Tamara Rakić

Characterization of oxidative and antioxidative events during dehydration and rehydration of resurrection plant Ramonda nathaliae

In order to investigate changes of oxidative status in relation to the activity of the various protective mechanisms in resurrection plant Ramonda nathaliae, we have analysed time and relative water content (RWC) related changes in lipid peroxidation and ion leakage, hydrogen peroxide accumulation, changes of pigment content and antioxidative enzyme activity, together with expression of dehydrins. The results indicate that enhanced oxidative status during dehydration, not previously reported for resurrection plants, could play an active role in inducing the desiccation adaptive response in R. nathaliae. A critical phase is shown to exist during dehydration (in the range of RWC between 50 and 70%) during which a significant increase in hydrogen peroxide accumulation, lipid peroxidation and ion leakage, accompanied by a general decline in antioxidative enzyme activity, takes place. This phase is designated as a transition characterized by change in the type of stress response. The initial response, relying mainly on the enzymatic antioxidative system, is suspended but more effective, desiccation specific protective mechanisms, such as expression of dehydrins, are then switched on. The expression of dehydrins in R. nathaliae could be inducible as well as constitutive. In order to cope with the oxidative stress associated with rapid rewatering, R. nathaliae reactivated antioxidative enzymes. We propose that controlled elevation of reactive oxygen species, such as hydrogen peroxide, could be an important mechanism enabling resurrection plants to sense dehydration and to trigger an adaptive programme at an appropriate stage during the dehydration/rehydration cycle.

One New Species of the Genus Edraianthus, and a Change in Taxonomic Status for Edraianthus serpyllifolius f. Pilosulus (Campanulaceae) from the Balkan Peninsula

Abstract A new species, Edraianthus pulevicii (Campanulaceae), endemic to the Durmitor mountain range (SE Dinaric Alps, Montenegro, Balkan Peninsula) is described and illustrated. Additionally, specific rank is proposed for E. serpyllifolius f. pilosulus from the Komovi Mts. (NW Prokletije mountain range, Montenegro, Balkan Peninsula), E. pilosulus. Both are morphologically similar to E. serpyllifolius s. s., all having spathulate leaves. Edraianthus pulevicii differs from E. serpyllifolius s. s. in having distinctly crenate as well as much longer and broader basal and cauline leaves with indumentum on the upper side, hairs on the leaf margin and leaf surface oriented towards the leaf base, broader and longer bracts, and revolute calyx lobes reflexed at the apex. Edraianthus pilosulus is morphologically similar to both E. serpyllifolius and E. pulevicii but differs from the former in having more or less dense hairs on the upper side of the leaves, and from the latter in having considerably smaller basal and cauline leaves with no or sparse crenation, hairs on the leaf margin and the leaf surface oriented towards the point, and narrower bracts. A new systematic treatment is supported by chloroplast DNA sequence data and AFLP fingerprinting data. A distribution map and a key to Edraianthus taxa with spathulate leaves are provided and the name Edraianthus sutjeskae is validated.

Resurrection plants of the genus Ramonda: prospective survival strategies – unlock further capacity of adaptation, or embark on the path of evolution?

Paleoendemic species of the monophyletic genus Ramonda (R. myconi, R. serbica and R.~nathaliae) are the remnants of the Tertiary tropical and subtropical flora in Europe. They are the rare resurrection plants of Northern Hemisphere temperate zone. Ramonda serbica and R. nathaliae are chorologically differentiated in the Balkan Peninsula and occupy similar habitats in calcareous, northward slopes in canyons and mountainsides. They remain well-hydrated during spring, late autumn and even in winter. In summer and early autumn when plants are subjected to drought and thermal stress, their desiccation tolerance comes into operation and they fall into anabiosis. Investigations revealed the permanent presence of ubiquitine and its conjugates, high amounts of oxalic acid and proline. Both species are homoiochlorophyllous. It enables them to rapidly resume photosynthesis upon rehydration, but also makes them susceptible to reactive oxygen species formation. Dehydration induces activation of antioxidative enzymes (ascorbate peroxidase, glutathione reductase, polyphenol oxidase), increase in amounts of AsA and GSH, phenolic acids, dehydrins, sucrose, and inorganic ions. Plasma membranes, characterized by high amount of cholesterol, are subjected to decrease in membrane fluidity mostly on account of increased level of lipid saturation. Cytogenetic analysis revealed that R. nathaliae is a diploid (2n = 48) and probably evolutionary older species, while R. serbica is a hexaploid (2n = 144). Two species live together in only two localities forming hybrid individuals (2n = 96). Polyploidization is the major evolutionary mechanism in the genus Ramonda that together with hybridization ability indicates that these relict species which have preserved an ancient survival strategy are not the evolutionary “dead end.”The species of the genus Ramonda are promising sources of data important for understanding the complex strategy of resurrection plants’ survival, appraised through a prism of their evolutionary and adaptive potential for multiple environmental stresses.

ECOPHYSIOLOGICAL AND ANATOMICAL CHARACTERISTICS OF THE SUBTROPICAL SHRUB ZANTHOXYLUM ACANTHOPODIUM (RUTACEAE) IN CONDITIONS OF A TEMPERATE CONTINENTAL CLIMATE (SERBIA)

The evergreen shrub Zanthoxylum acanthopodium DC. (Rutaceae), originating from warm temperate and subtropical Asia, has existed successfully in the Jevremovac Botanical Garden in Belgrade for more than 80 years. The seasonal pattern of water management in leaves, electrolyte leakage, essential oil composition, and leaf anatomy were examined in order to understand the resistance and viability of this subtropical shrub in the temperate continental cli- mate of Belgrade, Serbia.

Zinc accumulation, photosynthetic gas exchange, and chlorophyll a fluorescence in Zn-stressed Miscanthus × giganteus plants

Accumulation and distribution of zinc within Miscanthus × giganteus plants grown on elevated Zn concentrations and their photosynthetic performance were investigated. High concentrations of Zn in soils caused an increase of its concentrations in all plant organs. The bioconcentration factor, bioaccumulation factor, and translocation factor were lower than one indicating that M. × giganteus is an excluder plant species. Excessive Zn induced visible leaf damage, i.e. chlorosis and necrosis, only in the oldest leaves, pointing to Zn accumulation. Elevated amounts of Zn in leaves significantly lowered the photosynthetic rate, transpiration rate, stomatal conductance, intercellular CO2 concentrations, parameters of chlorophyll a fluorescence, and chlorophyll b content. Despite Zn excess in leaves, there was no severe reduction in the maximal quantum yield of PSII photochemistry, indicating a high photosynthetic capacity, high tolerance to elevated Zn concetrations, and ability of M. × giganteus to grow on Zn-contaminated soils.

Understanding biogeographical patterns in the western Balkan Peninsula using environmental niche modelling and geostatistics in polymorphic Edraianthus tenuifolius

Abstract The Balkan Peninsula represents one of the three southern European glacial refugia where biodiversity persisted throughout the climatically unstable Quaternary. This study considered the ‘refugia within refugia’ model, which assumes the environmental heterogeneity over time and space in larger refugia. To better understand patterns shaped during the Quaternary climatic oscillations, the present and last glacial maximum (LGM) environmental conditions and current morphological variability of Edraianthus tenuifolius, an endemic plant of the western Balkans with a well-known therphical structure, were considered. Potential present and LGM distributions were studied through environmental niche modelling using 161 data of occurrences and six bioclimatic variables, hindcasting the model to LGM conditions using three different global circulation models. To test the geographical variability of the reproductive region, 41 characters of 667 inflorescences from 35 populations within the distribution range were measured. Geographical patterns, using geostatistics together with univariate and multivariate statistical approaches, were analysed. The environmental niche model suggested the current potential distribution in correspondence to its known occurrences. The hindcast to LGM conditions suggested two separate areas of environmental suitability, one in the present-day northern Adriatic coast of Croatia (Istrian Peninsula, Kvarner) and another on the present-day south-eastern Adriatic coast (southern Dalmatia, Montenegro and northern Albania). Morphological variability showed a similar pattern, where southern populations separated from northern populations according to a major split in the central part of its distribution range (central Dalmatia). On other levels, stronger barriers were predicted to separate northern populations from the eastern Istrian Peninsula and the Kvarner area. The results suggested congruent biogeographical patterns to the already known phylogeographical structure. Both environmental niche modelling and the geographical variability of morphological characters suggested spatial partitioning, indicating the potential presence of two separate refugia during the LGM.