Abstract
The species of the aromatic plant family Apiaceae are mainly used as spices and foods,
but the family also includes medicinal and some poisonous plant species. Due to the
similar chemical compounds or aroma and morphology, the poisonous species are often
mistaken for the edible aromatic species. It is therefore imperative to correctly
identify the species present at the initial raw stage samples to ensure product safety
and efficacy. At the molecular level, plant species can be identified using DNA loci
either from nuclear or plastid genome with easily available universal oligonucleotides,
a technique called DNA barcoding. However, this is possible when single-species plant
material is present but may not work on a mixture of plants species. Another disadvantage
is that using universal oligonucleotides is of limited help, especially if the adulterating
material is present in low quantities. On the other hand, if using the species-specific
oligonucleotides, only single specific adulterating plant material could be detected
and, consequently, the unexpected adulterants may go undetected. Therefore, in the
current work, four degenerated oligonucleotides from ITS1 and ITS2 regions of the
nuclear genome were designed that can bind to a variety of Apiaceae genera only and
not to other genera belonging to different plant families. These family-specific oligonucleotides
were able to amplify a diagnostic PCR product from 16 Apiaceae species that, upon
sequencing, revealed the identity of the plant it was derived from. The size of these
products is around 140 bp for ITS1 and approximately 80 bp for the ITS2 region. The
size range of the amplified products falls in the category of a desired mini-barcode
size to be used for damaged/fragmented DNA and next generation sequencing.
Key words
Adulteration - Apiaceae - aromatic species - DNA mini-barcodes - family-specific oligonucleotides
- universal oligonucleotides
