scholarly journals Traceability of PDO Olive Oil “Terra di Bari” Using High Resolution Melting

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Cinzia Montemurro ◽  
Monica Marilena Miazzi ◽  
Antonella Pasqualone ◽  
Valentina Fanelli ◽  
Wilma Sabetta ◽  
...  
Keyword(s):  
High Resolution ◽  
Olive Oil ◽  
High Resolution Melting ◽  
Virgin Olive Oil ◽  
Cost Effective ◽  
Extra Virgin Olive Oil ◽  
Hrm Analysis ◽  
Olive Cultivars ◽  
Pcr Products ◽  
Set Up

The aim of the research was to verify the applicability of microsatellite (SSR) markers in High Resolution Melting (HRM) analysis for the identification of the olive cultivars used in the “Terra di Bari” PDO extra virgin olive oil. A panel of nine cultivars, widespread in Apulia region, was tested with seventeen SSR primer pairs and the PCR products were at first analysed with a Genetic Analyzer automatic sequencer. An identification key was obtained for the nine cultivars, which showed an unambiguous discrimination among the varieties constituting the “Terra di Bari” PDO extra virgin olive oil: Cima di Bitonto, Coratina, and Ogliarola. Subsequently, an SSR based method was set up with the DCA18 marker, coupled with HRM analysis for the distinction of the Terra di Bari olive oil from non-Terra di Bari olive oil using different mixtures. Thus, this analysis enabled the distinction and identification of the PDO mixtures. Hence, this assay provided a flexible, cost-effective, and closed-tube microsatellite genotyping method, well suited to varietal identification and authentication analysis in olive oil.

scholarly journals Genotyping of GATA4 Gene Variant (G296S) in Malaysian Congenital Heart Disease Subjects by Real-Time PCR High Resolution Melting Analysis

2013 ◽  
Vol 32 (2) ◽  
pp. 152-157
Author(s):  
Nora Fawzi ◽  
Ramachandran Vasudevan ◽  
Patimah Ismail ◽  
Mazeni Alwi ◽  
Ahmad Fazli Abdul Aziz ◽  
...  
Keyword(s):  
Congenital Heart Disease ◽  
Heart Disease ◽  
High Resolution ◽  
Congenital Heart ◽  
High Resolution Melting ◽  
New Technology ◽  
Cost Effective ◽  
Study Cohort ◽  
Hrm Analysis ◽  
Melting Analysis

Summary Background: Congenital heart disease (CHD) is the most common birth defect; however, the underlying etiology is unrecognized in the majority of cases. GATA-binding protein 4 (GATA4), a cardiac transcription factor gene, has a crucial role in the cardiogenesis process; hence, a number of heterozygote sequence variations were identified as a cause of CHD. G296S heterozygote variant is the most frequently reported GATA4 gene sequence alteration. This study aims to investigate the role of G296S variant of the GATA4 gene in Malaysian CHD subjects. Methods: We have investigated 86 Malaysian CHD subjects with cardiac septation defects for the presence of the GATA4 gene heterozygote variant (G296S) by the new technology of high resolution melting (HRM) analysis. Results: Genotyping of G296S (c.886G>A) by HRM analysis shows that all the sample genotypes were of the wild GG type genotype and the heterozygote mutant GA genotype was totally absent from this study cohort. Conclusions: The results of our study showed that the G296S variant of the GATA4 gene was not associated with the development of CHD in Malaysian subjects. The use of HRM analysis proved a cost-effective, high-throughput, specific and sensitive genotyping technique which eliminates the need for unnecessary sequencing.

Barcode DNA high-resolution melting (Bar-HRM) analysis as a novel close-tubed and accurate tool for olive oil forensic use

2013 ◽  
Vol 93 (9) ◽  
pp. 2281-2286 ◽  
Author(s):  
Ioannis Ganopoulos ◽  
Christos Bazakos ◽  
Panagiotis Madesis ◽  
Panagiotis Kalaitzis ◽  
Athanasios Tsaftaris
Keyword(s):  
High Resolution ◽  
Olive Oil ◽  
High Resolution Melting ◽  
Hrm Analysis ◽  
Barcode Dna

scholarly journals Phenolic Compounds and Triterpenes in Different Olive Tissues and Olive Oil By-Products, and Cytotoxicity on Human Colorectal Cancer Cells: The Case of Frantoio, Moraiolo and Leccino Cultivars (Olea europaea L.)

Foods ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2823
Author(s):  
Pujun Xie ◽  
Lorenzo Cecchi ◽  
Maria Bellumori ◽  
Diletta Balli ◽  
Lisa Giovannelli ◽  
...  
Keyword(s):  
Olive Oil ◽  
Virgin Olive Oil ◽  
Olive Tree ◽  
Extra Virgin Olive Oil ◽  
Dry Extract ◽  
Dependent Relationship ◽  
Colorectal Cancer Cells ◽  
Olive Cultivars ◽  
Hct 116 ◽  
Dose Dependent

Phenolic and triterpenoid compounds of the olive tree are recognized as having a key role in health promotion, thanks to their multiple protective actions in humans. To expand the source of these bioactive compounds, the phenolic and triterpenoid profiles of leaf, branch, destoned fruit, destoned pomace, shell, seed, and extra virgin olive oil from the Frantoio, Leccino, and Moraiolo olive cultivars were simultaneously characterized by HPLC-DAD-MS. Overall, 43 molecules were quantitated and expressed on the obtained dry extracts. Oleuropein was mainly concentrated in branches (82.72 g/kg), fruits (55.79 g/kg), leaves (36.71 g/kg), and shells (1.26 g/kg), verbascoside (4.88 g/kg) in pomace, and nüzhenide 11-methyl oleoside (90.91 g/kg) in seeds. Among triterpenoids, which were absent in shells, the highest amount of oleanolic acid was found in olive leaves (11.88 g/kg). HCT-116 colorectal cells were chosen to assess the cytotoxicity of the dry extract, using the phytocomplex from Frantoio, which was the richest in phenols and triterpenoids. The IC50 was also determined for 13 pure molecules (phenols and terpenoids) detected in the extracts. The greatest inhibition on the cell’s proliferation was induced by the branch dry extract (IC50 88.25 μg/mL) and by ursolic acid (IC50 24 μM). A dose-dependent relationship was observed for the tested extracts.

scholarly journals Identification of new avian Infectious Bronchitis virus variants in Iranian poultry flocks by High Resolution Melting curve analysis

2018 ◽  
Vol 69 (1) ◽  
pp. 783 ◽  
Author(s):  
A. MASOUDIAN ◽  
N. SHEIKHI ◽  
M. H. BOZORGMEHRI-FARD
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Melting Curve ◽  
Melting Curve Analysis ◽  
Nucleotide Sequencing ◽  
Infectious Bronchitis ◽  
Hrm Analysis ◽  
High Resolution Melting Curve ◽  
Performance Reduction ◽  
Pcr Products

Avian Infectious bronchitis (IB) is a common coronavirus infection of chickens and responsible for performance reduction and increasing mortality due to subsequent respiratory, renal and/or reproductive disorders. Classification of causative agent is necessary to plan successful vaccination strategies to prevent the infection due to poor inter-strains cross-reaction. To identify dominant circulating strains in Iran, a Real-time PCR combined with 3’ Un-Translated Region (3’ UTR) High Resolution Melting (HRM) analysis designed as a rapid and reliable method for IB Virus (IBV) detection and differentiation. Samples collected from 20-suspected flocks and after PCR products, HRM curves of samples as well as 6 commercial IB live vaccines with 2 standard strains, were analyzed as references. IBV genomes detected in 11 samples while according to HRM analysis and calculating Genotype Confidence Percentage (GCP), 6 positive specimens identified as 793/B field strains and the left 5 found as new IBV variant strains. Then obtained PCR products sent for nucleotide sequencing to determine genotype relativity. All five infectious agents, related to QX-like type and indicating circulation of new variants in Iran as a probable cause of vaccination failures and consequent economical losses.

scholarly journals Biophenolic Compounds Influence the In-Mouth Perceived Intensity of Virgin Olive Oil Flavours and Off-Flavours

Molecules ◽  
2020 ◽  
Vol 25 (8) ◽  
pp. 1969 ◽  
Author(s):  
Alessandro Genovese ◽  
Ferdinando Mondola ◽  
Antonello Paduano ◽  
Raffaele Sacchi
Keyword(s):  
Phenolic Compounds ◽  
Olive Oil ◽  
Volatile Compounds ◽  
Virgin Olive Oil ◽  
Extra Virgin Olive Oil ◽  
Olive Oils ◽  
Perceived Intensity ◽  
Panel Test ◽  
Headspace Concentration ◽  
Set Up

In this study, the influence of phenolic compounds on the sensory scores attributed to extra virgin olive oil (EVOO) by panel test was investigated. Two model olive oils (MOOs) with identical concentrations of volatile compounds, differing only in the amount of biophenols (297 vs. 511 mg kg−1), were analysed by two official panels and by SPME-GC/MS. Six other MOOs set up by the two previous models were also tested and analysed. They were formulated separately with the addition of three off-flavours (‘rancid’, ‘winey–vinegary’ and ‘fusty–muddy’). While high levels of EVOO phenolic compounds did not produce any effect on the headspace concentration of volatile compounds, they did affect the scores of both positive and negative sensory attributes of EVOO, due to the well-known in-mouth interactions between EVOO phenols, saliva and volatile compounds. In particular, a decrease of about 39% in the positive fruity score was found in the presence of a higher concentration of phenols. Regarding EVOO off-flavours, the higher level of phenolic compounds decreased by about 23% the score of ‘fusty–muddy’ defect and increased the score of ‘winey–vinegary’ defect about 733%. No important effect of EVOO phenolics on the perceived intensity of the ‘rancid’ defect was found. These findings could be helpful in explaining some discrepancies of panel test responses observed during extra virgin olive oil shelf life.

Characterization and Classification of Extra Virgin Olive Oil from Five Less Well-Known Greek Olive Cultivars

2016 ◽  
Vol 93 (6) ◽  
pp. 837-848 ◽  
Author(s):  
Ioanna Kosma ◽  
Maria Vavoura ◽  
Stavros Kontakos ◽  
Ioannis Karabagias ◽  
Michael Kontominas ◽  
...  
Keyword(s):  
Olive Oil ◽  
Virgin Olive Oil ◽  
Extra Virgin Olive Oil ◽  
Olive Cultivars

Combination of high resolution melting (HRM) analysis and SSR molecular markers speeds up plum genotyping: case study genotyping the Greek plum GeneBank collection

2016 ◽  
Vol 15 (4) ◽  
pp. 366-375 ◽  
Author(s):  
Georgios Merkouropoulos ◽  
Ioannis Ganopoulos ◽  
Athanasios Tsaftaris ◽  
Ioannis Papadopoulos ◽  
Pavlina Drogoudi
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Cost Effective ◽  
Ex Situ ◽  
Hrm Analysis ◽  
Prunus Salicina ◽  
Genebank Collection ◽  
Of Greece

AbstractIn a germplasm bank collection the conservation and characterization of genetic resources is a prerequisite in order to use the material in breeding projects aiming the creation of new cultivars. In the present study, 54 Prunus salicina domestica and Prunus domestica genotypes (including seven Greek cultivars), maintained in the ex situ National Genebank Collection of Greece, were classified using microsatellite (simple sequence repeat, SSR) markers on high resolution melting (HRM) analysis. The SSR primer pairs were chosen from the published literature as originally designed on Prunus species. This combined approach was used to genotype all plum accessions of the collection highlighting the benefits of either method (HRM and SSRs) for cultivar identification. Dendrograms for P. domestica and P. salicina and a combined one with all the genotypes assayed were produced. A total of 15 from the 19 P. domestica accessions analysed, including all the Greek accessions but ‘Avgati Skopelou’, were grouped into the same clade in the combined dendrogram, whereas the remaining four were dispersed into the P. salinica clades. Bayesian structure analysis confirmed that ‘Avgati Skopelou’ differs from the rest of the Greek plum cultivars since it was not grouped into the same cluster. The combination of HRM and SSRs, provided a considerably faster, cost-effective, closed-tube microsatellite genotyping method for molecular characterization of plum cultivars.

Microsatellite high-resolution melting (SSR-HRM) analysis for genotyping and molecular characterization of an Olea europaea germplasm collection

2014 ◽  
Vol 12 (3) ◽  
pp. 273-277 ◽  
Author(s):  
Aliki Xanthopoulou ◽  
Ioannis Ganopoulos ◽  
Georgios Koubouris ◽  
Athanasios Tsaftaris ◽  
Chrysa Sergendani ◽  
...  
Keyword(s):  
High Resolution ◽  
Microsatellite Markers ◽  
Molecular Characterization ◽  
Olea Europaea ◽  
High Resolution Melting ◽  
Germplasm Collection ◽  
Hrm Analysis ◽  
Olive Cultivars ◽  
Olea Europaéa

Olea europaea L. has been cultivated in the Mediterranean region for thousands of years and is of major economic importance. The origin of olive cultivars remains as complex to trace as their identification. Thus, their molecular characterization and discrimination will enable olive germplasm management. In addition, it would be a useful tool for authentication of olive products. High-resolution melting (HRM) analysis, coupled with five microsatellite markers, was integrated to facilitate molecular identification and characterization of main O. europaea cultivars collected from the National Olive Tree Germplasm Collection established in Chania, Greece. The five microsatellite loci used were highly informative and generated a unique melting curve profile for each of the 47 cultivars and for each microsatellite tested. In particular, three microsatellite markers (DCA03, DCA09 and DCA17), which generated 29 HRM profiles, were sufficient to genotype all the olive cultivars studied, highlighting their potential use for cultivar identification. Furthermore, this assay provided a flexible, cost-effective and closed-tube microsatellite genotyping method well suited for molecular characterization of olive cultivars.

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