Chemical and Biomolecular Characterization of Artemisia umbelliformis Lam., an Important Ingredient of the Alpine Liqueur “Genepì”

Abstract

Artemisia umbelliformis Lam., an important alpine plant used for the preparation of flavored beverages, showed a remarkable intraspecific variability, at both genomic and gene product (secondary metabolites) levels. The variability of A. umbelliformis Lam. currently cultivated in Piedmont (Italy, Au1) and in Switzerland (Au2) was investigated by combining the chemical analysis of essential oil and sesquiterpene lactones and the molecular characterization of the 5S-rRNA-NTS gene by PCR and PCR-RFLP. Marked differences were observed between the two plants. Au1 essential oil contained alpha- and beta-thujones as the main components, whereas Au2 contained 1,8-cineole, borneol, and beta-pinene. Au1 sesquiterpene lactone fractions contained cis-8-eudesmanolide derivatives and Au2 the trans-6-germacranolide costunolide. Specific A. umbelliformis Au1 and Au2 primers were designed on the sequence of the 5S-rRNA gene spacer region. Furthermore, a PCR-restriction fragment length polymorphism (PCR-RFLP) method was applied using RsaI and TaqI restriction enzymes. Chemical and biomolecular data contributed to the characterization of A. umbeliformis chemotypes.

PCR, sequencing and PCR–RFLP of the 5S‐rRNA‐NTS region as a tool for the DNA fingerprinting of medicinal and aromatic plants

“…[64] Two chemotypes of A. umbelliformis (with and without thujone) used to prepare a local liqueur, genepi, were studied and specifi c A. umbelliformis primers were designed on the sequence of the 5S-rRNA gene spacer region. When a PCR-RFLP method was applied, using RsaI and TaqI restriction enzymes, the two chemotypes were clearly distinguished [2] ( Figure 1, lanes 7-10). As recently presented at the 40th ISEO in Italy, our research is ongoing and we will soon report on new genotypic characterizations of medicinal, aromatic and food plants using the 5S-rRNA gene NTS region.…”

“…[1] This means that, according to environmental conditions, the same genotype may express diff erent chemical patterns or, conversely, that diff erent genotypes may respond to the same environmental pressure with the same phenotypic expression. [2] Chemotypes (or chemical phenotypes) are generally considered to be the phenotypical expression of a genotype, although diff erent chemotypes may derive from the same genotype.…”

“…[3] In this context, molecular genetic methods have recently been shown to be very eff ective in genotypic discrimination. [2] Molecular genetic methods have several advantages over classical morphological and chemical analyses. For instance, the genetic method requires genotype instead of phenotype, therefore DNA-based experiments have become widely employed techniques for rapid identifi cation of herbal medicines.…”

Analysis of the Plant Volatile Fraction

“…This approach involves the true quantitation of a marker, in general by official and/ or literature methods, as for instance is the case of the determination of a-and b-thujone (39 and 40, Figure 3.11) in bitter liqueurs, 71 or suspected allergens in cosmetics. 72 The marker concentration or absolute amount in a real-world sample is determined from its chromatographic area normalized versus an internal (or external) standard and calculated via a calibration curve constructed from amounts of pure marker standard in the operative concentration range.…”

“…The two chemotypes can be distinguished unequivocally with a total variability explained by the two principal components of about 74%. 71 These methods have not only increased the importance of the role played by data processing in the entire analytical process, but have also influenced analysis strategy and stimulated the introduction of new approaches. A usual online method to analyse the volatile fraction of a vegetable matrix combines headspace sampling for sample preparation, GC for separation and MS to identify and quantify as many components as possible (or at least the characterizing markers).…”