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Öğe Comparison of Serological and Molecular Detection Methods for Testing Individual and Composite Samples Using PPV-M and PPV-T Isolates(Int Soc Horticultural Science, 2015) Gazel, M.; Serce, C. Ulubas; Caglayan, K.Sharka disease of stone fruit trees caused by Plum pox virus (PPV) was first described in 1968 in a limited area of Turkey, but during the last decade the disease has progressively spread to a large part of the country. Although PPV-Rec and -D strains were found in Turkey, the most common PPV strains were detected as PPV-M and PPV-T. In this study, DAS-ELISA (5B-IVIA/AMR) monoclonal antibody) and Spot Real-time RT-PCR techniques have been evaluated in order to determine the best sampling time and ratio of PPV infected samples in non-infected-infected plant mixtures for detection of PPV-T and PPV-M strains. Dormant buds in winter and fresh leaves in spring from PPV-infected trees were used for testing in 2012. Six repetitions were performed by single (3 leaves or buds from infected plant) or composite plant mixture samples (3 leaves or buds from infected plant + 3 leaves from healthy plant, and the other composite samples, i.e., 3+6 to 3+27). All combinations and all repetitions of composite leaf samples of both strains were detected as positive in Spot Real-time RT-PCR. However, in DAS-ELISA, the number of PPV positive samples decreased for T and M strain in 6th composite (3 infected+12 healthy leaves) and in 9th composite (3 infected+21 healthy leaves) in spring, respectively. At least 3 repetitions in all combinations of composite samples for PPV-T and -M were found positive in dormant season by Spot Real-time RTPCR whereas it was negative only in the last composite sample (3 infected+27 healthy buds) of PPV-T by DAS-ELISA.Öğe Comparison of serological and molecular detection methods for testing individual and composite samples using PPV-M and PPV-T isolates(International Society for Horticultural Science, 2015) Gazel, M.; Ça?layan, K.; Ulubaş Serçe, Ç.Sharka disease of stone fruit trees caused by Plum pox virus (PPV) was first described in 1968 in a limited area of Turkey, but during the last decade the disease has progressively spread to a large part of the country. Although PPV-Rec and -D strains were found in Turkey, the most common PPV strains were detected as PPV-M and PPV-T. In this study, DAS-ELISA (5B-IVIA/AMR) monoclonal antibody) and Spot Real-time RT-PCR techniques have been evaluated in order to determine the best sampling time and ratio of PPV infected samples in non-infected-infected plant mixtures for detection of PPV-T and PPV-M strains. Dormant buds in winter and fresh leaves in spring from PPV-infected trees were used for testing in 2012. Six repetitions were performed by single (3 leaves or buds from infected plant) or composite plant mixture samples (3 leaves or buds from infected plant + 3 leaves from healthy plant, and the other composite samples, i.e., 3+6 to 3+27). All combinations and all repetitions of composite leaf samples of both strains were detected as positive in Spot Real-time RT-PCR. However, in DAS-ELISA, the number of PPV positive samples decreased for T and M strain in 6th composite (3 infected+12 healthy leaves) and in 9th composite (3 infected+21 healthy leaves) in spring, respectively. At least 3 repetitions in all combinations of composite samples for PPV-T and -M were found positive in dormant season by Spot Real-time RT-PCR whereas it was negative only in the last composite sample (3 infected+ 27 healthy buds) of PPV-T by DAS-ELISA.Öğe Detection of fig mosaic virus in viruliferous eriophyid mite Aceria ficus(2012) Caglayan, K.; Elci, E.; Ulubas Serce, C.; Kaya, K.; Gazel, M.; Medina, V.Fig leaves showing typical fig mosaic symptoms on cv. Bursa siyahi{dotless} (donor plant) were cut under a stereo microscope into small pieces each hosting 10 putatively viruliferous eriophyid mites (Aceria ficus Cotte) and placed directly on the top leaves of healthy Cucumis sativus, Chenopodium quinoa. C. amaranticolor, Nicotiana occidentalis, Catharanthus roseus, Fraxinus excelsior plants, and fig seedlings. Donor and test plants were analyzed by electron microscopy, RT-PCR and sequencing, whereas the mites (ErMs) underwent molecular assays using Fig mosaic virus (FMV)-specific primers. Mite-infested leaves of fig seedlings and C. roseus showed small yellowish spots after 10 days and 6 weeks, respectively, whereas no symptoms were observed in other test or control plants for three months. Electron microscopy observations showed the occurrence of double membrane bodies (DMBs) in the palisade cells of donor and mite-inoculated fig plants, but not in C. roseus. However, 302 bp RT-PCR products specific to FMV were amplified from donor and inoculated figs, C. roseus and ErMs. Nucleotide identity with the sequence of the FMV isolate in GenBank (accession No. AM941711.6) was 87%, 89% and 87% for donor plant (JQ708183), inoculated fig seedlings (JQ708184) and C. roseus (JQ408437, JQ408438), respectively. The sequences obtained from ErMs (JQ408432, JQ408436) showed 87% and 88% nucleotide identity with the reference FMV isolate, respectively. When dsRNA extracts were analyzed to confirm virus presence in inoculated periwinkles, a complex dsRNA profile was obtained, suggestive of infection by a multipartite virus or by multiple viruses. Sequence from RT-PCR amplicons of dsRNA (JX040436) showed 88% identity with those the reference FMV isolate (AM941716.1) and the donor plant (JQ708183). According to these results, Madagascar periwinkle (C. roseus) can be retained as a new experimental host for FMV and A. ficus appears to be able to transmit FMV from fig to periwinkle plants.Öğe DETECTION OF FIG MOSAIC VIRUS IN VIRULIFEROUS ERIOPHYID MITE ACERIA FICUS(Springer, 2012) Caglayan, K.; Elci, E.; Serce, C. Ulubas; Kaya, K.; Gazel, M.; Medina, V.Fig leaves showing typical fig mosaic symptoms on cv. Bursa siyahi (donor plant) were cut under a stereo microscope into small pieces each hosting 10 putatively viruliferous eriophyid mites (Aceria ficus Cotte) and placed directly on the top leaves of healthy Cucumis sativus, Chenopodium quinoa. C. amaranticolor, Nicotiana occidentalis, Catharanthus roseus, Fraxinus excelsior plants, and fig seedlings. Donor and test plants were analyzed by electron microscopy, RT-PCR and sequencing, whereas the mites (ErMs) underwent molecular assays using Fig mosaic virus (FMV)-specific primers. Mite-infested leaves of fig seedlings and C. roseus showed small yellowish spots after 10 days and 6 weeks, respectively, whereas no symptoms were observed in other test or control plants for three months. Electron microscopy observations showed the occurrence of double membrane bodies (DMBs) in the palisade cells of donor and mite-inoculated fig plants, but not in C. roseus. However, 302 bp RT-PCR products specific to FMV were amplified from donor and inoculated figs, C. roseus and ErMs. Nucleotide identity with the sequence of the FMV isolate in GenBank (accession No. AM941711.6) was 87%, 89% and 87% for donor plant (JQ708183), inoculated fig seedlings (JQ708184) and C. roseus (JQ408437, JQ408438), respectively. The sequences obtained from ErMs (JQ408432, JQ408436) showed 87% and 88% nucleotide identity with the reference FMV isolate, respectively. When dsRNA extracts were analyzed to confirm virus presence in inoculated periwinkles, a complex dsRNA profile was obtained, suggestive of infection by a multipartite virus or by multiple viruses. Sequence from RT-PCR amplicons of dsRNA (JX040436) showed 88% identity with those the reference FMV isolate (AM941716.1) and the donor plant (JQ708183). According to these results, Madagascar periwinkle (C. roseus) can be retained as a new experimental host for FMV and A. ficus appears to be able to transmit FMV from fig to periwinkle plants.Öğe Detection of Hop Stunt Viroid in Pomegranate (Punica granatum L.) Trees in the East Mediterranean Region of Turkey(Int Soc Horticultural Science, 2009) Gazel, M.; Caglayan, K.; Serce, C. Ulubas; Durgac, C.Hop stunt viroid (HSVd) infects several fruit tree species with agronomic importance, including pomegranate. Different pomegranate cultivars growing in Hatay, Adana and Icel provinces were surveyed for the presence of HSVd. The main symptoms observed were yellowing and some alteration of the leaves. This kind of symptoms showing samples and also symptomless leaf samples were collected in June-August 2006. In total 152 different pomegranate leaf samples were analyzed for the presence of HSVd. Total RNAs were isolated from pomegranate leaves and used as template in RT-PCR. HSVd was not detected in all tested pomegranate samples.Öğe Detection of Hop Stunt Viroid in Sweet and Sour Cherry Trees in Turkey by RT-PCR(Int Soc Horticultural Science, 2008) Gazel, M.; Ulubas, C.; Caglayan, K.During a survey of the phytosanitary status of sweet (Prunus avium L.) and sour (P. cerasus L.) cherry trees for viroid diseases in Turkey, 127 plant samples were collected from 36 orchards including cultivar collections of mother blocks and commercial grown trees. All samples were tested for the presence of Hop stunt viroid (HSVd) using RT-PCR methods. Total nucleic acids were extracted from about 500 mg leaf tissue of each sample according to Astrue et al. (1996). As a result of RTPCR tests, HSVd infection was detected in 21 samples. Of the 21 HSVd infected trees, 16 were sweet cherries (13.8% infection rate) and 5 were sour cherries (45.5% infection rate). The presence of HSVd infections in sweet and sour cherry trees represents a serious threat to the stone fruit industry of the country. Considering reported yield losses, it would be desirable to evaluate the impact of RSVd-infection on stone fruit production. This paper is the first report of HSVd in sweet and sour cherry in Turkey.Öğe Development of plants resistant to Plum pox virus by interspecific hybridization between peach and other Prunus species(Int Soc Horticultural Science, 2022) Tanriver, E.; Caglayan, K.; Elcicek, M.; Yegul, M.; Guler, P. Gok; Gazel, M.Sharka, caused by Plum pox virus (PPV), is one of the most important disease of peach [P. persica (L.) Batsch] and other Prunus species. PPV has been present in Turkey since 1968 however, contamination level is restricted in some regions depending on PPV strain. PPV-D and PPV-T strains were identified mostly in residential gardens, whereas PPV-M was mostly detected in the commercial orchards. The aim of this study is to identify genotypes that might be tolerant to PPV-M and to monitor the manner in which the resistance is transmitted to hybrid descendants by interspecific hybridization between peach and other Prunus species. The apricot varieties 'Stark Early Orange' and 'Harcot', the almond cultivar 'Ferraduel' and an accesion of P. davidiana identified as having less susceptibility to PPV were used in interspecific hybridizations to select hybrids to which this characteristic was transmitted. Commercial peach cultivars like 'J.H.Hale', 'Cresthaven', 'Flored', 'Persicum-Tan', 'UFO-3', 'Fantasia', 'Persicum-ATES', 'Pitarina-CS46', 'Crimson Baby' and some rootstocks like Garnem, 'GF 677' and 'Patrones-ARDA' were used in hybridization studies. The F1 individuals grown in insect-proof screen-house were submitted weekly to visual inspections and tested twice by DAS-ELISA and RT-PCR for the presence of PPV in one year period. According to the all evaluation criteria (morphological, phenological and phomological), 17 individuals were selected for phenotyping studies. They have been artificially inoculated by PPV-M strain for further scoring. According to the pomological trait assessments and phenotyping analysis for the presence of PPV, most of the peach genotypes showed symptoms and were DAS-ELISA and RT-PCR positive, indicating their susceptibility to PPV. On the basis of phenotyping studies, 4 PPV tolerant peach hybrids from the hybrid families in which P. davidiana and 'Ferraduel' were used as donors were selected for further breeding programs to observe their agronomic traits under the agro-ecological conditions of Adana-Turkiye.Öğe Evaluation of the susceptibility of different Prunus rootstocks to natural infection of plum pox virus-T(2013) Caglayan, K.; Serce, C.U.; Gazel, M.; Kaya, K.; Cengiz, F.C.; Vidal, E.; Cambra, M.Plum pox virus (PPV) has been observed in Turkey since 1968, but was not widespread except in apricot and plum trees in home gardens and ornamental parks in restricted areas. Susceptibility of six different Prunus rootstocks to strain PPV-T was assessed under natural inoculum pressure in the Izmir-Aegean region during 2010-2011. Aphid populations were monitored from the first week of April to the middle of June by the stickyplant method one year after the rootstock plantation was established. Aphids collected from different rootstocks were tested individually by squash real-time RT-PCR and all rootstocks were regularly tested by DASI-ELISA. The largest aphid populations were observed at the end of May and the most abundant aphid species as averages over the two years were Myzus persicae (20.15%), Hyalopterus pruni (18.64%), Aphis craccivora (9.04%) and Aphis gossypii (8.36%). In 2011, the highest percentage of viruliferous aphids was found in M. persicae (34.78%), followed by H. pruni (32.50%), Macrosiphum euphorbiae (25.00%), A. gossypii (23.80%), A. spiraecola (12.50%) and A. craccivora (10.00%). Of the six Prunus rootstocks tested, only Nemaguard and Myrobalan 29C were infected by PPV-T, infection rate in 2010 being 6.0% (Nemaguard) and 4.0% (Myrobalan 29C). The infection rate increased to 16.0% for Nemaguard and 14.0% for Myrobalan 29C in 2011. However, the other rootstocks, Prunus marianna GF8.1, Docera6, GF677 and Garnem tested negative for PPV-T throughout 2011. PPV isolates obtained from naturally infected apricot trees (inoculum source) and from infected rootstocks in the experimental plot were characterized as PPV-T and had more than 99.5% nucleotide sequence identity.Öğe EVALUATION OF THE SUSCEPTIBILITY OF DIFFERENT PRUNUS ROOTSTOCKS TO NATURAL INFECTION OF PLUM POX VIRUS-T(Springer, 2013) Caglayan, K.; Serce, C. U.; Gazel, M.; Kaya, K.; Cengiz, F. C.; Vidal, E.; Cambra, M.Plum pox virus (PPV) has been observed in Turkey since 1968, but was not widespread except in apricot and plum trees in home gardens and ornamental parks in restricted areas. Susceptibility of six different Prunus rootstocks to strain PPV-T was assessed under natural inoculum pressure in the Izmir-Aegean region during 2010-2011. Aphid populations were monitored from the first week of April to the middle of June by the sticky-plant method one year after the rootstock plantation was established. Aphids collected from different rootstocks were tested individually by squash real-time RT-PCR and all rootstocks were regularly tested by DASI-ELISA. The largest aphid populations were observed at the end of May and the most abundant aphid species as averages over the two years were Myzus persicae (20.15%), Hyalopterus pruni (18.64%), Aphis craccivora (9.04%) and Aphis gossypii (8.36%). In 2011, the highest percentage of viruliferous aphids was found in M. persicae (34.78%), followed by H. pruni (32.50%), Macrosiphum euphorbiae (25.00%), A. gossypii (23.80%), A. spiraecola (12.50%) and A. craccivora (10.00%). Of the six Prunus rootstocks tested, only Nemaguard and Myrobalan 29C were infected by PPV-T, infection rate in 2010 being 6.0% (Nemaguard) and 4.0% (Myrobalan 29C). The infection rate increased to 16.0% for Nemaguard and 14.0% for Myrobalan 29C in 2011. However, the other rootstocks, Prunus marianna GF8.1, Docera6, GF677 and Garnem tested negative for PPV-T throughout 2011. PPV isolates obtained from naturally infected apricot trees (inoculum source) and from infected rootstocks in the experimental plot were characterized as PPV-T and had more than 99.5% nucleotide sequence identity.Öğe Experimental transmission efficiency of Plum pox virus T by Myzus persicae Sulzer (Homoptera: Aphididae)(Int Soc Horticultural Science, 2017) Ates, S. Yalcin; Gazel, M.; Serce, C. Ulubas; Caglayan, K.[Abstract Not Available]Öğe First report of 'candidatus phytoplasma asteris' (group 16sri-b) infecting sweet cherries in Turkey(2013) Çaglayan, K.; Gazel, M.; Küçükgöl, C.; Paltrineri, S.; Contaldo, N.; Bertaccini, A.Five-year-old sweet cherry (Prunus avium L.) trees, exhibiting symptoms typical of phytoplasma diseases were observed in the Turkish province of Usak during 2011. The percentage of symptomatic plants, scattered in the orchards, was nearly 40%. Samples were collected during late spring and early summer from trees showing proliferation of branches, off season flowering and decline. In order to establish phytoplasma association with these symptoms, nucleic acid was extracted from leaf midribs of 10 symptomatic and five symptomless plants. Nested PCR assays using universal phytoplasma primers P1/P7 followed by R16F2n/R2 and by 16R758f/16R1232r (Duduk et al., 2013) provided positive responses for seven of the symptomatic samples. Restriction fragment length polymorphism (RFLP) analysis was performed on PCR products using restriction enzymes Tsp509I, Tru1I and AluI. Preliminary RFLP identification was confirmed by nested PCR assays with primers R16(I)F1/R1 (Lee et al., 1994) followed by RFLP analysis, that allowed phytoplasma classification in the 16SrI-B subgroup. Since all amplicons showed identical restriction profile, according to the enzymes and primers employed, one of them was sequenced in both directions using primers R16(I)F1 and R16(I)R1. The 1,006 nucleotide long sequence, deposited in GenBank under the accession No. KF476062, showed 99.0% identity with 16S rDNA from several phytoplasmas related to 'Candidatus Phytoplasma asteris', including strains associated with cherry little leaf (GenBank AY034089) and cherry bunchy leaf (Gen- Bank HM067754), that are affiliated to a different 16SrI subgroup (Jomantiene et al., 2011). This is the first report of 'Ca. P. asteris' infecting sweet cherries in Turkey.Öğe First Report of Grapevine Pinot gris virus in Grapevine in Turkey(Amer Phytopathological Soc, 2016) Gazel, M.; Caglayan, K.; Elci, E.; Ozturk, L.[Abstract Not Available]Öğe FIRST REPORT OF GRAPEVINE SYRAH VIRUS 1 IN GRAPEVINE IN TURKEY(Edizioni Ets, 2017) Caglayan, K.; Gazel, M.; Kocabag, H. D.[Abstract Not Available]Öğe First report of little cherry virus 1 in Turkey(2011) Ulubas Serçe, C.; Gazel, M.; Çaglayan, K.[No abstract available]Öğe FIRST REPORT OF LITTLE CHERRY VIRUS 1 IN TURKEY(Springer, 2011) Serce, C. Ulubas; Gazel, M.; Caglayan, K.[Abstract Not Available]Öğe First report of Olive latent virus 1 from olive trees in Turkey(Springer, 2007) Serce, C. Ulubas; Yalcin, S.; Gazel, M.; Caglayn, K.; Faggioli, F.[Abstract Not Available]Öğe Fourthy-Five Years of Sharka Disease in Turkey(Int Soc Horticultural Science, 2015) Caglayan, K.; Serce, C. Ulubas; Gazel, M.Sharka disease in Turkey has firstly been reported in 1968 in Edirne (Marmara region) which is located next to the Bulgarian border. Nowadays, new PPV outbreaks have been reported in Central Anatolia (Ankara, Kayseri), Aegean (Izmir) and Mediterranean regions (Adana, Mersin, Hatay). The distribution of PPV strains was mainly related to the geographical location and the period of PPV introduction in these regions. PPV-M was mainly detected in peach, nectarine and apricot which were recently imported from abroad to the Mediterranean region. PPV-T was detected in apricot and plums in Central Anatolia and in the Aegean Regions where PPV has been endemic and existing for years. These distributions might indicate that new outbreaks may be mainly due to latently infected material that has passed through the border control. Epidemiology and rootstock susceptibility to PPV has also been recently accomplished. A breeding program has been started in 2006 and its main aim is to obtain dried apricot cultivars resistant to PPV and well adapted to Turkish conditions.Öğe Incidence and genetic diversity of peach latent mosaic viroid isolates in Turkey(2008) Gazel, M.; Ulubas Serce, C.; Caglayan, K.; Luigi, M.; Faggioli, F.In 2004 and 2005, 51 peach and 5 nectarine trees from different peach growing areas of the East Mediterranean Region of Turkey were analyzed by RT-PCR for the presence of Peach latent mosaic viroid (PLMVd). Most of the samples were from germplasm collections and did not show specific symptoms. PLMVd was detected in three nectarine and eight peach trees. The nucleotide sequence of the eleven PLMVd isolates was determined. Like previously studied PLMVd isolates, Turkish isolates varied in length from 336 to 340 nucleotides. Multiple alignments and phylogenic analyses showed that 31 clones of 11 isolates clustered in PLMVd Group III. Three clones of isolate 99 showed the greatest variability among the isolates characterized in this work and 12 previously characterized isolates. mFold analysis of secondary structures of the three clones of isolate 99 showed that most of the variations are located in a stem involved in the pseudoknot structure, in loop 10 and in the hammerhead arm region. PLMVd isolate 99 showed a high identity with isolate 162.4 from peach cv. Early May Crest.Öğe INCIDENCE AND GENETIC DIVERSITY OF PEACH LATENT MOSAIC VIROID ISOLATES IN TURKEY(Springer, 2008) Gazel, M.; Serce, C. Ulubas; Caglayan, K.; Luigi, M.; Faggioli, F.In 2004 and 2005, 51 peach and 5 nectarine trees from different peach growing areas of the East Mediterranean Region of Turkey were analyzed by RT-PCR for the presence of Peach latent mosaic viroid (PLMVd). Most of the samples were from germplasm collections and did not show specific symptoms. PLMVd was detected in three nectarine and eight peach trees. The nucleotide sequence of the eleven PLMVd isolates was determined. Like previously studied PLMVd isolates, Turkish isolates varied in length from 336 to 340 nucleotides. Multiple alignments and phylogenic analyses showed that 31 clones of 11 isolates clustered in PLMVd Group III. Three clones of isolate 99 showed the greatest variability among the isolates characterized in this work and 12 previously characterized isolates. mFold analysis of secondary structures of the three clones of isolate 99 showed that most of the variations are located in a stem involved in the pseudoknot structure, in loop 10 and in the hammerhead arm region. PLMVd isolate 99 showed a high identity with isolate 162.4 from peach cv. Early May Crest.Öğe Investigation of resistance of apricot progeny to Plum pox virus through molecular markers(Int Soc Horticultural Science, 2017) Ates, S. Yalcin; Gazel, M.; Serce, C. Ulubas; Asma, B. M.; Caglayan, K.Plum pox virus (PPV) is the causal agent of sharka disease, which is mainly destructive on apricot (Prunus armeniaca L.), plum (Prunus domestica L.) and peach (Prunus persica L.). There are no efficient control methods except using PPV-free propagating materials and planting PPV-resistant or at least less-susceptible rootstocks. Therefore, lots of studies have been conducted in recent years on breeding of PPV-resistant plants. The objective of this study was the introduction and development of marker-assisted selection (MAS) for PPV resistance in F-1 and F-2 progenies of some Turkish apricot cultivars. Local apricot cultivars 'Adilcevaz 5', 'cologlu', 'Sekerpare' and 'cataloglul crossed with PPV-resistant 'Stark Early Orange' (SEO) were screened with molecular markers PGS1.21 and PGS2.23 co segregating with resistance to PPV in 2011. Of all combinations, seven of 20 progeny obtained from SEO x 12 of 34 progeny obtained from SEO x 'Adilcevaz 5, five of 10 progeny obtained from SEO x 'coloklu, 15 of 37 progeny obtained from SEO x `Sekerpare' and eight of 33 progeny obtained from SEO x 'cataloglu' exhibited resistant alleles.
