aphid Pentalonia nigronervosa Coq. The research was conducted to detect and to differentiate the virus bypolymerase chain reaction-restriction fragment length polymorphism (PCRRFLP) techniques. Infected plants were collected from Yogyakarta (Sleman, Yogyakarta city, Bantul, Gunung Kidul, and KulonProgo). Nucleon Phytopure DNA Extraction Kit method was used to extract the total DNA of infected plants. Universal primers of Common DNA region (S-CRF and S-CRR) and specific primers DNA-R (C1-CRF and CI-CRR) were used for PCR amplification. PCR products were analyzed by RFLP technique using the restriction enzyme of DraI. The results reconfirm previous reports that bunchy top disease of banana in Yogyakarta is caused by BBTV. The ABTV was not detected in this present study. Based on the RFLP analysis it was concluded that BBTV collected in this study could be divided into three groups. Group 1 consisted of BBTV isolate from Sleman and Yogyakarta city with two fragments DNA of 400 and 388 bp. Group 2 consisted of isolate BBTV from Kulon Progo and Gunung Kidul with three fragments DNA of 400, 388, and 323 bp. Group 3 consisted of isolate from Bantul with two fragments DNA of 723 and 376 bp. Further study on the complete characteristics of these groups is still needed.
Pemanfaatan Antibodi Monoklonal dalam Immunoassay untuk Deteksi Baculovirus oryctes Susamto Somowiyarjo; Hery Haryanto; Sri Wening
Jurnal Perlindungan Tanaman Indonesia Vol 6, No 2 (2000)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.12384

Abstract

The application of non-precoated Indirect Enzyme-Linked Immunosorbent Assay (ELISA) and Dot Immunobinding Assay (DIBA) employing monoclonal antibodies (MCA) against Yogyakarta isolate of Baculovirus oryctes Huger. was described. The MCA-Bv-4 having subclass of IgG2a and titer in vitro of 10^4 - 10^5 proved to be useful antibody for virus detection. The great potential of I-ELISA using MCA-Bv-4 has been it's specificity being able to discriminate between healthy an virus-infected coconut beetle (Oryctes rhinoceros L.). The assay could also differentiate between B. oryctes and Monodon baculovirus, the pathogen of shrimp disease. The best tissue for preparing virus antigen from crude extract was found to be the midgut of the beetle. Head, thorax, abdomen and tibia were not suitable for preparing the test antigen. The crude extract of beetle showed high endogenous enzyme activity to the substrate of DIBA, which precluded the detection of B. oryctes using DIBA. The MCA-Bv-4 could be used to improve the monitoring of the virus to support the program of biological control of coconut beetle using B. oryctes.
Pengendalian Diaphorina citri (Vektor Penyakit CVPD) dengan Metarrhizium anisopliae Kardi Raharjo; Susamto Somowiyarjo; F. X. Wagiman
Jurnal Perlindungan Tanaman Indonesia Vol 6, No 1 (2000)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.12404

Abstract

CVPD is transmitted by Diaphorina citri. Measures to control D. citri by using biological controlling agents have opportunity to reduce insecticide application. The objectives of the research are: to measure effectiveness of Metarrhiziurn anisopliae (Metch.) Sorok. in controlling D. citri, effect fructose and time application namely before and after insect infestation. The first phase of the research phase has been conducted in Temanggung, Completely Randomize Design (CRD) factorial with three time replication. Factor I: sterile water without fructose, concentration 10^6 conidia/ml without fructose, concentration 10^8 conidia/ml without fructose, concentration 10^10 conidia/ml: without fructose, sterile water + fructose 5 mg/ml, concentration 10^6 conidia/ml without fructose 5 mg/ml, concentration 10^8 conidia/ml + fructose (fungi application before insect infestation) and W1 (fungi application after insect infestation). Research phase II was carried out with the best treatment combination compare with control treatment in Temanggung and Bantul. The results of experiment showed that the initial die of D. citri caused by M. anisopliae infection are on 4-6 days after application. The application of M. anisopliae at concentration 10^10 conidia/ml without fructose, applied after insect infestation was most effective. The application after insect infestation was more effective compare with application before insect infestation especially on 4 days after application, but on 35th days after application there was no significant difference. Fructose has no effect to mortality of D. citri.
The Role of Extracellular Protein on the Pathogenicity of Xanthomonas campestris pv. citri Tri Joko; Siti Subandiyah; Susamto Somowiyarjo
Jurnal Perlindungan Tanaman Indonesia Vol 6, No 1 (2000)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.12405

Abstract

A research on the pathogenicity of Xanthomonas campestris pv. citri, the causal agent of citrus canker, has been carried out to study the growth characteristics of the pathogen on some media, physiological characteristics, and the role of extracellular protein on the bacterial pathogenicity. Extracellular protein of X. campestris pv. citri was extracted using ammonium sulfate precipitation. The extracted protein samples were electrophoresed on 10% polyacrilamide gel in the presence of sodium dodecyl sulfate at 15 mA/150 V for 1.5-2 hrs. Pathogenicity assay was conducted by infiltration of bacterial cell and extracellular protein suspension into citrus leaf tissues. The results showed that X. campestris pv. citri was able to grow well on all media. It possess specific protein with molecular weight of 25.71 KDa. Bacterial cell and extracellular protein of X. campestris pv. citri were able to produce typical symptoms of canker, while other closely related Xanthomonas campestris pathovars were only able to produce hypersensitive reaction on citrus leaves.
Pemanfaatan Enzyme-Linked Immunosorbent Assay untuk Mengukur Titer Virus dalam Bawang Putih Susamto Somowiyarjo; Endang Mugiastuti; Y. M. Sugi Maryudani
Jurnal Perlindungan Tanaman Indonesia Vol 6, No 1 (2000)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.12407

Abstract

Non-precoated Indirect ELISA had been developed by employing monoclonal antibodies against virus isolated from Sangga variety of garlic. The ELISA was used to measure the titer of virus in the plant. In comparation with biological assay using Chenopodium amaranticolor, ELISA was able to measure the virus titer faster and was more simpler. The highest titer of virus was obtained using the first leaf of garlic at age of 29-36 days after planting. Application of nitrogen at high dose and high temperature of garlic cultivation trends to increase the virus titer. The results of this experiment may be used to improve the method of sampling to detect virus in garlic tissues.
Pemanfaatan Antibodi Monoklonal dalam I-ELISA untuk Deteksi Penyebab Penyakit Busuk Pucuk Kelapa (Phytophthora palmivora) Susamto Somowiyarjo; Daisy Prapto Sriyanti; Mulyadi Mulyadi; Suryanti Suryanti; Y. M.S. Maryudani; Bambang Hadisutrisno
Jurnal Perlindungan Tanaman Indonesia Vol 5, No 2 (1999)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.12762

Abstract

Mice monoclonal antibodies (MCAs) against Phytophthora palmivora, Butl. (the pathogen of coconut bud rot) have been produced and tested. The selected hybridoma (Ab-PM.3) secreted monoclonal antibodies of IgM subclass and the titer in vitro of 10^2 - 1O^3, was employed to develop Indirect-Enzyme Linked lmmunosorbent Assay. This assay was able to differenciate coconut isolate of P. palmivora not only from cocoa and pepper isolate of the same species, but also from other fungus isolated from coconut bud rot diseased coconut (Fusarium sp., Colletotrichum sp., and Thielaviopsis sp.). This assay was able to differenciate P. pulmivora isolate from diseased coconut with those isolated from cocoa and pepper, and other fungus isolated from diseased coconut, thereby providing specific diagnostic tool for coconut bud rot which is needed for supporting the application of the concept of integrated pest mangement on coconut.
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

Abstract

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.
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

Abstract

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.
Co-Authors Bambang Hadisutrisno Daisy Prapto Sriyanti Endang Mugiastuti Fransiscus Xaverius Wagiman Hery Haryanto Kardi Raharjo Mulyadi Mulyadi Purwati Widiyati Rahma Ayu Priani SEDYO HARTONO Sri Wening Subandi Suharno Suharno Suryanti Suryanti Tri Joko Wuye Ria Andayani Y. B. Sumardiyono Y. M. Sugi Maryudani Y. M.S. Maryudani