A multidisciplinary approach to tackling invasive species: barcoding, morphology, and metataxonomy of the leafhopper Arboridia adanae.

The leafhopper genus Arboridia includes several species that feed on Vitis vinifera and cause leaf chlorosis. We report the first alien Arboridia infestation in Italy in 2021 in an Apulian vineyard. To confirm the taxonomic status of the species responsible for crop damage, and reconstruct its demographic history, we barcoded individuals from Apulia together with Arboridia spp. from Crete (Greece), A. adanae from Central Turkey and other specimens of the presumed sister species, A. dalmatina from Dalmatia (Croatia). Molecular phylogenies and barcoding gap analysis identified clades not associated with sampling locations. This result is incongruent with classical specimen assignment and is further supported by morphological analyses, which did not reveal significant differences among the populations. Therefore, we propose A. dalmatina as a junior synonym of A. adanae, which would become the only grapevine-related Arboridia species in the eastern Mediterranean. To further characterise A. adanae evolution, we performed a molecular clock analysis that suggested a radiation during the Pleistocene glaciations. Finally, to assess whether the Apulian individuals carried microorganisms of agricultural relevance, we sequenced their bacterial microbiota using 16S rRNA amplicon sequencing identifying three phytopathogens not generally associated with Arboridia activities as well as Wolbachia in one Apulian haplogroup. We discuss the agricultural implications of this infestation.

Gene expression differences consistent with water loss reduction underlie desiccation tolerance of natural Drosophila populations.

BACKGROUND: Climate change is one of the main factors shaping the distribution and biodiversity of organisms, among others by greatly altering water availability, thus exposing species and ecosystems to harsh desiccation conditions. However, most of the studies so far have focused on the effects of increased temperature. Integrating transcriptomics and physiology is key to advancing our knowledge on how species cope with desiccation stress, and these studies are still best accomplished in model organisms. RESULTS: Here, we characterized the natural variation of European D. melanogaster populations across climate zones and found that strains from arid regions were similar or more tolerant to desiccation compared with strains from temperate regions. Tolerant and sensitive strains differed not only in their transcriptomic response to stress but also in their basal expression levels. We further showed that gene expression changes in tolerant strains correlated with their physiological response to desiccation stress and with their cuticular hydrocarbon composition, and functionally validated three of the candidate genes identified. Transposable elements, which are known to influence stress response across organisms, were not found to be enriched nearby differentially expressed genes. Finally, we identified several tRNA-derived small RNA fragments that differentially targeted genes in response to desiccation stress. CONCLUSIONS: Overall, our results showed that basal gene expression differences across individuals should be analyzed if we are to understand the genetic basis of differential stress survival. Moreover, tRNA-derived small RNA fragments appear to be relevant across stress responses and allow for the identification of stress-response genes not detected at the transcriptional level.

High Resolution Assessment of Spatio-Temporal Changes in O2 Concentration in Root-Pathogen Interaction.

Fusarium wilt, caused by the fungus Fusarium oxysporum f. sp. lycopersici (Fol), is one of the most destructive soil-borne diseases of tomatoes. Infection takes place on the roots and the process starts with contact between the fungus and the roots hairs. To date, no detailed studies are available on metabolic activity in the early stages of the Fol and tomato root interaction. Spatial and temporal patterns of oxygen consumption could provide new insights into the dynamics of early colonization. Here, we combined planar optodes and spatial analysis to assess how tomato roots influence the metabolic activity and growth patterns of Fol. The results shows that the fungal metabolism, measured as oxygen consumption, increases within a few hours after the inoculation. Statistical analysis revealed that the fungus tends to growth toward the root, whereas, when the root is not present, the single elements of the fungus move with a Brownian motion (random). The combination of planar optodes and spatial analysis is a powerful new tool for assessing temporal and spatial dynamics in the early stages of root-pathogen interaction.

Leaf and canopy photosynthesis of a chlorophyll deficient soybean mutant.

The photosynthetic, optical, and morphological characteristics of a chlorophyll-deficient (Chl-deficient) "yellow" soybean mutant (MinnGold) were examined in comparison with 2 green varieties (MN0095 and Eiko). Despite the large difference in Chl content, similar leaf photosynthesis rates were maintained in the Chl-deficient mutant by offsetting the reduced absorption of red photons by a small increase in photochemical efficiency and lower non-photochemical quenching. When grown in the field, at full canopy cover, the mutants reflected a significantly larger proportion of incoming shortwave radiation, but the total canopy light absorption was only slightly reduced, most likely due to a deeper penetration of light into the canopy space. As a consequence, canopy-scale gross primary production and ecosystem respiration were comparable between the Chl-deficient mutant and the green variety. However, total biomass production was lower in the mutant, which indicates that processes other than steady state photosynthesis caused a reduction in biomass accumulation over time. Analysis of non-photochemical quenching relaxation and gas exchange in Chl-deficient and green leaves after transitions from high to low light conditions suggested that dynamic photosynthesis might be responsible for the reduced biomass production in the Chl-deficient mutant under field conditions.

Phenotypic differences determine drought stress responses in ecotypes of Arundo donax adapted to different environments.

Arundo donax has been identified as an important biomass and biofuel crop. Yet, there has been little research on photosynthetic and metabolic traits, which sustain the high productivity of A. donax under drought conditions. This study determined phenotypic differences between two A. donax ecotypes coming from stands with contrasting adaptation to dry climate. We hypothesized that the Bulgarian (BG) ecotype, adapted to drier conditions, exhibits greater drought tolerance than the Italian (IT) ecotype, adapted to a more mesic environment. Under well-watered conditions the BG ecotype was characterized by higher photosynthesis, mesophyll conductance, intrinsic water use efficiency, PSII efficiency, isoprene emission rate and carotenoids, whereas the IT ecotype showed higher levels of hydroxycinnamates. Photosynthesis of water-stressed plants was mainly limited by diffusional resistance to CO2 in BG, and by biochemistry in IT. Recovery of photosynthesis was more rapid and complete in BG than in IT, which may indicate better stability of the photosynthetic apparatus associated to enhanced induction of volatile and non-volatile isoprenoids and phenylpropanoid biosynthesis. This study shows that a large phenotypic plasticity among A. donax ecotypes exists, and may be exploited to compensate for the low genetic variability of this species when selecting plant productivity in constrained environments.

Current status, uncertainty and future needs in soil organic carbon monitoring.

Increasing human demands on soil-derived ecosystem services requires reliable data on global soil resources for sustainable development. The soil organic carbon (SOC) pool is a key indicator of soil quality as it affects essential biological, chemical and physical soil functions such as nutrient cycling, pesticide and water retention, and soil structure maintenance. However, information on the SOC pool, and its temporal and spatial dynamics is unbalanced. Even in well-studied regions with a pronounced interest in environmental issues information on soil carbon (C) is inconsistent. Several activities for the compilation of global soil C data are under way. However, different approaches for soil sampling and chemical analyses make even regional comparisons highly uncertain. Often, the procedures used so far have not allowed the reliable estimation of the total SOC pool, partly because the available knowledge is focused on not clearly defined upper soil horizons and the contribution of subsoil to SOC stocks has been less considered. Even more difficult is quantifying SOC pool changes over time. SOC consists of variable amounts of labile and recalcitrant molecules of plant, and microbial and animal origin that are often operationally defined. A comprehensively active soil expert community needs to agree on protocols of soil surveying and lab procedures towards reliable SOC pool estimates. Already established long-term ecological research sites, where SOC changes are quantified and the underlying mechanisms are investigated, are potentially the backbones for regional, national, and international SOC monitoring programs.

New spectral vegetation indices based on the near-infrared shoulder wavelengths for remote detection of grassland phytomass.

This article examines the possibility of exploiting ground reflectance in the near-infrared (NIR) for monitoring grassland phytomass on a temporal basis. Three new spectral vegetation indices (infrared slope index, ISI; normalized infrared difference index, NIDI; and normalized difference structural index, NDSI), which are based on the reflectance values in the H25 (863-881 nm) and the H18 (745-751 nm) Chris Proba (mode 5) bands, are proposed. Ground measurements of hyperspectral reflectance and phytomass were made at six grassland sites in the Italian and Austrian mountains using a hand-held spectroradiometer. At full canopy cover, strong saturation was observed for many traditional vegetation indices (normalized difference vegetation index (NDVI), modified simple ratio (MSR), enhanced vegetation index (EVI), enhanced vegetation index 2 (EVI 2), renormalized difference vegetation index (RDVI), wide dynamic range vegetation index (WDRVI)). Conversely, ISI and NDSI were linearly related to grassland phytomass with negligible inter-annual variability. The relationships between both ISI and NDSI and phytomass were however site specific. The WinSail model indicated that this was mostly due to grassland species composition and background reflectance. Further studies are needed to confirm the usefulness of these indices (e.g. using multispectral specific sensors) for monitoring vegetation structural biophysical variables in other ecosystem types and to test these relationships with aircraft and satellite sensors data. For grassland ecosystems, we conclude that ISI and NDSI hold great promise for non-destructively monitoring the temporal variability of grassland phytomass.

Temperature sensitivity of the turnover times of soil organic matter in forests.

Soils represent the largest carbon pool in the terrestrial biosphere, and climate change might affect the main carbon fluxes associated with this pool. These fluxes are the production of aboveground litter and root litter, and decomposition of the soil organic matter (SOM) pool by soil microorganisms. Knowledge about the temperature sensitivity of the decomposition of different SOM fractions is crucial in order to understand how climate change might affect carbon storage in soils. In this study, the temperature sensitivity of the turnover times of three different SOM fractions (labile, intermediate, and stabilized) was investigated for 11 forest sites along a temperature gradient. Carbon-14 isotope analyses of the SOM fractions combined with a model provided estimates of their turnover times. The turnover times of the labile SOM fraction were not correlated with mean annual soil temperature. Therefore it was not possible to estimate temperature sensitivity for the labile SOM fraction. Given considerable evidence elsewhere for significant temperature sensitivities of labile SOM, lack of temperature sensitivity here most likely indicates limitations of the applied methodology for the labile SOM fraction. The turnover times of the intermediate and the stabilized SOM fractions were both correlated with mean annual soil temperatures. The temperature sensitivity of the stabilized SOM fraction was at least equal to that of the intermediate SOM fraction and possibly more than twice as high. A correction for confounding effects of soil acidity and clay content on the temperature sensitivities of the intermediate and stabilized SOM fractions was included in the analysis. The results as observed here for the three SOM fractions may have been influenced by (1) modeling assumptions for the estimation of SOM turnover times of leaf and needle longevities, constant annual carbon inputs, and steady-state SOM pools, (2) the occurrence of summer drought at some sites, (3) differences between sites in quality of the SOM fractions, or (4) the relatively small temperature range. Our results suggested that a 1 degree C increase in temperature could lead to decreases in turnover times of 4-11% and 8-16%, for the intermediate and stabilized SOM fractions, respectively.

Soil Respiration in European Grasslands in Relation to Climate and Assimilate Supply.

Soil respiration constitutes the second largest flux of carbon (C) between terrestrial ecosystems and the atmosphere. This study provides a synthesis of soil respiration (R(s)) in 20 European grasslands across a climatic transect, including ten meadows, eight pastures and two unmanaged grasslands. Maximum rates of R(s) (R(s(max) )), R(s) at a reference soil temperature (10°C; R(s(10) )) and annual R(s) (estimated for 13 sites) ranged from 1.9 to 15.9 µmol CO(2) m(-2) s(-1), 0.3 to 5.5 µmol CO(2) m(-2) s(-1) and 58 to 1988 g C m(-2) y(-1), respectively. Values obtained for Central European mountain meadows are amongst the highest so far reported for any type of ecosystem. Across all sites R(s(max) ) was closely related to R(s(10) ).Assimilate supply affected R(s) at timescales from daily (but not necessarily diurnal) to annual. Reductions of assimilate supply by removal of aboveground biomass through grazing and cutting resulted in a rapid and a significant decrease of R(s). Temperature-independent seasonal fluctuations of R(s) of an intensively managed pasture were closely related to changes in leaf area index (LAI). Across sites R(s(10) ) increased with mean annual soil temperature (MAT), LAI and gross primary productivity (GPP), indicating that assimilate supply overrides potential acclimation to prevailing temperatures. Also annual R(s) was closely related to LAI and GPP. Because the latter two parameters were coupled to MAT, temperature was a suitable surrogate for deriving estimates of annual R(s) across the grasslands studied. These findings contribute to our understanding of regional patterns of soil C fluxes and highlight the importance of assimilate supply for soil CO(2) emissions at various timescales.

Major diffusion leaks of clamp-on leaf cuvettes still unaccounted: how erroneous are the estimates of Farquhar et al. model parameters?

Estimates of leaf gas-exchange characteristics using standard clamp-on leaf chambers are prone to errors because of diffusion leaks. While some consideration has been given to CO(2) diffusion leaks, potential water vapour diffusion leaks through chamber gaskets have been neglected. We estimated diffusion leaks of two clamp-on Li-Cor LI-6400 (Li-Cor, Inc., Lincoln, NE, USA) leaf chambers with polymer foam gaskets and enclosing either 2 or 6 cm(2) leaf area, and conducted a sensitivity analysis of the diffusion leak effects on Farquhar et al. photosynthesis model parameters - the maximum carboxylase activity of ribulose 1 x 5-bisphosphate carboxylase/oxygenase (Rubisco) (V(cmax)), capacity for photosynthetic electron transport (J(max)) and non-photorespiratory respiration rate in light (R(d)). In addition, net assimilation rate (A(n)) versus intercellular CO(2) (C(i)) responses were measured in leaves of Mediterranean evergreen species Quercus ilex L. enclosing the whole leaf chamber in a polyvinyl fluoride bag flushed with the exhaust air of leaf chamber, thereby effectively reducing the CO(2) and water vapour gradients between ambient air and leaf chamber. For the empty chambers, average diffusion leak for CO(2), K(CO2), (molar flow rate corresponding to unit CO(2) mole fraction difference) was ca. 0.40 micromol s(-1). K(CO2) increased ca. 50% if a dead leaf was clamped between the leaf chamber. Average diffusion leak for H(2)O was ca. 5- to 10-fold larger than the diffusion leak for CO(2). Sensitivity analyses demonstrated that the consequence of a CO(2) diffusion leak was apparent enhancement of A(n) at high CO(2) mole fraction and reduction at lower CO(2) mole fraction, and overall compression of C(i) range. As the result of these modifications, Farquhar et al. model parameters were overestimated. The degree of overestimation increased in the order of V(cmax) < J(max) < R(d), and was larger for smaller chambers and for leaves with lower photosynthetic capacity, leading to overestimation of all three parameters by 70-290% for 2 cm(2), and by 10-60% for 6 cm(2) chamber. Significant diffusion corrections (5-36%) were even required for leaves with high photosynthetic capacity measured in largest chamber. Water vapour diffusion leaks further enhanced the overestimation of model parameters. For small chambers and low photosynthetic capacities, apparent C(i) was simulated to decrease with increasing A(n) because of simultaneous CO(2) and H(2)O diffusion leaks. Measurements in low photosynthetic capacity Quercus ilex leaves enclosed in 2 cm(2) leaf chamber exhibited negative apparent C(i) values at highest A(n). For the same leaves measured with the entire leaf chamber enclosed in the polyvinyl fluoride bag, C(i) and A(n) increased monotonically. While the measurements without the bag could be corrected for diffusion leaks, the required correction in A(n) and transpiration rates was 100-500%, and there was large uncertainty in Farquhar et al. model parameters derived from 'corrected'A(n)/C(i) response curves because of uncertainties in true diffusion leaks. These data demonstrate that both CO(2) and water vapour diffusion leaks need consideration in measurements with clamp-on leaf cuvettes. As plants in natural environments are often characterized by low photosynthetic capacities, cuvette designs need to be improved for reliable measurements in such species.

Complex adjustments of photosynthetic potentials and internal diffusion conductance to current and previous light availabilities and leaf age in Mediterranean evergreen species Quercus ilex.

Mature non-senescent leaves of evergreen species become gradually shaded as new foliage develops and canopy expands, but the interactive effects of integrated light during leaf formation (Q(int)G), current light (Q(int)C) and leaf age on foliage photosynthetic competence are poorly understood. In Quercus ilex L., we measured the responses of leaf structural and physiological variables to Q(int)C and Q(int)G for four leaf age classes. Leaf aging resulted in increases in leaf dry mass per unit area (M(A)), and leaf dry to fresh mass ratio (D(F)) and decreases in N content per dry mass (N(M)). N content per area (N(A)) was independent of age, indicating that decreases in N(M) reflected dilution of leaf N because of accumulation of dry mass (NA = N(M) M(A)). M(A), D(F) and N(A) scaled positively with irradiance, whereas these age-specific correlations were stronger with leaf growth light than with current leaf light. Area-based maximum ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) carboxylase activity (V(cmax)A), capacity for photosynthetic electron transport (J(max)A) and the rate of non-photorespiratory respiration in light (R(d)A) were also positively associated with irradiance. Differently from leaf structural characteristics, for all data pooled, these relationships were stronger with current light with little differences among leaves of different age. Acclimation to current leaf light environment was achieved by light-dependent partitioning of N in rate-limiting proteins. Mass-based physiological activities decreased with increasing leaf age, reflecting dilution of leaf N and a larger fraction of non-photosynthetic N in older leaves. This resulted in age-dependent modification of leaf photosynthetic potentials versus N relationships. Internal diffusion conductance (g(m)) per unit area (g(m)A) increased curvilinearly with increasing irradiance for two youngest leaf age classes and was independent of light for older leaves. In contrast, g(m) per dry mass (g(m)M) was negatively associated with light in current-year leaves. Greater photosynthetic potentials and moderate changes in diffusion conductance resulted in greater internal diffusion limitations of photosynthesis in higher light. Both area- and mass-based g(m) decreased with increasing leaf age. The decrease in diffusion conductance was larger than changes in photosynthetic potentials, leading to larger CO2 drawdown from leaf internal air space to chloroplasts (delta(c)) in older leaves. The increases in diffusion limitations in older leaves and at higher light scaled with age- and light-dependent increases in MA and D(F). Overall, our study demonstrates a large potential of foliage photosynthetic acclimation to changes in leaf light environment, but also highlights enhanced structural diffusion limitations in older leaves that result from leaf structural acclimation to previous rather than to current light environment and accumulation of structural compounds with leaf age.