The association of protein-bound methionine sulfoxide with proteomic basis for aging in beech seeds.

BACKGROUND: European beech (Fagus sylvatica L.) trees produce seeds irregularly; therefore, it is necessary to store beech seeds for forestation. Despite the acquisition of desiccation tolerance during development, beech seeds are classified as intermediate because they lose viability during long-term storage faster than typical orthodox seeds. In this study, beech seeds stored for short (3 years) or long (20 years) periods under optimal conditions and displaying 92 and 30% germination capacity, respectively, were compared. RESULTS: Aged seeds displayed increased membrane damage, manifested as electrolyte leakage and lipid peroxidation levels. Analyses have been based on embryonic axes, which contained higher levels of reactive oxygen species (ROS) and higher levels of protein-bound methionine sulfoxide (MetO) in aged seeds. Using label-free quantitative proteomics, 3,949 proteins were identified, of which 2,442 were reliably quantified pointing to 24 more abundant proteins and 35 less abundant proteins in beech seeds under long-term storage conditions. Functional analyses based on gene ontology annotations revealed that nucleic acid binding activity (molecular function), ribosome organization or biogenesis and transmembrane transport (cellular processes), translational proteins (protein class) and membranous anatomical entities (cellular compartment) were affected in aged seeds. To verify whether MetO, the oxidative posttranslational modification of proteins that can be reversed via the action of methionine sulfoxide reductase (Msr) enzymes, is involved in the aging of beech seeds, we identified and quantified 226 MetO-containing proteins, among which 9 and 19 exhibited significantly up- and downregulated MetO levels, respectively, in beech seeds under long-term storage conditions. Several Msr isoforms were identified and recognized as MsrA1-like, MsrA4, MsrB5 and MsrB5-like in beech seeds. Only MsrA1-like displayed decreased abundance in aged seeds. CONCLUSIONS: We demonstrated that the loss of membrane integrity reflected in the elevated abundance of membrane proteins had a higher impact on seed aging progress than the MetO/Msr system. Proteome analyses enabled us to propose protein Sec61 and glyceraldehyde-3-phosphate dehydrogenase as potential longevity modulators in beech seeds.

Does long-term drought or repeated defoliation affect seasonal leaf N cycling in young beech trees?

Forest trees adopt effective strategies to optimize nitrogen (N) use through internal N recycling. In the context of more recurrent environmental stresses due to climate change, the question remains of whether increased frequency of drought or defoliation threatens this internal N recycling strategy. We submitted 8-year-old beech trees to 2 years of either severe drought (Dro) or manual defoliation (Def) to create a state of N starvation. At the end of the second year before leaf senescence, we labeled the foliage of the Dro and Def trees, as well as that of control (Co) trees, with 15N-urea. Leaf N resorption, winter tree N storage (total N, 15N, amino acids, soluble proteins) and N remobilization in spring were evaluated for the three treatments. Defoliation and drought did not significantly impact foliar N resorption or N concentrations in organs in winter. Total N amounts in Def tree remained close to those in Co tree, but winter N was stored more in the branches than in the trunk and roots. Total N amount in Dro trees was drastically reduced (-55%), especially at the trunk level, but soluble protein concentrations increased in the trunk and fine roots compared with Co trees. During spring, 15N was mobilized from the trunk, branches and twigs of both Co and Def trees to support leaf growth. It was only provided through twig 15N remobilization in the Dro trees, thus resulting in extremely reduced Dro leaf N amounts. Our results suggest that stress-induced changes occur in N metabolism but with varying severity depending on the constraints: within-tree 15N transport and storage strategy changed in response to defoliation, whereas a soil water deficit induced a drastic reduction of the N amounts in all the tree organs. Consequently, N dysfunction could be involved in drought-induced beech tree mortality under the future climate.

Long-term behaviour of Cs-137, Cs-133 and K in beech trees of French forests.

In the long-term after atmospheric deposit onto a forest ecosystem, Cs-137 becomes incorporated into the biogeochemical cycle of stable elements and progressively reaches a quasi-equilibrium state. This study aimed at determining to what extent Cs-137 activity distribution in tree vegetation could be predicted from that of stable caesium (Cs-133) and potassium (K), which are known to be stable chemical analogues and competitors for Cs-137 intake in tree organs. Field campaigns that focused on beech trees (Fagus sylvatica L.) were conducted in 2021 in three French forest stands with contrasted characteristics regarding either the contribution of global vs. Chornobyl fallouts, soil or climatic conditions. Decades after Cs-137 fallouts, it was found that more than 80% of the total radioactive inventory in the system remained confined in the top 20 cm mineral layers, while organic layers and beech vegetation (including roots) contributed each to less than 1.5%. The enhanced downward migration of Cs-137 in cambisol than podzol forest sites was presumably due to migration of clay particles and bioturbation. The distribution of Cs-137 and Cs-133 inventories in beech trees was very similar among sites but differed from that of K due a higher accumulation of Cs isotopes in roots (40-50% vs. < 25% for K). The aggregated transfer factor (Tag) of Cs-137 calculated for aerial beech organs were all lower than those reported in literature more than 20 years ago, this suggesting a decrease of bioavailability in soil due to ageing processes. Regarding their variability, Tags were generally lower by a factor 5 at the cambisol site, which was fairly well explained by a much higher value of RIP (radiocesium immobilisation potential). Cs-137 concentrations in trees organs normalized by the soil exchangeable fractions were linearly correlated to those of Cs-133 and the best fit was found for the linear regression model without intercept indicating that no more contribution of the foliar uptake could be observed on long term. Provided that the vertical distribution of caesium concentrations and fine root density are properly measured or estimated, Cs-133 was shown to be a much better proxy than K to estimate the root transfer of Cs-137.

Substantial uptake of nitrous oxide (N2O) by shoots of mature European beech.

Similar to soils, tree stems emit and consume nitrous oxide (N2O) from the atmosphere. Although tree leaves dominate tree surface area, they have been completely excluded from field N2O flux measurements and therefore their role in forest N2O exchange remains unknown. We explored the contribution of leaf fluxes to forest N2O exchange. We determined the N2O exchange of mature European beech (Fagus sylvatica) stems and shoots (i.e., terminal branches) and of adjacent forest floor, in a typical temperate upland forest in Germany. The beech stems, and particularly the shoots, acted as net N2O sinks (-0.254 ± 0.827 µg N2O m-2 stem area h-1 and -4.54 ± 1.53 µg N2O m-2 leaf area h-1, respectively), while the forest floor was a net source (2.41 ± 1.08 µg N2O m-2 soil area h-1). The unstudied tree shoots were identified as a significant contributor to the net ecosystem N2O exchange. Moreover, we revealed for the first time that tree leaves act as substantial N2O sinks. Although this is the first study of its kind, it is of global importance for the proper design of future flux studies in forest ecosystems worldwide. Our results demonstrate that excluding tree leaves from forest N2O flux measurements can lead to misinterpretation of tree and forest N2O exchange, and thus global forest greenhouse gas flux inventories.

Impact of various microplastics on the morphological characteristics and nutrition of the young generation of beech (Fagus sylvatica L.).

Microplastics have the capacity to accumulate in soil due to their high resistance to degradation, consequently altering soil properties and influencing plant growth. This study focused on assessing the impact of various types and doses of microplastics on beech seedling growth. In our experiment, we used polypropylene and styrene granules with diameter of 4.0 mm in quantities of 2.5% and 7%. The hypothesis was that microplastics significantly affect seedlings' nutritional status and growth characteristics. The research analysed seedlings' nutrition, root morphological features, above-ground growth, and enzymatic activity in the substrate. Results confirmed the importance of microplastics in shaping the nutritional status of young beech trees. Microplastic type significantly impacted N/P and Ca/Mg stoichiometry, while microplastic quantity influenced Ca/Al and Ca+K+Mg/Al stoichiometry. Notably, only in the case of root diameter were significantly thicker roots noted in the control variant, whereas microplastics played a role in shaping the leaves' characteristics of the species studied. The leaf area was significantly larger in the control variant compared to the variant with polypropylene in the amount of 2.5% and styrene in the amount of 7%. Additionally, the study indicates a significant impact of microplastics on enzyme activity. In the case of CB and SP, the activity was twice as high in the control variant compared to the variants with microplastics. In the case of BG, the activity in the control variant was higher in relation to the variants used in the experiment. Research on the impact of microplastics on the growth of beech seedlings is crucial for enhancing our understanding of the effects of environmental pollution on forest ecosystems. Such studies are integral in shaping forestry management practices and fostering a broader public understanding of the ecological implications of plastic pollution.