scholarly journals VIRUS NILAM: IDENTIFIKASI, KARAKTER BIOLOGI DAN FISIK, SERTA UPAYA PENGENDALIANNYA

2015 ◽  
Vol 34 (1) ◽  
pp. 1 ◽  
Author(s):  
Miftakhurohmah Miftakhurohmah ◽  
Rita Noveriza
Keyword(s):  
Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Tobacco Necrosis Virus ◽  
Rt Pcr ◽  
Cymbidium Mosaic Virus ◽  
Peanut Stripe Virus ◽  
Mild Mosaic Virus ◽  
Wilt Virus ◽  
Broad Bean Wilt Virus

Infeksi virus pada tanaman nilam dapat menyebabkan penurunan produksi dan kualitas minyak. Sembilan jenis virus diidentifikasi menginfeksi tanaman nilam, yaitu Patchouli mosaic virus (PatMoV), Patchouli mild mosaic virus (PatMMV), Telosma mosaic virus (TeMV), Peanut stripe virus (PStV), Patchouli yellow mosaic virus (PatYMV), Tobacco necrosis virus (TNV), Broad bean wilt virus 2 (BBWV2), Cucumber mosaic virus (CMV), dan Cymbidium mosaic virus (CymMV). Kesembilan virus tersebut memiliki genom RNA, tetapi panjang dan bentuk partikelnya berbeda. Deteksi dan identifikasi berdasarkan bagian partikel virus dapat dilakukan secara serologi dengan teknik ELISA dan secara molekuler dengan RT-PCR. Gejala awal tanaman nilam terserang virus yaitu mosaik atau belang pada daun pucuk dan pada gejala berat tanaman menjadi kerdil. Infeksi virus dapat bersifat tunggal, tetapi ada pula infeksi oleh beberapa virus. Virus menular secara mekanis dan sebagian melalui penyambungan dan vektor. TNV, BBWV2, dan CMV memiliki kisaran inang yang luas, sedangkan virus yang lain inangnya terbatas. Virus nilam umumnya memiliki titik panas inaktivasi dan titik batas pengenceran yang tinggi, sedangkan ketahanan in vitro tidak stabil. Pendekatan terbaik pengendalian virus ialah menggunakan bahan tanaman bebas virus atau tahan virus dan pengendalian vektor. Tanaman bebas virus dapat diperoleh melalui kultur meristem, sedangkan pengendalian vektor dapat menggunakan pestisida nabati atau kimia.

scholarly journals IDENTIFIKASI MOLEKULER BROAD BEAN WILT VIRUS 2 (BBWV2) DAN CYMBIDIUM MOSAIC VIRUS (CYMMV) ASAL TANAMAN NILAM (POGOSTEMON CABLIN BENTH.)

2016 ◽  
Vol 15 (2) ◽  
pp. 188
Author(s):  
Miftakhurohmah . ◽  
Gede Suastika ◽  
Tri Asmira Damayanti ◽  
Rita Noveriza
Keyword(s):  
South Korea ◽  
Mosaic Virus ◽  
Broad Bean ◽  
Rna Extraction ◽  
Rt Pcr ◽  
West Java ◽  
Cymbidium Mosaic Virus ◽  
Pogostemon Cablin ◽  
Wilt Virus ◽  
Broad Bean Wilt Virus

Molecular identification Broad Bean Wilt Virus 2 (BBWV2) and Cymbidium Mosaic Virus (CymMV) from patchouli plant (Pogostemon cablin Benth.). Several viruses have been reported to be associated with mosaic disease on patchouli plant in Indonesia. This study aims to identify the two viruses in patchouli cultivation in West Java by studying the molecular characterization. Mosaic symptomatic leaf samples taken from patchouli cultivation in Manoko (Bandung Barat District, West Java Province). RNA extraction was performed using Xprep Plant RNA mini kit. RNA amplification with RT-PCR technique using primers for the cp gene region of BBWV2 and CymMV. The PCR product was sent to PT. Science Genetics Indonesia to do sequencing, then analyzed nucleotide sequences. Results of RT-PCR were performed successfully obtained DNA bands with size accordance with the predictions of the primer design for BBWV2 and CymMV cp region. Further, based on nucleotide and amino acid sequence analyses, the two virus isolates were confirmed as BBWV2 and CymMV respectively. Phylogenetic analyses revealed that BBWV2 Manoko clustered with BBWV2 from Singapore (original host of Brazilian red-cloak), China (pepper) and South Korea (chili). Whereas, CymMV Manoko become one cluster with CymMV from India (Phaius sp.), Indonesia (Dendrobium), China (vanilla), Thailand (Oncidium), Hawai (Dendrobium) and South Korea Cymbidium).

scholarly journals DETEKSI SECARA SEROLOGI DAN MOLEKULER BEBERAPA JENIS VIRUS YANG BERASOSIASI DENGAN PENYAKIT MOSAIK TANAMAN NILAM (Pogostemon cablin Benth)

2020 ◽  
Vol 19 (3) ◽  
pp. 130
Author(s):  
MIFTAKHUROHMAH MIFTAKHUROHMAH ◽  
GEDE SUASTIKA ◽  
TRI ASMIRA DAMAYANTI
Keyword(s):  
Mosaic Virus ◽  
Indirect Elisa ◽  
Rt Pcr ◽  
Degenerate Primers ◽  
Cymbidium Mosaic Virus ◽  
Major Constraint ◽  
Pogostemon Cablin ◽  
Direct Elisa ◽  
Wilt Virus ◽  
Broad Bean Wilt Virus

<p>ABSTRAK<br />Penyakit mosaik pada tanaman nilam disebabkan oleh beberapa<br />jenis virus, yaitu Potyvirus, Potexvirus, Cucumber mosaic virus (CMV),<br />dan Broad bean wilt virus 2 (BBWV2). Penelitian ini bertujuan untuk<br />mengidentifikasi secara serologi dan molekuler virus-virus yang<br />berasosiasi dengan gejala mosaik pada nilam di KP. Manoko, KP. Cicurug<br />dan lahan petani di Cijeruk. Sampel daun nilam baik yang menunjukkan<br />gejala mosaik atau pun tidak diambil dari setiap lokasi penanaman<br />masing–masing sebanyak 30 sampel. Kejadian penyakit ditentukan melalui<br />deteksi serologi dengan Direct-ELISA dan Indirect-ELISA terhadap sampel<br />menggunakan empat antiserum, yaitu CMV, Cymbidium mosaic virus<br />(CymMV), Potyvirus, dan BBWV2. Deteksi molekuler dengan RT-PCR<br />dilakukan untuk mengonfirmasi virus baru yang ditemukan. Hasil<br />penelitian menunjukkan bahwa gejala infeksi virus yang ditemukan pada<br />nilam bervariasi, yaitu mosaik lemah, mosaik kuning hijau, mosaik dengan<br />penebalan, mosaik dengan malformasi daun, dan bintik kuning. Secara<br />serologi, kejadian virus pada setiap kebun bervariasi. Di KP Manoko,<br />Potyvirus dan BBWV2 lebih dominan (100%) dibandingkan CymMV. Di<br />KP Cicurug, kejadian Potyvirus dan CMV terlihat lebih dominan (83,3 dan<br />80%) dibandingkan BBWV2 dan CymMV, sedangkan di Cijeruk, BBWV2<br />lebih dominan (90%) dari Potyvirus (50%) dan CMV (13,3%). Hasil RT-<br />PCR dengan primer degenerate BBWV, diidentifikasi BBWV2 pada<br />sampel daun nilam dari Manoko, Cicurug, dan Cijeruk, sedangkan dengan<br />primer general Potexvirus, diidentifikasi CymMV hanya dari sampel daun<br />nilam dari asal Manoko. Hasil penelitian ini merupakan laporan pertama<br />tentang BBWV2 dan CymMV pada tanaman nilam di Jawa Barat yang<br />mengindikasikan bahwa virus merupakan kendala utama pada perbenihan<br />nilam yang harus segera diatasi.<br />Kata kunci: BBWV2, CymMV, mosaik, Pogostemon cablin Benth, PCR</p><p>ABSTRACT<br />Mosaic symptoms on patchouli plant are associated with several<br />viruses, i.e. Potyvirus, Potexvirus, CMV, and BBWV2. The objective of<br />the study was to detect virus(es) associated with mosaic symptoms on<br />patchouli at the the patchouli seed nurseries, in Manoko, Cicurug, and<br />Cijeruk. Thirty leaf samples either showing typical symptomatic mosaic or<br />asymptomatic were taken from each location. Serological testing by<br />Direct-ELISA and Indirect-ELISA using four antisera namely CMV,<br />Cymbidium mosaic virus (CymMV), Potyvirus, and BBWV2 was carried<br />out to test the incidence of each virus. Molecular detection by RT-PCR was<br />performed to confirm the new virus(es). The results showed that symptoms<br />of virus infection were found vary, i.e. weak mosaic, green yellow mosaic,<br />mosaic with thickening, mosaic with leaf malformations, and yellow spot.<br />Based on the serological detection, virus(es) incidence varied at each seed<br />nurseries. In Manoko, Potyvirus, and BBWV2 were more dominant<br />(100%) compared with CymMV. In Cicurug, Potyvirus and CMV were<br />more dominat (83.3 and 80%) compared with BBWV2 and CymMV.<br />While in Cijeruk, BBWV2 was the most dominant (90%) than Potyvirus<br />(50%) and CMV(13.3%). Result of RT-PCR with degenerate primers pair<br />of BBWV was succesfully identified BBWV2 from Manoko, Cicurug, and<br />Cijeruk samples, whereas by using Potexvirus general primary, CymMV<br />was identified only from Manoko samples. BBWV2 and CymMV were<br />first reported to infect patchouli in West Java. The result indicate that<br />virus(es) are the major constraint on patchouli seed that should be<br />managed immediately.<br />Key words: BBWV2, CymMV, mosaic, Pogostemon cablin Benth, PCR</p>

scholarly journals Widespread Occurrence of Wheat spindle streak mosaic virus in Belgium

Plant Disease ◽  
2006 ◽  
Vol 90 (6) ◽  
pp. 723-728 ◽  
Author(s):  
C. Vaïanopoulos ◽  
A. Legrève ◽  
C. Lorca ◽  
V. Moreau ◽  
S. Steyer ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Real Time ◽  
The United States ◽  
Yellow Mosaic Virus ◽  
Virus Concentration ◽  
Rt Pcr ◽  
Enzymelinked Immunosorbent Assay ◽  
Mild Mosaic Virus ◽  
First Time ◽  
Virus Quantification

In order to assess the occurrence of Wheat spindle streak mosaic virus (WSSMV) in Belgium, a reverse-transcription polymerase chain reaction (RT-PCR) was developed, targeting WSSMV isolates from Canada, France, Germany, Italy, and the United States. The primers also were designed for virus quantification by real-time RT-PCR with SYBR-Green. No cross-reaction with soilborne cereal viruses such as Barley mild mosaic virus, Barley yellow mosaic virus, Soilborne cereal mosaic virus, and Soil-borne wheat mosaic virus was observed. The RT-PCR and real-time quantitative RT-PCR allowed a more sensitive detection of WSSMV than enzymelinked immunosorbent assay. The incidence of WSSMV in Belgium was evaluated using a bioassay with wheat cvs. Cezanne and Savannah and rye cv. Halo, grown in 104 Belgian soils. The presence of WSSMV was detected from plants grown in 32% of the soils. The RT-PCR methods developed here, combined with large sampling, allowed WSSMV to be detected for the first time in Belgium. The real-time quantitative RT-PCR was developed as a tool for evaluating the resistance to WSSMV by quantifying the virus concentration in wheat cultivars.

scholarly journals First Report of Cucumber mosaic virus on Vigna marina in Taiwan

Plant Disease ◽  
2010 ◽  
Vol 94 (10) ◽  
pp. 1267-1267 ◽  
Author(s):  
T.-C. Deng ◽  
C.-H. Tsai ◽  
H.-L. Tsai ◽  
J.-Y. Liao ◽  
W.-C. Huang
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Broad Bean ◽  
Solomon Islands ◽  
Natural Host ◽  
Vector System ◽  
Rt Pcr ◽  
First Report ◽  
Wilt Virus ◽  
Broad Bean Wilt Virus

Vigna marina (Burm.) Merr., the dune bean or notched cowpea, is a tropical creeping vine that grows on sand dunes along the coastal regions of Taiwan. Although V. marina is a weed, some varieties are also grown for fodder and food. This legume is a natural host of Bean common mosaic virus in the Solomon Islands (1) and Alfalfa mosaic virus or Beet western yellows virus in Australia (2). In April 2009, plants of V. marina showing severe mosaic and chlorotic ringspots on the foliage were found in the coastal region of Hualien County in eastern Taiwan. Indirect ELISA on a single diseased plant showed positive results with antibodies against the cucumber isolate of Cucumber mosaic virus (CMV) but negative to Broad bean wilt virus-1, Broad bean wilt virus-2, and some potyviruses (Agdia Inc., Elkhart, IN). A pure isolate of CMV was obtained from V. marina through three successive passages of single lesion isolation in sap-inoculated Chenopodium quinoa. Results of mechanical inoculations showed that the CMV-V. marina isolate was successfully transmitted to C. amaranticolor, C. murale, C. quinoa, Chrysanthemum coronarium, Gomphrena globosa, Nicotiana benthamiana, N. tabacum cv. Vam-Hicks, Phaseolus limensis, P. lunatus, P. vulgaris, Tetragonia tetragonioides, V. marina, V. radiata, and V. unguiculata subsp. sesquipedalis. These results of artificial inoculations were confirmed by ELISA. Homologous reactions of the CMV-V. marina isolate with a stock polyclonal antiserum against the CMV-cucumber isolate (4) were observed in sodium dodecyl sulfate-immunodiffusion. To determine the specific CMV subgroup, total RNA was extracted from inoculated leaves of C. quinoa using the Total Plant RNA Extraction Miniprep System (Viogene, Sunnyvale, CA). A DNA fragment of 940 bp covering the 3′ end of the coat protein gene and C-terminal noncoding region of RNA-3 was amplified using the Cucumovirus-specific primers (3) after reverse transcription (RT)-PCR with AccuPower RT/PCR PreMix Kit (Bioneer, Daejeon, Korea). The product was gel purified by Micro-Elute DNA/Clean Extraction Kit (GeneMark Technology Co., Tainan, Taiwan) and cloned in yT&A Cloning Vector System (Yeastern Biotech Co., Taipei, Taiwan) for sequencing (Mission Biotech Co., Taipei, Taiwan) and the sequence was submitted to GenBank (No. HM015286). Pairwise comparisons of the sequence of CMV-V. marina isolate with corresponding sequences of other CMV isolates revealed the maximum (95 to 96%) nucleotide identities with CMV subgroup IB isolates (strains Nt9 and Tfn) compared with 94 to 95% identities with subgroup IA isolates (strains Y and Fny) or 77 to 78% identities with subgroup II (strains LS and Q). These results suggest that CMV is the causal agent for the mosaic disease of V. marina in Taiwan and the isolate belongs to subgroup I. To our knowledge, this is the first report of V. marina as a natural host of CMV. This strain of CMV with specific pathogenicity could threaten crop production in the coastal zones. In addition, V. marina associated with native coastal vegetation was injured by CMV infection, which might lead to ecological impacts on shoreline fading. References: (1) A. A. Brunt. Surveys for Plant Viruses and Virus Diseases in Solomon Islands. FAO, Rome, 1987. (2) C. Büchen-Osmond, ed. Viruses of Plants in Australia. Retrieved from http://www.ictvdb.rothamsted.ac.uk/Aussi/aussi.htm . September, 2002. (3) S. K. Choi et al. J. Virol. Methods 83:67, 1999. (4) S. H. Hseu et al. Plant Prot. Bull. (Taiwan) 29:233, 1987.

scholarly journals Detection of Banana Mild Mosaic Virus in Musa In Vitro Plants: High-Throughput Sequencing Presents Higher Diagnostic Sensitivity Than (IC)-RT-PCR and Identifies a New Betaflexiviridae Species

Plants ◽  
2022 ◽  
Vol 11 (2) ◽  
pp. 226
Author(s):  
Marwa Hanafi ◽  
Wei Rong ◽  
Lucie Tamisier ◽  
Chadi Berhal ◽  
Nicolas Roux ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Diagnostic Sensitivity ◽  
Rapid Screening ◽  
Mild Mosaic ◽  
Rt Pcr ◽  
In Vitro Plants ◽  
Mild Mosaic Virus

: The banana mild mosaic virus (BanMMV) (Betaflexiviridae, Quinvirinae, unassigned species) is a filamentous virus that infects Musa spp. and has a very wide geographical distribution. The current BanMMV indexing process for an accession requires the testing of no less than four plants cultivated in a greenhouse for at least 6 months and causes a significant delay for the distribution of the germplasm. We evaluated the sensitivity of different protocols for BanMMV detection from in vitro plants to accelerate the testing process. We first used corm tissues from 137 in vitro plants and obtained a diagnostic sensitivity (DSE) of only 61% when testing four plants per accession. After thermotherapy was carried out to eliminate BanMMV infection, the meristem was recovered and further grown in vitro. The same protocol was evaluated in parallel on the corm tissue surrounding the meristem, as a rapid screening to evaluate virus therapy success, and was compared to the results obtained following the standard protocol. The obtained results showed 28% false negatives when conducting testing from corm tissues, making this protocol unsuitable in routine processes. Furthermore, RT-PCR and high-throughput sequencing (HTS) tests were applied on tissues from the base (n = 39) and the leaves (n = 36). For RT-PCR, the average DSE per sample reached 65% from either the base or leaves. HTS was applied on 36 samples and yielded 100% diagnostic specificity (DSP) and 100% DSE, whatever the sampled tissue, allowing the identification of a new Betaflexiviridae species infecting Musa. These results suggest that a reliable diagnostic of BanMMV from in vitro plants using RT-PCR or HTS technologies might represent an efficient alternative for testing after greenhouse cultivation.

scholarly journals First Report of Barley yellow mosaic virus in Barley in Spain

Plant Disease ◽  
2005 ◽  
Vol 89 (1) ◽  
pp. 105-105 ◽  
Author(s):  
M. A. Achon ◽  
M. Marsiñach ◽  
C. Ratti ◽  
C. Rubies-Autonell
Keyword(s):  
Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Complete Agreement ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
First Report ◽  
Barley Yellow Mosaic Virus ◽  
Pcr Products ◽  
Mild Mosaic Virus ◽  
Polymerase Chain

Recently, the presence of Barley mild mosaic virus (BaMMV) and the weakly serological detection of Barley yellow mosaic virus (BaYMV) were reported in Spain (1); both viruses are members of the genus Bymovirus (family Potyviridae). Random and symptomatic surveys were conducted during February and March of 2003 in barley fields in northeastern Spain to determine the occurrence of BaMMV and BaYMV. Leaves from 316 samples collected in 15 fields were analyzed using enzyme-linked immunosorbent assay (ELISA) with commercial antisera specific for BaYMV and BaMMV (Loewe Biochemica, Munich) as well as antisera against both viruses (provided by T. Klumen). Positive ELISA samples were further analyzed using reverse transcription-polymerase chain reaction (RT-PCR) with specific primers that amplify 445 bp of BaMMV and 433 bp of BaYMV (2). Complete agreement was observed between the ELISA and RT-PCR results. Mixed infections of BaYMV and BaMMV were detected in 10 samples, BaYMV in 5 samples and BaMMV in 3 samples. Samples positive for both viruses that exhibited clear mosaic symptoms were collected in two fields. RT-PCR products from five BaYMV-infected samples were cloned and sequenced and showed 96 to 98% identity to BaYMV isolates previously reported from Europe (Genbank Accession Nos. AJ1515479-85 and X95695-7) and 92 to 95% identity with isolates reported from Asia (GenBank Accession Nos. AB023585-96, AJ132268, AJ224619-22, AJ224624-28, AF536944-46, AF536948-58, D01091, D00544, and Z24677). Sequence identity of Spanish isolates were 96 to 99%. To our knowledge, this is the first report of BaYMV infecting barley in Spain and illustrates the association of both Bymoviruses infecting barley. References: (1) M. A. Achon et al. Plant Dis. 87:1004, 2003. (2) D. Hariri et al. Eur. J. Plant Pathol. 106:365, 2000.

scholarly journals Towards plant resistance to viruses using protein-only RNase P

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Anthony Gobert ◽  
Yifat Quan ◽  
Mathilde Arrivé ◽  
Florent Waltz ◽  
Nathalie Da Silva ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Virus Replication ◽  
Yield Loss ◽  
Rnase P ◽  
Plant Viruses ◽  
Yellow Mosaic Virus ◽  
Resistance To Viruses ◽  
Target Plant

AbstractPlant viruses cause massive crop yield loss worldwide. Most plant viruses are RNA viruses, many of which contain a functional tRNA-like structure. RNase P has the enzymatic activity to catalyze the 5′ maturation of precursor tRNAs. It is also able to cleave tRNA-like structures. However, RNase P enzymes only accumulate in the nucleus, mitochondria, and chloroplasts rather than cytosol where virus replication takes place. Here, we report a biotechnology strategy based on the re-localization of plant protein-only RNase P to the cytosol (CytoRP) to target plant viruses tRNA-like structures and thus hamper virus replication. We demonstrate the cytosol localization of protein-only RNase P in Arabidopsis protoplasts. In addition, we provide in vitro evidences for CytoRP to cleave turnip yellow mosaic virus and oilseed rape mosaic virus. However, we observe varied in vivo results. The possible reasons have been discussed. Overall, the results provided here show the potential of using CytoRP for combating some plant viral diseases.

scholarly journals Metagenomic Analysis of Marigold: Mixed Infection Including Two New Viruses

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1254
Author(s):  
Hang Yin ◽  
Zheng Dong ◽  
Xulong Wang ◽  
Shuhao Lu ◽  
Fei Xia ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Mosaic Virus ◽  
Complete Genome ◽  
Plum Pox Virus ◽  
Cryptic Virus ◽  
Sequence Identity ◽  
Complete Genome Sequence Analysis ◽  
Wilt Virus ◽  
Broad Bean Wilt Virus

Marigold plants with symptoms of mosaic, crinkle, leaf curl and necrosis were observed and small RNA and ribo-depleted total RNA deep sequencing were conducted to identify the associated viruses. Broad bean wilt virus 2, cucumber mosaic virus, turnip mosaic virus, a new potyvirus tentatively named marigold mosaic virus (MMV) and a new partitivirus named as marigold cryptic virus (MCV) were finally identified. Complete genome sequence analysis showed MMV was 9811 nt in length, encoding a large polyprotein with highest aa sequence identity (57%) with the putative potyvirus polygonatumkingianum virus 1. Phylogenetic analysis with the definite potyviruses based on the polyprotein sequence showed MMV clustered closest to plum pox virus. The complete genome of MCV comprised of dsRNA1 (1583 bp) and dsRNA2 (1459 bp), encoding the RNA-dependent RNA polymerase (RdRp), and coat protein (CP), respectively. MCV RdRp shared the highest (75.7%) aa sequence identity with the unclassified partitivirus ambrosia cryptic virus 2, and 59.0%, 57.1%, 56.1%, 54.5% and 33.7% with the corresponding region of the definite delta-partitiviruses, pepper cryptic virus 2, beet cryptic virus 3, beet cryptic virus 2, pepper cryptic virus 1 and fig cryptic virus, respectively. Phylogenetic analysis based on the RdRp aa sequence showed MCV clustered into the delta-partitivirus group. These findings enriched our knowledge of viruses infecting marigold, but the association of the observed symptom and the identified viruses and the biological characterization of the new viruses should be further investigated.

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