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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.
Técnica de inmunoimpresión en membranas de nitrocelulosa: una detección rápida para estimar la incidencia de los virus PVX, PVY, PVS y PLRV que infectan a la papa (Solanum spp.) Tissue printing technique in nitrocelullose membranes: a rapid detection technique for estimating incidence of PVX, PVY, PVS and PLRV viruses infecting potato (Solanum spp.)  [cached]
Guzmán Mónica,Caro Marina,García Yenny
Revista Colombiana de Biotecnología , 2002,
Abstract: La técnica serológica de ELISA, se ha utilizado desde los a os setentas como técnica cuantitativa para la detección de diversos grupos de virus que infectan a las plantas. Más recientemente se ha implementando la técnica cualitativa de inmunoimpresión (IMI) en membrana de nitrocelulosa, en la detección de diferentes grupos virales. En el presente trabajo se adaptó la técnica de IMI para la detección de los virus PVX, PVY, PVS y PLRV que atacan los cultivos de diferentes especies y variedades de papa (Solanum sp.) como yema de huevo, capiro, morita, pastusa, monserrate, tuquere a, ICA Puracé, ICA Nari o. Estos cuatro virus pueden causar pérdidas entre el 30% y 60% en la producción, ya sea solos o actuando sinergísticamente, por lo cual pueden reducir sensiblemente los beneficios económicos de un país que como Colombia se caracteriza por ser un gran productor de papa, con más de 2.8 millones de toneladas al a o. La técnica IMI se comparó con la técnica ELISA (Enzyme Linked Immunosorbent Assay) realizada sobre las mismas muestras, lo que permitió confirmar la sensibilidad de la prueba para la detección de los virus. Sobre un total de 800.muestras analizadas por IMI procedentes de diferentes veredas del departamento de Nari o, se encontró una incidencia del 72% para PVY, 38.7%, para PVX, 85.6% para PVS y 91.1% para PLRV; estimaciones que fueron similares o superiores a las obtenidas por la técnica de ELISA. Los resultados son novedosos para Colombia tanto por la implementación de la fácil y sensible técnica IMI para la detección de estos cuatro grupos virales que infectan a la papa como por la estimación de su incidencia en Nari o, uno de los departamentos productores de papa más importantes para el país. La detección oportuna y ágil de los virus es útil para dar una respuesta efectiva para la erradicación de material contaminado, tanto en material de campo como si es necesario, proveniente de cultivo in vitro. Los resultados de este trabajo permiten sugerir que la implementación de la técnica IMI, puede ser muy útil para los programas de certificación de semillas de papa con amplios beneficios puesto que se mantiene la sensibilidad y la especificidad, se reducen costos y es además participativa pudiendo involucrar a técnicos y agricultores. The ELISA serological technique has been used since the 1970s as a quantative technique for the detection of many groups of virus which infect plants. The immune-impression (IMI) in nitrocelullose membrane qualitative technique has been implemented more recently for the detection of different viral groups. In this work,
马铃薯上PVY、PVS和PLRV的三重RT-PCR检测  [PDF]
张华鹏,张剑峰,刘俊莹,王聪聪
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。
Novos progenitores de batata imunes a PVY e PVX e resistentes à pinta-preta
Brune, Sieglinde;Melo, Paulo Eduardo de;ávila, Ant?nio Carlos de;
Horticultura Brasileira , 1999, DOI: 10.1590/S0102-05361999000200022
Abstract: viruses that occur frequently in potato crops in brazil are responsible for crop yield losses, mainly when caused by a combination of viruses. for pvy and pvx the type of resistance used is immunity, controlled by single and dominant genes. early blight caused by alternaria solani is one of the most important potato diseases in brazil, especially when high air humidity and warm weather conditions prevail. this disease can cause losses up to 73%. the use of resistant varieties is an efficient control measure, being simultaneously harmless to environment and humans and providing an expressive reduction in costs. genetic resistance has also the advantage that it can be used in association with other control methods in integrated disease management. in the potato breeding program, embrapa hortali?as annually evaluates the immunity to pvy and pvx, the resistance to early blight and agronomic traits. among the clones evaluated in 1991 and 1992, 'embrapa/cip-pp063' and 'embrapa/cip-pp084' stood out with good agronomical characteristics, immunity to pvy and pvx, and high levels of resistance to early blight. both are recommended for use as parents in breeding programs for resistance to early blight. clones embrapa/cip-pp063 and embrapa/cip-pp084 are available for cooperative work with other institutions, as in vitro plantlets or tubers.
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.
Aphid Transmission of Two Important Potato Viruses, PVY and PLRV by Myzus persicae (Sulz.) and Aphis gossypii (Glov.) in Hatay Province of Turkey
Erdal Sertkaya,Gulsen Sertkaya
Pakistan Journal of Biological Sciences , 2005,
Abstract: Transmission assays by using aphid vectors were established to determine the effects of two aphid species (Myzus persicae Sulz. and Aphis gossypii Glov. on four solanacaeous test plant species (Capsicum annum L., Lycopersicon esculentum L., Physalis floridana Rydb. and Solanum tuberosum L.). Transmission rates of PVY and PLRV from potato to other test plants were investigated by DAS-ELISA assays. All experiments were carried out at 26:22±2°C (day:night) under a 16:8 h (light:dark) photoperiod conditions. Late 4th instar or early adult M. persicae and A. gossypii apterae were used in the transmission experiments. The aphids were not subjected to preacquisition starving in the assays. Each aphid species were reared on PVY-infected source potato plants for 5 h as Acquisition Access Period (AAP). Then, aphid group including 10 individuals were taken from donor plants were transplanted on to healthy receptor test plants for 15 h as Inoculation Period (IP). For PLRV transmissions, two aphid species reared on source potato plants for 24 h as AAP were transferred on to each healthy test plants as a group including 10 individuals for 24 h as IP. After IP, the aphids were removed from plants mechanically. PVY and PLRV were transmitted in higher rates by M. persicae. PVY and PLRV were more efficiently transmitted from potato to pepper seedlings among the other inoculated solanaceous test plants by both aphid species. The lowest transmission was occurred in tomato (2/15) and P. floridana (3/15) by using A. gossypii. Seed transmission of viruses were not determined by DAS-ELISA in seedlings re-obtained from experimentally infected test plants.
Resistência de genótipos de batata ao vírus do enrolamento da folha da batata (PLRV) e ao vírus Y (PVY)
Daniels, Julio;Pereira, Arione da S.;
Horticultura Brasileira , 2004, DOI: 10.1590/S0102-05362004000300003
Abstract: potato leafroll virus (plrv) and potato virus y (pvy) are the main causes of degeneration of potato seeds in brazil. the field resistance of potato genotypes to these viruses was evaluated in rio grande do sul state, brazil, of 20 clones and cultivars, during three consecutive spring cropping seasons. virus detection was carried out by serological tests (das-elisa). using the grouping analysis, the genotypes were separated in three groups for plrv resistance (elvira, achat, bintje, monalisa, monte bonito, panda and araucária, resistant; baronesa, asterix, atlantic, 2cri-1149-1-78, c-1226-35-80, astrid, c-1714-7-94, a-1139-12-92, macaca, eliza and santo amor, susceptible; catucha and cristal, very susceptible) and in four groups for pvy resistance (asterix, astrid, catucha, cristal, macaca, monte bonito, a-1139-12-92, c-1226-35-80 and c-1714-7-94, resistant; baronesa, santo amor, monalisa, panda and 2cri-1149-1-78, intermediate resistance; bintje, atlantic, elvira and araucária, susceptible; achat and eliza, very susceptible).
Movement and Distribution of Potato Leafroll Virus Antigen in Resistant Potato Genotypes  [PDF]
W. Ahmed,P. E. Thomas
Pakistan Journal of Biological Sciences , 1999,
Abstract: Twelve potato clones with variable degrees of resistance to potato leafroll virus (PLRV) were co-infected with potato virus X (PVX) or potato virus Y (PVY) before or after inoculations with PLRV. In some PLRV resistant clones, PVX and PVY facilitated the movement, and also increased the concentration of PLRV antigen. PLRV could not be rub-transmitted in mixed infections with the sap-transmissible viruses, PVX and PVY. PLRV did not observably affect the movement and concentration of PVX and PVY in resistant potato clones. PLRV also did not affect the movement of PVX into non-inoculated leaf tissue of cultivars Saco and USDA seedling 41956, nor did it cause systemic movement of PVX in hypersensitive Gomphrena globosa. PLRV was detected in infected plants by enzyme-linked immunosorbent assay (ELISA) and tissue blotting on nitrocellulose membranes, using polyclonal antibodies. Tissue blotting proved to be a more sensitive method, especially for detection of low titer antigens from plant tissue. These studies indicate that an essential function of PLRV inhibited in resistant plants may be at least partially complemented by active function of PVX or PVY.
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