scholarly journals Influence of explant size, heat treatment, and variety on garlic (Allium sativum L) meristematic tissue growth

2021 ◽  
Vol 306 ◽  
pp. 01036
Author(s):  
Asih K. Karjadi ◽  
Nazly Aswani
Keyword(s):  
Heat Treatment ◽  
Growth And Development ◽  
Allium Sativum ◽  
Tissue Growth ◽  
Onion Yellow Dwarf Virus ◽  
Virus Elimination ◽  
Meristematic Tissue ◽  
Yellow Dwarf Virus ◽  
Das Elisa ◽  
Virus Testing

Garlic (Allium sativum L) is one of genus Allium sp which propagated vegetatively. In vegetative propagation, virus is one of the major diseases. However, it has been approved that OYDV (Onion Yellow Dwarf Virus) can be eliminated through unconventional propagation techniques. The aim of this research was to evaluate the effect of explant, variety and heat treatment on the growth and development of explants dealing with virus elimination. The research was conducted on the IVEGRI tissue culture laboratory from May to September 2018. The research was designed using combination between explant types e.g. meristem (E1) and shoot tip (E2), heat treatment (370C) for 1 month, and varieties e.g. Lumbu Hijau (V1); Tawang Mangu (V2); Lumbu Putih (V3). The composition of media consisted of MS + MS vitamine + Sucrose 30 g L-1+ IAA 2 mg L-1 + Kinetin 2 mgL-1 + GA30.01 mgL-1 + Gel gro 2 gL-1, pH 5.7 were used as media growth. The results concluded that explant size, heat treatment and varieties affected percentage of proliferation. The bigger the explant size used, the higher growth percentage resulted. From the average number of shoots, leaves and roots, it also suggested that explant type and varieties affected the explant growth and development. Moreover, it was shown that the culture contamination on 10 WAP ranged from 20% to 55% which generally caused by bacteria. The results of virus testing OYDV on plantlets using DAS ELISA Serology technique showed that the infected cultures ranged from 22.22% to 60 %.

scholarly journals Distribution of viruses in the shallot germplasm collection of the Czech Republic – Short Communication

2017 ◽  
Vol 44 (No. 1) ◽  
pp. 49-52 ◽  
Author(s):  
Kateřina Smékalová ◽  
Helena Stavělíková ◽  
Karel Dušek
Keyword(s):  
Germplasm Collection ◽  
Latent Virus ◽  
Onion Yellow Dwarf Virus ◽  
The Czech Republic ◽  
Leek Yellow Stripe Virus ◽  
Garlic Common Latent Virus ◽  
Shallot Latent Virus ◽  
Yellow Dwarf Virus ◽  
Yellow Stripe ◽  
Das Elisa

The Czech collection of shallot (Allium cepa var. ascalonicum) genetic resources (122 accessions) was surveyed for the presence of four different viruses, i.e. Onion yellow dwarf virus, Leek yellow stripe virus, Garlic common latent virus (GCLV), and Shallot latent virus, by DAS-ELISA. The shallot seems to be resistant against GCLV because none of the tested plants was infested by this virus. Other three viruses were found with an incidence ranging from 53% to 93% for genotypes and 48% to 87% for plants. Most of the tested shallot genotypes were simultaneously infected with two or three viruses. These results were compared with neighbouring collection of garlic where all four viruses were found widespread with an incidence ranging from 65% to 83% for genotypes and 39% to 61% for plants.  

scholarly journals Natural Infection of Verbena and Phlox by a Recently Described Member of the Carmovirus Genus

Plant Disease ◽  
2006 ◽  
Vol 90 (8) ◽  
pp. 1115-1115 ◽  
Author(s):  
F. M. Assis Filho ◽  
A. Harness ◽  
M. Tiffany ◽  
A. Gera ◽  
S. Spiegel ◽  
...  
Keyword(s):  
United States ◽  
Online Publication ◽  
Polyclonal Antibodies ◽  
Natural Infection ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Species ◽  
Virus Elimination ◽  
Das Elisa ◽  
Virus Testing

A novel carmovirus infecting angelonia (Angelonia angustifolia) was recently described independently by researchers in the United States, Israel, and Germany (1,2,4). Angelonia flower break virus (AnFBV) and Angelonia flower mottle virus were proposed as appropriate names for this carmovirus. The virus, causing stunting, mild leaf mottle, flower mottling, and flower breaking symptoms has been detected in naturally infected angelonia in the United States, Israel, and Germany (2,4). Here we report the first detection of natural infection of verbena (in the United States and Israel) and phlox (in the United States) by using a recently developed double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA; Agdia, Elkhart, IN). Prior to this report, verbena was considered insusceptible to carmovirus infection (3) and phlox was known as an experimental host for AnFBV (2). A comparative serological study including 27 virus species, demonstrated that DAS-ELISA did not cross-react with any viruses that commonly infect ornamentals or are related to carmoviruses, showing that the polyclonal antibodies are specific to AnFBV. Antibody specificity was confirmed by the carmovirus group PCR test (Agdia). Furthermore, reverse transcription-polymerase chain reaction with AnFBV specific primers (2) produced the expected 1172-bp band from all ELISA-positive samples tested. Between November 2005 and March 2006, AnFBV was detected in 181 of 567 verbena, 26 of 143 phlox, and 193 of 267 angelonia samples submitted to Agdia Testing Services by commercial ornamental propagators for virus testing. Most samples were asymptomatic, although a few exhibited mild leaf mottle. It should be noted that the number of AnFBV-infected samples might not accurately reflect the actual number of commercially produced plants infected with AnFBV because most of the samples analyzed originated from virus elimination programs. The detection of natural AnFBV infection of verbena, phlox, and angelonia suggests that AnFBV may be more widespread in the ornamental industry than previously thought. References: (1) S. Adkins et al. Phytopathology (Abstr.) 95(suppl.):S2, 2005. (2) S. Adkins et al. Phytopathology 96:460, 2006. (3) G. P. Martelli and M. Russo. Online publication. ICTVdB-The Universal Virus Database. 00.074.0.02, 2004. (4) S. Winter et al. New Disease Reports. Vol 12. Brit. Soc. Plant Pathol. Online publication, 2005.

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 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 Distribución viral en plantas de cebolla (Allium cepa L.) asintomáticas

2018 ◽  
Vol 3 (7) ◽  
pp. 1425-1434 ◽  
Author(s):  
Rodolfo Velásquez-Valle ◽  
Manuel Reveles-Hernández ◽  
Mario Domingo Amador-Ramírez
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
Onion Yellow Dwarf Virus ◽  
Leek Yellow Stripe Virus ◽  
Tomato Spotted Wilt ◽  
Allium Cepa L ◽  
Garlic Common Latent Virus ◽  
Yellow Dwarf Virus ◽  
Yellow Stripe ◽  
Wilt Virus ◽  
Das Elisa

Se estudió por medio de DAS- ELISA la distribución del Onion yellow dwarf virus (OYDV, virus del enanismo amarillo de la cebolla), Garlic common latent virus (GarCLV, virus latente común del ajo), Shallot latent v irus (SLV, virus latente del chalote), Leek yellow stripe virus (LYSV: virus de la franja amarilla del puerro), Tobacco etch virus (TEV, virus del jaspeado del tabaco) y Tomato spotted wilt virus (TSWV, virus de la marchitez manchada del jitomate) en hojas, psdeudotallo e inflorescencia de plantas de cebolla asintomáticas colectadas en diferentes épocas de 2010 en tres localidades del estado de Zacatecas, México. Se registró la presencia de esos virus en los órganos mencionados aunque resultó frecuente detectar infecciones mezcladas en hojas individuales donde las interacciones más comunes involucraban dos, tres y cuatro virus. No se encontró una tendencia que relacione la presencia viral con los órganos vegetales analizados o con la edad de las hojas; sin embargo, las infecciones con un solo virus parecen ser menos frecuentes en las hojas de edad intermedia.

scholarly journals Insiden Penyakit Virus Tular Umbi pada Tigabelas Varietas Bawang Merah Asal Jawa Barat dan Jawa Tengah

2016 ◽  
Vol 21 (2) ◽  
pp. 164
Author(s):  
Neni Gunaeni ◽  
Astri W Wulandari ◽  
Ati Srie Duriat ◽  
Agus Muharam
Keyword(s):  
Immunosorbent Assay ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Onion Yellow Dwarf Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Diseases ◽  
Double Antibody ◽  
Yellow Dwarf Virus ◽  
Yellow Stripe ◽  
Das Elisa

Penyakit virus tular umbi merupakan salah satu kendala dalam meningkatkan produksi bawang merah. Hal ini disebabkan oleh virus yang infeksinya bersifat sistemik. Apabila partikel virus berada dalam jaringan benih umbi, maka akan sulit untuk dikendalikan dan dapat membawa masalah baru pada pertanaman berikutnya. Penelitian bertujuan mengetahui insiden penyakit virus tular umbi pada 13 varietas bawang merah yang berasal dari Jawa Barat dan Jawa Tengah. Penelitian dilaksanakan di Rancaekek (elevasi 650 m dpl.) dan di Laboratorium Balai Penelitian Tanaman Sayuran Lembang (elevasi 1.250 m dpl.), sejak bulan Agustus sampai November 2004. Perlakuan terdiri atas 13 varietas bawang merah, yaitu: Lodra, Sumenep, Batu, Merah Maja, Merah Cigugur, Ciniru, Bima, Bima Curut, Bima Timor, Bima Arjuna, Kuning Tablet, Kuning Gombong, dan Philipina. Rancangan yang digunakan ialah acak kelompok dengan tiga ulangan. Hasil penelitian menunjukkan bahwa (1) insiden penyakit virus tular umbi pada masing-masing varietas bawang merah asal Jawa Barat dan Jawa Tengah berturut-turut yaitu varietas Lodra 84,67%, Sumenep 82,56-100%, Batu 39,86-78,67%, Merah Maja 95,25%, Merah Cigugur 100%, Ciniru 66,27%, Bima Curut 78,57%, Bima 100%, Bima Timor 57,98%, Bima Arjuna 47,96%, Kuning Tablet 57,48%, Kuning Gombong 97,92%, dan Philipina 97,92 %, (2) gejala infeksi virus pada daun umumnya berupa  klorosis, mosaik bergaris kuning vertikal terputus-putus, garis-garis hijau vertikal, dan ukuran daun menjadi kecil, (3) gejala-gejala tersebut bereaksi positif dengan OYDV(onion yellow dwarf virus) dan SYSV (shallot yellow stripe virus) berdasarkan uji DAS-ELISA (double antibody sandwich-enzyme linked immunosorbent assay). Informasi mengenai insiden virus tular umbi pada bawang merah ini sangat penting dalam rangka mengembangkan metode perbenihan bawang merah bebas virus. <br /><br />Virus disease is one of major problems in increasing shallots production, because its infection has a systemic character. If it is already in shallots bulb tissues, the virus is difficult to be controlled and will cause new problems to the next planting. The experiment was aimed to determine incidence of bulb-borne virus diseases on  thirteen varieties of shallots (Allium cepa var. ascalonicum) originated from West  and Central Java. The experiment was carried out at Indonesian Vegetable Research Institute Lembang (1,250 m asl.) and Rancaekek (650 m asl.), from  August to November 2004. The shallot varieties tested were Lodra, Sumenep, Batu, Merah Maja, Merah Cigugur, Ciniru, Bima, Bima Curut, Bima Timor, Bima Arjuna, Kuning Tablet, Kuning Gombong, and Philipina. A randomized complete block design with three replications were used in this experiment. The results of the experiment showed that  (1) incidence of virus diseases in shallots bulb on variety Lodra was 84.67%, Sumenep 82.56-100%, Batu 39.86-78.67%, Merah Maja 95.25%, Merah Cigugur 100%, Ciniru 66.27%, Bima Curut 78.57%, Bima 100%, Bima Timor 57.98%, Bima Arjuna 47.96%, Kuning Tablet 57.48%, Kuning Gombong 97.92%,  and Philipina 97.92 %, (2) the virus symptoms exhibited on infected shallots were  yellow stripe mosaic, chlorosis,  green stripe leaf,  and leaves became small, and (3) the symptoms were associated with OYDV (onion yellow dwarf virus) and SYSV (shallots yellow stripe virus) base on DAS-ELISA (double antibody sandwich-enzyme linked immunosorbent assay). Information on the incidence of viral diseases on shallots bulb is very important to develop the production technology of virus-free shallots bulb.<br /><br /><br />

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 EFEK ANTIVIRAL RIBAVIRIN DALAM PERTUMBUHAN DAN PERKEMBANGAN EKSPLAN BAWANG PUTIH CV. LUMBU HIJAU, CV. LUMBU KUNING DAN CV. TAWANGMANGU

Agrin ◽  
2019 ◽  
Vol 22 (2) ◽  
pp. 93
Author(s):  
Asih K. Karjadi ◽  
Neni Gunaeni
Keyword(s):  
Growth And Development ◽  
Allium Sativum ◽  
Visual Observation ◽  
High Concentration ◽  
Virus Diseases ◽  
Culture Techniques ◽  
Key Word ◽  
Das Elisa ◽  
Important Disease ◽  
Test Virus

Tanaman bawang putih (Allium sativum L) termasuk dalam genus Allium yang diperbanyak secara vegetatifmelalui umbi. Virus merupakan salah satu penyakit penting yang perlu dipecahkan pada pembiakan vegetatif ini.Teknik inkonvensional kultur jaringan yang dikombinasikan dengan kemoterapi dapat membantu menghilangkanpenyakit virus. Penelitian ini bertujuan untuk melihat pengaruh dari beberapa konsentrasi antiviral ribavirin dimedia MS terhadap pertumbuhan dan perkembangan shoot tip Bawang putih cv Lumbu Hijau, cv. Lumbu Kuning,cv. Tawangmangu. Percobaan dilakukan di laboratorium kultur jaringan, Balai Penelitian Tanaman Sayur(Balitsa), pada bulan Mei hingga Juli 2015. Sasaran penelitian adalah untuk menghasilkan tanaman bebas virusdengan menggunakan teknik kultur jaringan yang dikombinasikan dengan kemoterapi. Varibel yang diamatiadalah pertumbuhan dan perkembangan planlet bawang putih. Hasil dari penelitian (1) Kontaminasi kulturumumnya disebabkan oleh bakteri dan jamur dengan persentase 10 % sampai dengan 30%. (2) Penambahanantiviral ribavirin, semakin tinggi konsentrasi persentase tumbuh dan berkembang semakin rendah untuk ketigakultivar (3) Pengamatan secara visual penambahan antiviral ribavirin dan kultivar tidak berpengaruh pada jumlahtunas, rata-rata dari satu eksplan tumbuh satu tunas untuk ketiga kultivar (4). Penambahan antiviral ribavirin dankultivar tidak mempengaruhi pertumbuuhan daun, akar ketiga kultivar (5).Hasil pengujian virus dengan teknikDAS ELISA persentase kultur yang terinfeksi 54.55% sampai dengan 100 %.Kata kunci: bawang putih (Allium sativum L); antiviral ribavirin; kultivarABSTRAKThe garlic (Allium sativum L) belonging to the genus Allium, propagated in vegetative through bulb. Inthe plants propagated by vegetative technique, virus is an important disease to be solved. The tissue culturetechniques in combination with chemotheraphy could eliminate virus diseases. The experiment carried out in thelaboratory tissue culture, Balai Penelitian Tanaman Sayur (Balitsa) on May untill July 2015. The experiment aimsto observe the effect of several antiviral ribavirin concentration in MS medium on growth and development shoottip cv. Lumbu hijau , cv. Lumbu kuning , cv. Tawangmangu. It’s main goal is to produce virus-free plants usingtissue culture techniques combined with chemotheraphy. The variables observed were the growth and developmentof garlic plantlets. The results of the experiment are; (1). Culture contamination were generally caused by bacteriaand fungi with a percentage of 10% to 30%. (2) In the high concentration of antiviral ribavirin gave results ondecreasing growth and development of the three garlic cultivar (3) On visual observation, cultivar and antiviralribavirin has no effect on the number of shoots, each explants were growing one shoot. (4). The added of antiviralribavirin and cultivar does not affect on growth the three garlic cultivar . (5) The results of the test virus byserological test DAS ELISA techniques the percentage of infected culture were 54.55% to 100%.Key word : Garlic (Allium sativum L); Antiviral ribavirin; cultivar

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