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Distribution of PLRV, PVS, PVX and PVY (PVYN, PVYo and PVYc) in the Seed Potato Tubers in Turkey
H. Bostan,K. Haliloglu
Pakistan Journal of Biological Sciences , 2004,
Abstract: This study was conducted in order to determine the distribution ratio of PLRV, PVS and PVY (PVYN, PVYo and PVYc) in the seed potato tubers used for planting material in the important potato production regions of Turkey and observe the symptoms caused by single or mixed infection of these viruses under field condition. Firstly, over 880 leaf samples were tested by using virus-specific polyclonal antibodies. Secondly, 83 samples found to be infected with PVY in the result of first ELISA were retested by using PVYo, PVYN and PVYc-specific monoclonal antibodies. The ELISA results showed that seed potato tubers used for planting material was infected with at the rate of PLRV (14.2%), PVX (11.8%), PVS (4.6%) and PVY (17.7%). On the other hand, the result of monoclonal antibody for PVY-strains showed that the frequency of PVYN and PVYo were (13.4%, 4.3%) but PVYc was not found. Under field condition, plants infected with PLRV exhibited the rolling of young leaves, upright growth and pinky color but PVS did not cause any distinct symptoms. PVX alone or the combination of PVX with PLRV, PVS and PVY caused mild or severe mosaic symptoms on all cultivars. PVY induced yellowing of leaves, leaf drop streak, veinal necrosis on some plants from all cultivars, however, some plants did not develop any distinct symptoms in case of infected with PVY. The combination of PVY and PVX caused more severe mosaic, rugosity and reduced of leaf size. When plants infected with PVY and PLRV exhibited yellowing of leaves, leaf drop, dwarfing, rolling of leaves and rogositiy. However, some plants from Morfona and Granola cultivars died. On the other hand, the symptoms on plants infected with PVS and PLRV or PVS and PVY were similar to single infection of PLRV and PVY.
The feasibility of tetraplex RT-PCR in the determination of PVS, PLRV, PVX and PVY from dormant potato tubers
H Bostan, P Peker
African Journal of Biotechnology , 2009,
Abstract: Dormant potato tubers belonging to cvs., namely, Agria, Granola and Marfona known to be infected with potato viruses ( Potato leafroll virus, PLRV; Potato virus S, PVS; Potato virus X, PVX and Potato virus Y, PVY) were tested with uniplex RT-PCR and strong bands specific to each virus were obtained from cultivars. When cDNA synthesized for uniplex RT-PCR was used for tetraplex RT-PCR, the bands obtained from PVS, PLRV and PVX were too faint to be photographed and there is no any observed band for PVY. To improve the band density, the concentration of oligo dT primer in RT was increased from 20 to 40 ng in the subsequent experiments. The increasing of oligo dT primer concentration in RT increased the band density for PVS and PVX, but not PVY. Upon this, different amount of total RNA were tested in RT stage. The best result was obtained from 5 μl of total RNA and followed by 3.5 and 2.5 μl applications. In order to determine the effect of cDNA amount in PCR, 2 μl cDNA + 23 μl PCR, 5 μl cDNA + 20 μl PCR and 5 μl cDNA + 25 μl PCR mixture were compared. However, no distinct differences were observed among various cDNA amounts. As a result, instead of tetraplex RT-PCR, it is suggested the use of triplex RT-PCR for reliable detection of PLRV, PVS and PVX. However, uniplex PCR could be suggested for reliable detection of PVY from this study by using the same cDNA.
Obtaining PVX, PVY and PLRV-Free Micro Tuber from Granola, Pasinler 92 and Caspar Potato (Solanum tuberosum L.) Cultivars  [PDF]
Hidayet Bostan,Erkol Demirci
Pakistan Journal of Biological Sciences , 2004,
Abstract: This study was conducted to obtain virus-free propagation materials from Granola, Pasinler 92 and Caspar potato (Solanum tuberosum L.) cultivars infected with potato virus X (PVX), potato virus Y (PVY) and potato leaf roll virus (PLRV) by using meristem-tip culture. For in vitro propagation, it was tested the effect of different combinations and concentrations of benzylamino purine (BA) (0.0, 0.25, 0.50 mg L-1) and gibberellic acid (GA3) (0.0, 0.25, 0.50 mg L-1) on the number of shoot and node. On the other hand, it was evaluated the effect of BA (0.00, 5.00, 10.0 mg L-1) and CCC (chlorocholine chloride) (0.00, 500 mg L-1) on the tuberization under two photoperiodic regimes (light and dark). The MS salts and vitamins supplemented with 30 g L-1 sucrose was used as a medium and the media was solidified with 7.0 g L-1 agar and the ratio of sucrose added into media for micro tuber production had been increased from 3-8%. The highest number of shoots was obtained from 0.00/0.25, 0.25/0.50 and 0.00/0.00 mg L-1 BA/GA3 treatments for Granola, Pasinler 92 and Caspar cultivars as 1.52, 1.24 and 1.44, respectively. However, the highest number of node were determined on 0.00/0.50 for Granola (9.12), Pasinler 92 (8.76) and on 0.00/0.25 mg L-1 BA/GA3 treatments for Caspar (8.24). When the results were assayed according to total tuber number, the most micro-tubers for Granola, Pasinler 92 and Caspar cultivars were obtained from 5.00/5000 mg L-1 BA/CCC treatment as 5.6, 4.0, 4.8 per/bottle under dark treatments. All in vitro regenerated plant materials were tested by DAS-ELISA (double antibody sandwich enzyme-linked immunosorbent assay) to determine the presence and absence of viruses and PVX, PVY and PLRV viruses were eliminated from Granola (25, 40 and 60%), Pasinler 92 (16, 41.6 and 46.1%) and Caspar cultivars (28.5, 33.3 and 50%), respectively.
Incidencia y distribución altitudinal de 13 virus en cultivos de Solanum tuberosum (Solanaceae) en Costa Rica
Vásquez,Viviana; Montero-Astúa,Mauricio; Rivera,Carmen;
Revista de Biología Tropical , 2006,
Abstract: incidence and altitudinal distribution of 13 virus cultures in solanum tuberosum (solanaceae) from costa rica. a survey was conducted in 30 fields located at three different altitudes in cartago, costa rica?s main potato producing area. twenty plants were sampled per farm, for a total of 600 samples with 200 samples per altitude. elisa was used with commercial reagents to independently test for pvx, pvy, pvm, pva, pvs, plrv, pmtv, pamv, pvv, pvt, aplv, apmov and trsv. the presence of the following viruses was determined: pvx (77 %), pamv (62 %), plrv (42 %), trsv (42 %), pvt (39 %), pvv (37 %), pmtv (31 %), pvy (30 %), pvs (19 %), pvm (13 %), pva (8 %), and apmov (8 %). aplv was not detected in any sample. this is the first report in costa rica of the presence of the viruses pmtv, pamv, pvv, pvt and apmov. a high viral incidence in the tuber seed production area as well as a high rate of mixed infections is reported. rev. biol. trop. 54 (4): 1135-1141. epub 2006 dec. 29
华北农学报 , 2011, DOI: 10.7668/hbnxb.2011.05.009
Abstract: 根据PVY、PVS和PLRV外壳蛋白(Coatprotein,CP)基因序列的保守区域设计各自的引物,利用三重RT-PCR技术实现了在同一反应体系中同时扩增出3种病毒产物。PVY、PVS和PLRV扩增产物测序长度均与目的片段的长度相符,分别为781,181,364bp;各种病毒产物的测序结果同GeneBank中的序列比对后的同源性均高达95%以上。将3种病毒的RNA稀释后进行RT-PCR,PVY、PVS和PLRV的最低检测含量分别为4.5pg/μL~4.5fg/μL、3.7fg/μL和4.6fg/μL。三重RT-PCR为检测单独或复合感染PVY、PVS和PLRV的马铃薯材料,提供了一种方便、高效的分子学方法。
Influence of Aphids on the Epidemiology of Potato Virus Diseases (PVY, PVS and PLRV) in the High Altitude Areas of Turkey
Hidayet Bostan,Coskun Guclu,Erdogan Ozturk Isil Ozdemir,Havva Ilbagi
Pakistan Journal of Biological Sciences , 2006,
Abstract: Potato plants belonging to cv. Morfona were found infected with Potato leaf roll virus (PLRV), Potato virus Y (PVY) and Potato virus S (PVS) with the rates of 3.40, 6.47 and 5.30% in field conditions during the 2003 season. In order to determine the variations in the incidence rates of those potato viruses, harvested tubers from this field used for seed potato for the following seasons for the years of 2004 and 2005. Infection rates for 2004 and 2005 were determined as 7.66 and 22.33% for PLRV; 27.0 and 91.0% for PVY; 21.0 and 77.33% for PVS, respectively. While infection rates of those viruses were relatively low until 2003, the number of infected plants increased in following years. The results obtained from this study revealed that spreading rates of PVY and PVS were higher than PLRV as PVY higher than PVS. Winged aphid counts in the yellow-pan traps during the potato-growing seasons of 2004 and 2005 indicated that Myzus (Nectarosiphon) persicae (Sulzer, 1776), Therioaphis trifolii (Monell, 1882) and Aphis fabae (Scopoli, 1763) were the most abundant aphid species in Central District of Erzurum Province where field trails were established. Nevertheless there was not significant fluctuation of those aphid populations between 2004 and 2005. In addition to those aphids, Anoecia corni (Fabricius, Brachycaudus (Acaudus) cardui (Linnaeus), Cryptomyzus ribis (Linnaeus), Eulachnus rileyi (Williams, 1911), Hyperomyzus lactucae (Linnaeus, 1758) and Pterochloroides persicae (Cholodkovsky) species were collected from the pans but their number were very low.
A survey of viral status on potatoes grown in Eritrea and in vitro virus elimination of a local variety ‘Tsaeda embaba’
T Biniam, M Tadesse
African Journal of Biotechnology , 2008,
Abstract: Potato viruses are the major causes of yield loss and reduction in quality of seed tubers in Eritrea. A study was conducted to investigate the prevalence of viruses in potatoes (Solanum tuberosum L.) grown in Eritrea and to evaluate methods for their elimination. Leaf samples of two indigenous, (Tsaeda embaba and Keyih embaba) and three exotic varieties, (Ajiba, Spunta and Cosmos) were collected from fields growing potatoes in Maekel and Debub Administrative Zones and tested using the double antibody sandwich enzyme linked immunosorbent assay (DAS ELISA) technique. Five of the six most important potato viruses, PVX, PVY, PLRV, PVS and PVA, were detected in single and multiple infections. Virus elimination techniques were tested using in vitro plantlets of T. embaba established from field-grown tubers. Presence of PVX, PLRV and PVS was confirmed by ELISA test. The plantlets were then subjected to thermotherapy treatment for one and two weeks at 37oC. The treatment was successful in eliminating only PLRV but failed to eliminate PVX and PVS. When meristem culture was combined with thermotherapy treatment for one week all three viruses PVX, PLRV and PVS were eliminated with a success rate of 86, 83 and 100%, respectively.
Cloning and Sequencing of PLRV Coat Protein Gene and Construction of Vectors for.Transgenic Potato Resistant to Coinfection of PVY, PVX and PLRV

Cui Xiaojiang Peng Xuexian Shen Yufei Mang Keqiang,

生物工程学报 , 1994,
Abstract: The coat protein gene of potato leaf roll virus (Zhangjakou isolate) was cloned by PCR technology and sequenced. Comparisons of the nucleotide and deduced amino acid sequences of the PLRV coat protein gene with those of some reported isolates reveal over 97% homology between them. In order to acquire transgenic potato resistant to coinfection of PVY, PVX and PLRV, we managed to put the three virus coat protein genes driven by the enhanced 35S promoter respectively in one plant expression vector.
Detección serológica y caracterización molecular de Potato virus S (PVS, Carlavirus) en cultivos de papa de Colombia Detection and molecular characterization of Potato virus S (PVS, Carlavirus) from Colombia
José Fernando Gil,José Miguel Cotes,Mauricio Marín
Revista de Biología Tropical , 2013,
Abstract: El cultivo de papa en Colombia es afectado por diversos virus, que incluyen PVY, PLRV, PVX, PMTV y PVS; aunque se han realizado pocos estudios sobre la biología, distribución y patogenicidad de dichos virus en Colombia, siendo especialmente escasa la información referente al PVS. En este trabajo se evaluó mediante pruebas de ELISA, la presencia del PVS en cuatro departamentos de Colombia, así como sus niveles de variación, a partir de la secuenciación de una porción del gen de la cápside viral. Los resultados indicaron una detección promedio del virus en el 40% de las 320 muestras analizadas, con zonas como el Oriente cercano de Antioquia (49%) y Pasto (Nari o) (47%), donde se detectó en mayor proporción el virus. Los análisis de variación molecular indicaron la presencia de las dos razas de PVS (Ordinaria y Andina) en Colombia, siendo los aislamientos de PVS A los más diversos, al pre- sentar un rango de identidad del 88 al 99%. Estos hallazgos indican que es imperativo el fortalecimiento de los programas de certificación de semilla y vigilancia cuarentenaria en el país, especialmente para virus como el PVS, que aunque puede ser asintomático, causa pérdidas hasta del 20% en cultivos de papa. In Colombia, potato crops are affected by a wide variety of viruses such as PVY, PLRV, PVX, PMTV and PVS. Unfortunately, there are very few studies on the biology, distribution and pathogenicity of these viruses; this situation is even worse for the latent virus PVS. In this work, we evaluated the presence of PVS in four Colombian provinces (Antioquia, Boyacá, Cundinamarca, Nari o) by the use of ELISA. We also studied the degree of molecular variation by sequence comparison of a segment of the gene encoding for the viral coat protein. In average, PVS was detected in 40% of 320 analyzed samples of potato leaves; the highest levels were observed in the East of Antioquia (49%) and Pasto (Nari o) (47%), while in the other regions ranged between 35% and 42%. Analysis of sequence revealed the presence of two PVS strains in Colombia: three isolates were associated to PVS O (Ordinary) and twelve belonged to PVS A (Andean). A high diversity was observed among PVS A strains with percent identities in the range of 88-99%. These findings highlight the importance of strengthening seed certification programs and quarantine measures in Colombia for viruses like PVS, which can cause losses of up to 20% in potato crops and even higher in mixed virus infection.
Stock indexing and Potato virus Y elimination from potato plants cultivated in vitro
Nascimento, Luciana Cordeiro;Pio-Ribeiro, Gilvan;Willadino, Lilia;Andrade, Genira Pereira;
Scientia Agricola , 2003, DOI: 10.1590/S0103-90162003000300017
Abstract: potato cultivars (solanum tuberosum l.) have shown degeneration or run out caused by viruses after several cycles of propagation using seed tubers from commercial fields. this work reports the occurrence of single and mixed infections of four potato viruses in paraíba-brazil and presents a method for potato virus y (pvy) elimination, by using thermo-and chemotherapies. plants of potato cv. baraka were tested by direct antigen coating elisa. antisera against pvy, potato virus x (pvx), potato virus s (pvs), and potato leafroll virus (plrv) were used. materials with positive reaction to pvy were treated for virus elimination. single node cuttings (1.0 cm length) were excised and inoculated in murashige & skoog (ms) medium, supplemented with 1.0 mg l-1 of kinetin, 0.001 mg l-1 of naphthalene acetic acid (naa) and 0.1 mg l-1 of gibberellic acid (ga3). the thermotherapy at approximately 37oc, during 30 and 40 days, resulted in 20.0 and 37.5% pvy elimination, respectively. chemotherapy was undertaken with ribavirin (rbv), 5-azacytidine (aza), and 3-deazauridine (dzd). the rbv showed the highest rate of virus eradication, with 55.5% virus-free plants. simultaneous thermo and chemotherapy had higher efficiency for the elimination of pvy, reaching rates of healthy plants of 83.3% with rbv, 70.0% with aza, and 50.0% with dzd.
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