Occurrence of Morphological and Anatomical Adaptive Traits in Young and Adult Plants of the Rare Mediterranean Cliff Species Primula palinuri Petagna

Cliffs worldwide are known to be reservoirs of relict biodiversity. Despite the presence of harsh abiotic conditions, large endemic floras live in such environments. Primula palinuri Petagna is a rare endemic plant species, surviving on cliff sites along a few kilometres of the Tyrrhenian coast in southern Italy. This species is declared at risk of extinction due to human impact on the coastal areas in question. Population surveys have shown that most of the plants are old individuals, while seedlings and plants at early stages of development are rare. We followed the growth of P. palinuri plants from seed germination to the adult phase and analysed the morphoanatomical traits of plants at all stages of development. Our results showed that the pressure of cliff environmental factors has been selected for seasonal habitus and structural adaptive traits in this species. The main morphoanatomical modifications are suberized cell layers and accumulation of phenolic compounds in cell structures. These features are strictly related to regulation of water uptake and storage as well as defence from predation. However, we found them well established only in adult plants and not in juvenile individuals. These findings contribute to explain the rare recruitment of the present relict populations, identifying some of the biological traits which result in species vulnerability

Morphofunctional Traits and Pollination Mechanisms of Coronilla emerus L. Flowers (Fabaceae)

It is accepted that the papilionaceous corolla of the Fabaceae evolved under the selective pressure of bee pollinators. Morphology and function of different parts of Coronilla emerus L. flowers were related to their role in the pollination mechanism. The corolla has a vexillum with red nectar lines, a keel hiding stamens and pistil, and two wing petals fasten to the keel with two notched folds. Pollinators land on the complex of keel and wings, trigger the protrusion of pollen and finally of the stigma from the keel tip. Data on pollen viability and stigma receptivity prove that flowers are proterandrous. The results of hand-pollination experiments confirmed that insects are fundamental to set seed. Interaction with pollinators allows not only the transport of pollen but also the rupture of the stigmatic cuticle, necessary to achieve both allogamy and autogamy. Field observations showed that Hymenoptera, Lepidoptera, and Diptera visited the flowers. Only some of the Hymenoptera landed on the flowers from the front and elicited pollination mechanisms. Most of the insects sucked the nectar from the back without any pollen transfer. Finally, morphological and functional characteristics of C. emerus flowers are discussed in terms of floral larceny and reduction in pollination efficiency

From flower to honey bouquet: possible markers for the botanical origin of Robinia honey.

Flowers are complex structures devoted to pollinator attraction, through visual as well as chemical signals. As bees collect nectar on flowers to produce honey, some aspects of floral chemistry are transferred to honey, making chemical markers an important technique to identify the botanical and geographical origins of honey. We applied a new approach that considers the simultaneous analysis of different floral parts (petals, stamens + pistils, calyxes + nectarines, and nectar) and the corresponding unifloral honey. We collected fresh flowers of Robinia pseudoacacia L. (black locust), selected five samples of Robinia honey from different geographical origins, applied SPME-GC/MS for volatile analyses, and defined the chemical contribution added by different floral parts to the honey final bouquet. Our results show that honey blends products from nectar as well as other flower parts. Comparing honey and flower profiles, we detected compounds coming directly from flower parts but not present in the nectar, such as hotrienol and β-pinene. These may turn out to be of special interest when selecting floral markers for the botanical origin of honey

Changes in Morpho-Anatomical and Eco-Physiological Responses of Viburnum tinus L. var lucidum as Modulated by Sodium Chloride and Calcium Chloride Salinization

Salinity in water and soil is among the major constraints to the cultivation of ornamental crops since it can affect their growth and aesthetic value. A greenhouse experiment was carried out to assess whether the application of two different salts (80mM NaCl or 53.3mM CaCl2, with a final ionic concentration of 160 mM) could differently modulate the anatomical and physiological acclimation of an important ornamental species such as Viburnum tinus L. var. lucidum. Eco-physiological analyses (e.g., leaf gas exchange and chlorophyll a fluorescence emission) were performed and leaves were subjected to light microscopy analysis to quantify functional anatomical traits through digital image analysis. Results showed that the two iso-osmotic solutions induced different structuremediated physiological alterations in V. tinus plants. Photosynthesis was lowered by CaCl2 treatments (-58%) more than by NaCl (-37%), also due to the occurrence of photodamage apart from stomatal limitations. Neither Na+ nor Cl- exhibited toxic effects in leaf lamina structure which was reflected in the limited reduction in dry matter accumulation. Overall data were interpreted focusing on the coordination among leaf structural and functional traits suggesting that the fine control of functional anatomical traits contributes to physiological acclimation to both stressful condition

Wood growth in pure and mixed Quercus ilex L. forests: drought influence depends on site conditions.

Climate response of tree-species growth may be influenced by intra- and inter-specific interactions. The different physiological strategies of stress response and resource use among species may lead to different levels of competition and/or complementarity, likely changing in space and time according to climatic conditions. Investigating the drivers of inter- and intra-specific interactions under a changing climate is important when managing mixed and pure stands, especially in a climate change hot spot such as the Mediterranean basin. Mediterranean tree rings show intra-annual density fluctuations (IADFs): the links among their occurrence, anatomical traits, wood growth and stable isotope ratios can help understanding tree physiological responses to drought. In this study, we compared wood production and tree-ring traits in Quercus ilex L. dominant trees growing in two pure and two mixed stands with Pinus pinea at two sites in Southern Italy, on the basis of the temporal variation of cumulative basal area, intrinsic water use efficiency (WUEi), δ18O and IADF frequency in long tree-ring chronologies. The general aim was to assess whether Q. ilex trees growing in pure or mixed stands have a different wood production through time, depending on climatic conditions and stand structure. The occurrence of dry climatic conditions triggered opposite complementarity interactions for Q. ilex growing with P. pinea trees at the two sites. Competitive reduction was experienced at the T site characterized by higher soil water holding capacity (WHC), lower stand density and less steep slope than the S site; on the opposite, high competition occurred at S site. The observed difference in wood growth was accompanied by a higher WUEi due to a higher photosynthetic rate at the T site, while by a tighter stomatal control in mixed stand of S site. IADF frequency in Q. ilex tree rings was linked to higher WUEi, thus to stressful conditions and could be interpreted as strategy to cope with dry periods, independently from the different wood growth. Considering the forecasted water shortage, inter-specific competition should be reduced in denser stands of Q. ilex mixed with P. pinea. Such findings have important implications for forest management of mixed and pure Q. ilex forests

Stochastic changes affect Solanum wild species following autopolyploidization

Polyploidy is very common within angiosperms, and several studies are in progress to ascertain the effects of early polyploidization at the molecular, physiological, and phenotypic level. Extensive studies are available only in synthetic allopolyploids. By contrast, less is known about the consequences of autopolyploidization. The current study aimed to assess the occurrence and extent of genetic, epigenetic, and anatomical changes occurring after oryzaline-induced polyploidization of Solanum commersonii Dunal and Solanum bulbocastanum Dunal, two diploid (2n=2×=24) potato species widely used in breeding programmes. Microsatellite analysis showed no polymorphisms between synthetic tetraploids and diploid progenitors. By contrast, analysis of DNA methylation levels indicated that subtle alterations at CG and CHG sites were present in tetraploids of both species. However, no change occurred concurrently in all tetraploids analysed with respect to their diploid parent, revealing a stochastic trend in the changes observed. The morpho-anatomical consequences of polyploidization were studied in leaf main veins and stomata. With only a few exceptions, analyses showed no clear superiority of tetraploids in terms of leaf thickness and area, vessel number, lumen size and vessel wall thickness, stomata pore length and width, guard cell width, and stomatal density com- pared with their diploid progenitors. These results are consistent with the hypothesis that there are no traits system- atically associated with autopolyploidy

Timing of False Ring Formation in Pinus halepensis and Arbutus unedo in Southern Italy: Outlook from an Analysis of Xylogenesis and Tree-Ring Chronologies

Mediterranean tree rings are characterized by intra-annual density fluctuations (IADFs) due to partly climate-driven cambial activity. IADFs are used as structural signals to gain information on relations between environmental conditions and eco-physiological processes during xylogenesis, with intra-annual resolution. To reach an unbiased synchronization of the IADF position within tree rings and seasonal fluctuations in environmental conditions, it is necessary to know the timing of cambial activity and wood formation, which are species- and site-specific processes. We applied the microcoring technique to analyze xylogenesis in Pinus halepensis and Arbutus unedo. To the best of our knowledge, this is the first attempt to study xylogenesis in a hardwood species forming frequent IADFs. Both species co-occur at a site in southern Italy characterized by a Mediterranean climate. To facilitate tree-ring dating and identification of IADFs, we performed traditional dendroecological analysis. We analyzed xylogenesis during summer, which is considered a constraint for xylogenesis and a trigger for IADF formation. We followed the different phases of cell development in the current wood increment with the aim of evaluating whether and which type of IADFs were formed. We additionally analyzed the same phases again in September and in winter to verify the possible formation of IADFs in fall and whether cell production and differentiation was completed by the end of the calendar year. Both species formed the same type of IADFs (earlywood-like cells within latewood), due to temporary growth restoration triggered by rain events during the period of summer drought. At the end of the calendar year, no cells in the phases of enlargement and secondary cell wall deposition occurred. A. unedo was more sensitive than P. halepensis because IADFs were formed earlier in the season and were more frequent in the tree-ring series. The dendro-anatomical approach, combining analysis of tree-ring series and of xylogenesis, helped to detect the period of IADF formation in the two species. Results are discussed in functional terms, highlighting the environmental conditions triggering IADFs, and also in methodological terms, evaluating the applicability of xylogenesis analysis in Mediterranean woods, especially when the formation of IADFs is not uniform around the stem

How Leaf Vein and Stomata Traits Are Related with Photosynthetic Efficiency in Falanghina Grapevine in Different Pedoclimatic Conditions

The increase in severe drought events due to climate change in the areas traditionally suitable for viticulture is enhancing the need to understand how grapevines regulate their photosynthetic metabolism in order to forecast specific cultivar adaptive responses to the changing environment. This study aims at evaluating the association between leaf anatomical traits and eco-physiological adjustments of the 'Falanghina' grapevine under different microclimatic conditions at four sites in southern Italy. Sites were characterized by different pedoclimatic conditions but, as much as possible, were similar for plant material and cultivation management. Microscopy analyses on leaves were performed to quantify stomata and vein traits, while eco-physiological analyses were conducted on vines to assess plant physiological adaptation capability. At the two sites with relatively low moisture, photosynthetic rate, stomatal conductance, photosystem electron transfer rate, and quantum yield of PSII, linear electron transport was lower compared to the other two sites. Stomata size was higher at the site characterized by the highest precipitation. However, stomatal density and most vein traits tended to be relatively stable among sites. The number of free vein endings per unit leaf area was lower in the two vineyards with low precipitation. We suggest that site-specific stomata and vein traits modulation in Falanghina grapevine are an acclimation strategy that may influence photosynthetic performance. Overall in-depth knowledge of the structure/function relations in Falanghina vines might be useful to evaluate the plasticity of this cultivar towards site-specific management of vineyards in the direction of precision viticulture

Tree-rings in Mediterranean pines – can we ascribe them to calendar years?

A tree ring is defined as a band of xylem-cell layers produced in one year. In Mediterranean trees cambium often produces several bands of alternating early- and latewood during one calendar year, resulting in Intra-Annual Density Fluctuations (IADFs). We present some characteristics of growth layers in Pinus pinea (stone pine) growing on Vesuvius Mountain (near Naples, Italy). The trees showed uninterrupted wood production from January 2015 till the end of January 2016. The ring formed in this period contained five different layers of wood types. Therefore, the wood formed between January and March 2015 would be falsely ascribed to latewood of the tree ring 2014, and the wood formed between March 2015 and the end of January 2016 would be classified as tree ring 2015. We also showed differences in cambium activity in late autumn between years: in November 2015 and 2018 the cambium was still active and not active anymore, respectively. Furthermore, we present the structure of the wood formed in the year 2017 affected by a wildfire in July. Such cases where the growth layer cannot be exactly ascribed to a calendar year should be considered in dendrochronological studies of Pinus pinea and other Mediterranean species