scholarly journals Detection of viruses of Bangladeshi and Japanese garlic and their elimination through root meristem culture

2017 ◽  
Vol 28 (2) ◽  
pp. 55-63 ◽  
Author(s):  
MS Haque ◽  
K Hattori
Keyword(s):  
Root Meristem ◽  
Onion Yellow Dwarf Virus ◽  
Meristem Culture ◽  
Free Medium ◽  
Rt Pcr ◽  
Virus Elimination ◽  
Root Meristems ◽  
Yield And Quality ◽  
Yellow Dwarf Virus

A number of viruses cause considerable yield loss and quality deterioration in garlic. Root meristems of virus infected plants are known to be free from detectable viruses. This potentiality could be exploited to obtain virus free clones at a high frequency by culturing excised root meristems in vitro. We have developed efficient methods of direct and somatic embryo derived shoot regeneration from root meristems of garlic. The objectives of this work were to detect viruses infecting Bangladeshi and Japanese garlic clones and find an easy and efficient method of eliminating the viruses for the improvement of both yield and quality of garlic. At first, we confirmed the presence of detectable viruses in three Bangladeshi and one Japanese clones. The clones were infected with four different types of viruses: Garlic viruses (GarVs), Onion yellow dwarf virus (OYDV), Leek yellow stripe virus (LYSV), and Garlic common latent virus (GCLV). To eliminate those viruses, as per our previous method, root meristems were cultured on MS medium supplemented with 1.0 µM NAA and 10.0 µM BA. Shoot primordia developed from the cultured explants within 1 month. The regenerated individual shoot buds (2-5 mm) were separated from the mother explants and transferred to growth regulators free medium. RT-PCR confirmed that the viruses present in the mother garlic plants were absent in the shoots found after two-step culture. The regenerated shoots were rooted on growth regulator free medium and transferred to pots. Results indicated that the plants remained free from LYSV. Virus elimination through root meristem culture emerged as an efficient novel technique for the eradication of multiple viruses as confirmed by RT-PCR in this study. This technique has the potential for the production and supply of virus free propagules (plants/bulblets) for the yield and quality improvement of garlic.Progressive Agriculture 28 (2): 55-63, 2017

scholarly journals Production of Virus-Free Garlic Plants through Somatic Embryogenesis

Agronomy ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 876
Author(s):  
Snježana Kereša ◽  
Katarina Kurtović ◽  
Smiljana Goreta Ban ◽  
Darko Vončina ◽  
Ivanka Habuš Jerčić ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Onion Yellow Dwarf Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Virus Elimination ◽  
Leek Yellow Stripe Virus ◽  
Mother Plants ◽  
Yellow Dwarf Virus

The present study was conducted to establish a protocol for the regeneration of virus-free garlic plants through somatic embryogenesis of two Croatian garlic ecotypes. Basal parts of cloves from mother plants were cultured on a full Murashige and Skoog (MS) or modified MS medium (¼ of KNO3 and NH4NO3 and 2xMgSO4) containing 0.1 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) or 1 mg L−1 2,4-D + 0.5 mg L−1 kinetin (Kin) and representing four different treatments. Plants were regenerated in MS medium containing 0.1 mg L−1 2,4-D and rooted in a medium containing 0.05 mg L−1 1-naphthaleneacetic acid (NAA) + 0.005 mg L−1 6-(γ,γ-dimethylallylamino)purine (2iP). The presence of viruses (i.e., sanitary status) of the mother plants and regenerants was checked by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). The mother plants were infected with onion yellow dwarf virus (OYDV) and leek yellow stripe virus (LYSV). In addition, the presence of garlic common latent virus (GCLV) was confirmed in four mother plants. Embryogenic callus developed in all four treatments with success ranging from 55% to 81% depending on treatment and ecotype. Plant conversion was significantly higher in somatic embryos developed in media containing 0.1 mg L−1 2,4-D than those developed in media containing 1 mg L−1 2,4-D + 0.5 mg L−1 Kin. Virus elimination success ranged from 13.3% up to 62.5% depending on garlic ecotype and treatment. The overall rate of virus elimination by somatic embryogenesis for both treatments and ecotypes were 20.7%, 22.9%, and 30.5% for OYDV, GCLV, and LYSV, respectively. Based on these results, somatic embryogenesis has been shown to be equally or more successful in eliminating garlic viruses compared to other in vitro methods.

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
Keyword(s):  
Disease Incidence ◽  
Viral Disease ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Onion Yellow Dwarf Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Yellow Dwarf Virus

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.

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
Keyword(s):  
Early Detection ◽  
Latent Virus ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Onion Yellow Dwarf Virus ◽  
Rt Pcr ◽  
Double Infection ◽  
Specific Primers ◽  
Shallot Latent Virus ◽  
Yellow Dwarf Virus

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.

scholarly journals First Report of Shallot latent virus in Garlic in Argentina

Plant Disease ◽  
2010 ◽  
Vol 94 (7) ◽  
pp. 915-915 ◽  
Author(s):  
A. K. Torrico ◽  
E. E. Cafrune ◽  
V. C. Conci
Keyword(s):  
Latent Virus ◽  
Onion Yellow Dwarf Virus ◽  
Rt Pcr ◽  
Cp Gene ◽  
Virus Complex ◽  
Garlic Latent Virus ◽  
Shallot Latent Virus ◽  
Yellow Dwarf Virus ◽  
Garlic Virus ◽  
Das Elisa

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.

scholarly journals Identification and molecular characterization of onion yellow dwarf virus (OYDV) in Allium cepa crops in Poland

2021 ◽  
Vol 61 (3) ◽  
pp. 214-220
Keyword(s):  
Phylogenetic Analysis ◽  
Molecular Characterization ◽  
Coat Protein Sequence ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Onion Yellow Dwarf Virus ◽  
Rt Pcr ◽  
Pcr Products ◽  
Yellow Dwarf Virus

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.

scholarly journals The elimination of Plum pox virus in plum cv. Bluefree and apricot cv. Hanita by chemotherapy of in vitro cultures

2011 ◽  
Vol 38 (No. 2) ◽  
pp. 49-53 ◽  
Author(s):  
A. Hauptmanová ◽  
J. Polák
Keyword(s):  
In Vitro Cultures ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
Virus Elimination ◽  
Skoog Medium ◽  
Negative Results ◽  
Murashige Skoog ◽  
One Year

In vitro cultures of plum cv. Bluefree and apricot cv. Hanita infected with Plum pox virus (PPV) were used for the virus elimination by chemotherapy. Low ribavirin concentrations of 5 and 10 mg/l in Murashige-Skoog medium were applied in the treatment. Plum pox virus was completely eliminated by 5 mg/l of ribavirin in plum cv. Bluefree within twenty weeks and in apricot cv. Hanita in twelve weeks of the application. Plum pox virus was completely eliminated by 10 mg/l of ribavirin both in plum cv. Bluefree and apricot cv. Hanita within twelve weeks. The presence of PPV was not proved by RT-PCR. Clones of plum cv. Bluefree and apricot cv. Hanita were re-tested by RT-PCR one year after the termination of the ribavirin treatment and negative results confirmed the elimination of Plum pox virus.

scholarly journals HTS-Based Monitoring of the Efficiency of Somatic Embryogenesis and Meristem Cultures Used for Virus Elimination in Grapevine

Plants ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1782
Author(s):  
Mihaly Turcsan ◽  
Emese Demian ◽  
Tunde Varga ◽  
Nikoletta Jaksa-Czotter ◽  
Erno Szegedi ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
High Throughput Sequencing ◽  
Vitis Vinifera L ◽  
Meristem Culture ◽  
Rt Pcr ◽  
Virus Elimination ◽  
Mother Plants ◽  
Virus Diagnostics ◽  
Grapevine Pinot Gris Virus ◽  
First Time

Meristem culture and somatic embryogenesis are effective tools for virus elimination of vegetatively propagated crops including grapevine (Vitis vinifera L.). While both have been shown to be useful to eliminate the main grapevine viruses, their efficiency differs depending on the virus and grapevine variety. In our work, we investigated the efficiency of these two virus elimination methods using small RNA high-throughput sequencing (HTS) and RT-PCR as virus diagnostics. Field grown mother plants of four clones representing three cultivars, infected with different viruses and viroids, were selected for elimination via somatic embryogenesis (SE) and meristem culture (ME). Our results show for the first time that using SE, elimination in mother plants was effective for all viruses, i.e., grapevine rupestris vein feathering virus (GRVFV), grapevine Syrah virus 1 (GSyV-1), Grapevine virus T (GVT) and grapevine Pinot gris virus (GPGV). This study also confirms previous studies showing that SE is a possible strategy for the elimination of GFkV, GRSPaV, HSVd, and GYSVd-1. Our results demonstrate that the efficacy of virus elimination via SE is relatively high while the purging of viroids is lower. Our work provides evidence that the efficiency of SE is comparable to that of the technically difficult ME technique, and that SE will offer a more effective strategy for the production of virus-free grapevine in the future.

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