Flavonoids and other phenolic compounds in young needles of four pine species, Pinus peuce, P. nigra, P. mugo and P. sylvestris from the Macedonian flora were investigated. The amount of total ...phenols and total flavonoids were determined using Folin-Ciocalteau and aluminum chloride assay, respectively. The obtained results revealed that the total phenolic content (TPC) and total flavonoids content (TFC) varied among different pine species ranging from 9.8 to 14.0 mg GAE/g and from 3.3 to 7.2 mg CE/g of dried plant material, respectively. Qualitative analysis of flavonoids and other phenolic components was made by a LC-DAD/ESI-MSn optimized chromatographic method. A total of 17 phenolic components were identified and classified as: acids (2), procyanidins (2) and flavonoid glycosides (13). The most prevalent components were flavonoid glycosides, especially flavonols and methylated flavonols (9). Additionally, 3 components were found as acylated flavonol glycosides with ferulic and p-coumaric acid. The last one was found not only in esterified form but also in the free form. Only one flavone-apigenin glycoside was detected. Procyanidins were identified as catechin derivatives, both dimers and trimers.
Climate changes, their regional patterns, origin, and prediction are currently one of the most important scientific challenges. Tree-rings are among the most widely used proxies for past climate ...variation. However tree-ring width (TRW) from certain tree species and regions often do not contain robust climate signal. Other parameters such as Maximum latewood density (MXD) of conifer tree rings are more sensitive to summer temperatures, but the measurements have high costs. A potential surrogate for MXD is blue intensity (BI), which is based on the measurements of high-resolution images of the wood. The method has been tested and applied for several species. However it has not been tested up to now for Pinus heldreichii Christ (PIHE) and Pinus peuce Griseb. (PIPE). Those species are with limited distribution on the Balkan Peninsula and due to their longevity (frequently more than 500 years with potential to more than 1000 years) may serve as one of the best proxies for past climate variation in SE Europe.
We composed BI chronologies following standard procedures from PIHE and PIPE trees from subalpine locations in the Pirin Mts, Bulgaria. The correlation analysis with climate parameters revealed strong and significant positive correlations of PIHE BI (ΔBI and Latewood BI (LwBI) series) and summer temperatures. The highest were 0.74 (p < 0.05) with July-August average temperatures for the period 1933–1983. The correlation values of ΔBI remained higher than 0.6 for the whole period (1933–2014) and sub-periods both for average monthly and average maximum temperatures. LwBI values of the PIHE chronology had correlation coefficients above 0.52 (highest 0.64) with August and August-July temperatures. The PIPE BI chronologies were also positively correlated with summer temperatures, but showed lower values than PIHE with lower temporal stability. The most stable were the correlations with LwBI average August temperatures, which were above 0.52. Delta BI series displayed high correlations for the 1933–1983 period, but then decreased and this caused overall lower correlations with August temperatures. Our data shows that there is potential to develop long BI chronologies and proxy climate reconstructions from the studied species and in this way complement the knowledge of the past climate of SE Europe.
Pinus peuce (Macedonian pine) is considered a valuable ornamental tree that is frequently planted in parks and gardens, especially in Western Europe. This endemic pine is one of the most valuable ...conifer species in its native range, which currently consists of only two disjunct populations restricted to small mountainous areas of the Balkans and is listed as a near-threatened species. The reproduction of Macedonian pine by seed is limited, so in vitro propagation methods have emerged as a promising tool for large-scale propagation. The objective of this study was to develop an improved system for the micropropagation of P. peuce from juvenile plant material using a short-term liquid cytokinin pulse. For that, explants derived from 4-week-old seedlings were pulse-treated with different concentrations of N6-benzyladenine (BA) for 1 or 2 h to stimulate the induction of axillary buds. The highest axillary shoot formation was achieved with 222 µM BA pulse treatment, with an average number of ~six shoots per explant. Elongated shoots (≥10 mm) were detached from the explants and pulse-treated with 0.27 or 1.08 mM α-naphthaleneacetic acid (NAA) or 0.25 or 0.98 mM indole-3-butyric acid (IBA) for 1 or 2 h. IBA was more effective than NAA and led to a maximum rooting percentage (up to 40%) and the highest number of acclimatized plants (15–20%). Rooted plants were successfully transferred to ex vitro conditions.
This paper deals with relationships between vegetation of Macedonian pine (
) and soils developed on different parent materials on the territory of North Macedonia. We analysed the floristic ...composition at localities on limestone, on scree of dolomite marble and on scree of silicate. On limestone and scree of dolomite marble, rendzinas on hard limestone and dolomite have developed, and on silicate parent material brown forest soils. The vegetation was sampled according to the Braun-Blanquet approach. DCA and indicator values were used for ecological interpretation of the vegetation patterns. The mechanical and chemical properties of soil and textural classes were also processed. An evident increased presence of carbonates in the soil of scree of dolomite marble on Nidže Mountain was observed, unlike that on Shar Mountain which has formed on typical limestone. Although it is a forest community dominated by the same species, differences between the massifs, the precipitation regime, geology, differences in soil properties in relation to the appearance of carbonates and pH values, and other factors, result in differences in their floristic composition and are the reason for the distinction between the two groups. On silicate on Nidže Mountain, Macedonian pine forests have also developed on brown forest soils, with a different floristic composition to that of the other group on carbonate (dolomite and limestone).
Studies report the good potential of Pinus heldreichii (PIHE) and Pinus peuce (PIPE) for developing long chronologies from living trees and warn that the climate signal is weak in tree-ring widths of ...PIHE, and particularly PIPE. The goals of the study were to develop long chronologies, and to analyze the climate–growth relationship and potential for long climate reconstructions using tree-ring widths (TRW) and stable carbon isotopes ratios (δ13C) in tree rings at the northern edge of species distribution in the eastern part of Montenegro. The PIHE TRW chronology covers the period 1571–2013 (443 years) and the PIPE TRW chronology 1521–2013 (493 years). The temperature signal in PIHE TRW is weak and the precipitation signal is non-existent. PIPE has no climate signal in TRW. Both studied species have very similar δ13C chronologies, which allows us to merge isotope chronologies into a single composite δ13C Pinus chronology. The composite chronology has a strong signal related to average monthly temperature in June, July, and August and monthly values for cloudiness in July and August, with r > 0.6 and r < −0.6 for individual months, respectively. The climate signal was enhanced when June, July, and August values were merged into seasonal variables. The temporal stability of temperature, precipitation and cloudiness signals is consistent. The spatial extent of the δ13C chronology extends over a very large region, including all surrounding countries.
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