Mineral and organic fertilisation influence ammonia oxidisers and denitrifiers and nitrous oxide emissions in a long-term tillage experiment.

Nitrous oxide (N2O) emissions from different agricultural systems have been studied extensively to understand the mechanisms underlying their formation. While a number of long-term field experiments have focused on individual agricultural practices in relation to N2O emissions, studies on the combined effects of multiple practices are lacking. This study evaluated the effect of different tillage [no-till (NT) vs. conventional plough tillage (CT)] in combination with fertilisation [mineral (MIN), compost (ORG), and unfertilised control (CON)] on seasonal N2O emissions and the underlying N-cycling microbial community in one maize growing season. Rainfall events after fertilisation, which resulted in increased soil water content, were the main triggers of the observed N2O emission peaks. The highest cumulative emissions were measured in MIN fertilisation, followed by ORG and CON fertilisation. In the period after the first fertilisation CT resulted in higher cumulative emissions than NT, while no significant effect of tillage was observed cumulatively across the entire season. A higher genetic potential for N2O emissions was observed under NT than CT, as indicated by an increased (nirK + nirS)/(nosZI + nosZII) ratio. The mentioned ratio under NT decreased in the order CON > MIN > ORG, indicating a higher N2O consumption potential in the NT-ORG treatment, which was confirmed in terms of cumulative emissions. The AOB/16S ratio was strongly affected by fertilisation and was higher in the MIN than in the ORG and CON treatments, regardless of the tillage system. Multiple regression has revealed that this ratio is one of the most important variables explaining cumulative N2O emissions, possibly reflecting the role of bacterial ammonia oxidisers in minerally fertilised soil. Although the AOB/16S ratio aligned well with the measured N2O emissions in our experimental field, the higher genetic potential for denitrification expressed by the (nirK + nirS)/(nosZI + nosZII) ratio in NT than CT was not realized in the form of increased emissions. Our results suggest that organic fertilisation in combination with NT shows a promising combination for mitigating N2O emissions; however, addressing the yield gap is necessary before incorporating it in recommendations for farmers.

Temporal and spatial variability of phloem structure in Picea abies and Fagus sylvatica and its link to climate.

Using a unique 8-year data set (2010-2017) of phloem data, we studied the effect of temperature and precipitation on the phloem anatomy (conduit area, widths of ring, early and late phloem) and xylem-ring width in two coexisting temperate tree species, Picea abies and Fagus sylvatica, from three contrasting European temperate forest sites. Histometric analyses were performed on microcores taken from tree stems in autumn. We found high interannual variability and sensitivity of phloem anatomy and xylem-ring widths to precipitation and temperature; however, the responses were species- and site-specific. The contrasting response of xylem and phloem-ring widths of the same tree species to weather conditions was found at the two Slovenian sites generally well supplied with precipitation, while at the driest Czech site, the influence of weather factors on xylem and phloem ring widths was synchronised. Since widths of mean annual xylem and phloem increments were narrowest at the Czech site, this site is suggested to be most restrictive for the radial growth of both species. By influencing the seasonal patterns of xylem and phloem development, water availability appears to be the most important determinant of tissue- and species-specific responses to local weather conditions.

Seed Viability and Potential Germination Rate of Nine Endemic Boswellia Taxa (Burseraceae) from Socotra Island (Yemen).

The endemic Boswellia species (Burseraceae) on Socotra Island (Yemen) are of great local significance due to their various local ethnobotanical uses. However, despite the fact that these trees are endangered, little is known about their biology. We tested seed germination rates in controlled experiments (trials of 21 days) for two subsequent years and for nine endemic taxa of Boswellia occurring on Socotra Island. For this, seeds were collected island-wide from a wide range of localities and for several populations per species. We observed differences in germination among Boswellia species, among species and localities and among both years, which indicates that the development of seeds is strongly affected by external ecological factors. Although we noted a large variation in seed germination (relatively high in Boswellia socotrana), and half of the species showed relatively low mean daily germination, our study indicated that all endangered endemic Frankincense Tree taxa of Socotra harbor the potential for in situ conservation through recruitment, given that known impacts can be reduced in local replantation areas (e.g., grazing).

Climatic regulation of leaf and cambial phenology in Quercus pubescens: Their interlinkage and impact on xylem and phloem conduits.

Increased frequency and severity of stressful events affects the growth patterns and functioning of trees which adjust their phenology to given conditions. Here, we analysed environmental effects (temperature, precipitation, VPD and SWC) on the timing of leaf phenology, seasonal stem radial growth patterns, and xylem and phloem anatomy of Quercus pubescens in the sub-Mediterranean in the period 2014-2019, when various adverse weather events occurred, i.e. spring drought in 2015, summer fire in 2016 and summer drought in 2017. Results showed that the timings of leaf and cambium phenology do not occur simultaneously in Q. pubescens, reflecting different environmental and internal constraints. Although year-to-year variability in the timings of leaf and cambial phenology exists, their chronological sequence is fairly fixed. Different effects of weather conditions on different stages of leaf development in spring were observed. Common climatic drivers (i.e., negative effect of hot and dry summers and a positive effect of increasing moisture availability in winter and summer) were found to affect the widths of xylem and phloem increments with more pronounced effect on late formed parts. A legacy effect of the timing of leaf and cambial phenology of the previous growing season on the timing of phenology of the following spring was confirmed. Rarely available phloem data permitted a comprehensive insight into the interlinkage of the timing of cambium and leaf phenology and adjustment strategies of vascular tissues in Mediterranean pubescent oak to various environmental constraints, including frequent extreme events (drought, fire). Our results suggest that predicted changes in autumn/winter and spring climatic conditions for this area could affect the timings of leaf and stem cambial phenology of Q. pubescens in the coming years, which would affect stem xylem and phloem structure and hydraulic properties, and ultimately its performance.

Gentiana clusii Perr.&Song.: Enhanced production of secondary metabolites by in vitro propagation.

Shoot and root in vitro culture of endemic European species Gentiana clusii was established for the first time. The effects of different concentrations of benzyl adenine (BA), 6-phurphurylaminopurine (KIN), indole-3-butyric acid (IBA) and naphthalene acetic acid (NAA) on shoot propagation and rooting of G. clusii were investigated. The optimal in vitro conditions for shoot propagation and long-term maintenance were achieved using woody plant medium (WPM) supplemented with 0.5 mg l-1 KIN, and subsequent application of IBA at 0.5 mg l-1 significantly improved rooting of these shoots. Root culture was established from excised root tips cultured in ½ MS liquid media with increasing concentrations of IBA (0.1-1.0 mg l-1). A high root growth rate and considerable biomass yield were obtained by addition of 1.0 mg l-1 IBA. HPLC analysis revealed that in vitro culture considerably promoted the production of secondary metabolites in G. clusii. The selected protocol for shoot propagation (WPM + 0.5 mg l-1 KIN) increased the content of sweroside, gentiopicrin and norswertianin-1-O-primeveroside (N-1-P) for more than 2-fold compared with the wild plants. IBA promoted N-1-P and norswertianin production in root cultures; their contents were enhanced 6.4- and 18.6-fold, respectively, compared with the wild plants. The extract of these roots displayed the highest antioxidant capacity (IC50 = 66.57 µg ml-1). The established shoot and root propagation protocols facilitate in vitro conservation of G. clusii, and provides a promising tool for the large scale production of valuable secoiridoids and xanthones.

Post-fire effects on development of leaves and secondary vascular tissues in Quercus pubescens.

An increased frequency of fire events on the Slovenian Karst is in line with future climate change scenarios for drought-prone environments worldwide. It is therefore of the utmost importance to better understand tree-fire-climate interactions for predicting the impact of changing environment on tree functioning. To this purpose, we studied the post-fire effects on leaf development, leaf carbon isotope composition (δ13C), radial growth patterns and the xylem and phloem anatomy in undamaged (H-trees) and fire-damaged trees (F-trees) of Quercus pubescens Willd. with good resprouting ability in spring 2017, the growing season after a rangeland fire in August 2016. We found that the fully developed canopy of F-trees reached only half of the leaf area index values measured in H-trees. Throughout the season, F-trees were characterized by higher water potential and stomatal conductivity and achieved higher photosynthetic rates compared to unburnt H-trees. The foliage of F-trees had more negative δ13C values than those of H-trees. This reflects that F-trees less frequently meet stomatal limitations due to reduced transpirational area and more favourable leaf-to-root ratio. In addition, the growth of leaves in F-trees relied more on the recent photosynthates than on reserves due to the fire disturbed starch accumulation in the previous season. Cambial production stopped 3 weeks later in F-trees, resulting in 60 and 22% wider xylem and phloem increments, respectively. A novel approach by including phloem anatomy in the analyses revealed that fire caused changes in conduit dimensions in the early phloem but not in the earlywood. However, premature formation of the tyloses in the earlywood vessels of the youngest two xylem increments in F-trees implies that xylem hydraulic integrity was also affected by heat. Analyses of secondary tissues showed that although xylem and phloem tissues are interlinked changes in their transport systems due to heat damage are not necessarily coordinated.

Effect of soil water availability on intra-annual xylem and phloem formation and non-structural carbohydrate pools in stem of Quercus pubescens.

Non-structural carbohydrates (NSCs, i.e., starch and soluble sugars) are frequently quantified in the context of tree response to stressful events (e.g., drought), because they serve as a carbon reservoir for growth and respiration, as well as providing a critical osmotic function to maintain turgor and vascular transport under different environmental conditions. We investigated the impact of soil water availability on intra-annual leaf phenology, radial growth dynamics and variation in NSC amounts in the stem of pubescent oak (Quercus pubescens Willd.). from a sub-Mediterranean region. For this purpose, trees growing at two nearby plots differing in bedrock and, consequently, soil characteristics (F-eutric cambisol on eocene flysch bedrock and L-rendzic leptosol on paleogenic limestone bedrock) were sampled. Non-structural carbohydrates were analysed in outer xylem and living phloem (separately for non-collapsed and collapsed parts). Results showed that xylem and phloem increments were 41.6% and 21.2%, respectively, wider in trees from F plot due to a higher rate of cell production. In contrast, the amount of NSCs and of soluble sugars significantly differed among the tissue parts and sampling dates but not between the two plots. Starch amounts were the highest in xylem, which could be explained by the abundance of xylem parenchyma cells. Two clear seasonal peaks of the starch amount were detected in all tissues, the first in September-November, in the period of leaf colouring and falling, and the second in March-April, i.e., at the onset of cambial cell production followed by bud development. The amounts of free sugars were highest in inner phloem + cambium, at the sites of active growth. Soil water availability substantially influenced secondary growth in the stem of Q. pubescens, whereas NSC amounts seemed to be less affected. The results show how the intricate relationships between soil properties, such as water availability, and tree performance should be considered when studying the impact of stressful events on the growth and functioning of trees.

Chronological Sequence of Leaf Phenology, Xylem and Phloem Formation and Sap Flow of Quercus pubescens from Abandoned Karst Grasslands.

Intra-annual variations in leaf development, radial growth, including the phloem part, and sap flow have rarely been studied in deciduous trees from drought-prone environments. In order to understand better the chronological order and temporal course of these processes, we monitored leaf phenology, xylem and phloem formation and sap flow in Quercus pubescens from abandoned karst grasslands in Slovenia during the growing season of 2014. We found that the initial earlywood vessel formation started before bud opening at the beginning of April. Buds started to open in the second half of April and full leaf unfolding occurred by the end of May. LAI values increased correspondingly with leaf development. About 28% of xylem and 22% of phloem annual increment were formed by the time of bud break. Initial earlywood vessels were fully lignified and ready for water transport, indicating that they are essential to provide hydraulic conductivity for axial water flow during leaf development. Sap flow became active and increasing contemporarily with leaf development and LAI values. Similar early spring patterns of xylem sap flow and LAI denoted that water transport in oaks broadly followed canopy leaf area development. In the initial 3 weeks of radial growth, phloem growth preceded that of xylem, indicating its priority over xylem at the beginning of the growing season. This may be related to the fact that after bud break, the developing foliage is a very large sink for carbohydrates but, at the same time, represents a small transpirational area. Whether the interdependence of the chronological sequence of the studied processes is fixed in Q. pubescens needs to be confirmed with more data and several years of analyses, although the 'correct sequence' of processes is essential for synchronized plant performance and response to environmental stress.

Isotopic and elemental characterisation of Slovenian apple juice according to geographical origin: Preliminary results.

This study examined the applicability of stable isotope and multi-element data for determining the geographical origin of fresh apple juices. Samples included three apple cultivars (Idared, Golden Delicious and Topaz) harvested in 2011 and 2012 from five different geographical regions of Slovenia. Regional discrimination of the juice samples was most successful when using linear discriminant analysis (LDA) and taking into account the following parameters: δ(2)H and δ(18)O content of juice water; δ(15)N and δ(13)C content of the pulp, (D/H)I and (D/H)II in ethanol and the concentration of S, Cl, Fe, Cu, Zn and Sr. Overall prediction ability was 83.9%. The factors that best distinguished the different types of cultivar were the δ(2)H and δ(18)O content of fruit juice water; the δ(13)C and (D/H)I content of ethanol; and the concentration of S, Mg, K, Cu, and Ti. Prediction ability, taking into account all ten parameters, was 75.8%.

Traditional elderflower beverages: a rich source of phenolic compounds with high antioxidant activity.

Traditional elderflower beverages are very popular due to their pleasing taste and pro-health properties. Phenolics were quantified in three elderflower extracts (methanolic extraction and two water extracts prepared as fresh drinks according to local recipes) with HPLC-MS. Chromatographic patterns of methanolic and water extracts were quite similar; however, content levels of individual compounds differed between the two extraction solvents. Two feruloylquinic acids and two p-coumaroyl-caffeoylquinic acids and different flavonol glycosides have been identified in elderflowers for the first time. Hydroxycinnamic acids and flavonol glycosides were the major phenolic constituents in elderflowers. Methanolic extract contained higher levels of all phenolic groups compared to water-extracted samples. The yield of elderflower extracts depended on extract solution and time of extraction. Elderflower beverages were characterized by high total phenolic content (91.09 to 746.63 mg GAE/L). The results confirmed that elderflower beverages (sabesa, syrup) should be recommended as a good source of phenolics in human diet.

Natural radionuclides in trees grown on a uranium mill tailings waste pile.

OBJECTIVE: The purpose of the study was to investigate natural radionuclide uptake and allocation by trees. MATERIALS AND METHODS: Samples from six Scots pines (P. sylvestris), six Norway spruces (Picea abies) and one sycamore maple (Acer pseudoplatanus) tree, growing on the Borst uranium mill tailings waste pile in Slovenia were collected. (238)U, (230)Th, (226)Ra and (210)Pb activity concentrations in wood, shoots and 1-year-old needles or leaves were determined. Particular radionuclides were separated from the samples by appropriate radiochemical procedures and their activity concentrations measured with an alpha spectrometry system. In addition, concentration ratios for different plant parts were calculated. RESULTS AND CONCLUSIONS: Results showed that for all radionuclides, the highest activity concentrations were found in foliage, followed by shoots and wood. The activity concentrations in trees were from 0.01 to 5.4 Bq kg(-1) for (238)U, 0.03-11.3 Bq kg(-1) for (230)Th, 2.7-2,728 Bq kg(-1) for (226)Ra and 5.1-321 Bq kg(-1) for (210)Pb. All activity concentrations were calculated on dry weight basis. The calculated concentration ratios were from 1.05E-5 to 5.39E-3 for (238)U, 7.65E-6-2.88E-3 for (230)Th, 3.10E-4-3.16E-1 for (226)Ra and 6.70E-4-4.22E-2 for (210)Pb.