A total of 101 Pseudomonas syringae pv. strains, obtained from international culture collections or isolated diseased tissues herbaceous and woody plant species, were assessed by repetitive PCR using the BOX primer, for presence syrB gene. Representative strains also tested pathogenicity to lilac, pear, peach, corn bean, as well virulence lemon zucchini fruits. The unweighted pair‐group method arithmethic averages analysis (UPGMA) genomic fingerprints revealed 17 different patterns which...

A world‐wide collection of 61 Xanthomonas arboricola pv. juglandis strains, isolated from Persian walnut ( Juglans regia L.) or obtained international culture collections and bacterial plant diseases laboratories, were studied by means repetitive polymerase chain reaction (PCR) genomic fingerprinting using ERIC, BOX REP primer sets polyacrylamide gel electrophoresis. Cluster analyses performed UPGMA . Copper resistance, ability to hydrolize starch quinate metabolism the strains was also...

ABSTRACT Thirty-eight bacterial strains isolated from hazelnut (Corylus avellana) cv. Tonda Gentile delle Langhe showing a twig dieback in Piedmont and Sardinia, Italy, were studied by polyphasic approach. All assessed fatty acids analysis repetitive sequence-based polymerase chain reaction (PCR) fingerprinting using BOX ERIC primer sets. Representative also sequencing the 16S rDNA hrpL genes, determining presence of syrB gene, testing their biochemical nutritional characteristics,...

The last decade, smoke and smokeless products claiming to be tobacco-free, including herbal cigarettes shisha, became available on the European market gained popularity. This study proposes a new digital droplet PCR (ddPCR) method, designed based previously developed real-time (qPCR) method being currently used by U.S. Food Drug Administration (FDA) specifically detect presence of tobacco DNA in targeting sequence from Nicotiana tabacum nia-1 gene. To ensure harmonized reliable control...

Screening strategies for GMO detection in food and feed are a crucial aspect testing laboratories streamlining the analytical workflow reducing turnaround time costs. These can be more or less complex even targeted according to ingredients product, but whatever choice, good basic approach is generally based on search 35S promoter (P35S), nos-terminator (T-nos) FMV (P-FMV). In this study, we compare singleplex real PCR method P35S, T-nos P-FMV currently adopted by Italian National Reference...

ABSTRACT A total of 118 fluorescent pseudomonads associated with hazelnut decline, which has been occurring for many years in different areas northern Greece and Italy, were assessed by performing a repetitive PCR analysis enterobacterial intergenic consensus, box element, repetive extragenic palindromic primer sets, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) whole-cell protein extracts, carbon compound utilization analysis, an to determine the presence syrB gene....

A collection of 31 Xanthomonas arboricola pv. corylina strains isolated from Corylus maxima and C. avellana different countries were assessed by means repetitive PCR using ERIC, BOX REP primer sets analysis whole‐cell protein extracts; pathogenicity tests to three hazelnut ( ) cultivars; some key biochemical tests. From these studies, the X. clustered into five groups analysis, respectively, UPGMA cluster with two forming an outlier group these. The showed a high degree similarity. Strain...

A PCR-based DNA walking analysis was performed on a protease product suspected to contain new unauthorized genetically modified microorganism (GMM). Though the characterization of unnatural associations sequences between pUB110 shuttle vector and Bacillus amyloliquefaciens gene coding for protease, presence GMM shown. Based these interest, real-time PCR marker developed target specifically newly discovered GMM, namely protease2. The performance assessed in terms specificity sensitivity....

Common wheat is one of the most important staple food crops worldwide. However, unlike other such as maize or soybean, genetically modified (GM) not yet present in global market. Nonetheless, recent past, adventitious presence GM glyphosate-tolerant volunteers was reported open fields USA. The European Union Reference Laboratory for Food and Feed (EURL-GMFF) therefore called to develop a strategy detect unauthorised samples by using both taxon-specific screening tests. Two candidate common...

The comparison of five real-time polymerase chain reaction (PCR) methods targeted at maize (Zea mays) endogenous sequences is reported. PCR targets were the alcohol dehydrogenase (adh) gene for three and high-mobility group (hmg) other two. have been checked under repeatability conditions several dilution levels on both pooled DNA template from genetically modified (GM) certified reference materials (CRMs) single CRM extracts. Slopes R2 coefficients all curves obtained adopted regression...

Abstract Using a recently developed genetically modified microorganisms (GMM) detection strategy, unexpected contaminations of unauthorized GMM in commercialized microbial fermentation products have been reported. A first-line real-time PCR screening analysis was initially performed to determine the presence key targets frequently found (GM) bacteria. second-line subsequently applied identify specific GMM, including date GM Bacillus velezensis producing protease and B. subtilis vitamin B 2 ....

Abstract In order to point out possible genetic differences, 37 Pseudomonas avellanae strains isolated from northern Greece, Italy and two different areas of central Italy, were assessed by means Repetitive PCR (rep‐PCR) using Enterobacteriaceae Intergenic Consensus (ERIC) primers. After amplifications, the DNA fingerprints revealed three major groups. The Greece those similar showing, in common, products ranging approximately 250 1100 base pairs. However, can be differentiated presence...

The number and variety of genetically modified organisms (GMOs) used globally for the production food feed, potentially circulating in European Union (EU), is constantly increasing. This implies an additional effort EU enforcement laboratories to optimize available resources, contain costs time. A well established approach streamlining analytical workflow introduction a screening step, typically based on smart set real-time polymerase chain reaction (PCR) methods. multiplexing strategy,...

Pseudomonas avellanae is the causal agent of hazelnut ( Corylus avellana ) decline, both in northern Greece and central Italy, two lineages related to geographical origins pathogen have previously been identified. Forty strains, obtained from all areas where disease has so far observed, representing six different subpopulations lineages, were further assessed using insertion‐sequence PCR genomic fingerprinting. The data repetitive‐sequence ERIC BOX primer sets (IS50) analysed statistical...

A rapid polymerase chain reaction (PCR)‐based procedure was developed for the detection of Pseudomonas avellanae , causal agent hazelnut ( Corylus avellana ) decline in northern Greece and central Italy. The partial sequence 16S rRNA gene P. strain PD 2390, isolated Italy, compared with coding same syringae pv. type‐strain LMG 1247 t1 . Primers PAV 1 22 were chosen, after PCR, an amplification product 762 base pairs specifically produced only by 40 strains from No other bacterial species...

Pseudomonas avellanae is the main cause of hazelnut ( Corylus avellana L.) decline, so called ‘moria’, in central Italy where it has already killed more than 30 000 trees. Its current identification very long requiring biochemical, physiological and nutritional tests as well pathogenicity takes not less 6 months for its completion. In present study reliability repetitive polymerase chain reaction (rep‐PCR) technique a rapid accurate such pathogen was compared with traditional method. order...

Using high-throughput metagenomics on commercial microbial fermentation products, DNA from a new unauthorized genetically modified microorganism (GMM), namely the GM

Surveys in submediterranean forests of central Italy were carried out during 1996–98 to verify the possible presence bacterial canker caused by Pseudomonas avellanae wild hazelnut trees ( Corylus avellana L.). Wilted twigs noticed several times especially summer. In other cases, C. growing near orchards appeared completely wilted. Isolates that pathogenic , showing a different degree virulence, obtained both situations. Biochemical, physiological and nutritional tests as well comparison...

ISHS International Symposium on Molecular Markers for Characterizing Genotypes and Identifying Cultivars in Horticulture CHARACTERIZATION AND EVALUATION OF GENETIC DIVERSITY IN PEACH GERMPLASM USING RAPD RFLP MARKERS

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Due to the increasing number of authorized events in European Union, it is crucial for official laboratories enforce market control detect and quantify genetically modified organisms. In this study, an in-house validation quantitative duplex ddPCR methods was performed involving MON87701, MON87769, MON89788 CV-127-9 assays with respect lectin reference gene. Since provide accurate quantification, show less sensitivity PCR inhibitors are more suitable multiplexing compared real-time PCR,...