Molecular identification of honey entomological origin based on bee mitochondrial 16S rRNA and COI gene sequences

Kek, Siok Peng; Chin, Nyuk Ling; Tan, Sheau Wei; Yusof, Yus Aniza; Chua, Lee Suan

doi:10.1016/j.foodcont.2017.02.025

Cited by 42 publications

(36 citation statements)

References 25 publications

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“…However, as referred, the entomological source can indirectly give information about the geographical origin of honey. Despite this, only few studies have attempted the entomological identification of honey, either based on protein (Ramón-Sierra, Ruiz-Ruiz, & de la Luz Ortiz-Vázquez, 2015) or DNA analysis (Kek, Chin, Tan, Yusof, & Chua, 2017;Prosser & Hebert, 2017;Soares et al, 2018b). For this purpose, DNA-based methods are considered as being the most suitable tools since they allow unequivocal species identification.…”

Section: Introductionmentioning

confidence: 99%

Towards honey authentication: Differentiation of Apis mellifera subspecies in European honeys based on mitochondrial DNA markers

et al. 2019

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Honey is the natural sweet substance produced by Apis mellifera honeybees in Europe. Depending on the country/region, the A. mellifera subspecies native to Europe belong to three different lineages: A (A. m. iberiensis), M (A. m. iberiensis and A. m. mellifera) and C (A. m. ligustica and A. m. carnica). In this work, two DNAbased approaches were developed with the aim of entomological authentication of European honeys. A cytb specific PCR assay was proposed to identify A-lineage honeybees, while a second method based on real-time PCR coupled to high resolution melting analysis targeting the COI gene was developed to differentiate C-and Mlineages honeybees. The proposed methodologies were validated successfully with honeys of known origin and applied to the entomological authentication of 20 commercial samples from different European countries. The results highlight the predominance of honeys from C-lineage honeybees in Europe, except in Iberian Peninsula countries (honey from A-lineage honeybees).

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Section: Introductionmentioning

confidence: 99%

Towards honey authentication: Differentiation of Apis mellifera subspecies in European honeys based on mitochondrial DNA markers

et al. 2019

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“…These authors screened 6 samples of honey produced by A. mellifera and 1 by Melipona beecheii. The identification of bee species DNA on honey samples was also achieved by Kek, Chin, Tan, Yusof, and Chua (2017) based on mitochondrial DNA (mtDNA) sequences and phylogenetic analysis by means of forensically informative nucleotide sequencing. However, both reports rely on extensive sequencing analysis, which is not a cost-effective and highthroughput approach.…”

Section: Introductionmentioning

confidence: 99%

Novel diagnostic tools for Asian (Apis cerana) and European (Apis mellifera) honey authentication

Soares

Grazina

Mafra

et al. 2018

Food Research International

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Honey can be produced by different species of honeybees, with two being of economic importance due to their use in apiculture, namely Apis mellifera (known as European honeybee) and Apis cerana (known as Asian honeybee). Due to the decline of the wild populations of the Asian honeybee, this honey generally attains much higher market value, being prone to adulteration. This work aims at proposing new tools, based on the use of molecular markers, for the entomological authentication of honey. To this end, new species-specific primers were designed targeting the tRNA-cox2 intergenic region and allowing the detection of A. cerana DNA by qualitative polymerase chain reaction (PCR). Additionally, a novel real-time PCR method with high resolution melting analysis was developed to target the 16S rRNA gene of both bee species, allowing their discrimination in different clusters. The proposed methodologies were further applied with success in the authentication of Asian and European honey samples by the identification of honeybee DNA, demonstrating the usefulness of these simple and cost-effective new approaches.

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“…Schnell et al (2010) mentioned that amplicons of 100-150 bp are within the size range of genetic markers that can be used for species identification. The exclusion of Kelulut B in Table 3 was due to inability of DNA sequence match at 150 bp although a bigger amplicon size of 300 bp gave positive match (Kek et al, 2017b). Only one commercial sample (Y) from the three enabled the identification of the bee species (Apis mellifera).…”

Section: Sequencing Of Pcr Products For Honey Origin Identificationmentioning

confidence: 99%

“…Kek et al . () have also successfully used the mitochondrial large subunit ribosomal RNA (16S rRNA) gene, while Soares et al . () used mitochondrial tRNAleu‐cox2 (cytochrome c oxidase subunit II) intergenic region and the 16S rRNA gene for bee identification in honey samples.…”

Section: Introductionmentioning

confidence: 99%

“…Prosser & Hebert (2017) have identified bee source of Apis mellifera, a common European honey bee and also the stingless bee of Melipona beecheii in honey using the COI gene. Kek et al (2017b) have also successfully used the mitochondrial large subunit ribosomal RNA (16S rRNA) gene, while Soares et al (2018) used mitochondrial tRNAleu-cox2 (cytochrome c oxidase subunit II) intergenic region and the 16S rRNA gene for bee identification in honey samples. The 16S rRNA gene is one of the commonly used mitochondrial DNA (mtDNA) in bees' phylogenetic relationship studies (Cameron et al, 1992;Raffiudin & Crozier, 2007;Rasmussen & Cameron, 2010).…”

Section: Introductionmentioning

confidence: 99%

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Comparison of DNA extraction methods for entomological origin identification of honey using simple additive weighting method

Kek

Chin

Tan

et al. 2018

Int J of Food Sci Tech

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Summary Four different DNA protocols were compared to obtain the best method for DNA extraction of bees in honey samples. The efficiency was evaluated in terms of DNA concentration and purity, PCR amplification capacity targeting mitochondrial 16S rRNA gene, and execution time. The most efficient DNA extraction method was selected using simple additive weighting (SAW) analysis. The DNeasy method with silica membrane spin‐column format scored highest in the final ranking of SAW compared to CTAB‐based, Wizard and NucleoSpin methods. It yielded extracted DNA with sufficient concentration (second highest), acceptable purity (1.5–2.0), amplifiable for 16S rRNA gene region, and required least extraction time. This recommended DNA extraction protocol suggests that genomic methodologies using a 150 bp amplicon of the 16S rRNA gene of bees enables identification of the entomological origin of honey.

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Molecular identification of honey entomological origin based on bee mitochondrial 16S rRNA and COI gene sequences

Cited by 42 publications

References 25 publications

Towards honey authentication: Differentiation of Apis mellifera subspecies in European honeys based on mitochondrial DNA markers

Towards honey authentication: Differentiation of Apis mellifera subspecies in European honeys based on mitochondrial DNA markers

Novel diagnostic tools for Asian (Apis cerana) and European (Apis mellifera) honey authentication

Comparison of DNA extraction methods for entomological origin identification of honey using simple additive weighting method

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