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1

Zambon, Yuri, Alessandro Canel, Assunta Bertaccini, and Nicoletta Contaldo. "Molecular Diversity of Phytoplasmas Associated with Grapevine Yellows Disease in North-Eastern Italy." Phytopathology® 108, no. 2 (February 2018): 206–14. http://dx.doi.org/10.1094/phyto-07-17-0253-r.

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A 3-year survey was conducted in Northern Italy to verify the presence and diversity of phytoplasmas in selected vineyards showing symptoms of severe yellows. Symptomatic and asymptomatic grapevines were sampled, and insects were collected using yellow sticky traps. The phytoplasmas detected in grapevine samples were different according to the years: “flavescence dorée” (16SrV-C/D) was detected together with other phytoplasmas such as 16SrXII-A (‘Candidatus Phytoplasma solani’-related, bois noir), 16SrI-B (‘Ca. P. asteris’-related, aster yellows), 16SrX-B (‘Ca. P. prunorum’-related, European stone fruit yellows), and 16SrV-A (‘Ca. P. ulmi’-related, elm yellows). Moreover, phytoplasmas belonging to 16SrVII-A (‘Ca. P. fraxini’-related) and 16SrVI (‘Ca. P. trifolii’-related) subgroups were also identified. Identification of phytoplasmas was also carried out from insects and showed the presence of some of these phytoplasmas in Scaphoideus titanus and Orientus ishidae: 16SrXII-A, 16SrVII, and 16SrVI phytoplasmas were detected in specimens of both species, while 16SrXII-A and 16SrI-B phytoplasma strains were identified in Orientus ishidae and Hyalesthes obsoletus, and 16SrX-B in S. titanus. Direct sequencing of selected amplicons obtained from 16S rRNA, rp, and tuf genes from grapevine and insect samples confirmed the phytoplasma identification. The 16SrVII-A and 16SrVI phytoplasmas were never detected before in grapevine, S. titanus and Orientus ishidae in Europe and their epidemiological importance is being monitored.
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2

Gajardo, A., N. Fiore, S. Prodan, S. Paltrinieri, S. Botti, A. M. Pino, A. Zamorano, J. Montealegre, and A. Bertaccini. "Phytoplasmas Associated with Grapevine Yellows Disease in Chile." Plant Disease 93, no. 8 (August 2009): 789–96. http://dx.doi.org/10.1094/pdis-93-8-0789.

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An extensive survey was performed from 2002 to 2006 to detect and identify phytoplasmas associated with Chilean grapevines. Nested polymerase chain reaction assays using phytoplasma universal primer pairs P1/P7 and R16F2n/R2 detected phytoplasmas in 34 out of the 94 samples tested (36%). Restriction fragment length polymorphism (RFLP) analyses, cloning, and sequencing allowed identification of phytoplasmas belonging to ribosomal subgroups 16SrI-B, 16SrI-C, 16SrVII-A, and 16SrXII-A. The 16SrVII-A phytoplasma represents a new finding in grapevine; moreover, variability of the RFLP profile was observed in some of the 16SrXII-A phytoplasmas, indicating possible new ribosomal subgroups. Mixed phytoplasma infections and infections of phytoplasmas together with one or more viruses also occurred.
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3

Constable, F. E., J. R. Whiting, J. Jones, K. S. Gibb, and R. H. Symons. "The Distribution of Grapevine Yellows Disease Associated with the Buckland Valley Grapevine Yellows Phytoplasma." Journal of Phytopathology 151, no. 2 (February 2003): 65–73. http://dx.doi.org/10.1046/j.1439-0434.2003.00681.x.

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4

Iasur-Kruh, Lilach, Tirtza Zahavi, Roni Barkai, Shiri Freilich, Einat Zchori-Fein, and Vered Naor. "Dyella-Like Bacterium Isolated from an Insect as a Potential Biocontrol Agent Against Grapevine Yellows." Phytopathology® 108, no. 3 (March 2018): 336–41. http://dx.doi.org/10.1094/phyto-06-17-0199-r.

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Yellows diseases, caused by phytopathogenic bacteria of the genus Phytoplasma, are a major threat to grapevines worldwide. Because conventional applications against this pathogen are inefficient and disease management is highly challenging, the use of beneficial bacteria has been suggested as a biocontrol solution. A Dyella-like bacterium (DLB), isolated from the Israeli insect vector of grapevine yellows (Hyalesthes obsoletus), was suggested to be an endophyte. To test this hypothesis, the bacterium was introduced by spraying the plant leaves, and it had no apparent phytotoxicity to grapevine. Fluorescent in situ hybridization analysis showed that DLB is colonizing grapevine phloem. Because phytoplasmas inhabit the same niche, DLB interactions with this phytopathogen were examined. When the isolate was introduced to phytoplasma-infected Chardonnay plantlets, morphological disease symptoms were markedly reduced. The mode of DLB action was then tested using bioinformatics and system biology tools. DLB genome analysis suggested that the ability to reduce phytoplasma symptoms is related to inhibition of the pathogenic bacterium. These results provide the first step in examining the potential of DLB as a biological control agent against phytoplasmas in grapevine and, possibly, other agricultural crops.
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5

Davis, Robert E., Ellen L. Dally, Yan Zhao, and Tony K. Wolf. "Genotyping Points to Divergent Evolution of ‘Candidatus Phytoplasma asteris’ Strains Causing North American Grapevine Yellows and Strains Causing Aster Yellows." Plant Disease 102, no. 9 (September 2018): 1696–702. http://dx.doi.org/10.1094/pdis-10-17-1690-re.

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Grapevine yellows diseases occur in cultivated grapevine (Vitis vinifera L.) on several continents, where the diseases are known by different names depending upon the identities of the causal phytoplasmas. In this study, phytoplasma strains associated with grapevine yellows disease (North American grapevine yellows [NAGY]) in vineyards of Pennsylvania were characterized as belonging to 16S ribosomal RNA (rRNA) gene restriction fragment length polymorphism group 16SrI (aster yellows phytoplasma group), subgroup 16SrI-B (I-B), and variant subgroup I-B*. The strains (NAGYI strains) were subjected to genotyping based on analyses of 16S rRNA and secY genes, and to in silico three-dimensional modeling of the SecY protein. Although the NAGYI strains are closely related to aster yellows (AY) phytoplasma strains and are classified like AY strains in subgroup I-B or in variant subgroup I-B*, the results from genotyping and protein modeling may signal ongoing evolutionary divergence of NAGYI strains from related strains in subgroup 16SrI-B.
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6

Bendel, Nele, Andreas Backhaus, Anna Kicherer, Janine Köckerling, Michael Maixner, Barbara Jarausch, Sandra Biancu, et al. "Detection of Two Different Grapevine Yellows in Vitis vinifera Using Hyperspectral Imaging." Remote Sensing 12, no. 24 (December 18, 2020): 4151. http://dx.doi.org/10.3390/rs12244151.

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Grapevine yellows (GY) are serious phytoplasma-caused diseases affecting viticultural areas worldwide. At present, two principal agents of GY are known to infest grapevines in Germany: Bois noir (BN) and Palatinate grapevine yellows (PGY). Disease management is mostly based on prophylactic measures as there are no curative in-field treatments available. In this context, sensor-based disease detection could be a useful tool for winegrowers. Therefore, hyperspectral imaging (400–2500 nm) was applied to identify phytoplasma-infected greenhouse plants and shoots collected in the field. Disease detection models (Radial-Basis Function Network) have successfully been developed for greenhouse plants of two white grapevine varieties infected with BN and PGY. Differentiation of symptomatic and healthy plants was possible reaching satisfying classification accuracies of up to 96%. However, identification of BN-infected but symptomless vines was difficult and needs further investigation. Regarding shoots collected in the field from different red and white varieties, correct classifications of up to 100% could be reached using a Multi-Layer Perceptron Network for analysis. Thus, hyperspectral imaging seems to be a promising approach for the detection of different GY. Moreover, the 10 most important wavelengths were identified for each disease detection approach, many of which could be found between 400 and 700 nm and in the short-wave infrared region (1585, 2135, and 2300 nm). These wavelengths could be used further to develop multispectral systems.
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7

Magarey, P. A. "Grapevine Yellows, a Widespread, Apparently New Disease in Australia." Plant Disease 70, no. 7 (1986): 694d. http://dx.doi.org/10.1094/pd-70-694d.

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8

Welser, Mary Jean, and Martin C. Goffinet. "COMPARATIVE ANATOMY OF YELLOWS-INFECTED `CHARDONNAY' GRAPEVINES IN NEW YORK STATE, VIRGINIA, AND SOUTH AUSTRALIA." HortScience 41, no. 3 (June 2006): 495D—495. http://dx.doi.org/10.21273/hortsci.41.3.495d.

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Grapevine yellows is a destructive, worldwide disease of grapevines that is caused by a phytoplasma, a bacterium-like organism that infects and disrupts the vascular system of shoots. The North American form of grapevine yellows (NAGY) has been observed in New York State since the mid-1970s and in Virginia since the mid-1990s. Symptoms duplicate those of vines suffering from an Australian disease complex known as Australian grapevine yellows (AGY). We sought to determine if infected `Chardonnay' vines have common anatomical characteristics across the three regions. At each geographic site in late summer, 2003–04, leaf and internode samples were taken from younger green regions of shoots and from mature basal regions in the fruiting zone. These were processed for histology. The anatomy of each organ type was compared between locations on the shoot, between geographic locations, and between affected and normal shoots. The phloem was the only tissue universally affected in vines with NAGY or AGY symptoms. In stem internodes, both primary phloem and secondary phloem showed many senescent cells, abnormally proliferated giant cells, and hyperplasia. In affected secondary phloem there was disruption of the radial files of cells that normally differentiate from the cambium into mature phloem cell types. Normal bands of secondary phloem fibers (“hard phloem”) in internodes were weak or absent in affected vines. Leaves also had disrupted phloem organization but near-normal xylem organization in vines with symptoms. Leaves of infected vines frequently showed a disruption of sugar transport out of the leaf blades, manifested by a heavy buildup of starch in chloroplasts of mesophyll cells and bundle-sheath cells.
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9

Klejdysz, Tomasz, Agnieszka Zwolińska, Marcin Walczak, and Michał Kobiałka. "The first record of a potential pest Orientus ishidae (Matsumura, 1902) (Hemiptera: Cicadellidae) in Poland." Journal of Plant Protection Research 57, no. 2 (June 1, 2017): 107–12. http://dx.doi.org/10.1515/jppr-2017-0014.

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Abstract This study provides the first data on the occurrence of the mosaic leafhopper Orientus ishidae (Matsumura, 1902) (Hemiptera: Cicadellidae) in Poland. This species is native to Southeast Asia, adventive in Europe and feeds on cultivated plants. Orientus ishidae is a well-known carrier of Grapevine flavescence doree phytoplasma which causes the grapevine yellows disease. Symptoms of phytoplasma diseases of grapevine include deformations, leaf chlorosis and withering of plants. The appearance of this species in Poland might be caused by observed climate variations and insufficient plant health controls in the international trade of plants.
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10

Davis, Robert E., Ellen L. Dally, Yan Zhao, Ing-Ming Lee, Wei Wei, Tony K. Wolf, LeAnn Beanland, et al. "Unraveling the Etiology of North American Grapevine Yellows (NAGY): Novel NAGY Phytoplasma Sequevars Related to ‘Candidatus Phytoplasma pruni’." Plant Disease 99, no. 8 (August 2015): 1087–97. http://dx.doi.org/10.1094/pdis-11-14-1185-re.

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North American grapevine yellows (NAGY) disease has sometimes been attributed to infection of Vitis vinifera L. by Prunus X-disease phytoplasma (‘Candidatus Phytoplasma pruni’) but this attribution may not be fully adequate. In this study, phytoplasma strains related to ‘Ca. Phytoplasma pruni’ were found in NAGY-diseased grapevines in Maryland, Pennsylvania, Virginia, Ohio, Missouri, and New York State. Based on restriction fragment length polymorphism analysis of 16S ribosomal RNA gene (16S rDNA) sequences, the strains (termed NAGYIII strains) were classified in group 16SrIII (X-disease group) but they contained a recognition site for the restriction endonuclease MseI that is not present in the 16S rDNA of ‘Ca. Phytoplasma pruni’. The 16S rDNA of the strains differed by three or four nucleotides from that of ‘Ca. Phytoplasma pruni’, indicating that they belonged to two novel 16S rDNA sequevars, designated NAGYIIIα and NAGYIIIβ. Both sequevars differed from ‘Ca. Phytoplasma pruni’ by a single base in each of three regions corresponding to species-unique (signature) sequences described for ‘Ca. Phytoplasma pruni’. Phylogenetic analyses of 16S rRNA genes and SecY proteins, and single-nucleotide polymorphism analyses of secY and ribosomal protein genes, further distinguished the two grapevine sequevar lineages from one another and from ‘Ca. Phytoplasma pruni’. The NAGYIIIα and NAGYIIIβ sequevars also differed from ‘Ca. Phytoplasma pruni’ in regions of the folded SecY protein that are predicted to be near or exposed at the outer surface of the phytoplasma membrane. No evidence indicated that diseased grapevines contained any phytoplasma strain conforming to ‘Ca. Phytoplasma pruni’ sensu stricto. Because the NAGYIII sequevars have not been reported in X-disease, a question is raised as to whether NAGYIII and Prunus X-disease are caused by different phytoplasma genotypes.
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11

Sharon, Rakefet, Maor Tomer, Tamar Sokolsky, Carmit Sofer-Arad, and Tirtza Zahavi. "Trap plants reduces grapevine yellows disease incidence in commercial vineyards." Phytopathogenic Mollicutes 5, no. 1s (2015): S107. http://dx.doi.org/10.5958/2249-4677.2015.00045.6.

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12

Pierro, R., A. Passera, A. Panattoni, P. Casati, A. Luvisi, D. Rizzo, P. A. Bianco, F. Quaglino, and A. Materazzi. "Molecular Typing of Bois Noir Phytoplasma Strains in the Chianti Classico Area (Tuscany, Central Italy) and Their Association with Symptom Severity in Vitis vinifera ‘Sangiovese’." Phytopathology® 108, no. 3 (March 2018): 362–73. http://dx.doi.org/10.1094/phyto-06-17-0215-r.

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Bois noir (BN) is the most widespread disease of the grapevine yellows complex in the Euro-Mediterranean area. BN is caused by ‘Candidatus Phytoplasma solani’ (BNp), transmitted from herbaceous plants to grapevine by polyphagous insect vectors. In this study, genetic diversity among BNp strains and their prevalence and possible association with grapevine symptom severity were investigated in a Sangiovese clone organic vineyard in the Chianti Classico area (Tuscany). Field surveys over 2 years revealed a range of symptom severity on grapevine and an increase of BN incidence. A TaqMan allelic discrimination assay detected only tufB type b among BNp strains, suggesting the prevalence of the bindweed-related ecology. Nucleotide sequence analyses of vmp1 and stamp genes identified 12 vmp1 and 16 stamp sequence variants, showing an overall positive selection for such genes. The prevalent genotype was Vm43/St10, reported for the first time in this study and closely related to strains identified only in the French Eastern Pyrenees. BNp strains identified in the examined vineyard and mostly grouped in separate bindweed-related phylogenetic clusters showed statistically significant differences in their distribution in grapevines exhibiting distinct symptom severity. These results suggest the possible occurrence of a range of virulence within BNp strain populations in the Chianti Classico area.
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13

Duduk, B., M. Ivanovic, N. Dukic, S. Botti, and A. Bertaccini. "First Report of an Elm Yellows Subgroup 16SrV-C Phytoplasma Infecting Grapevine in Serbia." Plant Disease 87, no. 5 (May 2003): 599. http://dx.doi.org/10.1094/pdis.2003.87.5.599c.

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During a 2002 survey in Serbia, samples of grapevine (Vitis vinifera) were collected from plants showing typical phytoplasma-like symptoms: leaf roll, leaf redness, vein chlorosis and necrosis, and absence of lignification. The material was collected from one viticultural region (Zupa Aleksandrovac), where the disease was recorded in 2000 and showed an increasing percentage of symptomatic plants every year. Total nucleic acid was extracted separately from leaf midveins and stem bark collected from 10 symptomatic and 2 asymptomatic plants. Phytoplasma infection was detected using polymerase chain reaction (PCR) assays with universal primer pair P1/P7 for the amplification of phytoplasma 16S rRNA gene, and primer pair FD9f2/FD9r followed by FD9f3/FD9r2 in nested PCR for specific amplification of the FD9 nonribosomal DNA fragment of the EY-group (1). Phytoplasmas were detected in 9 of 10 midvein extracts from symptomatic grapevines (three of cv. Plovdina, two of cv. Smederevka, and four of cv. Gamé). Also, 6 of 10 bark preparations representing stem collections from the same plants were positive (two samples of cv. Plovdina, both samples of cv. Smederevka, and two samples of cv. Gamé). Both collections of midveins and bark tissues from asymptomatic plants were negative. Fragments amplified with universal P1/P7 primers (16S-23S rDNA) were analyzed by restriction fragment length polymorphism with TruI and TaqI restriction enzymes. The phytoplasmas produced identical restriction profiles to those of 16SrV Elm Yellows group and 16SrV-C Flavescence doreé subgroup (2). To our knowledge, this is the first report of phytoplasma infecting grapevines in Serbia, and the first survey in progress to verify the presence of Scaphoideus titanus to determine if this grapevine yellows could be defined as Flavescence dorée. References: (1) E. Angelini et al. Vitis 40:79, 2001. (2) M. Martini et al. Mol. Cell. Probes 16:197, 2002.
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14

Osler, R. "Symptom Expression and Disease Occurrence of a Yellows Disease of Grapevine in Northeastern Italy." Plant Disease 77, no. 5 (1993): 496. http://dx.doi.org/10.1094/pd-77-0496.

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15

Credi, R. "Mycoplasma–like Organisms Associated with a Grapevine Yellows Disease Occurring in Italy." Journal of Phytopathology 141, no. 2 (June 1994): 113–20. http://dx.doi.org/10.1111/j.1439-0434.1994.tb01451.x.

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16

Pierro, Roberto, Alessandra Panattoni, Alessandro Passera, Alberto Materazzi, Andrea Luvisi, Augusto Loni, Marco Ginanni, Andrea Lucchi, Piero Attilio Bianco, and Fabio Quaglino. "Proposal of A New Bois Noir Epidemiological Pattern Related to ‘Candidatus Phytoplasma Solani’ Strains Characterized by A Possible Moderate Virulence in Tuscany." Pathogens 9, no. 4 (April 7, 2020): 268. http://dx.doi.org/10.3390/pathogens9040268.

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Bois noir (BN), associated with ‘Candidatus Phytoplasma solani’ (CaPsol), is the most widespread disease of the grapevine yellows complex worldwide. In this work, BN epidemiology was investigated in a case study vineyard where an unusual CaPsol strain, previously detected only in other host plants, was found to be prevalent in grapevine. Experimental activities included: symptom observation; sampling of symptomatic vines, Auchenorrhyncha specimens, and weeds; molecular detection and typing of CaPsol strains; statistical analyses for determining possible relationships between CaPsol relative concentration, strain type, and symptom severity. Among insects, Reptalus quinquecostatus was the most abundant and was found to be highly infected by CaPsol, while Hyalesthes obsoletus, the main CaPsol vector, was not caught. Moreover, R. quinquecostatus harbored CaPsol strains carrying uniquely the stamp sequence variant St10, also identified as prevalent in vines and in the majority of weeds, and all the secY variants identified in the vineyard. Statistical analyses revealed that CaPsol strains carrying the St10 variant are not associated with severe symptoms, suggesting their possible moderate virulence. Based on such evidence, a new BN epidemiological pattern related to these CaPsol strains and involving grapevine, R. quinquecostatus, and/or weeds is proposed. Furthermore, the possible presence of other players (vectors and weeds) involved in CaPsol transmission to grapevines was highlighted.
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17

Zambon, Yuri, Nicoletta Contaldo, Romolo Laurita, Alessandro Canel, Matteo Gherardi, Vittorio Colombo, and Assunta Bertaccini. "Plasma activated water as a possible sustainable strategy towards grapevine yellows disease management." Phytopathogenic Mollicutes 9, no. 1 (2019): 163. http://dx.doi.org/10.5958/2249-4677.2019.00082.3.

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18

Maixner, M., U. Ahrens, and E. Seemüller. "Detection of Mycoplasmalike Organisms associated with a Yellows Disease of Grapevine in Germany." Journal of Phytopathology 142, no. 1 (September 1994): 1–10. http://dx.doi.org/10.1111/j.1439-0434.1994.tb00001.x.

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19

Prince, James P. "Molecular Detection of Diverse Mycoplasmalike Organisms (MLOs) Associated with Grapevine Yellows and Their Classification with Aster Yellows, X-Disease, and Elm Yellows MLOs." Phytopathology 83, no. 10 (1993): 1130. http://dx.doi.org/10.1094/phyto-83-1130.

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20

Botti, S., and A. Bertaccini. "First Report of Phytoplasmas in Grapevine in South Africa." Plant Disease 90, no. 10 (October 2006): 1360. http://dx.doi.org/10.1094/pd-90-1360b.

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In April 2006, grapevine plants with typical symptoms of yellows (GY) were observed in some South African vineyards. The affected plants showed premature yellowing or reddening and downward rolling of leaves. In some cases, these symptoms were associated with extensive lack of cane lignification that was undistinguishable from yellows symptoms reported in grapevine in the major viticultural areas of the world. Nucleic acids were extracted separately from 0.1 g of fresh leaf midribs and cane phloem scrapes from three symptomatic and three asymptomatic grapevine plants, cv. Shiraz, and from three symptomatic plants, cv. Cabernet, collected from three different locations using Qiagen (Milan, Italy) DNAeasy Plant Mini Kit. A nested polymerase chain reaction (PCR) assay was employed for phytoplasma detection with 2.5 μl of the extracted DNA. Direct and nested PCR assays were performed with P1/P7 (2) and R16F2/R2 (1) universal primer pairs, respectively, obtaining the expected products only from phloem scrapes of the symptomatic plant samples cv. Shiraz. Restriction fragment length polymorphism (RFLP) analyses of R16F2/R2 amplicons with TruI and Tsp509I restriction enzymes, discriminating among phytoplasma ribosomal group and subgroups, showed profiles corresponding to those of “Candidatus Phytoplasma aurantifolia” (ribosomal subgroup 16SrII-B) in all three positive samples. A Stolbur phytoplasma profile (ribosomal subgroup 16SrXII-A) was also observed in one of those samples, indicating the presence of mixed phytoplasma infection (1). Sequencing of the obtained amplicons confirmed the RFLP phytoplasma identification; in particular 16SrXII-A could be the same phytoplasma associated with the ‘Bois Noir’ disease reported in grapevine; the 1601-bp sequence of 16SrII-B phytoplasma showed 98% similarity to U15442, i.e., to the phytoplasma associated with lime witches'-broom disease in Oman (“Ca. P. aurantifolia”) confirming RFLP results. To our knowledge, this is the first report of phytoplasmas in grapevine in South Africa. References: (1) I.-M. Lee et al. Phytopathology 85:728, 1995. (2) B. Schneider et al. Pages 369–380 in: Molecular and Diagnostic Procedures in Mycoplasmology Vol. I. Academic Press Inc., 1995.
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21

Salem, N. M., F. Quaglino, A. Abdeen, P. Casati, D. Bulgari, A. Alma, and P. A. Bianco. "First Report of ‘Candidatus Phytoplasma solani’ Strains Associated with Grapevine Bois Noir in Jordan." Plant Disease 97, no. 11 (November 2013): 1505. http://dx.doi.org/10.1094/pdis-04-13-0428-pdn.

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During a survey carried out in Jordanian vineyards in August and October 2012, grapevine (Vitis vinifera L.) plants showing typical grapevine yellows (GY) disease symptoms, including leaf discoloration and curling, berry shriveling, and irregular maturation of wood, were observed. In the same vineyards, bindweed (Convolvulus arvensis L.) plants showing stunting and leaf chromatic alteration were found, suggesting the involvement of phytoplasmas in the disease etiology. Using a CTAB method, total DNA was extracted from leaf veins of 25 symptomatic and two asymptomatic grapevines, and from five symptomatic and two asymptomatic bindweeds for PCR analysis. DNAs from periwinkle (Catharanthus roseus (L.) G. Don) plants infected by ‘Ca. Phytoplasma asteris’ strain SAY (group 16SrI), ‘Ca. Phytoplasma solani’ strain STOL (group 16SrXII), and ‘Ca. Phytoplasma ulmi’ strain EY1 (group 16SrV), were used as positive controls. DNAs from healthy periwinkle and reactions without template DNA were employed as negative controls. 16S rDNA nested PCRs, carried out using the primer pairs P1/P7, followed by R16F2n/R16R2 (1), yielded an amplicon of the expected size (1,250-bp) in three grapevine and in five bindweed samples, and in positive controls. Amplicons were not produced with DNA from 22 symptomatic grapevines (probably because samples were collected late in the growing season and phytoplasma distribution in plants was non-uniform [2]); nor from asymptomatic plants and negative controls. PCR products were sequenced by commercial services in Italy (Primm, Milan) and Korea (Macrogen Inc., Soul). Representative 16S rDNA nucleotide sequences were deposited in NCBI GenBank with accessions KC835139 (from grapevine) and KC835140 (from bindweed). The 16S rDNA nucleotide sequences of phytoplasmas identified in grapevine and bindweed in Jordan shared >99.5% sequence identity with ‘Ca. Phytoplasma solani’ reference strain STOL (AF248959), and carried identical STOL-unique signature sequences and distinguishing sequence blocks (3). Phylogenetic and in silico RFLP analyses confirmed the affiliation of phytoplasma strains identified in grapevine and bindweed in Jordan to the species ‘Ca. Phytoplasma solani’ (subgroup 16SrXII-A), opening an avenue to future studies on the dissemination and impact of Bois noir (BN) in Jordan. These studies may add new information about BN, previously reported in neighboring countries (4). Further studies will investigate the role of Hyalesthes obsoletus Signoret, a polyphagous Cixiidae responsible for the BN phytoplasma transmission in Europe, and other possible insect vector(s) in the BN spread in Jordan. References: (1) I.-M. Lee et al. Int. J. Syst. Bact. 48:1153, 1998. (2) F. E. Constable et al. Plant Pathol. 52:267, 2003. (3) F. Quaglino et al. Int. J. Syst. Evol. Microb. 63:2879. (4) E. Choueiri et al. Plant Dis. 86:697, 2002.
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22

Quaglino, F., D. Maghradze, N. Chkhaidze, P. Casati, O. Failla, and P. A. Bianco. "First Report of ‘Candidatus Phytoplasma solani’ and ‘Ca. P. convolvuli’ Associated with Grapevine Bois Noir and Bindweed Yellows, Respectively, in Georgia." Plant Disease 98, no. 8 (August 2014): 1151. http://dx.doi.org/10.1094/pdis-01-14-0026-pdn.

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A survey carried out in Georgian vineyards, located in the Khaketi region, in September 2013, showed the presence of vines of the cultivar Chardonnay with typical grapevine yellows (GY) symptoms including leaf discoloration and curling, berry shriveling, and irregular maturation of wood. In the same vineyards, bindweed (Convolvulus arvensis L.) plants showing shoot proliferation and leaf yellowing were found, suggesting the involvement of phytoplasmas in the disease etiology. Total DNA was extracted by a CTAB method from leaf veins of 18 symptomatic and two asymptomatic grapevines, and from four symptomatic and two asymptomatic bindweeds, and analyzed by PCR assays. Moreover, DNA extracted from ‘Candidatus Phytoplasma asteris’ strain SAY (group 16SrI), ‘Ca. P. solani’ strain STOL (group 16SrXII), and ‘Ca. P. ulmi’ strain EY1 (group 16SrV) were used as positive controls. DNA extracted from healthy periwinkle and a reaction mixture without template were employed as negative controls. Nested PCRs targeting the 16S rDNA, carried out using the primer pairs P1/P7 followed by R16F2n/R16R2 (1), produced a band of the expected size (1,250 nt) in all the symptomatic grapevine and bindweed plants, and in the positive controls. No amplification was observed with DNA from asymptomatic plants nor the negative controls. PCR products were sequenced by a commercial sequencing service (Primm, Milan, Italy). The 16S rDNA nucleotide sequences of phytoplasmas identified in all grapevines and in two bindweed samples shared >99.5% sequence identity with ‘Ca. P. solani’ reference strain STOL (GenBank Accession No. AF248959), and carried identical STOL-unique signature sequence and distinguishing sequence blocks (3). Moreover, nucleotide sequences of phytoplasmas identified in the other two bindweed samples shared >99.6% sequence identity with ‘Ca. P. convolvuli’ reference strain BY-S57/11 (JN833705) (2). RFLP and phylogenetic analyses confirmed the affiliation of the phytoplasma strains identified in grapevine and bindweed plants in Georgia to the species ‘Ca. P. solani’ (subgroup 16SrXII-A) and ‘Ca. P. convolvuli’ (subgroup 16SrXII-H). Representative 16S rDNA nucleotide sequences were deposited in NCBI GenBank website with accession nos. KF996535 and KF996536 (‘Ca. P. solani’ from grapevine and bindweed, respectively), and KF996537 (‘Ca. P. convolvuli’). Future studies will focus on investigating the spread and impact of ‘Ca. P. solani’-associated bois noir (BN) in Georgia. In particular, the identification of ‘Ca. P. solani’ in bindweeds suggested the presence of the insect Hyalesthes obsoletus, a polyphagous cixiidae responsible for BN phytoplasma transmission in vineyards in Europe. Accurate surveys and molecular analyses will be performed for identifying the insect vector(s) of the BN associated phytoplasma strains in Georgia. Additional studies will be performed to study the spread and impact of ‘Ca. P. convolvuli,’ identified only in Italy, Germany, Serbia, and Bosnia and Herzegovina (2), throughout the Caucasian countries. References: (1) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (2) M. Martini et al. Int. J. Syst. Evol. Microbiol. 62:2910, 2013. (3) F. Quaglino et al. Int. J. Syst. Evol. Microbiol. 63:2879, 2013.
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Johannesen, Jes, Benjamin Lux, Kristina Michel, Alfred Seitz, and Michael Maixner. "Invasion biology and host specificity of the grapevine yellows disease vector Hyalesthes obsoletus in Europe." Entomologia Experimentalis et Applicata 126, no. 3 (March 2008): 217–27. http://dx.doi.org/10.1111/j.1570-7458.2007.00655.x.

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Green, Margaret J., Dan A. Thompson, and Donald J. MacKenzie. "Easy and Efficient DNA Extraction from Woody Plants for the Detection of Phytoplasmas by Polymerase Chain Reaction." Plant Disease 83, no. 5 (May 1999): 482–85. http://dx.doi.org/10.1094/pdis.1999.83.5.482.

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A simple and efficient procedure for the extraction of high-quality DNA from phytoplasma-infected woody and herbaceous plants for polymerase chain reaction (PCR) detection is described. This procedure does not require phenol, chloroform, or alcohol for the precipitation of nucleic acids. Herbaceous and woody plant material are extracted in an identical manner with no additional purification or enrichment steps required. The method utilizes commercially available microspin-column matrices, and the extraction of total DNA can be achieved in less than 1 h. The method has been used to successfully purify phytoplasma DNA from whole leaves, leaf petioles and midribs, roots, and dormant wood from a diverse selection of plant material. The phytoplasmas detected by PCR include pear decline, western X-disease, peach yellow leaf roll, peach rosette, apple proliferation, Australian grapevine yellows, and Vaccinium witches'-broom.
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Engelbrecht, M., J. Joubert, and J. T. Burger. "First Report of Aster Yellows Phytoplasma in Grapevines in South Africa." Plant Disease 94, no. 3 (March 2010): 373. http://dx.doi.org/10.1094/pdis-94-3-0373a.

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For many years phytoplasma diseases have caused serious losses in most of the major grape-growing regions of the world, except South Africa, where a mixed phytoplasma infection was first reported in 2006 (1). During the early growing season of 2006, symptoms consistent with phytoplasma disease were observed in vineyards in the Olifants River Valley. Symptoms included yellowing of leaves, incomplete lignification of shoots, shortening of internodes, and the abortion of growth tips and immature bunches. Symptomatic shoots and leaves from grapevine cultivars (Merlot, Shiraz, Cabernet Sauvignon, Ruby Cabernet, Pinotage, Corinth, Chardonnay, Columbar, Chenin blanc, Sauvignon blanc, Sultana, and Regal) were collected during the early growing season (November) of 2006, 2007, and 2008. Total DNA was extracted from 32 of these samples (from single plants in the same vineyards over the 3 years) with the Invisorb Spin Plant Mini Kit (Invitek, Berlin, Germany) and tested by nested PCR using two universal primer pairs, P1/P7 and R16F2n/R16R2 (3). The first round of PCR of the 2006 samples yielded 1.8-kb fragments for 17 of the samples, while the nested PCR yielded an additional seven positive samples, confirming the necessity of nested PCR for reliable diagnosis. A similar trend was observed in the 2007 and 2008 PCR test results. All asymptomatic plants, which were included as negative controls, and water controls were negative by nested PCR. Twenty-four 1,245-bp amplicons, generated by nested PCR, were excised from gels, purified with a NucleoSpin Extract II Kit (Macherey-Nagel, Düren, Germany) and directly sequenced. Sequence data was compiled with the BioEdit Version 7.0.4.1 sequence alignment editor software (2), aligned using ClustalW Version 1.4 (4), and a consensus sequence was generated (GenBank Accession No. GQ365729). A BLAST search of the NCBI GenBank database using the individual sequences revealed high sequence identities (≥99%) with the aster yellows phytoplasma group (16SrI) and specifically with the subgroup 16SrI-B. In a comparison of the sequences of the 1.2-kb PCR fragments of 24 local samples with each other, sequence identities of ~99% were observed. These results clearly illustrate that all vines screened were infected with the same phytoplasma. Single nucleotide differences observed between different isolates may indicate the presence of closely related sequence variants of this phytoplasma. Aster yellows occurs worldwide and has been reported to infect grapevine–South Africa can now be added to this list. During the three seasons of our study, the area in which symptomatic vineyards were observed increased significantly, indicating spread by a biological vector. Moreover, infected vineyards were noticed in two other South African grape-growing regions. In contrast to the previous report, which reported a mixed infection of phytoplasmas of groups 16SrXII-A and 16SrII-B (1), PCR screening and sequencing of more than 40 individual samples from these areas confirmed these all to be infected with aster yellows phytoplasma only. To our knowledge, this is the first report of the detection and identification of an aster yellows phytoplasma causing grapevine yellows disease in South Africa. References: (1) S. Botti and A. Bertaccini. Plant Dis. 90:1360, 2006. (2) T. A. Hall. Nucleic Acids. Symp. Ser. 41:95, 1999. (3) I.-M. Lee et al. Phytopathology 83:834, 1993. (4) J. D. Thompson et al. Nucleic Acids Res. 22:4673, 1994.
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Chen, K. H. "Comparison of Monoclonal Antibodies, DNA Probes, and PCR for Detection of the Grapevine Yellows Disease Agent." Phytopathology 83, no. 7 (1993): 915. http://dx.doi.org/10.1094/phyto-83-915.

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27

Kessler, Sébastien, Santiago Schaerer, Nicolas Delabays, Ted C. J. Turlings, Valeria Trivellone, and Patrik Kehrli. "Host plant preferences of Hyalesthes obsoletus, the vector of the grapevine yellows disease ‘bois noir’, in Switzerland." Entomologia Experimentalis et Applicata 139, no. 1 (March 10, 2011): 60–67. http://dx.doi.org/10.1111/j.1570-7458.2011.01107.x.

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IMO, Miriam, Julia LUNEBURG, Thomas HANKELN, Alfred SEITZ, and Jes JOHANNESEN. "Highly polymorphic di- and trinucleotide microsatellite markers for the grapevine yellows disease vector Hyalesthes obsoletus (Auchenorrhyncha: Cixiidae)." European Journal of Entomology 108, no. 1 (January 3, 2011): 161–63. http://dx.doi.org/10.14411/eje.2011.019.

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29

Olivier, C. Y., D. T. Lowery, L. W. Stobbs, C. Vincent, B. Galka, J. Saguez, L. Bittner, et al. "First Report of Aster Yellow Phytoplasmas (‘Candidatus Phytoplasma asteris’) in Canadian Grapevines." Plant Disease 93, no. 6 (June 2009): 669. http://dx.doi.org/10.1094/pdis-93-6-0669a.

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In North America, elm yellows, aster yellows (AY), and X-disease phytoplasmas have been detected in American grapevines (1), and recently, Bois noir was detected in Canadian vineyards from British Columbia (BC) and Ontario (ON) (2). Typical symptoms of grapevine yellows (GY) include leaf rolling and chlorosis, uneven or total lack of lignification of canes, flower abortion or berry withering, and stunting. In 2006 and 2007, independent surveys were conducted by the Canadian Food Inspection Agency (CFIA) and Agriculture and Agri-Food Canada (AAFC) to detect phytoplasmas in Canadian vineyards containing different cultivars in BC, ON, Québec (QC), Nova Scotia, New Brunswick, and Prince Edward Island. The CFIA collected and tested 651 fresh leaf samples from recently imported grapevines and older grapevines in the same or neighboring blocks displaying symptoms typical of those associated with disease caused by phytoplasmas. Many vineyards were surveyed only once. AAFC collected and tested 3,485 samples from symptomatic and asymptomatic grapevines from established vineyards in ON, BC, and QC. The same vineyards were sampled in ON and BC both years; QC vineyards were only sampled in 2007. AAFC-collected leaf samples were freeze dried and stored at –20°C before processing. CFIA samples were tested by a modified real-time PCR assay and TaqMan probe targeting the 16S ribosomal RNA gene that detects a wide range of known phytoplasmas (2). Positive samples were confirmed by conventional PCR using the phytoplasma-specific primers P1/P7 (3) and the resulting ~1,800-bp fragment was cloned and sequenced as previously described (2). DNA extracted by AAFC was amplified by nested PCR technology with universal phytoplasma specific primer pairs P1/P6 and R16R2/R16F2 (3) and the resulting 1,200-bp fragment was cloned and sequenced. Two plants, one located in ON in 2006 and the other in BC in 2007, were found to be infected with an AY-like phytoplasma by the CFIA. The phytoplasmas detected in both infected plants had a 99.9% nt sequence identity with AY phytoplasma sequences from GenBank (Accession Nos. AF222063 and AY665676, respectively), with the BC isolate also showing 100% identity to a strain of AY, ash witches'-broom phytoplasma (GenBank Accession No. AY566302). AAFC detected phytoplasma DNA in both years in a total of 17 symptomatic plants and 21 asymptomatic plants from different vine varieties in ON, BC, and QC. Positive samples were found to have a 99.0% nt sequence identity to AY subgroup 16SrI-A (GenBank Accession No. AY180956). Sequences were exchanged for confirmation of phytoplasma identity and were deposited in Genbank under Accession Nos. FJ659844 and FJ824597. Phytoplasma strains were identified for all plants in which phytoplasmas were detected. Results show that AY is present in vineyards in the provinces of ON, BC, and QC. To our knowledge, this is the first report of AY being detected in grapevines in Canada. References: (1) E. Boudon-Padieu. Bull. O I V, 79:299, 2003. (2) M. Rott et al. Plant Dis. 91:1682, 2007. (3) E. Tanne et al. Phytopathology 91:741, 2001.
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Bianco, P. A., A. Alma, P. Casati, G. Scattini, and A. Arzone. "Transmission of 16SrV phytoplasmas by Scaphoi-deus titanus Ball in northern Italy." Plant Protection Science 37, No. 2 (January 1, 2001): 49–56. http://dx.doi.org/10.17221/8365-pps.

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Flavescence dorée (FD) has been defined as the disease that is caused by a phytoplasmas belonging to group 16SrV (elm yellows) and transmitted to plants by the insect Scaphoideus titanus Ball. We investigated transmission of FD agent by S. titanus in a vineyard located in Veneto region to determine which phytoplasma(s) may be transmitted in this region. Group 16SrV-C phytoplasma was detected and identified in field-collected S. titanus adults, in plant of grapevine (Vitis vinifera L. cv. Gamay) and broadbean (Vicia faba cv. Arlŕ) that had been fed upon by the insects. No evidence of experimental transmission of phytoplasma belonging to 16SrI-B subgroup phytoplasma by S. titanus has been observed.
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Panassiti, Bernd, Michael Breuer, Stacey Marquardt, and Robert Biedermann. "Influence of environment and climate on occurrence of the cixiid planthopperHyalesthes obsoletus, the vector of the grapevine disease ‘bois noir’." Bulletin of Entomological Research 103, no. 6 (April 29, 2013): 621–33. http://dx.doi.org/10.1017/s0007485313000163.

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AbstractSpecies distribution models (SDMs), which are well established in many fields of biological research, are still uncommon in the agricultural risk analysis of pest insects. To exemplify the use of SDMs, we investigated the influence of environmental factors on the occurrence ofHyalesthes obsoletusSignoret (Hemiptera: Cixiidae). The planthopper is the only known vector of the grapevine yellows disease ‘bois noir’. The study was conducted in 145 locations in the Baden region of southwest Germany. The planthopper was surveyed on host plant patches, consisting of stinging nettle and/or bindweeds. We used a stratified modelling framework where (1) species presence–absence data were related to an extensive environmental dataset using logistic regressions; and (2) different types of average models were developed based on an information theoretic method. The results show that the incidence ofH. obsoletusis associated to above- as well as below-ground environmental factors, particularly to the amount of fine soil and average annual precipitation. This result was consistent across all average models. The relative importance of other environmental variables was dependent upon the average model under consideration and thus may vary according to their intended use, either the explanation of habitat requirements or the prediction and mapping of occurrence risks. The study showed that SDMs offer a quantification of species’ habitat requirements and thus, could represent a valuable tool for pest management purposes. By providing examples of current issues of grapevine pests in viticulture, we discuss the use of SDMs in agricultural risk analysis and highlight their advantages and caveats.
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Beanland, LeAnn, Robert Noble, and Tony K. Wolf. "Spatial and Temporal Distribution of North American Grapevine Yellows Disease and of Potential Vectors of the Causal Phytoplasmas in Virginia." Environmental Entomology 35, no. 2 (April 1, 2006): 332–44. http://dx.doi.org/10.1603/0046-225x-35.2.332.

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Ján, Tancik, and Seljak Gabriel. "Occurrence of Scaphoideus titanus Ball and some other Auchenorrhyncha in the vineyards of western Slovakia." Plant Protection Science 53, No. 2 (November 14, 2016): 96–100. http://dx.doi.org/10.17221/40/2016-pps.

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A study of Auchenorrhyncha was carried out in 2014 and 2015 in 7 vineyard plots with different varieties and pest management strategies in the Nitra wine region and Lesser Carpathian wine region in western Slovakia. The aim of this study was to obtain information related to the presence of potential vector insects associated with grapevine yellows phytoplasmas from the Flavescence dorée and Bois noir groups. Insects were collected by sweeping with an entomological net. Thirty species of Auchenorrhyncha were identified as belonging to 6 families. Cicadellidae were the most abundant, comprising 20 species. Scaphoideus titanus was collected at 4 localities. Identification of the phytoplasma vector is critical to the national strategy for assessment and control of vectors spreading the phytoplasma disease in Slovakian vineyards. The first finding of Metcalfa pruinosa was noticed in vineyards in Slovakia.
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34

Davis, Robert E. "Restriction Fragment Length Polymorphism Analyses and Dot Hybridizations Distinguish Mycoplasmalike Organisms Associated withFlavescence Doreeand Southern European Grapevine Yellows Disease in Italy." Phytopathology 83, no. 9 (1993): 772. http://dx.doi.org/10.1094/phyto-83-772.

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35

Lherminier, J., G. Prensier, E. Boudon-Padieu, and A. Caudwell. "Immunolabeling of grapevine flavescence dorée MLO in salivary glands of Euscelidius variegatus: a light and electron microscopy study." Journal of Histochemistry & Cytochemistry 38, no. 1 (January 1990): 79–85. http://dx.doi.org/10.1177/38.1.2294149.

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Flavescence dorée (FD), a grapevine yellows disease, is caused by a mycoplasma-like organism (MLO). A colloidal gold indirect immunolabeling technique identified MLO in salivary glands of a vector leafhopper, Euscelidius variegatus. After aldehyde fixation, tissue samples were prepared by cryoultramicrotomy or embedding in acrylic resins. Double fixation with aldehydes and osmium retroxide, followed by embedding in epon, was also performed. Thin or semi-thin serial sections were treated with polyclonal anti-FD-MLO rabbit antibodies, then with gold-conjugated anti-rabbit IgG. Labeling was revealed using the silver enhancement technique for light microscopy. MLO in frozen thin sections of glands were efficiently labeled. Optimal results were obtained with 4% paraformaldehyde-0.1% glutaraldehyde fixation and low-temperature embedding in LR White resin. Both scattered MLO and unusual dense forms of MLO were easily detected with the electron-dense gold probe. This method distinguished MLO from other membrane-limited bodies and provided a good tool for studying infection in large regions of FD-infected tissues by light microscopy.
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Arnaud, Guillaume, Sylvie Malembic-Maher, Pascal Salar, Patrick Bonnet, Michael Maixner, Carmine Marcone, Elisabeth Boudon-Padieu, and Xavier Foissac. "Multilocus Sequence Typing Confirms the Close Genetic Interrelatedness of Three Distinct Flavescence Dor�e Phytoplasma Strain Clusters and Group 16SrV Phytoplasmas Infecting Grapevine and Alder in Europe." Applied and Environmental Microbiology 73, no. 12 (April 27, 2007): 4001–10. http://dx.doi.org/10.1128/aem.02323-06.

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ABSTRACT Vineyards of southern France and northern Italy are affected by the flavescence dor�e (FD) phytoplasma, a quarantine pathogen transmitted by the leafhopper of Nearctic origin Scaphoideus titanus. To better trace propagation of FD strains and identify possible passage between the vineyard and wild plant compartments, molecular typing of phytoplasma strains was applied. The sequences of the two genetic loci map and uvrB-degV, along with the sequence of the secY gene, were determined among a collection of FD and FD-related phytoplasmas infecting grapevine, alder, elm, blackberry, and Spanish broom in Europe. Sequence comparisons and phylogenetic analyses consistently indicated the existence of three FD phytoplasma strain clusters. Strain cluster FD1 (comprising isolate FD70) displayed low variability and represented 17% of the disease cases in the French vineyard, with a higher incidence of the cases in southwestern France. Strain cluster FD2 (comprising isolates FD92 and FD-D) displayed no variability and was detected both in France (83% of the cases) and in Italy, whereas the more-variable strain cluster FD3 (comprising isolate FD-C) was detected only in Italy. The clonal property of FD2 and its wide distribution are consistent with diffusion through propagation of infected-plant material. German Palatinate grapevine yellows phytoplasmas (PGY) appeared variable and were often related to some of the alder phytoplasmas (AldY) detected in Italy and France. Finally, phylogenetic analyses concluded that FD, PGY, and AldY were members of the same phylogenetic subclade, which may have originated in Europe.
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Chen, K. H., and T. A. Chen. "A novel method for cloning DNA of plant-pathogenic mycoplasmalike organisms." Canadian Journal of Microbiology 41, no. 8 (August 1, 1995): 753–57. http://dx.doi.org/10.1139/m95-104.

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A novel method was developed for cloning the DNA from a representative of plant-pathogenic mycoplasmalike organisms (MLOs). This procedure utilized random amplified polymorphic DNA (RAPD) and basic recombinant DNA techniques. It consisted of amplification of total DNA from diseased plants using one oligonucleotide primer with arbitrary sequence and separation of RAPD products in agarose gels. Unique RAPD band(s) of MLO origin was (were) then recovered from the gel and cloned into the specifically designed vector pCRTM II. With this method, a DNA fragment of the SA2 isolate of grapevine yellows MLO was cloned. Southern blot hybridizations revealed that most of the DNA in the unique RAPD band was derived from MLO. Results from dot-blot hybridizations used for screening showed that approximately 60% of transformants harbored MLO-specific recombinant plasmids. Our approach is relatively simple, quite efficient, and not limited by the amount of diseased material available. It does not depend on DNA sequence information for primer design and does not rely on restriction endonucleases for cloning. In addition, it can be used directly for disease diagnosis and for differentiation of closely related MLOs. Our system may serve as a model for cloning DNAs of other fastidious plant pathogens.Key words: mycoplasmalike organism, DNA, cloning, RAPD, detection.
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Wei, W., E. Pérez-López, L. Bermúdez-Díaz, R. E. Davis, C. Granda-Wong, and Y. Zhao. "First Report of a New Grapevine Yellows Disease in Peru and its Association With Infection by a ‘Candidatus Phytoplasma brasiliense’-Related Phytoplasma Strain." Plant Disease 101, no. 3 (March 2017): 502. http://dx.doi.org/10.1094/pdis-08-16-1152-pdn.

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Eichmeier, Aleš, Eliška Peňázová, Jana Čechová, and Akila Berraf-Tebbal. "Survey and Diversity of Grapevine Pinot gris virus in Algeria and Comprehensive High-Throughput Small RNA Sequencing Analysis of Two Isolates from Vitis vinifera cv. Sabel Revealing High Viral Diversity." Genes 11, no. 9 (September 22, 2020): 1110. http://dx.doi.org/10.3390/genes11091110.

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Grapevine Pinot gris virus (GPGV) is a putative causal agent of grapevine leaf mottling and deformation disease that has been reported worldwide throughout the grapevine-growing regions. Fifty-four grapevines collected from five Algerian grapevine-growing regions were tested for the presence of GPGV in phloem tissues. Eight of the tested grapevines were infected by GPGV. Viromes of two selected Vitis vinifera cv. Sabel grapevines infected by GPGV and showing virus-like symptoms were analyzed by small RNA sequencing. Phylogenetic analyses of the partial coding sequence (cds) of the RNA-dependent RNA polymerase (RdRp) domain showed that all Algerian GPGV isolates were grouped with some already-described asymptomatic isolates. This study provides the first survey of the occurrence of GPGV in Algeria. Moreover, Grapevine fleck virus, Grapevine rupestris stem pitting-associated virus, Grapevine virus B, Grapevine rupestris vein feathering virus, Hop stunt viroid and Grapevine yellow speckle viroid 1 were detected in Algeria for the first time.
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Shahryari, F., T. Allahverdipour, and Z. Rabiei. "Phytoplasmas associated with grapevine yellows disease in Iran: first report of a 'Candidatus Phytoplasma trifolii'-related strain and further finding of a 'Ca. P. solani'-related strain." New Disease Reports 40 (December 2, 2019): 17. http://dx.doi.org/10.5197/j.2044-0588.2019.040.017.

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Naidu, Rayapati, Adib Rowhani, Marc Fuchs, Deborah Golino, and Giovanni P. Martelli. "Grapevine Leafroll: A Complex Viral Disease Affecting a High-Value Fruit Crop." Plant Disease 98, no. 9 (September 2014): 1172–85. http://dx.doi.org/10.1094/pdis-08-13-0880-fe.

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Grapevine (Vitis spp.) is one of the most widely grown fruit crops in the world. It is a deciduous woody perennial vine for which the cultivation of domesticated species began approximately 6,000 to 8,000 years ago in the Near East. Grapevines are broadly classified into red- and white-berried cultivars based on their fruit skin color, although yellow, pink, crimson, dark blue, and black-berried cultivars also exist. Grapevines can be subject to attacks by many different pests and pathogens, including graft-transmissible agents such as viruses, viroids, and phytoplasmas. Among the virus and virus-like diseases, grapevine leafroll disease (GLD) is by far the most widespread and economically damaging viral disease of grapevines in many regions around the world. The global expansion of the grape and wine industry has seen a parallel increase in the incidence and economic impact of GLD. Despite the fact that GLD was recognized as a potential threat to grape production for several decades, our knowledge of the nature of the disease is still quite limited due to a variety of challenges related to the complexity of this virus disease, the association of several distinct GLD-associated viruses, and contrasting symptoms in red- and white-berried cultivars. In view of the growing significance of GLD to wine grape production worldwide, this feature article provides an overview of the state of knowledge on the biology and epidemiology of the disease and describes management strategies currently deployed in vineyards.
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Jović, Jelena, Slavica Marinković, Miljana Jakovljević, Oliver Krstić, Tatjana Cvrković, Milana Mitrović, and Ivo Toševski. "Symptomatology, (Co)occurrence and Differential Diagnostic PCR Identification of ‘Ca. Phytoplasma solani’ and ‘Ca. Phytoplasma convolvuli’ in Field Bindweed." Pathogens 10, no. 2 (February 3, 2021): 160. http://dx.doi.org/10.3390/pathogens10020160.

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Field bindweed (Convolvulus arvensis) is one of the major natural plant hosts and reservoirs of ‘Candidatus Phytoplasma solani’ (‘Ca. P. solani’), the causal agent of plant diseases in diverse agricultural crops, including Bois noir (BN) disease of grapevine. Phylogenetically, the most closely related phytoplasma to ‘Ca. P. solani’, the ‘Ca. P. convolvuli’, induces disease in field bindweed that is known by its symptoms as bindweed yellows (BY). The occurrence, coinfection and symptoms association of the two phytoplasmas in shared host plants were the subject of this study. Specific primers for the amplification of the elongation factor Tu gene (tuf) were developed for the identification of ‘Ca. P. convolvuli’ (by conventional nested PCR), as well as primers for simultaneous detection of ‘Ca. P. solani’ and ‘Ca. P. convolvuli’ by duplex SYBR Green real-time PCR. Among symptomatic bindweed plants, 25 and 41% were infected with a single phytoplasma species, ‘Ca. P. solani’ and ‘Ca. P. convolvuli’, respectively, while 34% were infected with both phytoplasmas. None of the non-symptomatic control plants carried phytoplasma, while non-symptomatic plants from our previous epidemiological studies in BN-affected vineyards were confirmed to be infected solely with ‘Ca. P. solani’. Stamp gene typing revealed Rqg50 and Rqg31 ‘Ca. P. solani’ genotypes in plants coinfected with ‘Ca. P. convolvuli’, while three diverse genotypes (Rqg50, GGY and Rpm35) were identified in a single locality with symptomatic bindweeds infected solely with ‘Ca. P. solani’. Variations in symptoms and their association with each of the phytoplasmas are described and documented. The symptom of bushy appearance could be single out as specific for ‘Ca. P. convolvuli’ infection, while occurrence of ‘Ca. P. solani’ could not be unequivocally associated with specific alterations in infected bindweeds. The results are discussed in the context of the epidemiological and ecological complexity of ‘Ca. P. solani’-induced diseases and the relationship between the two phytoplasma relatives in shared host plant.
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Schneider, B., A. Padovan, S. De La Rue, R. Eichner, R. Davis, A. Bernuetz, and K. Gibb. "Detection and differentiation of phytoplasmas in Australia: an update." Australian Journal of Agricultural Research 50, no. 3 (1999): 333. http://dx.doi.org/10.1071/a98106.

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Phytoplasmas were found in 33 plant species that were not described as host plants in an earlier Australian survey. Plants displayed characteristic symptoms of little leaf, proliferation, and floral abnormalities. Restriction fragment length polymorphism analysis revealed 13 different restriction patterns. The majority of phytoplasmas showed a restriction pattern identical to that of either the tomato big bud (TBB) or sweet potato little leaf V4 (SPLL-V4) phytoplasma. Phytoplasmas from 6 plant species showed a restriction pattern similar to that of the pigeonpea little leaf (PLL) phytoplasma. One phytoplasma from garden bean displayed a restriction pattern identical to that found in papaya dieback and Australian grapevine yellows (AGY) phytoplasmas. Seven new restriction fragment patterns have been detected and sequence analysis of the 16S/23S spacer region revealed that 3 of these phytoplasmas are related to the faba bean phyllody (FBP) group. The spacer region of a graminaceous phytoplasma was most similar to phytoplasmas from the sugarcane white leaf group. Another graminaceous phytoplasma was identical to a phytoplasma from Indonesia. The spacer region of a phytoplasma from poinsettia (PoiBI) was identical to the western X-disease phytoplasma from North America and Europe. The spacer region of a phytoplasma in stylosanthes contained no tRNAIle. Full-length 16S rRNA gene sequences from selected new phytoplasmas were determined to corroborate results obtained from the spacer region analyses. Three of these phytoplasmas (galactia little leaf, vigna little leaf, and stylosanthes little leaf) are, along with the PoiBI phytoplasma and the graminaceous phytoplasmas, members of phytoplasma groups that have not been reported before in Australia.
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44

Terlizzi, F., A. R. Babini, and R. Credi. "First Report of Stolbur Phytoplasma (16SrXII-A) on Strawberry in Northern Italy." Plant Disease 90, no. 6 (June 2006): 831. http://dx.doi.org/10.1094/pd-90-0831a.

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Strawberry (Fragaria × ananassa Duch.) is one of the most important small-fruit crops in northern Italy. During the autumn of 2003, in nurseries located in Ravenna Province (Emilia-Romagna Region), a disease characterized by pronounced stunting and a very poor root system was observed in plants of the cv. Tethis. Older leaves of diseased plants were rolled upward and displayed a marked premature purple discoloration; new leaves showed size reduction, shortened petioles, chlorosis, and were generally cupped. Some of these plants were potted and kept in greenhouse conditions; the following spring, they exhibited typical floral abnormalities as virescent and phylloid petals. Flowers were fully or partly sterile, producing small and deformed fruits; new foliage was dwarfed, asymmetrical, and pale green with chlorotic margins. Later, the affected plants expressed a quick decline consisting of growth cessation, bronzing of mature leaves, wilting, and death. This strawberry yellows-type disease was suggestive of a phytoplasmal infection. Symptoms were identical to “marginal chlorosis”, a stolbur-associated disease occurring in France (4). To acquire more information, field inspections were extended to the 2004 and 2005 seasons. Additional cultivars (Alba, Aromas, Camarosa, Gemma, Maya, NF 20, Queen Elisa, Roxana, and Selva) affected by a similar disorder were identified in strawberry nurseries and production fields from different sites of Ravenna and Forlì-Cesena provinces. Total DNA extracted from collected plants was tested using nested polymerase chain reaction (nPCR) performed with universal phytoplasma primers P1/P7, followed by phytoplasma-specific primer pair R16F2/R2 or group 16SrI and 16SrXII-specific primer pair R16(I)F1/R1 (1,2). Results from nPCR revealed that 21 of 23 diseased nursery plants were infected by a phytoplasma. On the contrary, no positive reaction was obtained with diseased strawberry plants collected from production fields. Subsequent restriction fragment length polymorphism analysis of the nPCR-amplified product R16(I)F1/R1 with enzyme MseI indicated that all diseased plants contained the same phytoplasma belonging to the phytoplasma subgroup 16SrXII-A. Subsequently, these results were confirmed by nPCR using group 16SrXII specific primer pair fSTOL/rSTOL (1). The fragments amplified from three samples were sequenced (GenBank Accession Nos. DQ350615-DQ350617) and showed 99.6 to 99.8% nucleotide sequence identity with a grapevine stolbur isolate (GenBank Accession No. AJ964960). In addition, all samples were assayed using nPCR with primer pair fTuf1/rTuf1 and primers fTufAy/rTufAy, specific for groups 16SrI and 16SrXII (1). Results showed the presence of an expected 945-bp product from infected samples. Sequencing of five amplicons (GenBank Accession Nos. DQ418456-DQ418460) shared 99.4 to 99.9% nucleotide sequence homology with a periwinkle stolbur isolate (GenBank Accession No. L46370). Before now, stolbur phytoplasma has been found to be associated with a strawberry plant showing phyllody symptoms in southern Italy (3). Our report is a wider demonstration of this pathogen infecting strawberry in major cultivations areas of northern Italy. References: (1) M. Langer et al. Extended abstracts ICVG 14:66, 2003. (2) I. M. Lee et al. Phytopathology 84:559, 1994. (3) M. Pastore et al. J. Plant. Pathol. 85:314, 2003. (4) L. Zreik et al. Acta Hortic. 551:101, 2001.
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45

Martin, M. T., L. Martin, M. J. Cuesta, and P. García-Benavides. "First Report of Cylindrocarpon pauciseptatum Associated with Grapevine Decline from Castilla y León, Spain." Plant Disease 95, no. 3 (March 2011): 361. http://dx.doi.org/10.1094/pdis-10-10-0750.

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During a survey for grapevine decline, 10 young grapevines (cvs. Tempranillo and Verdejo) with low vigor and little foliage were collected between June 2008 and August 2009. Small pieces of vascular and brown wood were placed onto malt extract agar supplemented with 0.25 g/liter of chloramphenicol and incubated at 25°C. Fifteen resulting colonies were transferred to potato dextrose agar in petri dishes (90 mm). Colonies with white mycelium covered the dishes after 10 days of incubation at 25°C in darkness; mycelium gradually became yellowish with some brownish aspect. Macroconida were predominantly three septate (40 to 45 to 50 × 8.6 to 9 to 9.5 μm with a length and width ratio of 4.7 to 5 to 5.4), straight, and cylindrical with both ends broadly rounded. Chlamydospora and ovoidal microconidia were observed on synthetic nutrient-poor agar (1). Cylindrocarpon pauciseptatum was not easy to distinguish from other Cylindrocarpon species based on morphological characteristics. Ribosomal internal transcribed spacer region sequences of single-spore cultures confirmed the morphological identification and revealed 100% genetic identity with other isolates of C. pauciceptatum present in GenBank (EF607090), a sequence of the fragment was deposited with Accession No. EU983277. Pathogenicity tests were conducted with two isolates. The inoculations were done on 110R rootstock wood of four different young plants and 15 detached canes of current-season growth (cv. Tempranillo). Plants were inoculated with an agar plug containing C. pauciceptatum; controls were treated with agar only. Grapevines were maintained in a greenhouse at 20 to 25°C. After 3 to 4 months, C. pauciceptatum was reisolated from brown tissues and internal vascular lesions in 45% of inoculated samples, fulfilling Koch's postulates. Control plants were asymptomatic and C. pauciceptatum was not recovered. To our knowledge, this is the first report implicating C. pauciceptatum as a cause of grapevine black foot disease in Spain with potentially significant impact on grapevine nurseries. Reference: (1) H. J. Schroers et al. Mycol. Res. 112:82, 2008.
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46

Koltunow, A. M., L. R. Krake, S. D. Johnson, and M. A. Rezaian. "Two Related Viroids Cause Grapevine Yellow Speckle Disease Independently." Journal of General Virology 70, no. 12 (December 1, 1989): 3411–19. http://dx.doi.org/10.1099/0022-1317-70-12-3411.

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47

Téliz, Daniel, Blanca B. Landa, Hava F. Rapoport, Fernando Pérez Camacho, Rafael M. Jiménez-Díaz, and Pablo Castillo. "Plant-Parasitic Nematodes Infecting Grapevine in Southern Spain and Susceptible Reaction to Root-Knot Nematodes of Rootstocks Reported as Moderately Resistant." Plant Disease 91, no. 9 (September 2007): 1147–54. http://dx.doi.org/10.1094/pdis-91-9-1147.

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Incidence and nematode population densities of plant-parasitic nematodes were determined in 64 samples of soil and grapevine roots collected from commercial vineyards in southern Spain between October 2003 and May 2005. In addition, a histopathological study was done of root-stock roots naturally infected by root-knot nematodes (Meloidogyne spp.). Nematodes infecting the rootstocks were identified according to conventional procedures, and the Meloidogyne spp. were furthermore identified by sequence characterized amplified region-polymerase chain reaction (SCAR-PCR) and isozyme esterase analyses. The most important plant-parasitic nematodes detected, in order of decreasing frequency of total soil infestation and root infection (percentage of samples), were Mesocriconema xenoplax (34.4%), Meloidogyne incognita (26.6%), Meloidogyne javanica (14.1%), Xiphinema index (12.5%), Xiphinema italiae (10.9%), Pratylenchus vulnus (6.3%), and Meloidogyne arenaria (1.6%). No disease symptoms were observed on aboveground plant parts of the infected grapevines, except for plants in some fields where soil was infested with the virus-vector nematodes X. index and X. italiae. Those grapevines showed a yellow mosaic pattern in leaves early in the growing season and the internode shortening characteristic of infections by Grapevine fanleaf virus. Rootstocks infected by root-knot nematodes (Meloidogyne spp.) showed distorted feeder roots and large- to moderate-sized root galls, present either singly or in clusters. Histopathology of galled roots showed a typical susceptible response to infection by root-knot nematodes: cellular alterations were induced in the cortex, endodermis, pericycle, and vascular system, including giant-cell formation and severe distortion of vascular tissues. Most Meloidogyne egg masses ocurred on the surface of the galled root tissues, a position that could facilitate dispersion of the nematode eggs and juveniles and the occurrence of secondary infections. Some of the grapevine rootstocks surveyed in this study (Paulsen 1103, Richter 110, Rupestris du Lot, and SO4) had previously been reported to be resistant to Meloidogyne spp.; however, the population densities of these nematodes found in soil and roots sampled in the present study, as well as the compatible host-parasite relationship revealed by histopathology, indicate a susceptible response to Meloidogyne spp. from southern Spain.
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48

Tessitori, Matilde, Rosa La Rosa, and Cristina Marzachì. "Flavescence Dorée and Bois Noir Diseases of Grapevine Are Evolving Pathosystems." Plant Health Progress 19, no. 2 (January 1, 2018): 136–38. http://dx.doi.org/10.1094/php-10-17-0057-mr.

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Grapevine yellows (GYs) are mainly spread in European viticultural areas. Phytosanitary measures worldwide are based on data obtained on classical pathosystems, but the latest findings suggest that the epidemiological pattern of these diseases is evolving and adapting to different ecological conditions. Risk assessment of introduction in GY-free areas has to be reconsidered in view of the broadening of the range of pathways of entry and means of diffusion in the field. The purpose of this review is to make available an update on findings of recent years to provide useful tools to prevent further spread of these serious diseases of grapevine.
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Szalárdi, Tímea, Antal Nagy, and Gábor Tarcali. "Examination of the American grapevine leafhopper (Scaphoideus titanus Ball) in Debrecen and Micske (Misca, West Romania)." Acta Agraria Debreceniensis, no. 62 (November 2, 2014): 77–81. http://dx.doi.org/10.34101/actaagrar/62/2171.

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Grapevine flavescence dorée (FD) was detected first in Hungary in 2013 in Zala County (South-West-Hungary). The disease is a serious danger for grapevine growing and grapevine propagating production. In 2014, the pathogen has been found in several new places in Hungary, viz. in Vas and Fejér Counties, and it was also detected in the former location in Zala County. The american grapevine leafhopper (Scaphoideus titanus) is the main vector of the disease. This pest was detected first in Hungary in 2006 and then it has spread all over the country. Since we have not detailed distribution data of this pest in surroundings of Debrecen, therefore we made observations in this region in 2014. The presence of the pest was confirmed by yellow sticky cards in two locations in Debrecen and another site in West Romania near to Hungarian border. We found that S. titanus is present in each sampled sites that cause serious potential danger for the appearance and spread of Grapevine flavescence dorée (FD) in this region.
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Credi, R. "Occurrence of Anomalous Mycoplasma-like Organisms in Grapevine Yellows-diseased Phloem." Journal of Phytopathology 142, no. 4 (December 1994): 310–16. http://dx.doi.org/10.1111/j.1439-0434.1994.tb00027.x.

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