Conditions Promoting Mycorrhizal Parasitism Are of Minor Importance for Competitive Interactions in Two Differentially Mycotrophic Species.

Interactions of plants with arbuscular mycorrhizal fungi (AMF) may range along a broad continuum from strong mutualism to parasitism, with mycorrhizal benefits received by the plant being determined by climatic and edaphic conditions affecting the balance between carbon costs vs. nutritional benefits. Thus, environmental conditions promoting either parasitism or mutualism can influence the mycorrhizal growth dependency (MGD) of a plant and in consequence may play an important role in plant-plant interactions. In a multifactorial field experiment we aimed at disentangling the effects of environmental and edaphic conditions, namely the availability of light, phosphorus and nitrogen, and the implications for competitive interactions between Hieracium pilosella and Corynephorus canescens for the outcome of the AMF symbiosis. Both species were planted in single, intraspecific and interspecific combinations using a target-neighbor approach with six treatments distributed along a gradient simulating conditions for the interaction between plants and AMF ranking from mutualistic to parasitic. Across all treatments we found mycorrhizal association of H. pilosella being consistently mutualistic, while pronounced parasitism was observed in C. canescens, indicating that environmental and edaphic conditions did not markedly affect the cost:benefit ratio of the mycorrhizal symbiosis in both species. Competitive interactions between both species were strongly affected by AMF, with the impact of AMF on competition being modulated by colonization. Biomass in both species was lowest when grown in interspecific competition, with colonization being increased in the less mycotrophic C. canescens, while decreased in the obligate mycotrophic H. pilosella. Although parasitism-promoting conditions negatively affected MGD in C. canescens, these effects were small as compared to growth decreases related to increased colonization levels in this species. Thus, the lack of plant control over mycorrhizal colonization was identified as a possible key factor for the outcome of competition, while environmental and edaphic conditions affecting the mutualism-parasitism continuum appeared to be of minor importance.

High intraspecific ability to adjust both carbon uptake and allocation under light and nutrient reduction in Halimium halimifolium L.

The allocation of recently assimilated carbon (C) by plants depends on developmental stage and on environmental factors, but the underlying mechanisms are still a matter of debate. In the present study, we investigated the regulation of C uptake and allocation and their adjustments during plant growth. We induced different allocation strategies in the Mediterranean shrub Halimium halimifolium L. by a reduction of light (Low L treatment) and nutrient availability (Low N treatment) and analyzed allocation parameters as well as morphological and physiological traits for 15 months. Further, we conducted a (13)CO2 pulse-labeling and followed the way of recently assimilated carbon to eight different tissue classes and respiration for 13 days. The plant responses were remarkably distinct in our study, with mainly morphological/physiological adaptions in case of light reduction and adjustment of C allocation in case of nutrient reduction. The transport of recently assimilated C to the root system was enhanced in amount (c. 200%) and velocity under nutrient limited conditions compared to control plants. Despite the 57% light reduction the total biomass production was not affected in the Low L treatment. The plants probably compensated light reduction by an improvement of their ability to fix C. Thus, our results support the concept that photosynthesis is, at least in a medium term perspective, influenced by the C demand of the plant and not exclusively by environmental factors. Finally, our results indicate that growing heterotrophic tissues strongly reduce the C reflux from storage and structural C pools and therefore enhance the fraction of recent assimilates allocated to respiration. We propose that this interruption of the C reflux from storage and structural C pools could be a regulation mechanism for C translocation in plants.

The effects of mineral nitrogen limitation, competition, arbuscular mycorrhiza, and their respective interactions, on morphological and chemical plant traits of Plantago lanceolata.

Plants are sessile organisms that suffer from a multitude of challenges such as abiotic stress or the interactions with competitors, antagonists and symbionts, which influence their performance as well as their eco-physiological and biochemical responses in complex ways. In particular, the combination of different stressors and their impact on plant biomass production and the plant's ability to metabolically adjust to these challenges are less well understood. To study the effects of mineral nitrogen (N) availability, interspecific competition and the association with arbuscular mycorrhizal fungi (AMF) on biomass production, biomass allocation patterns (root/shoot ratio, specific leaf area) and metabolic responses, we chose the model organism Plantago lanceolata L. (Plantaginaceae). Plants were grown in a full factorial experiment. Biomass production and its allocation patterns were assessed at harvest, and the influence of the different treatments and their interactions on the plant metabolome were analysed using a metabolic fingerprinting approach with ultra-high performance liquid chromatography coupled with time-of-flight-mass spectrometry. Limited supply of mineral N caused the most pronounced changes with respect to plant biomass and biomass allocation patterns, and altered the concentrations of more than one third of the polar plant metabolome. Competition also impaired plant biomass production, yet affected the plant metabolome to a much lesser extent than limited mineral N supply. The interaction of competition and limited mineral N supply often caused additive changes on several traits. The association with AMF did not enhance biomass production, but altered biomass allocation patterns such as the root/shoot ratio and the specific leaf area. Interestingly, we did not find significant changes in the plant metabolome caused by AMF. A targeted analysis revealed that only limited mineral N supply reduced the concentrations of one of the main target defence compounds of P. lanceolata, the iridoid glycoside catalpol. In general, the interaction of competition and limited mineral N supply led to additive changes, while the association with AMF in any case alleviated the observed stress responses. Our results show that the joint analysis of biomass/allocation patterns and metabolic traits allows a more comprehensive interpretation of plant responses to different biotic and abiotic challenges; specifically, when multiple stresses interact.

Dynamic carbon allocation into source and sink tissues determine within-plant differences in carbon isotope ratios.

Organs of C3 plants differ in their C isotopic signature (δ13C). In general, leaves are 13C-depleted relative to other organs. To investigate the development of spatial δ13C patterns, we induced different C allocation strategies by reducing light and nutrient availability for 12 months in the Mediterranean shrub Halimium halimifolium L. We measured morphological and physiological traits and the spatial δ13C variation among seven tissue classes during the experiment. A reduction of light (Low-L treatment) increased aboveground C allocation, plant height and specific leaf area. Reduced nutrient availability (Low-N treatment) enhanced C allocation into fine roots and reduced the spatial δ13C variation. In contrast, control and Low-L plants with high C allocation in new leaves showed a high δ13C variation within the plant (up to 2.5‰). The spatial δ13C variation was significantly correlated with the proportion of second-generation leaves from whole-plant biomass (R2=0.46). According to our results, isotope fractionation in dark respiration can influence the C isotope composition of plant tissues but cannot explain the entire spatial pattern seen. Our study indicates a foliar depletion in 13C during leaf development combined with export of relatively 13C-enriched C by mature source leaves as an important reason for the observed spatial δ13C pattern.

Potential advantages of highly mycotrophic foraging for the establishment of early successional pioneer plants on sand.

Adaptive traits ensuring efficient nutrient acquisition, such as extensive fine root systems, are crucial for establishment of pioneer plants on bare sand. Some successful pioneer species of temperate, European sand ecosystems are characterised as obligate mycorrhizals, thus likely substituting fine roots with arbuscular mycorrhizal fungi (AMF). However, it is not clear whether AM fungal-mediated acquisition of scarce and immobile nutrients such as phosphorus (P) is an advantageous strategy on bare sand over foraging via roots. We compared the foraging performance of three obligately mycorrhizal forbs and two facultatively mycorrhizal grasses, regarding the influence of AMF on their capacity to acquire P from bare sand. Comparison of mycorrhizal and non-mycorrhizal individuals revealed a markedly higher AM fungal-dependency for P acquisition and growth in the forbs than in the grasses. Periodical soil core sampling, allowing for assessment of root and hyphal growth rates, revealed hyphal growth to markedly enlarge the total absorptive surface area (SA) in the forbs, but not in the grasses. Correlations between SA growth and P depletion suggest an AM fungal-induced enhanced capacity for rapid soil P exploitation in the forbs. Our study showed that AM fungal-mediated foraging may be an advantageous strategy over root-mediated foraging in sand pioneer plants.

Adjustments in epidermal UV-transmittance of leaves in sun-shade transitions.

Epidermal UV transmittance (TUV ) and UV-absorbing compounds were measured in sun and shade leaves of Populus tremuloides and Vicia faba exposed to contrasting light environments under field conditions to evaluate UV acclimation potentials and regulatory roles of photosynthetically active radiation (PAR) and UV in UV-shielding. Within a natural canopy of P. tremuloides, TUV ranged from 4 to 98% and showed a strong nonlinear relationship with mid-day horizontal fluxes of PAR [photon flux density (PFD) = 6-1830 µmol m⁻² s⁻¹]; similar patterns were found for V. faba leaves that developed under a comparable PFD range. A series of field transfer experiments using neutral-density shade cloth and UV blocking/transmitting films indicated that PAR influenced TUV during leaf development to a greater degree than UV, and shade leaves of both species increased their UV-shielding when exposed to full sun; however, this required the presence of UV, with both UV-A and UV-B required for full acclimation. TUV of sun leaves of both species was largely unresponsive to shade either with or without UV. In most, but not all cases, changes in TUV were associated with alterations in the concentration of whole-leaf UV-absorbing compounds. These results suggest that, (1) moderate-to-high levels of PAR alone during leaf development can induce substantial UV-protection in field-grown plants, (2) mature shade leaves have the potential to adjust their UV-shielding which may reduce the detrimental effects of UV that could occur following sudden exposures to high light and (3) under field conditions, PAR and UV play different roles in regulating UV-shielding during and after leaf development.

Evaluating high time-resolved changes in carbon isotope ratio of respired CO2 by a rapid in-tube incubation technique.

Recent insights into fractionation during dark respiration and rapid dynamics in isotope signatures of leaf- and ecosystem-respired CO(2) indicate the need for new methods for high time-resolved measurements of the isotopic signature of respired CO(2) (delta(13)C(res)). We present a rapid and simple method to analyse delta(13)C(res) using an in-tube incubation technique and an autosampler for small septum-capped vials. The effect of storage on the delta(18)O and delta(13)C ratios of ambient CO(2) concentrations was tested with different humidity and temperatures. delta(13)C ratios remained stable over 72 h, whereas delta(18)O ratios decreased after 24 h. Storage at 4 degrees C improved the storage time for delta(18)O. Leaves or leaf discs were incubated in the vials, flushed with CO(2)-free air and respired CO(2) was automatically sampled within 5 min on a microGas autosampler interfaced to a GV-Isoprime isotope ratio mass spectrometer. Results were validated by simultaneous on-line gas-exchange measurements of delta(13)C(res) of attached leaves. This method was used to evaluate the short-term (5-60 min) and diurnal dynamics of delta(13)C(res) in an evergreen oak (Quercus ilex) and a herb (Tolpis barbata). An immediate depletion of 2-4 per thousand from the initial delta(13)C(res) value occurred during the first 30 min of darkening. Q. ilex exhibited further a substantial diurnal enrichment in delta(13)C(res) of 8 per thousand, followed by a progressive depletion during the night. In contrast, T. barbata did not exhibit a distinct diurnal pattern. This is in accordance with recent theory on fractionation in metabolic pathways and may be related to the different utilisation of the respiratory substrate in the fast-growing herb and the evergreen oak. These data indicate substantial and rapid dynamics (within minutes to hours) in delta(13)C(res), which differed between species and probably the growth status of the plant. The in-tube incubation method enables both high time-resolved analysis and extensive sampling across different organs, species and functional types.

Phenotypic plasticity of an invasive acacia versus two native Mediterranean species.

The phenotypic plasticity and the competitive ability of the invasive Acacia longifolia v. the indigenous Mediterranean dune species Halimium halimifolium and Pinus pinea were evaluated. In particular, we explored the hypothesis that phenotypic plasticity in response to biotic and abiotic factors explains the observed differences in competitiveness between invasive and native species. The seedlings' ability to exploit different resource availabilities was examined in a two factorial experimental design of light and nutrient treatments by analysing 20 physiological and morphological traits. Competitiveness was tested using an additive experimental design in combination with 15N-labelling experiments. Light and nutrient availability had only minor effects on most physiological traits and differences between species were not significant. Plasticity in response to changes in resource availability occurred in morphological and allocation traits, revealing A. longifolia to be a species of intermediate responsiveness. The major competitive advantage of A. longifolia was its constitutively high shoot elongation rate at most resource treatments and its effective nutrient acquisition. Further, A. longifolia was found to be highly tolerant against competition from native species. In contrast to common expectations, the competition experiment indicated that A. longifolia expressed a constant allocation pattern and a phenotypic plasticity similar to that of the native species.

Spatial and age-dependent modifications of photosynthetic capacity in four Mediterranean oak species.

Drought is one of the most important limitations of photosynthesis in Mediterranean climates. However, Mediterranean sclerophyllous species with long-lived leaves also support extensive and dynamic canopies, with potentially large spatial and age-dependent gradients. We studied within-canopy and temporal patterns in foliage structure, chemistry and photosynthesis in the evergreen species Quercus coccifera L., Q. ilex L. subsp. ballota (Desf.) Samp. in Bol. and Q. suber L. and in the semi-deciduous marcescent species Q. faginea Lam. to determine the role of within-canopy shading and leaf age on foliage functioning. There was a 2.5-fold within-canopy gradient in leaf dry mass per unit area (MA) that was accompanied by a 3-fold range in area-based leaf nitrogen (N) content, the capacity for photosynthetic electron transport (Jmax) and maximum Rubisco carboxylase activity (Vcmax), while the fractional investments of leaf nitrogen in electron transport (FB) and in Rubisco (FR) were relatively constant within the canopy. Leaf aging led to increased MA, larger or constant mass-based N content, larger phosphorous (P) and structural carbon contents, but decreased movable cation contents. Age-dependent increases in MA and N per dry mass meant that Jmax and Vcmax per area were weakly related to leaf age, with a trend of decreasing values in older leaves. However, Jmax and Vcmax per unit dry mass decreased 4-fold across the range of leaf age, primarily owing to decreases in apparent N investments in photosynthetic machinery. This decrease in apparent N investments in photosynthetic machinery was possibly the result of a larger fraction of N bound to cell walls, or of an enhanced CO2 diffusion resistance from the outer surface of cell walls to the chloroplasts in older leaves with thicker and more lignified cell walls. The age-dependent variation in apparent fractional investments of N in photosynthetic machinery reduced the generality of leaf nitrogen v. photosynthesis relationships. Photosynthetic characteristics qualitatively fitted the same patterns with leaf age in all species, but at a common leaf age, area-based leaf photosynthetic potentials depended on species-specific values of MA. These data collectively demonstrate important canopy and age-dependent controls on leaf structure, chemistry and photosynthetic potentials that should be included in larger-scale photosynthesis simulations in Mediterranean climates.

Characteristic patterns of chronic and dynamic photoinhibition of different functional groups in a Mediterranean ecosystem.

To analyse characteristic patterns of dynamic and chronic photoinhibition within a plant community, a new technique is proposed, which is based on the long- and short-term recovery time of maximum photochemical efficiency of PSII (F v/F m) after environmental stress. Chronic photoinhibition was determined as a sustainable decrease in predawn F v/F m, occurring during periods of prolonged stress, whereas dynamic photoinhibition was assessed from the fully reversible diurnal decline in F v/F m. Applied to a Mediterranean macchia ecosystem, this definition allowed the characterization of typical annual patterns of chronic and dynamic photoinhibition. Both types of photoinhibition were highest during summer drought. However, differences emerged among the ten dominant macchia species regarding their susceptibility to chronic photoinhibition during different seasons. Chronic and dynamic photoinhibition were dependent on leaf orientation. Semi-deciduous species avoided enhanced chronic photoinhibition through a reduction of excessive light interception by vertical foliage orientation during summer, whereas evergreen sclerophylls did not exhibit pronounced structural photoprotective mechanisms. Chronic and total photoinhibition were significantly correlated with predawn and midday water potentials, respectively, and a grouping of the macchia species into three functional groups is proposed according to this relationship.