scholarly journals Pengaruh Elektroterapi dan Termoterapi secara in Vitro terhadap Eliminasi Onion yellow dwarf virus

2018 ◽  
Vol 13 (6) ◽  
pp. 199 ◽  
Author(s):  
Siti Shofiya Nasution ◽  
Diny Dinarti ◽  
Sri Hendrastuti Hidayat

Infection of Onion yellow dwarf virus (OYDV) are reported causing problems in garlic production. Planting virus-free bulbs might help reduce viral disease incidence in the field. This research was aimed to develop method for eliminating OYDV from garlic bulbs using combination of electrotherapy (0, 5, 10, 15, and 20 mA each for 10 minutes) and thermotherapy (23, 28, 33, 38°C each for 4 weeks). Two garlic cultivars, i.e. Sangga Sembalun and Lumbu Hijau were used as seed bulbs for OYDV elimination tests. Virus infection was confirmed using transcription-polymerase chain reaction (RT-PCR).  The result showed that thermotherapy at 33 °Cwas the best method to eliminate OYDV in garlic although the efficiency was not the same for all cultivars. The efficiency reached up to 60% for cv. Lumbu Hijau, whereas for cv. Sangga Sembalun only reached up to 40%. Electrotherapy alone or in combination with thermotherapy were not able to produce OYDV-free plantlets.

2003 ◽  
Vol 93 (11) ◽  
pp. 1386-1392 ◽  
Author(s):  
Boovaraghan Balaji ◽  
Dennis B. Bucholtz ◽  
Joseph M. Anderson

Reliable detection and quantification of barley and cereal yellow dwarf viruses (YDVs) is a critical component in managing yellow dwarf diseases in small grain cereal crops. The method currently used is enzyme-linked immunosorbent assay (ELISA), using antisera against the coat proteins that are specific for each of the various YDVs. Recently, quantitative real-time reverse-transcription polymerase chain reaction (Q-RT-PCR) has been used to detect bacterial and viral pathogens and to study gene expression. We applied this technique to detect and quantify YDVs using primers specific for Barley yellow dwarf virus-PAV (BYDV-PAV) and Cereal yellow dwarf virus-RPV (CYDV-RPV) coat protein genes because of the higher sensitivity of RT-PCR and the advantage of using a real-time PCR instrument. This Q-RT-PCR was used to detect BYDV and CYDV, and to examine disease development in a resistant wheatgrass, a resistant wheat line, a susceptible wheat line, and a susceptible oat line. BYDV-PAV and CYDV-RPV were detected as early as 2 and 6 h, respectively, in susceptible oat compared with detection by ELISA at 4 and 10 days postinoculation. BYDV-PAV RNA accumulated more rapidly and to a higher level than CYDV-RPV RNA in both oat and wheat, which may account for PAV being more prevalent and causing more severe viral disease than CYDV. Q-RT-PCR is reproducible, sensitive, and has the potential to be used for examining yellow dwarf disease and as a rapid diagnostic tool for YDVs.


2021 ◽  
Vol 25 (1) ◽  
pp. 40
Author(s):  
Nurenik Nurenik ◽  
Sedyo Hartono ◽  
Sri Sulandari ◽  
Susamto Somowiyarjo ◽  
Argawi Kandito

Viruses have been a problem on garlic cultivations in various countries. There are several viruses reported infecting garlic. Genera Potyvirus and Carlavirus are the most common viruses found infecting garlic. Mixed infection on garlic is often designated as a “garlic viral complex”. These viruses can be transmitted through imported garlic seeds. Therefore, it is necessary to conduct early detection of garlic seeds to prevent the epidemic of these viruses. This study aimed to detect Onion yellow dwarf virus (OYDV) and Shallot latent virus (SLV) on garlic. Garlic samples were obtained from Enrekang, Magelang, Temanggung, Tawangmangu, and Yogyakarta. Total RNA was extracted from the samples and subsequently used for RT-PCR using two pairs of specific primers SLV-F/SLV-R and OYDV-F/OYDV-R. Primary pair SLV-F/SLV-R in amplicons sized 276 bp, while OYDV-F/OYDV-R in amplicons sized 112 bp. RT-PCR results showed that OYDV was found in all samples tested in this study. Meanwhile, double infections (OYDV and SLV) were found in eight out of ten samples tested. These results indicated that double infections on garlic were common in Indonesia.


2021 ◽  
Vol 61 (3) ◽  
pp. 214-220

Onion yellow dwarf virus is distributed worldwide significantly reducing yield of crops from the Allium genus. The aim of the study was the detection and molecular characterization of newly identified OYDV isolates infecting onions in Poland. The virus was detected by transmission electron microscopy and RT-PCR techniques using two pairs of diagnostic primers: OYDV-NibCPF1/R1 and OYDV-CPF2/R2. The specificity of obtained RT-PCR products was confirmed by Sanger sequencing and received viral coat protein sequence was used for phylogenetic analysis. The phylogenetic analysis was carried out using CP sequences of the new Polish onion isolate obtained in this study and 37 other sequences of OYDV retrieved from GenBank. The analysis revealed that the Polish OYDV isolate is the most similar to the OYDV isolates derived from onions from Argentina and Germany, which may indicate their common origin. Moreover, it was observed that the Polish onion and garlic isolates are very diverse and belong to different phylogroups.


2010 ◽  
Vol 21 (1) ◽  
pp. 64-68 ◽  
Author(s):  
Sandeep Kumar ◽  
V. K. Baranwal ◽  
Subodh Joshi ◽  
Meenakshi Arya ◽  
S. Majumder

Plant Disease ◽  
2003 ◽  
Vol 87 (4) ◽  
pp. 446-446 ◽  
Author(s):  
N. L. Robertson

The Matanuska-Susitna Valley is one of the most fertile regions in Alaska for growing cool-season vegetables. Barley (Hordeum vulgare) and oat (Avena sativa) crops are also sown for animal feed and green manure. The most damaging and widely distributed viral disease of small grains worldwide is barley yellow dwarf (BYD), caused by several species from two genera in the family Luteoviridae: luteovirus (Barley yellow dwarf virus [BYDV-MAV and BYDV-PAV]) and polerovirus (Cereal yellow dwarf virus [CYDV-RPV, formerly BYDV-RPV]) and three unassigned species (BYDV-RMV, BYDV-SGV, and BYDV-GPV) (2,4). Even though barley and oat have been grown in Alaska for more than 50 years, BYD has not been documented in small grains in this region. During September 2001, barley plants with bright yellow leaves were collected from five barley fields near Palmer. Three plants from each field were assayed using a reverse transcription-polymerase chain reaction (RT-PCR) protocol targeting members of the luteoviridae (3). The resulting ≈530-bp PCR product and its restriction fragment length polymorphism (RFLP) produced by digestion with NdeII implied that plants were infected with BYDV-PAV. In September 2002, three of the five sites were surveyed again for BYDV. Two of the fields (BF-1 and BF-2) had been replanted with barley and the other (OF-3) was planted with oats. Leaf samples from 36 symptomatic barley plants from each field and 60 symptomatic oat plants were randomly collected and stored at -80°C. In 2002, in addition to RT-PCR and RFLP analyses, enzyme-linked immunosorbent assays (ELISA) using Agdia kits (Agdia, Elkhart, IN) for BYDV-PAV, CYDV-RPV, and BYDV-SGV were also performed (1). First, RT-PCR and RFLP were completed on all samples using 0.5 g of tissue. Of samples from BF-1, BF-2, and OF-3, 61, 100, and 70%, respectively, generated luteoviridae-specific fragments. The RFLP profiles from barley were all PAV-like, whereas 71% of oat samples were PAV-like, and 29% were of an unknown pattern. No bands were observed from apparently healthy field plants. ELISA (0.2 g of tissue) was performed on all PCR-positive samples, resulting in 22, 97, and 33% detection for BYDV-PAV from BF-1, BF-2, and OF-3, respectively. An additional 29% of oat samples (OF-3) tested positive for CYDV-RPV, whereas none of the barley plants tested positive. One oat plant had a mixed infection with both PAV and RPV profiles, and all oat plants with the unidentified RFLP pattern were serologically positive for RPV. No BYDV-SGV was detected in either barley or oats. The PCR assay was clearly more sensitive than ELISA, especially for plants that had mature and necrotic tissue, which were predominately found in BF-1 and OF-3. Based on these direct tests on the coat protein's nucleic acid (PCR) and serology (ELISA), it is concluded that two distinct viruses, BYDV-PAV and CYDV-RPV, were found in oats, whereas only the former was found in barley. To my knowledge, this is the first report of luteovirus and polerovirus infection in small grains in Alaska. References: (1) M. F. Clark and A. N. Adams. J. Gen. Virol. 34, 475, 1977. (2) C. J. D'Arcy and P. A. Burnett. Barley Yellow Dwarf: 40 Years of Progress. The American Phytopathological Society, St. Paul, MN, 1995. (3) N. L. Robertson and R. French. J. Gen. Virol. 72,1473, 1991. (4) M. H. V. van Regenamortel et al. Virus Taxonomy. Seventh Report of the International Committee on Taxonomy of Viruses. Academic Press, NY, 2000.


2017 ◽  
Vol 8 (1) ◽  
pp. 22
Author(s):  
Aqlima , ◽  
Bambang S. Purwoko ◽  
Sri Hendrastuti Hidayat ◽  
Diny Dinarti

<p>ABSTRACT<br />Meristem tip culture is culture of isolated meristem with 1-2 leaf primordia on suitable medium. This method is generally used to obtain free virus plant. Optimation of plant growth regulators (PGRs) was done to accelerate explant growth without callus formation and to avoid somaclonal variation in meristem tip culture. The aims of this study were to achieve the best combination of PGR for meristem tip growth and to evaluate meristem tip culture potential for Onion yellow dwarf virus (OYDV) elimination in shallot. This study used combination of PGRs 0.25 mg L-1 (2-ip, BAP, GA3, kinetin) with or without 0.1 mg L-1 IAA and medium without PGR. This research consisted of two experiments conducted separately. In experiment I, cv. Bima Brebes was used and experiment II cv. Tiron was used. Each experiment was arranged in completely randomized block design with single factor (PGR combination) that has 8 combination levels and 3 replications. The result showed that medium without PGR was the most efficient for meristem tip growth. Primary shoot was growing without callus formation. RT-PCR analysis showed that all of the tested samples were still infected by OYDV. Meristem tip culture method did not eliminate OYDV in both cultivars.<br />Keywords: Auxin, cytokinin, GA3, OYDV, RT-PCR</p><p>ABSTRAK<br />Kultur meristem tip merupakan kultur meristem yang diisolasi 1-2 primordia daun dan pada media yang sesuai. Metode ini umum digunakan untuk mendapatkan tanaman bebas virus. Optimasi terhadap zat pengatur tumbuh (ZPT) dilakukan untuk mempercepat pertumbuhan eksplan tanpa disertai pembentukan kalus untuk menghindari terjadinya variasi somaklonal pada kultur meristem tip. Penelitian ini bertujuan untuk mendapatkan kombinasi ZPT terbaik bagi pertumbuhan meristem tip dan untuk mengevaluasi potensi kultur meristem tip dalam mengeliminasi virus Onion yellow dwarf virus (OYDV) pada tanaman bawang merah. Penelitian ini menggunakan 0.25 mg L-1 (2-ip, BAP, GA3, kinetin) dengan penambahan atau tanpa 0.1 mg L-1 IAA serta media tanpa ZPT. Penelitian ini terdiri atas 2 percobaan terpisah. Percobaan 1 menggunakan cv. Bima Brebes dan Percobaan 2 menggunakan cv. Tiron. Masing-masing percobaan disusun berdasarkan rancangan kelompok lengkap teracak (RKLT) dengan 1 faktor, yaitu kombinasi ZPT yang terdiri atas 8 taraf kombinasi dan 3 ulangan. Hasil yang didapat menunjukkan bahwa media tanpa penambahan ZPT<br />merupakan media yang paling efisien untuk pertumbuhan tunas meristem tip. Tunas utama tumbuh tanpa disertai pembentukan kalus. Hasil analisis RT-PCR menunjukkan bahwa seluruh sampel yang dideteksi masih terinfeksi OYDV. Metode kultur meristem tip belum dapat mengeliminasi virus OYDV pada kedua kultivar bawang merah.<br />Kata kunci: Auksin, GA3, OYDV, RT-PCR, sitokinin</p>


Blood ◽  
1997 ◽  
Vol 90 (2) ◽  
pp. 865-872 ◽  
Author(s):  
Ellen L.W. Kittler ◽  
Stefan O. Peters ◽  
Rowena B. Crittenden ◽  
Michelle E. Debatis ◽  
Hayley S. Ramshaw ◽  
...  

Using a murine bone marrow transplantation model, we evaluated the long-term engraftment of retrovirally transduced bone marrow cells in nonmyeloablated hosts. Male bone marrow was stimulated in a cocktail of interleukin-3 (IL-3), IL-6, IL-11, and stem cell factor (SCF ) for 48 hours, then cocultured on the retroviral producer line MDR18.1 for an additional 24 hours. Functional transduction of hematopoietic progenitors was detected in vitro by reverse transcriptase-polymerase chain reaction (RT-PCR) amplification of multiple drug resistance 1 (MDR1) mRNA from high proliferative potential-colony forming cell (HPP-CFC) colonies. After retroviral transduction, male bone marrow cells were injected into nonablated female mice. Transplant recipients received three TAXOL (Bristol-Myers, Princeton, NJ) injections (10 mg/kg) over a 14-month period. Transplant recipient tissues were analyzed by Southern blot and fluorescence in situ hybridization for Y-chromosome–specific sequences and showed donor cell engraftment of approximately 9%. However, polymerase chain reaction amplification of DNAs from bone marrow, spleen, and peripheral blood showed no evidence of the transduced MDR1 gene. RT-PCR analysis of total bone marrow RNA showed that transcripts from the MDR1 gene were present in a fraction of the engrafted donor cells. These data show functional transfer of the MDR1 gene into nonmyeloablated murine hosts. However, the high rates of in vitro transduction into HPP-CFC, coupled with the low in vivo engraftment rate of donor cells containing the MDR1 gene, suggest that the majority of stem cells that incorporated the retroviral construct did not stably engraft in the host. Based on additional studies that indicate that ex vivo culture of bone marrow induces an engraftment defect concomitantly with progression of cells through S phase, we propose that the cell cycle transit required for proviral integration reduces or impairs the ability of transduced cells to stably engraft.


Author(s):  

Abstract A new distribution map is provided for Onion yellow dwarf virus. Potyviridae: Potyvirus. Hosts: onion (Allium cepa) and garlic (Allium sativum). Information is given on the geographical distribution in Europe (Austria, Croatia, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Italy, mainland Italy, Sicily, Lithuania, Moldova, Netherlands, Poland, Portugal, Romania, Russia, Far East, Siberia, Serbia, Slovenia, Spain, UK, England and Wales and Ukraine), Asia (China, Henan, Hubei, Hunan, Jiangsu, Shandong, Yunnan, Zhejiang, India, Delhi, Gujarat, Haryana, Madhya Pradesh, Maharashtra, Indian Punjab, Rajasthan, Uttar Pradesh, Indonesia, Java, Iran, Israel, Japan, Pakistan, Taiwan, Turkey, Vietnam and Yemen), Africa (Egypt, Morocco, Nigeria and Sudan), North America (Canada, British Columbia, Nova Scotia, Ontario, Mexico, USA, California, Colorado, Florida, Idaho, Iowa, Minnesota, Nevada, New York, Oregon, Texas, Vermont, Washington and West Virginia), Central America and Caribbean (Cuba), South America (Argentina, Brazil, Bahia, Goias, Minas Gerais, Parana, Rio Grande do Sul, Santa Catarina, Sao Paulo, Chile, Ecuador and Urugay) and Oceania (Australia and New Zealand).


Sign in / Sign up

Export Citation Format

Share Document