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IDENTIFICATION AND CHARACTERIZATION OF CARMOVIRUS ON CARNATION (Dianthus caryophyllus L.) IN WEST JAVA, INDONESIA Diningsih, Erniawati; Suastika, Gede; Damayanti, Tri Asmira; Susanto, Slamet
AGRIVITA, Journal of Agricultural Science Vol 37, No 2 (2015)
Publisher : Faculty of Agriculture University of Brawijaya and Indonesian Agronomic Assossiation

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Abstract

Carnation has a highly economic demand of cut flower in Indonesia.  Field observations in West Java Indonesia was conducted in order to find the typical mottle symptoms that was a suspect caused by a virus disease. Identification of the virus was respectively conducted by performing ELISA test with four anti sera and characterizations held by bioassay, observing of virion particles, detecting of nucleic acid by RT-PCR and nucleotide sequencing. Total of 403 samples were collected from plants with or no virus-like symptoms. Among those all tested, 83% were found to be infected by Carnation mottle virus (CarMV), but negatively against Carnation ringspot virus (CRSV), Carnation laten virus (CLV), and Carnation vein mottle virus (CVMV) antisera. By mechanical inoculation, the virus was able to infect systemically Cenopodium quinoa and locally infect on others. However on Phalaenopsis sp and Gomprena globosa, there was symptompless found. The isometric CarMV particles size was approximately 30 nm. RT-PCR using specific primers of CP gene of CarMV successfully amplified a DNA sized 1000 bp.  CarMV West Java Indonesian (Idn-WJ) isolates possessed the highest nucleotide and amino acid homology with CarMV from Spain and was in the same cluster with CarMV from China, Taiwan and Israel.Keyword: Dianthus caryhophyllus, RT-PCR, sequence, Tombusviridae
DETEKSI SECARA SEROLOGI DAN MOLEKULER BEBERAPA JENIS VIRUS YANG BERASOSIASI DENGAN PENYAKIT MOSAIK TANAMAN NILAM (Pogostemon cablin Benth) MIFTAKHUROHMAH, MIFTAKHUROHMAH; SUASTIKA, GEDE; DAMAYANTI, TRI ASMIRA
853-8212
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

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Abstract

ABSTRAKPenyakit mosaik pada tanaman nilam disebabkan oleh beberapajenis virus, yaitu Potyvirus, Potexvirus, Cucumber mosaic virus (CMV),dan Broad bean wilt virus 2 (BBWV2). Penelitian ini bertujuan untukmengidentifikasi secara serologi dan molekuler virus-virus yangberasosiasi dengan gejala mosaik pada nilam di KP. Manoko, KP. Cicurugdan lahan petani di Cijeruk. Sampel daun nilam baik yang menunjukkangejala mosaik atau pun tidak diambil dari setiap lokasi penanamanmasing–masing sebanyak 30 sampel. Kejadian penyakit ditentukan melaluideteksi serologi dengan Direct-ELISA dan Indirect-ELISA terhadap sampelmenggunakan empat antiserum, yaitu CMV, Cymbidium mosaic virus(CymMV), Potyvirus, dan BBWV2. Deteksi molekuler dengan RT-PCRdilakukan untuk mengonfirmasi virus baru yang ditemukan. Hasilpenelitian menunjukkan bahwa gejala infeksi virus yang ditemukan padanilam bervariasi, yaitu mosaik lemah, mosaik kuning hijau, mosaik denganpenebalan, mosaik dengan malformasi daun, dan bintik kuning. Secaraserologi, kejadian virus pada setiap kebun bervariasi. Di KP Manoko,Potyvirus dan BBWV2 lebih dominan (100%) dibandingkan CymMV. DiKP Cicurug, kejadian Potyvirus dan CMV terlihat lebih dominan (83,3 dan80%) dibandingkan BBWV2 dan CymMV, sedangkan di Cijeruk, BBWV2lebih dominan (90%) dari Potyvirus (50%) dan CMV (13,3%). Hasil RT-PCR dengan primer degenerate BBWV, diidentifikasi BBWV2 padasampel daun nilam dari Manoko, Cicurug, dan Cijeruk, sedangkan denganprimer general Potexvirus, diidentifikasi CymMV hanya dari sampel daunnilam dari asal Manoko. Hasil penelitian ini merupakan laporan pertamatentang BBWV2 dan CymMV pada tanaman nilam di Jawa Barat yangmengindikasikan bahwa virus merupakan kendala utama pada perbenihannilam yang harus segera diatasi.Kata kunci: BBWV2, CymMV, mosaik, Pogostemon cablin Benth, PCRABSTRACTMosaic symptoms on patchouli plant are associated with severalviruses, i.e. Potyvirus, Potexvirus, CMV, and BBWV2. The objective ofthe study was to detect virus(es) associated with mosaic symptoms onpatchouli at the the patchouli seed nurseries, in Manoko, Cicurug, andCijeruk. Thirty leaf samples either showing typical symptomatic mosaic orasymptomatic were taken from each location. Serological testing byDirect-ELISA and Indirect-ELISA using four antisera namely CMV,Cymbidium mosaic virus (CymMV), Potyvirus, and BBWV2 was carriedout to test the incidence of each virus. Molecular detection by RT-PCR wasperformed to confirm the new virus(es). The results showed that symptomsof virus infection were found vary, i.e. weak mosaic, green yellow mosaic,mosaic with thickening, mosaic with leaf malformations, and yellow spot.Based on the serological detection, virus(es) incidence varied at each seednurseries. In Manoko, Potyvirus, and BBWV2 were more dominant(100%) compared with CymMV. In Cicurug, Potyvirus and CMV weremore dominat (83.3 and 80%) compared with BBWV2 and CymMV.While in Cijeruk, BBWV2 was the most dominant (90%) than Potyvirus(50%) and CMV(13.3%). Result of RT-PCR with degenerate primers pairof BBWV was succesfully identified BBWV2 from Manoko, Cicurug, andCijeruk samples, whereas by using Potexvirus general primary, CymMVwas identified only from Manoko samples. BBWV2 and CymMV werefirst reported to infect patchouli in West Java. The result indicate thatvirus(es) are the major constraint on patchouli seed that should bemanaged immediately.Key words: BBWV2, CymMV, mosaic, Pogostemon cablin Benth, PCR
Preparasi RNA Virus Mosaik Bergaris dari Tanaman Tebu Menggunakan Metode Tabung PCR Tri Asmira Damayanti; Lilik Koesmihartono Putra
Jurnal Fitopatologi Indonesia Vol. 8 No. 1 (2012)
Publisher : The Indonesian Phytopathological Society (Perhimpunan Fitopatologi Indonesia)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (583.464 KB) | DOI: 10.14692/jfi.8.1.22

Abstract

Sugarcane streak mosaic virus (SCSMV) is a new emerging virus infecting sugarcane in Indonesia. The virus can be found almost in every sugarcane plantations in Java and it was known as sett-borne virus. Attempt to get virus-free seedling meet difficulties due to lacking an easy, and accurate detection method. SCSMV antiserum is not available yet commercially. Nucleic acid detection by RT-PCR also hampered difficulties in releasing RNA virus from sugarcane tissues. Here we reported the simple direct tube method with minor modification as the convenient way to provide total RNA template from infected sugarcane leaf, stalk and sheath for RT-PCR detection of SCSMV.Key words: RNA extraction, RT-PCR, SCSMV, tube PCR
Penyakit Mosaik pada Koro Pedang Tri Asmira Damayanti
Jurnal Fitopatologi Indonesia Vol. 11 No. 1 (2015)
Publisher : The Indonesian Phytopathological Society (Perhimpunan Fitopatologi Indonesia)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (603.001 KB) | DOI: 10.14692/jfi.11.1.29

Abstract

Recently, a mosaic disease was observed on jack bean cultivation in Dramaga, Bogor,West Java. The symptom on infected plants, i.e. combination of pale and dark green area was more obvious on young leaves. Vein banding, leaf malformation, and growth inhibition was also observed in infected plants. The symptom was not very obvious in older leaves. Serological detection gave positive reaction to Bean common mosaic virus (BCMV) antiserum, but reaction was negative when detected using specific primer to coat protein (CP) of BCMV strain black eye by reverse transcription polymerase chain reaction. DNA fragments were successfully amplified by RT-PCR using universal primer of CP gene and degenerated primer for cylindrical inclusion gen (CI) of Potyvirus. DNA analysis showed the highest homology (86.4–91.5%) to several isolates of BCMV-infecting different crops from several countries based on partial sequence of CP, whereas based on partial sequence of CI the highest homology (87.3%) was to BCMV-RU1-P. This is the first report of BCMV infection on jack bean in Bogor, West Java, Indonesia.
DETEKSI SECARA SEROLOGI DAN MOLEKULER BEBERAPA JENIS VIRUS YANG BERASOSIASI DENGAN PENYAKIT MOSAIK TANAMAN NILAM (Pogostemon cablin Benth) MIFTAKHUROHMAH MIFTAKHUROHMAH; GEDE SUASTIKA; TRI ASMIRA DAMAYANTI
Jurnal Penelitian Tanaman Industri Vol 19, No 3 (2013): September 2013
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jlittri.v19n3.2013.130-138

Abstract

ABSTRAKPenyakit mosaik pada tanaman nilam disebabkan oleh beberapajenis virus, yaitu Potyvirus, Potexvirus, Cucumber mosaic virus (CMV),dan Broad bean wilt virus 2 (BBWV2). Penelitian ini bertujuan untukmengidentifikasi secara serologi dan molekuler virus-virus yangberasosiasi dengan gejala mosaik pada nilam di KP. Manoko, KP. Cicurugdan lahan petani di Cijeruk. Sampel daun nilam baik yang menunjukkangejala mosaik atau pun tidak diambil dari setiap lokasi penanamanmasing–masing sebanyak 30 sampel. Kejadian penyakit ditentukan melaluideteksi serologi dengan Direct-ELISA dan Indirect-ELISA terhadap sampelmenggunakan empat antiserum, yaitu CMV, Cymbidium mosaic virus(CymMV), Potyvirus, dan BBWV2. Deteksi molekuler dengan RT-PCRdilakukan untuk mengonfirmasi virus baru yang ditemukan. Hasilpenelitian menunjukkan bahwa gejala infeksi virus yang ditemukan padanilam bervariasi, yaitu mosaik lemah, mosaik kuning hijau, mosaik denganpenebalan, mosaik dengan malformasi daun, dan bintik kuning. Secaraserologi, kejadian virus pada setiap kebun bervariasi. Di KP Manoko,Potyvirus dan BBWV2 lebih dominan (100%) dibandingkan CymMV. DiKP Cicurug, kejadian Potyvirus dan CMV terlihat lebih dominan (83,3 dan80%) dibandingkan BBWV2 dan CymMV, sedangkan di Cijeruk, BBWV2lebih dominan (90%) dari Potyvirus (50%) dan CMV (13,3%). Hasil RT-PCR dengan primer degenerate BBWV, diidentifikasi BBWV2 padasampel daun nilam dari Manoko, Cicurug, dan Cijeruk, sedangkan denganprimer general Potexvirus, diidentifikasi CymMV hanya dari sampel daunnilam dari asal Manoko. Hasil penelitian ini merupakan laporan pertamatentang BBWV2 dan CymMV pada tanaman nilam di Jawa Barat yangmengindikasikan bahwa virus merupakan kendala utama pada perbenihannilam yang harus segera diatasi.Kata kunci: BBWV2, CymMV, mosaik, Pogostemon cablin Benth, PCRABSTRACTMosaic symptoms on patchouli plant are associated with severalviruses, i.e. Potyvirus, Potexvirus, CMV, and BBWV2. The objective ofthe study was to detect virus(es) associated with mosaic symptoms onpatchouli at the the patchouli seed nurseries, in Manoko, Cicurug, andCijeruk. Thirty leaf samples either showing typical symptomatic mosaic orasymptomatic were taken from each location. Serological testing byDirect-ELISA and Indirect-ELISA using four antisera namely CMV,Cymbidium mosaic virus (CymMV), Potyvirus, and BBWV2 was carriedout to test the incidence of each virus. Molecular detection by RT-PCR wasperformed to confirm the new virus(es). The results showed that symptomsof virus infection were found vary, i.e. weak mosaic, green yellow mosaic,mosaic with thickening, mosaic with leaf malformations, and yellow spot.Based on the serological detection, virus(es) incidence varied at each seednurseries. In Manoko, Potyvirus, and BBWV2 were more dominant(100%) compared with CymMV. In Cicurug, Potyvirus and CMV weremore dominat (83.3 and 80%) compared with BBWV2 and CymMV.While in Cijeruk, BBWV2 was the most dominant (90%) than Potyvirus(50%) and CMV(13.3%). Result of RT-PCR with degenerate primers pairof BBWV was succesfully identified BBWV2 from Manoko, Cicurug, andCijeruk samples, whereas by using Potexvirus general primary, CymMVwas identified only from Manoko samples. BBWV2 and CymMV werefirst reported to infect patchouli in West Java. The result indicate thatvirus(es) are the major constraint on patchouli seed that should bemanaged immediately.Key words: BBWV2, CymMV, mosaic, Pogostemon cablin Benth, PCR
Molecular Characterization of Begomovirus on Cucumber in Java Listihani Listihani; Tri Asmira Damayanti; Sri Hendrastuti Hidayat; Suryo Wiyono
Jurnal Perlindungan Tanaman Indonesia Vol 23, No 2 (2019)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.41402

Abstract

A survey on several cucumber cultivation areas in West Java, Central Java, Yogyakarta, and East Java found many plants showing typical Begomovirus symptoms such as yellow mosaic, cupping, and vein banding. This study was aimed to determine disease frequency, detection and molecular characterization of the causal virus of those symptoms on cucumber in Java. Sampling was conducted by purposive sampling by collecting 50 symptomatic plants from each location in West Java (Indramayu, Subang, and Bogor), Central Java (Brebes and Klaten), Yogyakarta (Kulon Progo), and East Java (Nganjuk, Kediri, and Tulungagung). The detection and disease frequency was determined based on DIBA test using a specific antiserum of Tomato leaf curl New Delhi virus (ToLCNDV) and Squash leaf curl virus (SLCV). The identification of nucleic acid was conducted by PCR using specific primer of ToLCNDV and SLCV, DNA cloning, and sequencing. The results of serological detection showed the disease frequency of ToLCNDV and SLCV ranged from 92.77-100% and 78.33-93.3%, respectively. PCR using specific primer of ToLCNDV successfully amplified the coat protein gene at a size of 600 bp from all samples. Homology nucleotide and amino acid sequences among ToLCNDV Java isolate ranging from 95.6-99.2% and 99.7-100%. ToLCNDV isolates Java had highest nucleotide and amino acid sequences similarity with cucumber isolate from Klaten, Indonesia (AB613825) ranging from 96.1-98.1% and 99.7-100%, and was considered as “Indonesia” strain. SLCV not amplified on all samples by PCR using specific primer, indicating it might not present yet on cucumber in Java.
EFISIENSI PENULARAN VIRUS MOSAIK BENGKUANG DENGAN Aphis craccivora Koch. DAN A. gossypii Glover. Tri Asmira Damayanti; Endah Muliarti; Dewi Sartiami
Agrovigor Vol 3, No 2 (2010): SEPTEMBER
Publisher : Universitas Trunojoyo Madura

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21107/agrovigor.v3i2.262

Abstract

Yambean mosaic virus is the most important virus infecting yam bean in Indonesia. The virus were transmitted either mechanically or via aphid.  This study to test the transmission efficiency of the virus via A. craccivora and A. gossypii by using different number of aphid such 1, 3, 5, 7, and 10 for each treatment. To determine the transmission efficiency, incubation period, type of symptom and incidence were used as parameter. Transmission of virus by A. craccivora showed incidence range 90 to 100%, significant differences in incubation time of 1 aphid compared to other treatments and showed severe leaf mosaic, vein-banding and severe leaf malformation such as string. However, the incidence of transmission of virus by A. gossypii was range 70 to 100%, with longer incubation period in compare with A. craccivora. There was no significant differences of incubation period among treatments by A. gossypii. The infected plants showed leaf malformation, vein-banding, wrinkle and blotch on the leaves. Based on these, both aphids species could transmitted virus efficiently, and among them A. craccivora considerate has higher ability as efficient insect vector to transmit the virus in compare with A. gossypii.
Deteksi dan Identifikasi Potyvirus pada Ubi Jalar di Tana Toraja, Sulawesi Selatan Laras Anjarsari; Gede Suastika; Tri Asmira Damayanti
Jurnal Fitopatologi Indonesia Vol. 9 No. 6 (2013)
Publisher : The Indonesian Phytopathological Society (Perhimpunan Fitopatologi Indonesia)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (603.803 KB) | DOI: 10.14692/jfi.9.6.193

Abstract

Typical viral symptoms including chlorotic and uneven interveinal yellowing on leaves without leaf malformation was observed on sweet potato field in Tana Toraja, South Sulawesi. To identify the causal of the disease, reverse transcription-polymerase chain reaction (RT-PCR) and DNA sequencing were carried out to detect the virus from infected plants. RT-PCR using universal primer for C1 gene of Potyvirus was successfully amplified approximately 683bp DNA fragment. The nucleotide sequences of this C1 gene fragment showed 98% homology to Sweet potato virus G (SPVG). Amplification using specific primer for coat protein (CP) gene of SPVG followed by DNA sequencing confirmed the association of SPVG from infected plants. Further nucleotide analysis shwowed that SPVG isolate from Tana Toraja had 99.2% homology to isolate from Japan. This is the firstt report of SPVG infection on sweet potato in South Sulawesi.
Utilization of Root-Colonizing Bacteria to Protect Hot-Pepper Against Tobacco Mosaic Tobamovirus TRI ASMIRA DAMAYANTI; HENDRA PARDEDE; NISA RACHMANIA MUBARIK
HAYATI Journal of Biosciences Vol. 14 No. 3 (2007): September 2007
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (56.048 KB) | DOI: 10.4308/hjb.14.3.105

Abstract

Tobacco Mosaic Tobamovirus (TMV) is one of many important viruses infecting Solanaceous plants including hot pepper in Indonesia. To accomplish and improve the effectiveness of virus management, we used root-colonizing bacteria (rhizobacteria) isolated from healthy hot pepper. Eight rhizobacteria isolates were selected and their capacity in enhancing plant growth and inducing systemic resistance (ISR) against TMV in greenhouse trials were evaluated. The rhizobacteria was applied as seed treatment and soil drench. Bacterized-seedling showed a better growth vigor, fitness and a milder symptom than non-bacterized control plants. The protective effect of rhizobacteria was more pronounced after challenging inoculation by TMV, especially for plants treated by isolates I-6, I-16, and I-35. However, TMV accumulation was slightly affected by bacterial treatment. The rhizobacteria might improved ISR by increasing peroxidase enzyme activity but this depends on the species. Based on whole results, isolate I-35 was the potential plant growth promotion rhizobacteria (PGPR). The I-35 was identified as Bacillus cereus based on morphological characteristics and nucleotide sequences of 16S r-RNA. Key words: root-colonizing bacteria, TMV, ISR
Penyakit Bercak Kuning Ubi Jalar di Bogor, Jawa Barat Anastasya Hondo; Kartika Catur Damaiyanti; Muhammad Fikri Hafizh; Niky Elfa Amanatillah; Tri Asmira Damayanti
Jurnal Fitopatologi Indonesia Vol. 14 No. 2 (2018)
Publisher : The Indonesian Phytopathological Society (Perhimpunan Fitopatologi Indonesia)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (357.9 KB) | DOI: 10.14692/jfi.14.2.69

Abstract

Yellow Spot Disease on Sweet Potato in Bogor, West JavaYellow spot symptom on sweet potato red cultivar was observed in Cikarawang and Leweung kolot area, Bogor regency. Symptomatic leaf samples gave positive reaction in serological detection by DIBA method using polyclonal antiserum for Potyvirus. DNA fragment of ± 700 bp in length was successfully amplified by RT-PCR using degenerate primer for cylindrical inclusion gene (CI) of Potyvirus.  Further nucleotide sequence analysis indicated high homology to Sweet potato feathery  mottle virus (SPFMV). The homology of SPFMV Cikarawang (Crkw) isolate ranging from 89–98% to several isolates from other countries with the highest homology to Japan and Spain isolates. SPFMV Crkw isolate was in the same cluster with the Japan and Spain isolates based on phylogenetic analysis of both nucleotide and amino acid sequences. This is the first report of SPFMV on sweet potato in Bogor, West Java, Indonesia.