DNA barcoding is a standard and efficient method, frequently used for identification, discrimination and discovery of new species. Although this approach is very useful for classifying the world's biodiversity, little is known about its usefulness in barcoding at lower taxonomic level and its discrimination rate for closely related species, like conifers. In this study, we compared the genetic variation of eight chloroplast DNA barcode regions (matK, rbcL, trnH-psbA, trnL-trnF, rpl20-rps18, trnV, ycf1, ycf2) in 17 conifers - three closely related pines from Pinus mugo complex and 14 more distant conifers representing two genera and four sections of the Pinaceae family. The discrimination rate for a single and for multiple DNA barcode regions analyzed in this study was estimated using the Tree-Building and PWG-Distance methods. The usefulness of the DNA barcoding approach for analyzing and resolving taxonomic inconsistency among closely related and more phylogenetically distant conifers was evaluated and discussed.
Dracaena draco, which belongs to the genus Dracaena, is an endemic succulent of the Canary Islands. Although it is one of the most popular and widely grown ornamental plants in the world, little is known about its genomic variability. Next generation sequencing, especially in combination with advanced bioinformatics analysis, is a new standard in taxonomic and phylogenetic research. Therefore, in this study, the complete D. draco chloroplast genome (cp) was sequenced and analyzed in order to provide new genomic information and to elucidate phylogenetic relationships, particularly within the genus Dracaena. The D. draco chloroplast genome is 155,422 bp, total guanine-cytosine (GC) content is 37.6%, and it has a typical quadripartite plastid genome structure with four separate regions, including one large single copy region of 83,942 bp length and one small single copy region of 18,472 bp length, separated by two inverted repeat regions, each 26,504 bp in length. One hundred and thirty-two genes were identified, 86 of which are protein-coding genes, 38 are transfer RNAs, and eight are ribosomal RNAs. Seventy-seven simple sequence repeats were also detected. Comparative analysis of the sequence data of various members of Asparagales revealed mutational hotspots potentially useful for their genetic identification. Phylogenetic inference based on 16 complete chloroplast genomes of Asparagales strongly suggested that Dracaena species form one monophyletic group, and that close relationships exist between D. draco, D. cochinchinensis and D. cambodiana. This study provides new and valuable data for further taxonomic, evolutionary and phylogenetic studies within the Dracaena genus.
Headspace solid-phase microextraction (HS-SPME) coupled to GC/MS analysis was used to identify the constituents of pine-needle volatiles differentiating three closely-related pine species within the Pinus mugo complex, i.e., P. uncinata Ramond ex DC., P. uliginosa G.E.Neumann ex Wimm., and P. mugo Turra. Moreover, chemosystematic markers were proposed for the three analyzed pine species. The major constituents of the pine-needle volatiles were α-pinene (28.4%) and bornyl acetate (10.8%) for P. uncinata, δ-car-3-ene (21.5%) and α-pinene (16.1%) for P. uliginosa, and α-pinene (20%) and δ-car-3-ene (18.1%) for P. mugo. This study is the first report on the application of the composition of pine-needle volatiles for the reliable identification of closely-related pine species within the Pinus mugo complex.
Damage caused to pine forests by industrial pollution is observed even several decades after emissions have stopped down. A simple morphological feature – the leaf fluctuating asymmetry – was used for assessing the condition of Scots pine (Pinus sylvestris) stands growing in a heavily degraded area. In 2011, a study was performed on the developmental instability of needles in four naturally reforested Polish populations of P. sylvestris. Studies were conducted within the protective zone at a zinc smelter, a copper smelter, a cement plant and train tracks. All selected areas manifested a high extent of anthropogenic pollution‐induced environmental degradation until the end of the 90s. Currently, a reduced level of environmental pollution is recorded at these sites. Control group was a natural population of Scots pine from the region of National Park of Wielkopolska. The results confirm the usefulness of fluctuating asymmetry as a highly sensitive indicator of non‐specific stress. Also, it was shown that in areas degraded by human activity, poor condition of Scots pine persists that the stress factor has been eliminated. This tendency occurs particularly to areas contaminated by heavy metals.
Pinus mugo (dwarf mountain pine) is an important component of European mountain ecosystems. However, little is known about the present genetic structure and population differentiation of this species at the DNA level, possibly due to a lack of nuclear microsatellite markers (SSR) developed for Pinus mugo. Therefore in this study we transferred microsatellite markers originally developed for Pinus sylvestris and Pinus taeda to Pinus mugo. This cross-species amplification approach is much faster and less expensive than isolation and characterization of new microsatellite markers. The transfer rates from the source species to Pinus mugo were moderately low (26%). There were no differences in microsatellite repeat motifs between the source species and Pinus mugo. Nuclear microsatellite markers successfully transferred to Pinus mugo can be applied to various genetic studies on this species, due to the high level of their polymorphism and high value of polymorphic information content.
The levels of variation and genetic diversity of offspring of randomly selected old mother trees in four marginal populations of the Pinus cembra in the Tatra Mountains were analyzed. Twenty-four isozyme loci were analyzed (nine of them were monomorphic). The analyzed offspring of Swiss stone pine showed highly diverse polymorphism at the levels of both provenances and individual families (the offspring of one mother tree). The mean observed heterozygosity was low and very similar to that of other Carpathian populations. The genetic diversity (mean Fst = 11%) between the four provenances was higher than that observed for populations from the Carpathian Mountains and the Alps. The genetic uniqueness (high genetic richness and diversity) of the analyzed Tatra populations of P. cembra as a whole and particular tree stands requires protection because of their valuable contribution to the species total genetic diversity (gene pool).
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