Identification of sweet chesnut pollen in bee pollen pellet using using molecular analysis.

Authors

  • Jana Žiarovská Slovak University of Agriculture in Nitra, Faculty of Agrobiology and Food Resources, Department of Genetics and Plant Breeding, Tr. A. Hlinku 2, 949 76 Nitra
  • Olga Grygorieva Institute of Biodiversity Conservation and Biosafety; Tr. A. Hlinku 2, 949 76 Nitra
  • Lucia Zeleňáková Slovak University of Agriculture, Faculty of Biotechnology and Food Sciences, Department of Food Hygiene and Safety, Tr. A. Hlinku 2, 949 76 Nitra
  • Milan Bežo Slovak University of Agriculture, Faculty of Agrobiology and Food Resources, Department of Genetics and Plant Breeding, Tr. A. Hlinku 2, 949 76 Nitra
  • Ján Brindza Slovak University of Agriculture, Faculty of Agrobiology and Food Resources, Department of Genetics and Plant Breeding, Tr. A. Hlinku 2, 949 76 Nitra

DOI:

https://doi.org/10.5219/497

Keywords:

Castanea sativa, pollen, pollen pellet, identification, PCR

Abstract

Castanea sativa posses many characteristics that are used by human for different purposes, not only as a part of the food. One of them is the utilization of the sweet chesnut pollen for its pharmacological benefits. Actually, no information about the DNA based identification of the sweet chesnut exist. Here, an identification of Castanea sativa based on the specific DNA fragment amplification is described for the first time. Sweet chesnut identification was performed in the very complex sample of bee pollen pellets that were identified as to contain sweet chesnut pollen grains by morphological analysis. First, bioinformatic analysis was performed to find a Castanea sativa conservative part of galactol synthase gene. BLAST alignment of the CDS of GolS1 gene was performed by BLASTtn against plants nucleotide sequences in the NCBI database to ensure for the specifity or existing nucleotide differences. Then, specific primers were subsequently designed and PCR amplification was performed. All the PCRs have run in duplicates for pollen pellet sample and two independent samples of Castanea sativa pure pollen. Restriction cleavage of the PCR amplified fragment was performed to confirm the specifity of the obtained PCR product with the positive confirmation as the predicted three restriction fragments were obtained that fully correspond by the length to those from virtual clevage. Restriction endonuclease Hpy166II was used in restriction cleavage analysis. Castanea sativa pollen grains were confirmed reliable in multifloral pollen pellet by PCR and this approach has the potential to be used effectively for the authentication purposes of sweet chesnut.

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References

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Published

2015-09-11

How to Cite

Žiarovská, J. ., Grygorieva, O. ., Zeleňáková, L. ., Bežo, M. ., & Brindza, J. . (2015). Identification of sweet chesnut pollen in bee pollen pellet using using molecular analysis. Potravinarstvo Slovak Journal of Food Sciences, 9(1), 352–357. https://doi.org/10.5219/497

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