Juniperus excelsa s. str. in crimea – differentiation and history inferred from genetic and morphological markers

Juniperus excelsa s. str. (Greek juniper) in Crimea is a relic species on the limits of its range, and represents the Mediterranean flora in the Sub-Mediterranean part of the peninsula. Its origin and history in this area remains unresolved. We aimed to analyze phylogeny and potential demographic expansion of the juniper in the Crimea as well as to study its morphological differentiation. We analyzed plant material from 59 trees inhabiting eight populations. Genetic variation assessments were based on the four non-coding chloroplast DNA (cpDNA) fragments and the nuclear internal transcribed spacer region ITS1-5,8S-ITS2 (ITS). To examine the morphological differentiation, eight measured/counted traits of cones, seeds, and shoots were chosen and eight ratios were calculated. Morphological parameters were compared using ANOVA, Student’s t test, discrimination analysis and Kruskal-Wallis and U Mann-Whitney tests. Two cpDNA fragments were polymorphic and, in total, 10 cpDNA haplotypes were found. Haplotype diversity (Hd) ranged from 0.0 to 0.9. Based on both cpDNA and ITS sequences variation, phylogenetic analyses revealed a close relationship of the Crimean junipers to the individuals from other parts of the species range. In general, our molecular results confirmed the low level of genetic differentiation of J. excelsa individuals inhabiting different parts of the species range, likely resulting from a common ancestry. Only slight morphological differences were found between populations with different geographic location or habitat. The analyzes showed the distinctness of the populations from the southern part of the coast. Some unique morphological and molecular features of southern coastal populations imply that they are remnants of Late Pleistocene abundant forests. We suggest that the recent fragmentation of the Juniperus populations in the Crimean Peninsula could have arisen during the Atlantic period of the Holocene.

Essential Oil Composition and Bioactivity of Two Juniper Species from Bulgaria and Slovakia

Juniperus excelsa M. Bieb and J. sabina L. contain essential oil (EO), while J. sabina also contains podophyllotoxin, which is used as a precursor for anti-cancer drugs. Two studies were conducted. The first assessed the variability in the EO profile and podophyllotoxin concentration of the two junipers, depending on the location and tree gender. The main EO constituents of J. excelsa were α-cedrol, α-limonene and α-pinene, while the constituents in J. sabina were sabinene, terpinen-4-ol, myrtenyl acetate and α-cadinol. The podophyllotoxin yield of 18 J. sabina accessions was 0.07–0.32% (w/w), but this was not found in any of the J. excelsa accessions. The second study assessed the effect of hydrodistillation (Clevenger apparatus) and steam distillation (in a semi-commercial apparatus) on the EO profile and bioactivity. The extraction type did not significantly alter the EO composition. The EO profiles of the two junipers and their accessions were different and may be of interest to the industry utilizing juniper leaf EO. Breeding and selection programs could be developed with the two junipers (protected species) in order to identify chemotypes with (1) a high EO content and desirable composition, and (2) a high concentration of podophyllotoxin in J. sabina. Such chemotypes could be established as agricultural crops for the commercial production of podophyllotoxin and EO.

Monitoring Carbon Stock and Land-Use Change in 5000-Year-Old Juniper Forest Stand of Ziarat, Balochistan, through a Synergistic Approach

The Juniper forest reserve of Ziarat is one of the biggest Juniperus forests in the world. This study assessed the land-use changes and carbon stock of Ziarat. Different types of carbon pools were quantified in terms of storage in the study area in tons/ha i.e., above ground, soil, shrubs and litter. The Juniper species of this forest is putatively called Juniperus excelsa Beiberstein. To estimate above-ground biomass, different allometric equations were applied. Average above ground carbon stock of the forest was estimated as 8.34 ton/ha, 7.79 ton/ha and 8.4 ton/ha using each equation. Average carbon stock in soil, shrubs and litter was calculated as 24.35 ton/ha, 0.05 ton/ha and 1.52 ton/ha, respectively. Based on our results, soil carbon stock in the Juniper forest of Ziarat came out to be higher than the living biomass. Furthermore, the spatio-temporal classified maps for Ziarat showed that forest area has significantly decreased, while agricultural and barren lands increased from 1988 to 2018. This was supported by the fact that estimated carbon stock also showed a decreasing pattern between the evaluation periods of 1988 to 2018. Furthermore, the trend for land use and carbon stock was estimated post 2018 using a linear prediction model. The results corroborate the assumption that under a business as usual scenario, it is highly likely that the Juniperus forest will severely decline.