scholarly journals First Report of Bois Noir Phytoplasma in Grapevine in Canada

Plant Disease ◽  
2007 ◽  
Vol 91 (12) ◽  
pp. 1682-1682 ◽  
Author(s):  
M. Rott ◽  
R. Johnson ◽  
C. Masters ◽  
M. Green
Keyword(s):  
Tissue Sample ◽  
Hot Water ◽  
Taqman Probe ◽  
Test Procedures ◽  
Specific Primers ◽  
Bois Noir ◽  
Flavescence Dorée ◽  
Nursery Stock ◽  
Stolbur Phytoplasma ◽  
Infected Grapevine

During the summer and fall of 2006, a survey was done to detect European phytoplasmas of quarantine significance in Canadian vineyards. This survey was developed as one of the 2006 import requirements for grapevine nursery stock from Europe. This addresses the increased concerns regarding inadvertent phytoplasma introductions. Grapevines imported in 2006 and established grapevines were observed for symptoms typical of those associated with diseases caused by phytoplasmas on grapevine. Samples were tested from 155 grapevines. One plant, located in the lower Okanagan Valley, British Columbia, tested positive by a modified real-time PCR assay and TaqMan probe targeting the 16S region of the ribosomal RNA gene (1), which detects a wide variety of known phytoplasmas. The sample was further analyzed and found to be positive by conventional PCR with the phytoplasma-specific primers, P1/P7 (3), and Stolbur specific primers, STOL11f2/r1 (2). Additional PCR tests with primers specific to flavescence doree (FD9f/r) (2) and western X disease (P1/W INT) (3) were negative. These phytoplasmas are also known to infect grapevine. The approximate 1,800-bp fragment obtained with P1/P7 was sequenced (GenBank Accession No. EU086529) and found to have 99.7% nucleotide sequence identity to the Stolbur STOL #11 isolate (GenBank Accession No. AF248959) originally isolated from eastern Europe. This was the highest match to any available phytoplasma sequence obtained and indicates that the phytoplasma in the British Columbian sample is an isolate of bois noir, a pest of quarantine significance to Canada. Additional phylogenetic analysis using CLUSTAL W (Lasergene; DNASTAR, Madison, WI) confirmed this result. The presence and identity of the phytoplasma was confirmed from a second tissue sample that was analyzed by PCR and sequenced using the same test procedures as for the first sample, with identical results. The bois noir phytoplasma belongs to the stolbur group (16SrVII) with the principal vector being a cixiid planthopper. Stolbur phytoplasmas cause diseases in other crops, but bois noir disease is caused by a specific member of that group and is the only stolbur phytoplasma known to infect grapevines in Europe. The infected grapevine was from a lot of 1,965 plants of Grenache clone 70 on rootstock 3309 clone 143 that was imported from Europe in 2006. All plants in this importation have been destroyed. This phytoplasma has not been detected in any other grapevines in Canada. Additional import conditions requiring hot water treatment of European vines have been implemented for 2007. Further survey work for phytoplasma in grapevine will continue. References: (1) N. M. Christensen et al. Mol. Plant-Microbe Interact. 17:1175, 2004. (2) X. D. Daire et al. Eur. J. Plant Pathol. 103:507, 1997. (3) C. D. Smart et al. Appl. Environ. Microbiol. 62:2988, 1996.

scholarly journals Phytoplasma diseases of grapevine and the possible measures to control them

2011 ◽  
Vol 17 (3) ◽  
Author(s):  
M. Kölber
Keyword(s):  
Feeding Habits ◽  
Economic Loss ◽  
Hot Water ◽  
Control Measures ◽  
Convolvulus Arvensis ◽  
Preventive Control ◽  
Bois Noir ◽  
Flavescence Dorée ◽  
Scaphoideus Titanus ◽  
Stolbur Phytoplasma

Phytoplasmas are a special group of phloem-living pathogens in several plant species. Grapevine yellows (GY) is a term for phytoplasma diseases occurring on Vitis vinifera and inducing the same or very similar symptoms and causing severe losses worldwide. Flavescence Dorée (16SrV) phytoplasma (FD, species name: ‘Candidatus Phytoplasma vitis’) is considered a quarantine pest in several countries due to its epidemic character and high economic loss it provokes. The leafhopper Scaphoideus titanus is the univoltine and monophagous vector of FD. Bois noir disease caused by stolbur (16SrXII-A) phytoplasma (species name: ‘Candidatus Phytoplasma solani’) is described under different disease names in different countries. Hyalesthes obsoletus (Cixiidae) is the only proved polyphagous vector of BN. However, distribution of BN disease is increasing also on those areas where H. obsoletus is not prevalent or only in a very low number. Therefore the presence of other vectors cannot be concluded. The ‘Tuf-a’ type Stolbur phytoplasma is associated with stinging nettle (Urtica dioica) and the tuf-b type one to field bindweed (Convolvulus arvensis). There are only preventive control measures against phytoplasmas: the use of pathogen-free propagating material, hot water treatment of propagating material, as well as control of vectors and weeds. S. titanus can be efficiently controlled by insecticide treatments. However, in case of H. obsoletus, insecticides are not effective due to the biological characters and feeding habits of the vector.Weed control can reduce H. obsoletus specimen and their abundance to a certain extent. Extensive research is needed on wild hosts of GY phytoplasmas especially on BN phytoplasma and its vectors to the better understanding of their epidemiology.

scholarly journals Developing, Modifying, and Validating a TaqMan Real-Time PCR Technique for Accurate Identification of Leishmania Parasites Causing Most Leishmaniasis in Iran

2021 ◽  
Vol 11 ◽  
Author(s):  
Reza Fotouhi-Ardakani ◽  
Seyedeh Maryam Ghafari ◽  
Paul Donald Ready ◽  
Parviz Parvizi
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Leishmania Major ◽  
Taqman Probe ◽  
Amino Acid Permease ◽  
Specific Primers ◽  
Accurate Identification ◽  
Parasitic Load ◽  
Species Specific

Many laboratory methods are used to diagnose leishmaniasis because it is characterized by varied symptoms and caused by different Leishmania species. A quantitative real-time PCR method based on a TaqMan probe was developed and modified for accurate identification of human cutaneous leishmaniasis (caused by Leishmania major or Leishmania tropica) from endemic areas of Iran. Two gene regions of amino acid permease 3 (AAP3) and cytochrome oxidase II (COII) were considered. Six new sets of species-specific primers and probes were designed. A total of 123 samples were examined and employed to evaluate and validate real-time PCR. According to parasitic load of the genesig®Leishmania Advanced Standard Kit, a serial dilution of purified plasmid (2–2×107 copies/reaction) was prepared under the same conditions for both genes. Specific primers and probes were able to detect three and six parasite copies in AAP3 and COII genes, respectively, and were able to detect three copies of parasites for L. major and L. tropica. The sensitivities of the reference kit and our method were 98.7 and 98.1%, respectively, and specificity was 100% for detecting parasite genomes in all assays. Designed primers and probes performed well in terms of efficiency and regression coefficient. For AAP3 and COII genes, respectively, the linear log range was 7 and the correlation coefficient (R2) was 0.749 and 0.996 for the reference kit using the standard generated curve and 0.98 and 0.96 with serial dilutions of parasite DNA. This research detected L. major and L. tropica definitely and opens the horizon for the other scientists in the multiplex reactions in designing and optimization of the conditions in silico and in vivo.

scholarly journals Recovery of Vitis vinifera L. cv. ‘Kékfrankos’ from ‘bois noir’ disease

2019 ◽  
Vol 156 (3) ◽  
pp. 987-991
Author(s):  
Anikó Mátai ◽  
Péter Teszlák ◽  
Gábor Jakab
Keyword(s):  
Gas Exchange ◽  
Vitis Vinifera ◽  
Photosynthetic Performance ◽  
Photochemical Quenching ◽  
Vitis Vinifera L ◽  
Bois Noir ◽  
Flavescence Dorée ◽  
Exchange Performance ◽  
Concentration Changes ◽  
Non Photochemical Quenching

AbstractInvestigation of diseases caused by phytoplasmas, a group of cell-wall-less gram-positive bacteria has received significant attention in plant pathology. Grapevine is a host of two, genetically distinct phytoplasmas: Line Flavescence dorée (FD) phytoplasma associated to ‘flavescence dorée’ and ‘Candidatus Phytoplasma solani’ responsible for ‘bois noir’ (BN) disease. In the current study, we focused on BN diseased grapevines (Vitis vinifera L. cv. ‘Kékfrankos’), measured their photosynthetic performance and leaf hydrogen peroxide (H2O2) concentration. The latter is generally considered as a key molecule in the process of ‘recovery’ which is a spontaneous and unpredictable long-term remission of disease symptoms. This phenomenon also occurred during the time of our experiment. Infection resulted in reduced gas exchange performance and maximum quantum efficiency of PSII with an increased regulated non-photochemical quenching of PSII and H2O2 concentration. Changes in gas exchange seem to be a systemic response, while reduced photochemistry is a local response to ‘Ca. P. solani’ infection. H2O2 accumulation in BN phytoplasma infected plants, unlike in FD disease, was found to be a typical response to the appearance of a biotic stressor.

scholarly journals Identification and Epidemic Distribution of Two Flavescence Dorée—Related Phytoplasmas in Veneto (Italy)

Plant Disease ◽  
1999 ◽  
Vol 83 (10) ◽  
pp. 925-930 ◽  
Author(s):  
Marta Martini ◽  
Ermanno Murari ◽  
Nicola Mori ◽  
Assunta Bertaccini
Keyword(s):  
16S Rdna ◽  
Fragment Length Polymorphism ◽  
The European Union ◽  
Direct Pcr ◽  
Nested Polymerase Chain Reaction ◽  
Flavescence Dorée ◽  
Scaphoideus Titanus ◽  
Polymerase Chain ◽  
Elm Yellows ◽  
Infected Grapevine

Grapevine yellows associated with phytoplasmas of the elm yellows group (16SrV), better known as flavescence dorée (FD), is a serious quarantine problem in some important grapevine growing regions in the European Union. A survey was carried out in 1997 to 1998 in Veneto region (Italy) where a serious outbreak of FD was in progress. Phytoplasma identification by nested polymerase chain reaction (PCR)/restriction fragment length polymorphism (RFLP) analyses on 275 grapevine samples and on batches of Scaphoideus titanus was carried out. RFLP analyses of the 16S rDNA/spacer region with TaqI detected the presence of two distinct elm yellows phytoplasma subgroups designated 16SrV-C and 16SrV-D in 77 FD-infected grapevine samples. Only phytoplasmas of the 16SrV-D subgroup were detected in S. titanus. In 1997, the two phytoplasma subgroups appeared to be located in two diverse geographic areas; but in 1998, the 16SrV-D type also was detected in the provinces where in 1997 only 16SrV-C type was identified. The sequencing of a 400-bp fragment at the 3′ end of 16S rDNA plus spacer region allowed a specific primer construction that was successfully employed for detection of both FD types in grapevine by direct PCR.

scholarly journals Occurrence of Scaphoideus titanus Ball and some other Auchenorrhyncha in the vineyards of western Slovakia

2016 ◽  
Vol 53 (No. 2) ◽  
pp. 96-100
Author(s):  
Tancik Ján ◽  
Seljak Gabriel
Keyword(s):  
Management Strategies ◽  
National Strategy ◽  
Bois Noir ◽  
Potential Vector ◽  
Flavescence Dorée ◽  
First Finding ◽  
Scaphoideus Titanus ◽  
Phytoplasma Vector ◽  
Wine Region ◽  
And Control

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.

scholarly journals Molecular Diversity of Phytoplasmas Associated with Grapevine Yellows Disease in North-Eastern Italy

2018 ◽  
Vol 108 (2) ◽  
pp. 206-214 ◽  
Author(s):  
Yuri Zambon ◽  
Alessandro Canel ◽  
Assunta Bertaccini ◽  
Nicoletta Contaldo
Keyword(s):  
Molecular Diversity ◽  
Direct Sequencing ◽  
Northern Italy ◽  
Bois Noir ◽  
Aster Yellows ◽  
Flavescence Dorée ◽  
Scaphoideus Titanus ◽  
North Eastern ◽  
Elm Yellows ◽  
Hyalesthes Obsoletus

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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