The taxonomic significance of ddRADseq based microsatellite markers in the closely related species of Heracleum (Apiaceae).

Many studies on Heracleum have shown poor correspondence between observed molecular clusters and established taxonomic classification amongst closely related species. This might reflect both unresolved taxonomy but perhaps also a lack of good genetic markers. This lack of appropriate and cost effective species-specific genetic markers hinders a resolved relationship for the species complex, and this in turn causes profound management challenges for a genus that contains both endemic species, with important ecological roles, and species with an invasive potential. Microsatellites are traditionally considered markers of choice for comprehensive, yet inexpensive, analyses of genetic variation, including examination of population structure, species identity, linkage map construction and cryptic speciation. In this study, we have used double digest restriction site associated DNA sequencing (ddRADseq) to develop microsatellite markers in Heracleum rechingeri. Genomic DNA from three individuals were digested with Sbf1 and Nde1 and size selected for library construction. The size-selected fragments were sequenced on an Ion Torrent sequencer and a total of 54 microsatellite sequences were bioinformatically confirmed. Twenty five loci were then tested for amplification, resulting in 19 of these being successfully amplified across eight species, comprising both the so-called thick-stemmed species (H. persicum, H. rechingeri, H. gorganicum and H. lasiopetalum), and thin-stemmed species (H. anisactis, H. pastinasifolium and H. transcaucasicum). Both Bayesian and distance-based clustering, and principal coordinate analyses clearly separated these into two groups. Surprisingly, three H. pastinacifolium populations were not separated from populations of the morphologically similar endemic species, H. anisactis, suggesting lack of genetic differentiation. Likewise, high genetic similarity was found between H. persicum and H. rechingeri populations, questioning taxonomic separation at the species level between these taxa. Further analyses are needed to re-evaluate the taxonomic significance of observed morphological variability currently applied to distinguish these sister taxa. Nevertheless, our results represent progress in the effort to develop cost-efficient molecular tools for species discrimination in this genus.

Large Scale Geographical Mapping of Essential Oil Volatiles in Heracleum (Apiaceae): Identification of Novel Compounds and Unraveling Cryptic Variation.

Conspecific populations of plants in their native range are expected to show considerable variation due to long-term ecological and evolutionary factors. We investigated the levels of secondary metabolites in Heracleum including H. persicum a valuable medicinal plant to depict the magnitude of cryptic variation and the potential significance of novel chemical traits. The essential oil volatiles from fruits of 34 populations from different species of Heracleum in Iranian distribution range and a native of H. sphondylium and an invasive population of H. persicum from Norway were analyzed with GC/MS. Out of 48 compounds identified, a contrasting pattern in the level of two major compounds, octyl acetate and hexyl butyrate was found among all studied species. Interestingly, a significant geographic pattern was observed; the hexyl butyrate/octyl acetate ratio was high (range 1.8 - 3.2) in the northwestern Iranian populations of H. persicum compared to that in northern and central populations (range 0.3 - 0.9). Four populations from Zagros mountains also exhibited a unique composition. Anethole was found in two populations of H. persicum from central Zagros, which has not been previously reported for essential oil of fruits of Heracleum so far. The results suggest high efficiency of large scale sampling from distribution range of species in identifying novel compounds. The unique pattern of geographic structuring also provides novel information to unravel cryptic variation in Heracleum.

Essential oils of three cow parsnips - composition and activity against nosocomial and foodborne pathogens and food contaminants.

Although some widespread, native cow parsnips (Heracleum L. spp., Apiaceae) had broad medicinal and culinary applications throughout history, the knowledge about their volatile constituents is insufficient. This work investigates the composition and bioactivities of H. sphondylium L. (HSPH), H. sibiricum L. (HSIB) and H. montanum Schleich. ex Gaudin (HMON) essential oils. The composition was tested by GC and GC-MS. (Z)-ß-Ocimene was the most abundant in HSPH (28.9%) and HMON (20.4%) root oils, while in HSIB root oil, ß-pinene (26.2%), methyl eugenol (22.3%) and elemicin (25.6%) prevailed. Leaf and flower oils were dominated by various sesquiterpenes (germacrene D, ß-sesquiphellandrene, (E)-ß-farnesene and/or (E)-caryophyllene) and/or phenylpropanoids (apiole, methyl eugenol, elemicin and/or (Z)-isoelemicin). Octyl acetate (57.5-67.1%) was the main constituent of all fruit oils. The antimicrobial activity was screened by a microdilution method against eight bacteria and eight fungi. The strongest antimicrobial effect, in several cases better than the activity of antibiotics, was shown by HSPH (MICs = 0.12-3.30 mg mL-1) and HMON (MICs = 0.10-1.30 mg mL-1) flower oils against bacteria, and HSIB fruit oil against fungi (MICs = 0.15-0.40 mg mL-1). The MTT test revealed that the oils were not or weakly cytotoxic against human malignant HeLa, LS174 and/or A549 cells (except HSPH root oil; IC50 = 5.72-24.31 µg mL-1) and that tested oils were not toxic against human normal MRC-5 cells (at 200.00 µg mL-1). Significant activity observed against microorganisms that are the common cause of foodborne diseases, food contamination and/or hospital-acquired infections justifies certain traditional uses of the investigated plants and represents a good basis for further research of these Heracleum oils.

Ethnobotany of Heracleum persicum Desf. ex Fisch., an invasive species in Norway, or how plant names, uses, and other traditions evolve.

BACKGROUND: Heracleum persicum was introduced to Norway as an ornamental in the 1830's. Towards the end of the 19th century, it started spreading outside gardens, later to become a frequent sight in the major towns and settlements of North Norway - and a veritable pest plant. During the last 100 years or so, a substantial ethnobotanical tradition related to the species has evolved, demonstrating that folk knowledge is not only forgotten and lost, but also charting new terrain. METHODS: This survey is based on data extracted from all relevant publications, including botanical literature, travel accounts, newspaper notes, etc., as far as they have come to my attention. In addition, information on vernacular names and various uses of the H. persicum in Norway has been extracted from my own, substantial archive of interviews, questionnaires, and correspondence related to the ethnobotany of Norway. RESULTS: Where extant, H. persicum tends to be known to everyone, even by city dwellers who otherwise generally neglect plants. People tend to love or hate it, and in Tromsø, the largest town of northern Norway, the species has become more or less emblematic of the city. Both here and in other areas of northern Norway, it is referred to by a variety of vernacular names, partly borrowed from other species, partly derived from the Latin genus name, and partly coined for this species only. In the latter group, tromsøpalme ('the palm of Tromsø') has proved by far the most popular invention. It was seemingly first used (and coined) by German soldiers during the World War II occupation of Norway, but now largely replaces other vernacular names. The plant is still popular with children, who frequently play in and with it, whereas adults have been more prone to speculate on its origins - and how to get rid of it. Salt is the most popular "herbicide" for this purpose. CONCLUSIONS: Over the years, H. persicum has accumulated at least twenty different vernacular names in Norway, and a variety of other traditions. By necessity, all these traditions are less than 180 years old, showing that even modern and urban societies may produce a substantial body of plant lore, which certainly merits ethnobotanical attention.