scholarly journals Double Infection of Onion yellow dwarf virus and Shallot latent virus in Garlic from Several Regions in Indonesia

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.

Plant Disease ◽  
2010 ◽  
Vol 94 (7) ◽  
pp. 915-915 ◽  
Author(s):  
A. K. Torrico ◽  
E. E. Cafrune ◽  
V. C. Conci

Because of exclusively agamic propagation, garlic is commonly infected with a virus complex mainly composed of species within the genera Potyvirus, Allexivirus, and Carlavirus. This virus complex causes leaf striping that ranges from various shades of green to yellow and results in yield losses (2,4). Onion yellow dwarf virus, Leek yellow stripe virus (potyviruses), Garlic virus A, Garlic virus C (allexiviruses), and Garlic common latent virus (carlavirus) have been detected in Argentina previously (1,2). Recently, Shallot latent virus (SLV; another carlavirus) was detected in 25 of 30 garlic plants (cv. Morado) growing in four different fields near Córdoba, Argentina by double-antibody sandwich (DAS)-ELISA using BIOREBA (Reinach, Switzerland) antibodies. To confirm the presence of the virus, DAS-ELISA-positive plants were also analyzed by one-step reverse transcription (RT)-PCR using the Access RT-PCR system (Promega, Madison, WI) with specific primers reported by Tsuneyoshi et al. (3). RNA extractions were performed from 100 mg of leaves with the Qiagen RNeasy Plant Mini Kit (Qiagen, Valencia, CA). Primers used were Car-V1 (5′-AAACCTTTTGGTTCACTTTAGG-3′); Car-V2 (5′-AGGTGCATTGTTATCATTACTGG-3′); and Car-Cp3 (5′-GCGTGCTATATTTAAGTTGCATAC-3′). Primer pairs Car-V1/Car-Cp3 and Car-V2/Car-Cp3 were used for the amplification of the coat protein (CP) gene of SLV and an isolate of SLV formerly known as Garlic latent virus, respectively. Fragments of 992 bp and 1,079 bp were amplified with these primer pairs, respectively. The RT-PCR products were cloned with the TOPO TA Cloning Kit in the 3.9-kb pCR-TOPO vector (Qiagen). The nucleotide sequences of both fragments were determined and were found to be identical (GenBank No. GU355922) showing 94.2% nt sequence identity with the CP gene of an isolate of SLV from Indonesian garlic (GenBank No. AB004686) formerly referred to as Garlic latent virus (3). Consequently, the Argentinean virus is now considered a garlic isolate of SLV. References: (1) E. Cafrune et al. Plant Dis. 90:898, 2006. (2) V. C. Conci. Virus y Fitoplasmas de Ajo. Page 267 in: 50 Temas Sobre Producción de Ajo. Vol. 3. J. L. Burba, ed. Ediciones INTA, Mendoza, Argentina. 1997. (3) T. Tsuneyoshi et al. Arch. Virol. 143:1093, 1998. (4) D. G. A. Walkey and D. N. Antill. J. Hortic. Sci. 64:53, 1989.


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.


Plant Disease ◽  
2014 ◽  
Vol 98 (6) ◽  
pp. 858-858 ◽  
Author(s):  
M. Chodorska ◽  
E. Paduch-Cichal ◽  
E. Kalinowska ◽  
M. S. Szyndel

Garlic (Allium sativum L.) is vegetatively propagated and can be affected by a virus complex (1) consisting of two potyviruses, Onion yellow dwarf virus (OYDV) and Leek yellow stripe virus (LYSV), and two carlaviruses, Garlic common latent virus (GCLV) and Shallot latent virus (SLV) (2). OYDV, GCLV, and SLV are economically important viral pathogens of bulb garlic crops in many garlic-growing areas of the world. A general mosaic and yellowing of leaves of four garlic cultivars (Blanko, Harnaś, Jarus, and Mega) was observed in 11 garlic-production fields in the Lodz, Mazowieckie, Małopolska, and Pomorskie regions of Poland in July 2012. ELISA was carried out with extracts from 29 collected garlic leaf samples to detect OYDV, GCLV, and SLV using commercial antiserum (DSMZ, Braunschweig, Germany). Results indicated that 6 samples (20.7%) were infected with OYDV, 25 samples (86.2%) were infected with GCLV, and 23 samples (79.3%) were infected with SLV. The presence of these viruses in garlic leaf samples was confirmed by reverse transcription (RT)-PCR using total RNA extracted using the Spectrum Plant Total RNA kit (Sigma-Aldrich, Munich, Germany) and primers, designed in this study, specific to the whole coat protein gene of OYDV (OYDVF 5′-TAGGGTTGGATTATGATTTCTCGA-3′ and OYDVR 5′-TAGTGGTACACCACATTTCGT-3′), GCLV (GCLVF 5′-TTATAGGGACGGCACAAAATCAATCA-3′ and GCLVR 5′-AATAGCACTCCTAGAACAACCATT-3′) and SLV (SLVF 5′-AATYATTTACAATCGTCCAGCTA-3′ and SLVR 5′-ATAATATCAATCAAATMCACACAATT-3′). Amplicons of the expected size were obtained for each virus. The amplified products were purified and sequenced in both directions. Sequence information of the CP genes of 9 OYDV, 12 GCLV, and 7 SLV isolates has been submitted to NCBI-GenBank with accession numbers KF862683 to KF862710. Sequence analysis showed that the coat protein gene of OYDV shared 86% identity with the coat protein gene of OYDV isolate MS/SW1 from Australia (GenBank Accession No. HQ258894). Comparison of the coat protein gene sequences of Polish GCLV isolates with those available in GenBank showed 85 to 91% sequence identities. Multiple sequence alignment revealed 84% nucleotide identity between the Polish isolate of SLV and an SLV isolate from Chinese garlic (AF314147) formerly referred to as Garlic latent virus (3). To the best of our knowledge, this is the first report of OYDV, GCLV, and SLV in garlic plants in Poland. The accurate identification of viruses present in garlic plants will help to use the appropriate strategies to reduce viral incidence in garlic-growing areas. References: (1) J. Chen et al. Arch Virol 146:1841, 2001. (2) A. M. G. King et al. Virus Taxonomy: Ninth Report of the International Committee on Taxonomy of Viruses. Elsevier Academic Press, San Diego, CA, 2011. (3) T. Tsuneyoshi et al. Arch. Virol. 143:1093, 1998.


1991 ◽  
Vol 57 (1) ◽  
pp. 65-69 ◽  
Author(s):  
Isamu SAKO ◽  
Wataru NAKASONE ◽  
Kiyotugu OKADA ◽  
Satoshi T. OHKI ◽  
Takeshi OSAKI ◽  
...  

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>


2020 ◽  
Vol 21 (4) ◽  
Author(s):  
Heri Harti ◽  
SRI HENDRASTUTI HIDAYAT ◽  
SOBIR ◽  
SURYO WIYONO

Abstract. Harti H, Hidayat SH, Sobir, Wiyono S. 2020. Detection of major viruses infecting shallot and molecular characterization of Onion yellow dwarf virus from several locations in Indonesia. Biodiversitas 21: 1697-1701. Research was conducted to identify main viruses infecting shallot in several regions in Indonesia and to further characterize genetic variation of Onion yellow dwarf virus (OYDV). Field survey was conducted in Central Java (Brebes), East Java (Probolinggo), West Sumatera (Alahan Panjang), West Nusa Tenggara (Bima), and South Sulawesi (Enrekang). Virus detection from field samples was conducted by dot immunobinding assay. This detection confirmed that infection of OYDV, Shallot yellow stunt virus (SYSV), Shallot latent virus (SLV), and Garlic common latent virus (GarCLV) has occurred in all field with incidence ranged from 20 to 93.5%, 2 to 93%, 21.5 to 80%, and 2 to 80.5%, respectively. The specific primers of Nib gene successfully amplified DNA fragments of OYDV from all locations. Sequencing of DNA fragments revealed that the amplified product was 351 bp. Sequence analysis indicated that the present OYDV isolates from Indonesia shared homology from 82 to 95%; and they had homology from 81 to 95% with OYDV isolates from other countries. The similarity of OYDV isolates from different geographical locations reflected the movements of seed bulbs among and within countries. The phylogenetic tree also revealed that OYDV isolates from different countries did not group together indicating their diverse origin.


Sign in / Sign up

Export Citation Format

Share Document