Exploiting DNA mini-barcodes as molecular markers to authenticate saffron (Crocus sativus L.)

Villa, Caterina; Costa, Joana; Meira, Liliana; Oliveira, M. Beatriz P.P.; Mafra, Isabel

doi:10.1016/j.foodcont.2016.01.008

Cited by 38 publications

(19 citation statements)

References 32 publications

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“…Besides, the combined approach of matK, as a mini-barcode candidate, with HRM analysis was already demonstrated for Crocus spp. differentiation (Villa, Costa, Meira, Oliveira, & Mafra, 2016). So, the new primers (Lav1-F/Lav1-R) targeting the matK sequence were used to amplify DNA from the voucher specimens under study.…”

Section: End-point Pcrmentioning

confidence: 99%

Botanical authentication of lavender (Lavandula spp.) honey by a novel DNA-barcoding approach coupled to high resolution melting analysis

et al. 2018

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a b s t r a c tMonofloral honeys (such as, lavender honey) are highly appreciated by the consumers due to their flavour, taste and properties. However, since they are considered prime products, they are often targets of adulteration. This work exploits DNA barcoding combined with high resolution melting (HRM) analysis to establish the botanical origin of honey, using lavender honey as a case study. The plastidial matK gene was targeted as a candidate barcode for Lavandula species identification. The method allowed differentiating the species in three clusters with confidence levels >99%, being the results well correlated with the sequencing analysis. It was successfully applied to identify the botanical origin of several lavender honeys, which were grouped in the cluster of the most common species in Portugal (L. stoechas subsp., L. penduculata and L. viridis). The proposed method represents a simple, specific and cost-effective tool to authenticate the botanical origin of honey.

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Section: End-point Pcrmentioning

confidence: 99%

Botanical authentication of lavender (Lavandula spp.) honey by a novel DNA-barcoding approach coupled to high resolution melting analysis

et al. 2018

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“…Therefore, the Bar-HRM method proposed in the present study could be applied in cases where DNA barcoding has failed. Previous reports have shown the success of these combined methods for species authentication in medicinal plants or for commercial herbal products [ 24 , 25 , 31 , 32 , 33 ].…”

Section: Discussionmentioning

confidence: 99%

“…Several studies have reported the successful application of HRM technology using various DNA barcodes in medicinal plant authentication [ 15 , 21 , 22 , 23 ]. Previous publications also claimed that the combination of Bar-HRM analysis is low in cost, simple, highly sensitive and precise in authenticating various plant species and herbal products [ 24 , 25 , 26 , 27 , 28 ]. However, the application of HRM analysis in the authentication of herbal products is still in its initial stages; moreover, the classification and identification of herbal products are limited, especially in developing countries.…”

Section: Introductionmentioning

confidence: 99%

Authenticity Testing and Detection of Eurycoma longifolia in Commercial Herbal Products Using Bar-High Resolution Melting Analysis

Fadzil

Wagiran

Salleh

et al. 2018

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The present study demonstrated High Resolution Melting (HRM) analysis combined with DNA barcode (Bar-HRM) as a fast and highly sensitive technique for detecting adulterants in Eurycoma longifolia commercial herbal products. Targeting the DNA barcoding of the chloroplastic region-ribulose biphosphate carboxylase large chain (rbcL) and the nuclear ribosomal region- internal transcribed spacer 2 (ITS2), PCR amplification and HRM analysis using saturated Eva green dye as the source of fluorescence signals, was accomplished by employing a real-time cycler. The results were further validated by sequencing to identify unknown sequence from Genbank database and to generate phylogenetic tree using neighbour joint (NJ) analysis. Both of the DNA markers exhibited a distinguishable melting temperature and shape of the normalised curve between the reference and the adulterants. In the case of species identification, ITS2 was more successful in differentiating between species. Additionally, detection of admixture sample containing small traces of targeted E. longifolia DNA (w/v) can be detected as low as 5% for rbcL and less than 1% for ITS2, proving the sensitivity and versatility of the HRM analysis. In conclusion, the Bar-HRM analysis is a fast and reliable technique that can effectively detect adulterants in herbal products. Therefore, this will be beneficial for regulatory agencies in order to regulate food safety issues.

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“…The trnL region discriminated 13 species from the Leguminaceae family (Madesis, Ganopoulos, Anagnostis, & Tsaftaris, 2012) and five species used for fruit juice (Faria, Magalhães, Nunes, & Oliveira, 2013). The matK region was used to discriminate five species of Crocus sativus (Villa, Costa, Meira, Oliveira, & Mafra, 2016) and three different Lavandula species in lavender honey (Soares et al, 2018). A combination of the rbcL and trnL regions (two individual simplex reactions) was used to identify eight broomrape species (Rolland, Dupuy, Pelleray, & Delavault, 2016).…”

Section: Introductionmentioning

confidence: 99%

High-resolution melting of multiple barcode amplicons for plant species authentication

et al. 2019

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In recent years, species identification in herbs has attracted considerable attention due to several cases of fraud; hence inexpensive high-throughput authentication methods are highly welcomed. Species authentication is often performed through DNA analysis and several specific regions (barcodes) are considered suitable. Each barcode (Bar) possesses different qualities in terms of universality and discrimination power. A multiplexed format where information can be extracted simultaneously from several barcode regions is seemingly appropriate to ensure the power of both universality and discrimination. In this approach, we amplified DNA from five different barcode regions in a multiplexed PCR format followed by high-resolution melting (HRM). This multiplexed Bar-HRM approach was first applied to plants spanning the plant kingdom and then gradually narrowing down the genetic variability within the Lamiaceae and the Solanaceae families to finally reach closely related cultivars. Universality was demonstrated through distinct melting profiles obtained for species originating from 29 different families spanning the angiosperms, gymnosperm, mosses, and liverwort (Marchantiophyta). Discrimination power was retained for species, sub-species, and a few cultivars through the application of multivariate statistics to the high-resolution melting profiles. This preliminary investigation has shown the potential to discriminate a vast amount of species within the whole plant kingdom. It requires no a priori knowledge of the species' DNA sequence and occurs in a closed system within 2.5 h at a reduced cost per sample compared to other DNA based approaches.

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Exploiting DNA mini-barcodes as molecular markers to authenticate saffron (Crocus sativus L.)

Cited by 38 publications

References 32 publications

Botanical authentication of lavender (Lavandula spp.) honey by a novel DNA-barcoding approach coupled to high resolution melting analysis

Botanical authentication of lavender (Lavandula spp.) honey by a novel DNA-barcoding approach coupled to high resolution melting analysis

Authenticity Testing and Detection of Eurycoma longifolia in Commercial Herbal Products Using Bar-High Resolution Melting Analysis

High-resolution melting of multiple barcode amplicons for plant species authentication

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