Distinguishing between Bread Wheat and Spelt Grains Using Molecular Markers and Spectroscopy

Curzon, Arie Y; Chandrasekhar, K.; Nashef, Y. K.; Abbo, Shahal; Bonfil, David J.; Reifen, Ram; Bar-el, S.; Avneri, Asaf; Ben‐David, Roi

doi:10.1021/acs.jafc.9b00131

Cited by 14 publications

(9 citation statements)

References 16 publications

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“…Enzyme-linked immunosorbent assay (ELISA) [9], chromatographic methods [9][10][11][12][13], near-infrared spectroscopy (NIRS) [14,15] and real-time PCR [16][17][18][19][20][21][22] and other molecular techniques [23] were used for the specific detection of cereal species in food. Compared to other analytical parameters, such as proteins and fats, DNA is sufficiently stable, depending on the degree of processing, to be detected even in more processed foods [24].…”

Section: Introductionmentioning

confidence: 99%

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Development and validation of a method for quantification of common wheat, durum wheat, rye and barley by droplet digital PCR

Schulze

Geuthner

Mäde

2021

Eur Food Res Technol

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Food fraud is becoming a prominent topic in the food industry. Thus, valid methods for detecting potential adulterations are necessary to identify instances of food fraud in cereal products, a significant component of human diet. In this work, primer–probe systems for real-time PCR and droplet digital PCR (ddPCR) for the detection of these cereal species: bread wheat (together with spelt), durum wheat, rye and barley for real-time PCR and ddPCR were established, optimized and validated. In addition, it was projected to validate a molecular system for differentiation of bread wheat and spelt; however, attempts for molecular differentiation between common wheat and spelt based on the gene GAG56D failed because of the genetic variability of the molecular target. Primer–probe systems were further developed and optimized on the basis of alignments of DNA sequences, as well as already developed PCR systems. The specificity of each system was demonstrated on 10 (spelt), 11 (durum wheat and rye) and 12 (bread wheat) reference samples. Specificity of the barley system was already proved in previous work. The calculated limits of detection (LOD95%) were between 2.43 and 4.07 single genome copies in real-time PCR. Based on the “three droplet rule”, the LOD95% in ddPCR was calculated to be 9.07–13.26 single genome copies. The systems were tested in mixtures of flours (rye and common wheat) and of semolina (durum and common wheat). The methods proved to be robust with regard to the tested conditions in the ddPCR. The developed primer–probe systems for ddPCR proved to be effective in quantitatively detecting the investigated cereal species rye and common wheat in mixtures by taking into account the haploid genome weight and the degree of milling of a flour. This method can correctly detect proportions of 50%, 60% and 90% wholemeal rye flour in a mixture of wholemeal common wheat flour. Quantitative results depend on the DNA content, on ploidy of cereal species and are also influenced by comminution. Hence, the proportion of less processed rye is overestimated in higher processed bread wheat and adulteration of durum wheat by common wheat by 1–5% resulted in underestimation of common wheat.

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Section: Introductionmentioning

confidence: 99%

“…The barley system 1, based on the hordein gene and developed by zeltner et al, showed good specificity not only for Hordeum vulgare, but also for Hordeum murinum [21]. In addition, several systems claimed to be able to distinguish bread wheat from spelt [14,16,22].…”

Section: Introductionmentioning

confidence: 99%

Development and validation of a method for quantification of common wheat, durum wheat, rye and barley by droplet digital PCR

Schulze

Geuthner

Mäde

2021

Eur Food Res Technol

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“…Spelt Fatty acids profile [14] Triticum species PCR-RFLP (Q-locus) [15] Spelt RLP-LOC-CE, Real-time PCR (γ-gliadin) [16] Farro della Garfagnana in cereal mixtures padlock probe ligation and multiplex microarray [17] Spelt LC-MS peptide markers identification [18] Einkorn, emmer and spelt tubulin-based polymorphism (TBP) [19] Spelt PCR (γ-gliadin, Q-locus); NIR [20] Italian emmer landraces Spectroscopy and chemometrics [21] Spelt Duplex droplet digital PCR (Q-locus) [22] Most of the proposed assays are DNA-based methods used for the identification and quantification of spelt. Moreover, assays developed by Voorhuijzen et al [17] and by Foschi et al [21] are focused on the traceability of accessions specifically cultivated in Italian environments.…”

Section: Analytical Target Methods Referencementioning

confidence: 99%

“…γ-gliadin polymorphisms were exploited in the analytical protocols developed by Mayer et al [ 16 ] and by Curzon et al [ 20 ].…”

Section: Introductionmentioning

confidence: 99%

A Digital PCR Assay to Quantify the Percentages of Hulled vs. Hulless Wheat in Flours and Flour-Based Products

Morcia

Bergami

Scaramagli

et al. 2021

Biology

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Several food products, made from hulled wheats, are now offered by the market, ranging from grains and pasta to flour and bakery products. The possibility of verifying the authenticity of wheat species used at any point in the production chain is relevant, in defense of both producers and consumers. A chip digital PCR assay has been developed to detect and quantify percentages of hulless (i.e., common and durum wheat) and hulled (i.e., einkorn, emmer and spelt) wheats in grains, flours and food products. The assay has been designed on a polymorphism in the miRNA172 target site of the AP2-5 transcription factor localized on chromosome 5A and involved in wheat spike morphogenesis and grain threshability. The assay has been evaluated even in a real-time PCR system to assess its applicability and to compare the analytical costs between dPCR and real-time PCR approaches.

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“…22 Fingerprinting methods involving spectroscopy techniques have been developed that overcome the limits of targeted approaches and allow multiple wheat species authentication such as the DNA-based fingerprinting assay based on TBP 21 and an integrative near-infrared spectroscopic model based on Q gene classification for discrimination between spelt and bread wheat. 23 However, the practical application of these models may be hampered, especially in the case of multi-cereal mixtures, due to complexity of the models. In an overview of the most promising biomarkers and analytical tools for authentication of organic food products, Capuano et al 1 noted that a single marker was insufficient to confirm authenticity.…”

Section: Introductionmentioning

confidence: 99%

Identification of biomarkers in hydrosoluble extracts from spelt and wheat cultivated in different production systems

et al. 2020

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BACKGROUND: In the present paper, a method for differentiation between common and spelt wheat grown in different farming systems (biodynamic, ecological, integrated, conventional), based on biomarkers identified from aqueous flour extracts (nitrogen and 14 soluble carbohydrates) was employed. RESULTS:The identification and determination of soluble carbohydrate content were carried out using gas chromatographymass spectrometry, with the UV spectrum generated by mass spectrometry for comparison with the WILEY database. Soluble carbohydrates were determined in the peak area between 21.92 and 43.63 min −1 retention time. The obtained data set was analyzed by multivariate statistical techniques. It was revealed that common wheat exerted a much more pronounced tendency than spelt wheat to be influenced by the farming system. CONCLUSION: This differentiation was particularly well visualized after subjecting the data set to principal component analysis (PCA) and cluster analysis (CA). In the PCA graph, all spelt samples were positioned closer to the corresponding control sample, in contrast to the wheat samples, which were distributed over a huge area in the factor space. CA showed that the spelt samples grown under different farming systems were highly similar and grouped into one cluster. Common wheat samples cultivated under organic, biodynamic and integrated system were similar and represented the second cluster, whereas that cultivated under the conventional system was clearly separated from other samples.

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Distinguishing between Bread Wheat and Spelt Grains Using Molecular Markers and Spectroscopy

Cited by 14 publications

References 16 publications

Development and validation of a method for quantification of common wheat, durum wheat, rye and barley by droplet digital PCR

Development and validation of a method for quantification of common wheat, durum wheat, rye and barley by droplet digital PCR

A Digital PCR Assay to Quantify the Percentages of Hulled vs. Hulless Wheat in Flours and Flour-Based Products

Identification of biomarkers in hydrosoluble extracts from spelt and wheat cultivated in different production systems

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