scholarly journals Diagnosis of Pierce's Disease Using Biomarkers Specific to Xylella fastidiosa rRNA and Vitis vinifera Gene Expression

2010 ◽  
Vol 100 (10) ◽  
pp. 1089-1099 ◽  
Author(s):  
H.-K. Choi ◽  
F. Goes da Silva ◽  
H.-J. Lim ◽  
A. Iandolino ◽  
Y.-S. Seo ◽  
...  
Keyword(s):  
Xylella Fastidiosa ◽  
In Silico Analysis ◽  
Control Measures ◽  
Pierce's Disease ◽  
Gene Transcript ◽  
Rt Pcr ◽  
Breeding Programs ◽  
Pierce’S Disease ◽  
Polymerase Chain ◽  
Pcr Techniques

Pierce's disease (PD), caused by Xylella fastidiosa, represents one of the most damaging diseases of cultivated grape. Management of PD in the vineyard often relies on the removal of infected individuals, which otherwise serve as a source of inoculum for nearby healthy vines. Effective implementation of such control measures requires early diagnosis, which is complicated by the fact that infected vines often harbor high titers of the pathogen in advance of visual symptom development. Here, we report a biomarker system that simultaneously monitors Xylella-induced plant transcripts as well as Xylella ribosomal (r)RNA. Plant biomarker genes were derived from a combination of in silico analysis of grape expressed sequence tags and validation by means of reverse-transcriptase polymerase chain reaction (RT-PCR). Four genes upregulated upon PD infection were individually multiplexed with an X. fastidiosa marker rRNA and scored using either real-time RT-PCR or gel-based conventional RT-PCR techniques. The system was sufficiently sensitive to detect both host gene transcript and pathogen rRNA in asymptomatic infected plants. Moreover, these plant biomarker genes were not induced by water deficit, which is a component of PD development. Such biomarker genes could have utility for disease control by aiding early detection and as a screening tool in breeding programs.

scholarly journals Real-Time Polymerase Chain Reaction for One-Hour On-Site Diagnosis of Pierce's Disease of Grape in Early Season Asymptomatic Vines

2002 ◽  
Vol 92 (7) ◽  
pp. 721-728 ◽  
Author(s):  
N. W. Schaad ◽  
D. Opgenorth ◽  
P. Gaush
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Real Time Pcr ◽  
Xylella Fastidiosa ◽  
Molecular Techniques ◽  
Plant Diseases ◽  
Pierce's Disease ◽  
Chain Reaction ◽  
Pierce’S Disease ◽  
Polymerase Chain

Molecular-based techniques, such as polymerase chain reaction (PCR), can reduce the time needed for diagnosis of plant diseases when compared with classical isolation and pathogenicity tests. However, molecular techniques still require 2 to 3 days to complete. To the best of our knowledge, we describe for the first time a real-time PCR technique using a portable Smart Cycler for one-hour on-site diagnosis of an asymptomatic plant disease. Pierce's disease (PD) of grape, caused by the fastidious bacterium Xylella fastidiosa, causes serious losses in grapes in California and the southeastern United States. The disease has been difficult to diagnose because typical leaf scorching symptoms do not appear until late (June and after) in the season and the organism is very difficult to isolate early in the season. Sap and samples of macerated chips of secondary xylem from trunks of vines were used in a direct real-time PCR without extraction of DNA. Using two different sets of primers and probe, we diagnosed PD in 7 of 27 vines (26%) from four of six vineyards sampled 10 to 12 days after bud break in Kern, Tulare, and Napa counties of California. The diagnosis was confirmed by isolation of Xylella fastidiosa from two of the original PCR positive samples and later from symptomatic leaf petioles of four out of four vines from one vineyard that were originally PCR positive.

scholarly journals Draft Genome Sequence of Xylella fastidiosa subsp. fastidiosa Strain Stag’s Leap

2016 ◽  
Vol 4 (2) ◽  
Author(s):  
J. Chen ◽  
F. Wu ◽  
Z. Zheng ◽  
X. Deng ◽  
L. P. Burbank ◽  
...  
Keyword(s):  
Disease Resistance ◽  
Genome Sequence ◽  
Gene Function ◽  
Xylella Fastidiosa ◽  
Draft Genome ◽  
Draft Genome Sequence ◽  
Pierce's Disease ◽  
Pierce’S Disease ◽  
Knockout Mutants ◽  
Fastidiosa Strain

Xylella fastidiosa subsp. fastidiosa causes Pierce’s disease of grapevine. Presented here is the draft genome sequence of the Stag’s Leap strain, previously used in pathogenicity/virulence assays to evaluate grapevine germplasm bearing Pierce’s disease resistance and a phenotypic assessment of knockout mutants to determine gene function.

scholarly journals Use of immunodot blot and multiplex reverse transcriptase-polymerase chain reaction in dengue virus detection in macerates of Aedes aegypti larvae

2012 ◽  
Vol 3 (1) ◽  
pp. 13
Author(s):  
Aline T.A. Chagas ◽  
Michelle D. Oliveira ◽  
Jose M.S. Mezencio ◽  
Eduardo A.M. Silva ◽  
Leandro L. Oliveira ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Reverse Transcriptase ◽  
Epidemiological Surveillance ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Denv Serotypes ◽  
Polymerase Chain ◽  
Pcr Techniques

The <em>Dengue virus</em> is the main arbovirus that affects man in terms of morbidity and mortality. The detection of the virus is very important for epidemiological surveillance, so here we propose to standardize and compare the immunodot blot (IDB) and multiplex reverse transcriptase-polymerase chain reaction (M-RT-PCR) techniques to detect and characterize the dengue virus (DENV) serotypes in samples of <em>Aedes aegypti</em> larvae. Thus, the IDB and M-RT-PCR techniques were standardized using macerated samples of larvae collected in nature. The use of monoclonal antibodies in IDB has not shown great results, but DENV detection through this method was possible using polyclonal antibodies. The distinction of serotypes 1, 2 and 3 was carried out by M-RT-PCR.

scholarly journals A serological and molecular investigation of American cutaneous leishmaniasis in dogs, three years after an outbreak in the Northwest of Paraná State, Brazil

2009 ◽  
Vol 25 (1) ◽  
pp. 97-104 ◽  
Author(s):  
Gustavo Kiyoshi Massunari ◽  
Evandra Maria Voltarelli ◽  
Demilson Rodrigues dos Santos ◽  
Ademar Rodrigues dos Santos ◽  
Luiz Paschoal Poiani ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cutaneous Leishmaniasis ◽  
Control Measures ◽  
Transmission Control ◽  
Chain Reaction ◽  
American Cutaneous Leishmaniasis ◽  
Molecular Investigation ◽  
Polymerase Chain ◽  
Pcr Techniques

Classic and molecular (polymerase chain reaction - PCR) techniques were used to diagnose American cutaneous leishmaniasis in 149 dogs from an area in the northwest of Paraná State, Brazil, where an American cutaneous leishmaniasis outbreak occurred in 2002. The results were compared to a set of previously obtained results. Twenty-five dogs had positive indirect immunofluorescence (IIF) (titers > 40), including two animals with suggestive lesions. The percentage of dogs with positive IIF was similar to that found in a previous study. The cultures of the lesion, blood and bone marrow were negative for Leishmania. A direct search for the parasite in the lesions proved negative, although PCR tests were positive. The PCR did not detect the DNA of Leishmania (Viannia) in the blood, even for those that had positive PCR in a previous study. The follow up of the 27 dogs showed that the majority of them had maintained the same levels of antibodies that had been detected previously. There was a reduction in the number of dogs with lesions, probably due to the transmission control measures that were adopted after the outbreak.

scholarly journals Grape Cultivar and Sap Culture Conditions Affect the Development of Xylella fastidiosa Phenotypes Associated with Pierce's Disease

PLoS ONE ◽  
2016 ◽  
Vol 11 (8) ◽  
pp. e0160978 ◽  
Author(s):  
Lingyun Hao ◽  
Paulo A. Zaini ◽  
Harvey C. Hoch ◽  
Thomas J. Burr ◽  
Patricia Mowery
Keyword(s):  
Xylella Fastidiosa ◽  
Culture Conditions ◽  
Pierce's Disease ◽  
Grape Cultivar ◽  
Pierce’S Disease

Accuracy of real-time polymerase chain reaction to detect Schistosoma mansoni – infected individuals from an endemic area with low parasite loads

Parasitology ◽  
2020 ◽  
Vol 147 (10) ◽  
pp. 1140-1148
Author(s):  
Fernanda do Carmo Magalhães ◽  
Samira Diniz Resende ◽  
Carolina Senra ◽  
Carlos Graeff-Teixeira ◽  
Martin Johannes Enk ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Schistosoma Mansoni ◽  
Real Time ◽  
Diagnostic Methods ◽  
Control Measures ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Cross Sectional ◽  
Intestinal Schistosomiasis ◽  
Polymerase Chain

AbstractDue to the efforts to control schistosomiasis transmission in tropical countries, a large proportion of individuals from endemic areas present low parasite loads, which hinders diagnosis of intestinal schistosomiasis by the Kato-Katz (KK) method. Therefore, the development of more sensitive diagnostic methods is essential for efficient control measures. The aim was to evaluate the accuracy of a real-time polymerase chain reaction (RT-PCR) to detect Schistosoma mansoni DNA in fecal samples of individuals with low parasite loads. A cross-sectional population-based study was conducted in a rural community (n = 257) in Brazil. POC-CCA® was performed in urine and feces were used for RT-PCR. In addition, fecal exams were completed by 18 KK slides, saline gradient and Helmintex techniques. The combined results of the three parasitological tests detected schistosome eggs in 118 participants (45.9%) and composed the consolidated reference standard (CRS). By RT-PCR, 117 out of 215 tested samples were positive, showing 91.4% sensitivity, 80.2% specificity and good concordance with the CRS (kappa = 0.71). RT-PCR identified 86.9% of the individuals eliminating less than 12 eggs/g of feces, demonstrating much better performance than POC-CCA® (50.8%). Our results showed that RT-PCR is a valuable alternative for the diagnosis of intestinal schistosomiasis in individuals with very low parasite loads.

Radicinin from Cochliobolus sp. inhibits Xylella fastidiosa , the causal agent of Pierce’s Disease of grapevine

2015 ◽  
Vol 116 ◽  
pp. 130-137 ◽  
Author(s):  
Thomas J. Aldrich ◽  
Philippe E. Rolshausen ◽  
M. Caroline Roper ◽  
Jordan M. Reader ◽  
Matthew J. Steinhaus ◽  
...  
Keyword(s):  
Xylella Fastidiosa ◽  
Causal Agent ◽  
Pierce's Disease ◽  
Pierce’S Disease

scholarly journals Citrus and Coffee Strains of Xylella fastidiosa Induce Pierce's Disease in Grapevine

Plant Disease ◽  
2002 ◽  
Vol 86 (11) ◽  
pp. 1206-1210 ◽  
Author(s):  
W.-B Li ◽  
C. -H. Zhou ◽  
W. D. Pria ◽  
D. C. Teixeira ◽  
V. S. Miranda ◽  
...  
Keyword(s):  
United States ◽  
Sequence Data ◽  
Xylella Fastidiosa ◽  
Experimental System ◽  
The United States ◽  
Full Genome Sequence ◽  
Pierce's Disease ◽  
Plant Colonization ◽  
Pierce’S Disease ◽  
Citrus Groves

Xylella fastidiosa causes citrus variegated chlorosis (CVC) disease in Brazil and Pierce's disease of grapevines in the United States. Both of these diseases cause significant production problems in the respective industries. The recent establishment of the glassy-winged sharpshooter in California has radically increased the threat posed by Pierce's disease to California viticulture. Populations of this insect reach very high levels in citrus groves in California and move from the orchards into the vineyards, where they acquire inoculum and spread Pierce's disease in the vineyards. Here we show that strains of X. fastidiosa isolated from diseased citrus and coffee in Brazil can incite symptoms of Pierce's disease after mechanical inoculation into seven commercial Vitis vinifera varieties grown in Brazil and California. Thus, any future introduction of the CVC strains of X. fastidiosa into the United States would pose a threat to both the sweet orange and grapevine industries. Previous work has clearly shown that the strains of X. fastidiosa isolated from Pierce's disease- and CVC-affected plants are the most distantly related of all strains in the diverse taxon X. fastidiosa. The ability of citrus strains of X. fastidiosa to incite disease in grapevine is therefore surprising and creates an experimental system with which to dissect mechanisms used by X. fastidiosa in plant colonization and disease development using the full genome sequence data that has recently become available for both the citrus and grapevine strains of this pathogen.

Nutritional requirements of Xylella fastidiosa, which causes Pierce's disease in grapes

2000 ◽  
Vol 46 (3) ◽  
pp. 291-293 ◽  
Author(s):  
C J Chang ◽  
R C Donaldson
Keyword(s):  
Amino Acids ◽  
Potato Starch ◽  
Xylella Fastidiosa ◽  
Pierce's Disease ◽  
Inorganic Salts ◽  
Nutritional Requirements ◽  
Phenol Red ◽  
Isoleucine Leucine ◽  
Pierce’S Disease ◽  
Starting Point

A defined medium (XF-26) containing 3 inorganic salts, 2 tricarboxylic acids, 17 amino acids, potato starch, phenol red, and agar was used as the starting point for the study. Deletions of one or more ingredients were performed to prepare various media. A medium was considered able to support growth of Xylella fastidiosa strains responsible for Pierce's disease in grapes, only after 10 serial passages had been completed. Of 3 inorganic salts, K2HPO4 and MgSO4·7H2O were essential, and (NH4)2HPO4 was nonessential for growth. Of the Krebs cycle intermediates, all (citrate, alpha-ketoglutarate, succinate, fumarate, malate, and oxaloacetate) but isocitrate supported growth of cultivated strains, whereas only citrate alone or citrate plus succinate supported the primary isolation of PD bacterium. Of 17 amino acids, 6 uncharged polar R groups (asparagine, cysteine, glutamine, glycine, serine, and threonine) supported growth, whereas 8 nonpolar R groups (alanine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, and valine) or 3 positively charged polar groups (arginine, histidine, and lysine) did not. Starch proved to be nonessential.Key words: Xylella fastidiosa, nutritional requirements.

Sign in / Sign up

Export Citation Format

Share Document