Distinguishing carbon gains from photosynthesis and heterotrophy in C3-hemiparasite-C3-host pairs.

BACKGROUND AND AIMS: Previous carbon stable isotope (13C) analyses have shown for very few C3-hemiparasites utilizing C4- or CAM-hosts the use of two carbon sources, autotrophy and heterotrophy. This 13C approach, however, failed for the frequently occurring C3-C3 parasite-host pairs. Thus, we used hydrogen stable isotope (2H) natural abundances as a substitute for 13C within a C3-Orobanchaceae sequence graded by haustoria complexity and C3-Santalaceae. METHODS: Parasitic plants and their real or potential host plants as references were collected in Central European lowland and alpine mountain meadows and forests. Parasitic plants included the xylem-feeding holoparasite Lathraea squamaria parasitizing on the same carbon nutrient source (xylem-transported organic carbon compounds) as potentially Pedicularis, Rhinanthus, Bartsia, Melampyrum and Euphrasia hemiparasites. Reference plants were used for an autotrophy-only isotope baseline. A multi-element stable isotope natural abundance approach was applied. KEY RESULTS: Species-specific heterotrophic carbon gain ranging from 0 to 51 % was estimated by a 2H mixing-model. The sequence in heterotrophic carbon gain mostly met the morphological grading by haustoria complexity: Melampyrum- < Rhinanthus- < Pedicularis-type. CONCLUSION: Due to higher transpiration and lower water-use efficiency, depletion in 13C, 18O and 2H compared to C3-host plants should be expected for tissues of C3-hemiparasites. However, 2H is counterbalanced by transpiration (2H-depletion) and heterotrophy (2H-enrichment). Progressive 2H-enrichment can be used as a proxy to evaluate carbon gains from hosts.

Partial mycoheterotrophy is common among chlorophyllous plants with Paris-type arbuscular mycorrhiza.

BACKGROUND AND AIMS: An arbuscular mycorrhiza is a mutualistic symbiosis with plants as carbon providers for fungi. However, achlorophyllous arbuscular mycorrhizal species are known to obtain carbon from fungi, i.e. they are mycoheterotrophic. These species all have the Paris type of arbuscular mycorrhiza. Recently, two chlorophyllous Paris-type species proved to be partially mycoheterotrophic. In this study, we explore the frequency of this condition and its association with Paris-type arbuscular mycorrhiza. METHODS: We searched for evidence of mycoheterotrophy in all currently published 13C, 2H and 15N stable isotope abundance patterns suited for calculations of enrichment factors, i.e. isotopic differences between neighbouring Paris- and Arum-type species. We found suitable data for 135 plant species classified into the two arbuscular mycorrhizal morphotypes. KEY RESULTS: About half of the chlorophyllous Paris-type species tested were significantly enriched in 13C and often also enriched in 2H and 15N, compared with co-occurring Arum-type species. Based on a two-source linear mixing model, the carbon gain from the fungal source ranged between 7 and 93 % with ferns > horsetails > seed plants. The seed plants represented 13 families, many without a previous record of mycoheterotrophy. The 13C-enriched chlorophyllous Paris-type species were exclusively herbaceous perennials, with a majority of them thriving on shady forest ground. CONCLUSIONS: Significant carbon acquisition from fungi appears quite common and widespread among Paris-type species, this arbuscular mycorrhizal morphotype probably being a pre-condition for developing varying degrees of mycoheterotrophy.

Stable C and N isotope natural abundances of intraradical hyphae of arbuscular mycorrhizal fungi.

Data for stable C and N isotope natural abundances of arbuscular mycorrhizal (AM) fungi are currently sparse, as fungal material is difficult to access for analysis. So far, isotope analyses have been limited to lipid compounds associated with fungal membranes or storage structures (biomarkers), fungal spores and soil hyphae. However, it remains unclear whether any of these components are an ideal substitute for intraradical AM hyphae as the functional nutrient trading organ. Thus, we isolated intraradical hyphae of the AM fungus Rhizophagus irregularis from roots of the grass Festuca ovina and the legume Medicago sativa via an enzymatic and a mechanical approach. In addition, extraradical hyphae were isolated from a sand-soil mix associated with each plant. All three approaches revealed comparable isotope signatures of R. irregularis hyphae. The hyphae were 13C- and 15N-enriched relative to leaves and roots irrespective of the plant partner, while they were enriched only in 15N compared with soil. The 13C enrichment of AM hyphae implies a plant carbohydrate source, whereby the enrichment was likely reduced by an additional plant lipid source. The 15N enrichment indicates the potential of AM fungi to gain nitrogen from an organic source. Our isotope signatures of the investigated AM fungus support recent findings for mycoheterotrophic plants which are suggested to mirror the associated AM fungi isotope composition. Stable isotope natural abundances of intraradical AM hyphae as the functional trading organ for bi-directional carbon-for-mineral nutrient exchanges complement data on spores and membrane biomarkers.

Mucoromycotina Fine Root Endophyte Fungi Form Nutritional Mutualisms with Vascular Plants.

Fungi and plants have engaged in intimate symbioses that are globally widespread and have driven terrestrial biogeochemical processes since plant terrestrialization >500 million years ago. Recently, hitherto unknown nutritional mutualisms involving ancient lineages of fungi and nonvascular plants have been discovered, although their extent and functional significance in vascular plants remain uncertain. Here, we provide evidence of carbon-for-nitrogen exchange between an early-diverging vascular plant (Lycopodiella inundata) and Mucoromycotina (Endogonales) fine root endophyte fungi. Furthermore, we demonstrate that the same fungal symbionts colonize neighboring nonvascular and flowering plants. These findings fundamentally change our understanding of the physiology, interrelationships, and ecology of underground plant-fungal symbioses in modern terrestrial ecosystems by revealing the nutritional role of Mucoromycotina fungal symbionts in vascular plants.

Picky carnivorous plants? Investigating preferences for preys' trophic levels - a stable isotope natural abundance approach with two terrestrial and two aquatic Lentibulariaceae tested in Central Europe.

BACKGROUND AND AIMS: Stable isotope two-source linear mixing models are frequently used to calculate the nutrient-uptake efficiency of carnivorous plants from pooled prey. This study aimed to separate prey into three trophic levels as pooled prey limits statements about the contribution of a specific trophic level to the nutrition of carnivorous plants. Phytoplankton were used as an autotrophic reference for aquatic plants as the lack of suitable reference plants impedes calculation of their efficiency. METHODS: Terrestrial (Pinguicula) and aquatic (Utricularia) carnivorous plants alongside autotrophic reference plants and potential prey from six sites in Germany and Austria were analysed for their stable isotope natural abundances (δ15N, δ13C). A two-source linear mixing model was applied to calculate the nutrient-uptake efficiency of carnivorous plants from pooled prey. Prey preferences were determined using a Bayesian inference isotope mixing model. KEY RESULTS: Phytophagous prey represented the main contribution to the nutrition of Pinguicula (approx. 55 %), while higher trophic levels contributed a smaller amount (diverse approx. 27 %, zoophagous approx. 17 %). As well as around 48 % nitrogen, a small proportion of carbon (approx. 9 %) from prey was recovered in the tissue of plants. Aquatic Utricularia australis received 29 % and U. minor 21 % nitrogen from zooplankton when applying phytoplankton as the autotrophic reference. CONCLUSIONS: The separation of prey animals into trophic levels revealed a major nutritional contribution of lower trophic level prey (phytophagous) for temperate Pinguicula species. Naturally, prey of higher trophic levels (diverse, zoophagous) are rarer, resulting in a smaller chance of being captured. Phytoplankton represents an adequate autotrophic reference for aquatic systems to estimate the contribution of zooplankton-derived nitrogen to the tissue of carnivorous plants. The autonomous firing of Utricularia bladders results in the additional capture of phytoplankton, calling for new aquatic references to determine the nutritional importance of phytoplankton for aquatic carnivorous plants.

Silver stress differentially affects growth of phototrophic and heterotrophic chrysomonad flagellate populations.

Silver ions are among the predominant anthropogenic introduced pollutants in aquatic systems. As silver has effects on species at all trophic levels the community composition in aquatic habitats can be changed as a result of silver stress. The response of planktonic protists to environmental stressors is particularly important as they act both as producers and consumers in complex planktonic communities. Chrysomonad flagellates are of major interest, since this group includes heterotrophic, mixotrophic and phototrophic taxa, and therefore allows analysis of silver stress in organisms with contrasting nutritional strategies independent of a potential taxonomic bias. In a series of lab experiments, we compared the response of different trophic chrysophyte strains to low (5 µg L-1), medium (10 µg L-1) and high (20 µg L-1) nominal Ag concentrations in combination with changes in temperature and light intensity (phototrophs), temperature and food concentration (heterotrophs), or a combination of the above settings (mixotrophs). All tested strains were negatively affected by silver in their growth rates. The phototrophic strains reacted strongly to silver stress, whereas light intensity and temperature had only minor effects on growth rates. For heterotrophic strains, high food concentration toned down the effect of silver, whereas temperatures outside the growth optimum had a combined stress effect. The mixotrophic strains reacted differently depending on whether their nutritional mode was dominated by heterotrophy or by phototrophy. The precise response pattern across all variables was uniquely different for every single species we tested. The present work contributes to a deeper understanding of the effects of environmental stressors on complex planktonic communities. It indicates that silver will negatively impact planktonic communities and may create shifts in their composition and functioning.

Spatial and temporal variation of macro-, meso- and microplastic abundance on a remote coral island of the Maldives, Indian Ocean.

Plastic debris is ubiquitous in the marine environment and the world's shores represent a major sink. However, knowledge about plastic abundance in remote areas is scarce. Therefore, plastic abundance was investigated on a small island of the Maldives. Plastic debris (>1mm) was sampled once in natural long-term accumulation zones at the north shore and at the high tide drift line of the south shore on seven consecutive days to quantify daily plastic accumulation. Reliable identification of plastic debris was ensured by FTIR spectroscopy. Despite the remoteness of the island a considerable amount of plastic debris was present. At both sites a high variability in plastic abundance on a spatial and temporal scale was observed, which may be best explained by environmental factors. In addition, our results show that snapshot sampling may deliver biased results and indicate that future monitoring programs should consider spatial and temporal variation of plastic deposition.