A Single-Pore Residue Renders the Arabidopsis Root Anion Channel SLAH2 Highly Nitrate Selective.

In contrast to animal cells, plants use nitrate as a major source of nitrogen. Following the uptake of nitrate, this major macronutrient is fed into the vasculature for long-distance transport. The Arabidopsis thaliana shoot expresses the anion channel SLOW ANION CHANNEL1 (SLAC1) and its homolog SLAC1 HOMOLOGOUS3 (SLAH3), which prefer nitrate as substrate but cannot exclude chloride ions. By contrast, we identified SLAH2 as a nitrate-specific channel that is impermeable for chloride. To understand the molecular basis for nitrate selection in the SLAH2 channel, SLAC1 and SLAH2 were modeled to the structure of HiTehA, a distantly related bacterial member. Structure-guided site-directed mutations converted SLAC1 into a SLAH2-like nitrate-specific anion channel and vice versa. Our findings indicate that two pore-occluding phenylalanines constrict the pore. The selectivity filter of SLAC/SLAH anion channels is determined by the polarity of pore-lining residues located on alpha helix 3. Changing the polar character of a single amino acid side chain (Ser-228) to a nonpolar residue turned the nitrate-selective SLAH2 into a chloride/nitrate-permeable anion channel. Thus, the molecular basis of the anion specificity of SLAC/SLAH anion channels seems to be determined by the presence and constellation of polar side chains that act in concert with the two pore-occluding phenylalanines.

SLAH3-type anion channel expressed in poplar secretory epithelia operates in calcium kinase CPK-autonomous manner.

Extrafloral nectaries secrete a sweet sugar cocktail that lures predator insects for protection from foraging herbivores. Apart from sugars and amino acids, the nectar contains the anions chloride and nitrate. Recent studies with Populus have identified a type of nectary covered by apical bipolar epidermal cells, reminiscent of the secretory brush border epithelium in animals. Border epithelia operate transepithelial anion transport, which is required for membrane potential and/or osmotic adjustment of the secretory cells. In search of anion transporters expressed in extrafloral nectaries, we identified PttSLAH3 (Populus tremula × Populus tremuloides SLAC1 Homologue3), an anion channel of the SLAC/SLAH family. When expressed in Xenopus oocytes, PttSLAH3 displayed the features of a voltage-dependent anion channel, permeable to both nitrate and chloride. In contrast to the Arabidopsis SLAC/SLAH family members, the poplar isoform PttSLAH3 is independent of phosphorylation activation by protein kinases. To understand the basis for the autonomous activity of the poplar SLAH3, we generated and expressed chimera between kinase-independent PttSLAH3 and kinase-dependent Arabidopsis AtSLAH3. We identified the N-terminal tail and, to a lesser extent, the C-terminal tail as responsible for PttSLAH3 kinase-(in)dependent action. This feature of PttSLAH3 may provide the secretory cell with a channel probably controlling long-term nectar secretion.

Segregation of nitrogen use between ammonium and nitrate of ectomycorrhizas and beech trees.

Here, we characterized nitrogen (N) uptake of beech (Fagus sylvatica) and their associated ectomycorrhizal (EM) communities from NH4+ and NO3- . We hypothesized that a proportional fraction of ectomycorrhizal N uptake is transferred to the host, thereby resulting in the same uptake patterns of plants and their associated mycorrhizal communities. 15 N uptake was studied under various field conditions after short-term and long-term exposure to a pulse of equimolar NH4+ and NO3- concentrations, where one compound was replaced by 15 N. In native EM assemblages, long-term and short-term 15 N uptake from NH4+ was higher than that from NO3- , regardless of season, water availability and site exposure, whereas in beech long-term 15 N uptake from NO3- was higher than that from NH4+ . The transfer rates from the EM to beech were lower for 15 N from NH4+ than from NO3- . 15 N content in EM was correlated with 15 N uptake of the host for 15 NH4+ , but not for 15 NO3- -derived N. These findings suggest stronger control of the EM assemblage on N provision to the host from NH4+ than from NO3- . Different host and EM accumulation patterns for inorganic N will result in complementary resource use, which might be advantageous in forest ecosystems with limited N availability.

Ectomycorrhizal Communities on the Roots of Two Beech (Fagus sylvatica) Populations from Contrasting Climates Differ in Nitrogen Acquisition in a Common Environment.

Beech (Fagus sylvatica), a dominant forest species in Central Europe, competes for nitrogen with soil microbes and suffers from N limitation under dry conditions. We hypothesized that ectomycorrhizal communities and the free-living rhizosphere microbes from beech trees from sites with two contrasting climatic conditions exhibit differences in N acquisition that contribute to differences in host N uptake and are related to differences in host belowground carbon allocation. To test these hypotheses, young trees from the natural regeneration of two genetically similar populations, one from dryer conditions (located in an area with a southwest exposure [SW trees]) and the other from a cooler, moist climate (located in an area with a northeast exposure [NE trees]), were transplanted into a homogeneous substrate in the same environment and labeled with (13)CO2 and (15)NH4 (+). Free-living rhizosphere microbes were characterized by marker genes for the N cycle, but no differences between the rhizospheres of SW or NE trees were found. Lower (15)N enrichment was found in the ectomycorrhizal communities of the NE tree communities than the SW tree communities, whereas no significant differences in (15)N enrichment were observed for nonmycorrhizal root tips of SW and NE trees. Neither the ectomycorrhizal communities nor the nonmycorrhizal root tips originating from NE and SW trees showed differences in (13)C signatures. Because the level of (15)N accumulation in fine roots and the amount transferred to leaves were lower in NE trees than SW trees, our data support the suggestion that the ectomycorrhizal community influences N transfer to its host and demonstrate that the fungal community from the dry condition was more efficient in N acquisition when environmental constraints were relieved. These findings highlight the importance of adapted ectomycorrhizal communities for forest nutrition in a changing climate.

What Drove Program Participants to Initially Attend Congregate Meals? Socialization, Health, and Other Reasons.

The Congregate Nutrition Services support efforts to keep older Americans independent and engaged in their communities. We examined participants' self-reported reasons for initially attending the congregate meals program and whether reasons differed by participant characteristics. Descriptive statistics and tests of differences were used to compare participants (n = 1,072). Individuals attended congregate meals for several reasons, with the top two being socialization (36.3%) and age- or health-related reasons (18.7%). Those attending for socialization were less likely to be lower income, have food insecurity, or live with 3+ ADL limitations while participants who first attended due to age or health-related reasons were more likely to be low income, food insecure, and from historically marginalized populations. Health and social service professionals and community organizations could expand data collection on older adults in their communities and partner with congregate meal providers to encourage participation for individuals with unmet nutritional, health, and socialization needs.

Prevention and Treatment of Malnutrition in Older Adults Living in Long-Term Care or the Community: An Evidence-Based Nutrition Practice Guideline.

Malnutrition in older adults can decrease quality of life and increase risk of morbidities and mortality. Accurate and timely identification of malnutrition, as well as subsequent implementation of effective interventions, are essential to decrease poor outcomes associated with malnutrition in older adults. The Academy of Nutrition and Dietetics Evidence Analysis Center conducted a systematic review of the literature to develop an evidence-based nutrition practice guideline for the prevention and treatment of malnutrition in older adults. The objective of this guideline was to provide evidence-based recommendations to identify, prevent, or treat protein-energy malnutrition in older adults (mean age ≥65 years) living in long-term care and community settings. This guideline provides 11 nutrition recommendations to inform shared decision making among dietitians, members of the health care team, family members or caregivers, and older adults living in long-term care or the community to prevent or treat malnutrition. Topics include dietitian effectiveness, nutrition assessment tools, oral nutrition supplements, food fortification, and home-delivered and congregate meals. Guideline implementation should include consideration of the importance of comprehensive individualized nutrition care for older adults. Future research is needed to address gaps that were identified related to the validity, reliability, and feasibility of nutrition assessment tools, as well as the effectiveness of dietitian interventions on outcomes of interest in older adults living in long-term care and the community.

Rhizospheric NO affects N uptake and metabolism in Scots pine (Pinus sylvestris L.) seedlings depending on soil N availability and N source.

We investigated the interaction of rhizospheric nitric oxide (NO) concentration (i.e. low, ambient or high) and soil nitrogen (N) availability (i.e. low or high) with organic and inorganic N uptake by fine roots of Pinus sylvestris L. seedlings by (15) N feeding experiments under controlled conditions. N metabolites in fine roots were analysed to link N uptake to N nutrition. NO affected N uptake depending on N source and soil N availability. The suppression of nitrate uptake in the presence of ammonium and glutamine was overruled by high NO. The effects of NO on N uptake with increasing N availability showed different patterns: (1) increasing N uptake regardless of NO concentration (i.e. ammonium); (2) increasing N uptake only with high NO concentration (i.e. nitrate and arginine); and (3) decreasing N uptake (i.e. glutamine). At low N availability and high NO nitrate accumulated in the roots indicating insufficient substrates for nitrate reduction or its storage in root vacuoles. Individual amino acid concentrations were negatively affected with increasing NO (i.e. asparagine and glutamine with low N availability, serine and proline with high N availability). In conclusion, this study provides first evidence that NO affects N uptake and metabolism in a conifer.

Responses of native and invasive woody seedlings to combined competition and drought are species-specific.

Woody species invasions are a major threat to native communities with intensified consequences during increased periods of summer drought as predicted for the future. Competition for growth-limiting nitrogen (N) between native and invasive tree species might represent a key mechanism underlying the invasion process, because soil water availability and N acquisition of plants are closely linked. To study whether the traits of invasive species provide an advantage over natives in Central Europe in the competition for N under drought, we conducted a greenhouse experiment. We analyzed the responses of three native (i.e., Fagus sylvatica L., Quercus robur L. and Pinus sylvestris L.) and two invasive woody species (i.e., Prunus serotina Ehrh. and Robinia pseudoacacia L.) to competition in terms of their organic and inorganic N acquisition, as well as allocation of N to N pools in the leaves and fine roots. In our study, competition resulted in reduced growth and changes in internal N pools in both native and invasive species mediated by the physiological characteristics of the target species, the competitor, as well as soil water supply. Nitrogen acquisition, however, was not affected by competition indicating that changes in growth and N pools were rather linked to the remobilization of stored N. Drought led to reduced N acquisition, growth and total soluble protein-N levels, while total soluble amino acid-N levels increased, most likely as osmoprotectants as an adaptation to the reduced water supply. Generally, the consequences of drought were enhanced with competition across all species. Comparing the invasive competitors, P. serotina was a greater threat to the native species than R. pseudoacacia. Furthermore, deciduous and coniferous native species affected the invasives differently, with the species-specific responses being mediated by soil water supply.

Tree species rather than type of mycorrhizal association drive inorganic and organic nitrogen acquisition in tree-tree interactions.

Mycorrhizal fungi play an important role for the nitrogen (N) supply of trees. The influence of different mycorrhizal types on N acquisition in tree-tree interactions is, however, not well understood, particularly with regard to the competition for growth-limiting N. We studied the effect of competition between temperate forest tree species on their inorganic and organic N acquisition in relation to their mycorrhizal type (i.e., arbuscular mycorrhiza or ectomycorrhiza). In a field experiment, we quantified net N uptake capacity from inorganic and organic N sources using 15N/13C stable isotopes for arbuscular mycorrhizal tree species (i.e., Acer pseudoplatanus L., Fraxinus excelsior L., and Prunus avium L.) as well as ectomycorrhizal tree species (i.e., Carpinus betulus L., Fagus sylvatica L., and Tilia platyphyllos Scop.). All species were grown in intra- and interspecific competition (i.e., monoculture or mixture). Our results showed that N sources were not used complementarily depending on a species' mycorrhizal association, but their uptake rather depended on the competitor, indicating species-specific effects. Generally, ammonium was preferred over glutamine and glutamine over nitrate. In conclusion, our findings suggest that the inorganic and organic N acquisition of the studied temperate tree species is less regulated by mycorrhizal association but rather by the availability of specific N sources in the soil as well as the competitive environment of different tree species.

Girdling affects ectomycorrhizal fungal (EMF) diversity and reveals functional differences in EMF community composition in a beech forest.

The relationships between plant carbon resources, soil carbon and nitrogen content, and ectomycorrhizal fungal (EMF) diversity in a monospecific, old-growth beech (Fagus sylvatica) forest were investigated by manipulating carbon flux by girdling. We hypothesized that disruption of the carbon supply would not affect diversity and EMF species numbers if EM fungi can be supplied by plant internal carbohydrate resources or would result in selective disappearance of EMF taxa because of differences in carbon demand of different fungi. Tree carbohydrate status, root demography, EMF colonization, and EMF taxon abundance were measured repeatedly during 1 year after girdling. Girdling did not affect root colonization but decreased EMF species richness of an estimated 79 to 90 taxa to about 40 taxa. Cenococcum geophilum, Lactarius blennius, and Tomentella lapida were dominant, colonizing about 70% of the root tips, and remained unaffected by girdling. Mainly cryptic EMF species disappeared. Therefore, the Shannon-Wiener index (H') decreased but evenness was unaffected. H' was positively correlated with glucose, fructose, and starch concentrations of fine roots and also with the ratio of dissolved organic carbon to dissolved organic nitrogen (DOC/DON), suggesting that both H' and DOC/DON were governed by changes in belowground carbon allocation. Our results suggest that beech maintains numerous rare EMF species by recent photosynthate. These EM fungi may constitute biological insurance for adaptation to changing environmental conditions. The preservation of taxa previously not known to colonize beech may, thus, form an important reservoir for future forest development.

Allocation of nitrogen to chemical defence and plant functional traits is constrained by soil N.

Plants have evolved a vast array of defence mechanisms to avoid or minimize damage caused by herbivores and pathogens. The costs and benefits of defences are thought to vary with the availability of resources, herbivore pressure and plant functional traits. We investigated the resource (nitrogen) and growth cost of deploying cyanogenic glycosides in seedlings of Eucalyptus cladocalyx (Myrtaceae). To do this, we grew the plants under a range of soil N conditions, from levels that were limiting for growth to those that were saturating for growth, and we measured correlations between foliar chemical and performance attributes. Within each N treatment, we found evidence that, for every N invested in cyanogenic glycosides, additional N is added to the leaf. For the lowest N treatment, the additional N was less than one per cyanogenic glycoside, rising to some two Ns for the other treatments. The interaction between cyanogenic glycosides and both condensed tannins and total phenolic compounds was also examined, but we did not detect correlations between these compounds under constant leaf N concentrations. Finally, we did not detect a correlation between net assimilation rate, relative growth rate and cyanogenic glycoside concentrations under any soil N treatment. We conclude that the growth cost of cyanogenic glycosides was likely too low to detect and that it was offset to some degree by additional N that was allocated alongside the cyanogenic glycosides.

The presence of amino acids affects inorganic N uptake in non-mycorrhizal seedlings of European beech (Fagus sylvatica).

To investigate the impact of organic N compounds for inorganic nitrogen uptake in the rhizosphere, we fed ammonium nitrate with or without amino acids (i.e., glutamine or arginine) to the roots of non-mycorrhizal beech (Fagus sylvatica L.) seedlings under controlled conditions at different levels of N availability. Uptake of individual N sources was determined from ¹5N (inorganic N) and ¹5N ¹³C (organic N) accumulation in the roots. In addition, gene fragments encoding proteins involved in N uptake and metabolism were cloned from beech for gene expression analyses by quantitative real-time PCR in the roots. Generally, ammonium was preferred over nitrate as N source. Organic N sources were taken up by beech roots as intact molecules. Uptake of organic N was significantly higher than inorganic N uptake, thus contributing significantly to N nutrition of beech. Depending on the level of N availability, inorganic N uptake was negatively affected by the presence of organic N sources. This result indicates an overestimation of the contribution of inorganic N uptake to N nutrition of beech in previous studies. Apparently, association with mycorrhizal fungi is not essential for organic N uptake by beech roots. Gene expression analyses showed that transcriptional regulation of the amino acid transporters FsCAT3, FsCAT5, FsAAT and FsAAP and the ammonium transporter FsAMT1.2 in the roots is involved in N nutrition of beech.

Woody legumes: a (re)view from the South.

This review is focused on woody legumes from the southern continents. We highlight that the evolution of the Caesalpinioideae and Mimosoideae with old soils, with variable supplies of water and also with fire has produced a suite of advantageous physiological characteristics. These include good potential for nitrogen fixation and mechanisms for acquiring P. The latter includes the ability to form cluster roots and produce extracellular phosphatase enzymes. Further, many of the species in these subfamilies are known to synthesize in significant amounts osmotically compatible solutes, such as pinitol and other cyclitols/polyols, that help them cope with even severe drought conditions. In many cases, these species regenerate prolifically after fire from seed. Such species and their beneficial characters can now be better exploited to help sequester carbon, provide key nutrients such as nitrogen and phosphorus for companion crops and other plants and provide feedstocks for a range of industries, including energy industries.

Mechanisms of Carbon Sequestration in Highly Organic Ecosystems - Importance of Chemical Ecology.

Organic matter decomposition plays a major role in the cycling of carbon (C) and nutrients in terrestrial ecosystems across the globe. Climate change accelerates the decomposition rate to potentially increase the release of greenhouse gases and further enhance global warming in the future. However, fractions of organic matter vary in turnover times and parts are stabilized in soils for longer time periods (C sequestration). Overall, a better understanding of the mechanisms underlying C sequestration is needed for the development of effective mitigation policies to reduce land-based production of greenhouse gases. Known mechanisms of C sequestration include the recalcitrance of C input, interactions with soil minerals, aggregate formation, as well as its regulation via abiotic factors. In this Minireview, we discuss the mechanisms behind C sequestration including the recently emerging significance of biochemical interactions between organic matter inputs that lead to C stabilization.

A Brief Intervention for Malnutrition among Older Adults: Stepping Up Your Nutrition.

Despite a multitude of nutritional risk factors among older adults, there is a lack of community-based programs and activities that screen for malnutrition and address modifiable risk among this vulnerable population. Given the known association of protein and fluid consumption with fall-related risk among older adults and the high prevalence of falls among Americans age 65 years and older each year, a brief intervention was created. Stepping Up Your Nutrition (SUYN) is a 2.5 h workshop developed through a public/private partnership to motivate older adults to reduce their malnutrition risk. The purposes of this naturalistic workshop dissemination were to: (1) describe the SUYN brief intervention; (2) identify participant characteristics associated with malnutrition risk; and (3) identify participant characteristics associated with subsequent participation in Stepping On (SO), an evidence-based fall prevention program. Data were analyzed from 429 SUYN participants, of which 38% (n = 163) subsequently attended SO. As measured by the SCREEN II®, high and moderate malnutrition risk scores were reported among approximately 71% and 20% of SUYN participants, respectively. Of the SUYN participants with high malnutrition risk, a significantly larger proportion attended a subsequent SO workshop (79.1%) compared to SUYN participants who did not proceed to SO (65.8%) (χ2 = 8.73, p = 0.013). Findings suggest SUYN may help to identify malnutrition risk among community-dwelling older adults and link them to needed services like evidence-based programs. Efforts are needed to expand the delivery infrastructure of SUYN to reach more at-risk older adults.

Species-Specific Outcome in the Competition for Nitrogen Between Invasive and Native Tree Seedlings.

The outcome of competition for nitrogen (N) between native and invasive tree species is a major concern when considering increasing anthropogenic N deposition. Our study investigated whether three native (i.e., Fagus sylvatica, Quercus robur, and Pinus sylvestris) and two invasive woody species (i.e., Prunus serotina and Robinia pseudoacacia) showed different responses regarding morphological and physiological parameters (i.e., biomass and growth indices, inorganic vs. organic N acquisition strategies, and N allocation to N pools) depending on the identity of the competing species, and whether these responses were mediated by soil N availability. In a greenhouse experiment, tree seedlings were planted either single or in native-invasive competition at low and high soil N availability. We measured inorganic and organic N acquisition using 15N labeling, total biomass, growth indices, as well as total soluble amino acid-N and protein-N levels in the leaves and fine roots of the seedlings. Our results indicate that invasive species have a competitive advantage via high growth rates, whereas native species could avoid competition with invasives via their higher organic N acquisition suggesting a better access to organic soil N sources. Moreover, native species responded to competition with distinct species- and parameter-specific strategies that were partly mediated by soil N availability. Native tree seedlings in general showed a stronger response to invasive P. serotina than R. pseudoacacia, and their strategies to cope with competition reflect the different species' life history strategies and physiological traits. Considering the responses of native and invasive species, our results suggest that specifically Q. robur seedlings have a competitive advantage over those of R. pseudoacacia but not P. serotina. Furthermore, native and invasive species show stronger responses to higher soil N availability under competition compared to when growing single. In conclusion, our study provides insights into the potential for niche differentiation between native and invasive species by using different N forms available in the soil, the combined effects of increased soil N availability and competition on tree seedling N nutrition, as well as the species-specific nature of competition between native and invasive tree seedlings which could be relevant for forest management strategies.

Responses to competition for nitrogen between subtropical native tree seedlings and exotic grasses are species-specific and mediated by soil N availability.

Competitive interactions between native tree seedlings and exotic grasses frequently hinder forest restoration. We investigated the consequences of competition with exotic grasses on the growth and net nitrogen (N) uptake capacity of native rainforest seedlings used for reforestation depending on soil N availability and N source. Tree seedlings and grasses were grown in the greenhouse in different competition regimes (one tree species vs one grass species) and controls (grass monocultures or single tree seedlings) at low and high soil N. After 8 weeks, we quantified net N uptake capacity using 15N-labelled organic (i.e., glutamine and arginine) and inorganic (i.e., ammonium and nitrate) N sources and biomass indices. Depending on soil N availability, we observed different species-specific responses to growth and N acquisition. Tree seedlings generally increased their net N uptake capacity in response to competition with grasses, although overall seedling growth was unaffected. In contrast, the responses to competition by the grasses were species-specific and varied with soil N availability. The different N acquisition strategies suggest the avoidance of competition for N between trees and grasses. Overall, the results highlight that quantifying underlying mechanisms of N acquisition complements the information on biomass allocation as a measure of responses to competition, particularly with varying environmental conditions.

Tannins and Their Complex Interaction with Different Organic Nitrogen Compounds and Enzymes: Old Paradigms versus Recent Advances.

Tannins, an abundant group of plant secondary compounds, raise interest in different fields of science, owing to their unique chemical characteristics. In chemical ecology, tannins play a crucial role in plant defense against pathogens, herbivores, and changing environmental conditions. In the food industry and in medicine, tannins are important because of their proven positive effect on human health and disease treatment. Such wide interests fueled studies on tannin chemistry, especially on their flagship ability to precipitate proteins. In this Review, we expand the basic knowledge on tannin chemistry to the newest insights from the field. We focus especially on tannin reactions with different non-protein organic N compounds, as well as the complex interactions of tannins with enzymes, resulting in either an increase or decrease in enzyme activity.

Defoliating Insect Mass Outbreak Affects Soil N Fluxes and Tree N Nutrition in Scots Pine Forests.

Biotic stress by mass outbreaks of defoliating pest insects does not only affect tree performance by reducing its photosynthetic capacity, but also changes N cycling in the soil of forest ecosystems. However, how insect induced defoliation affects soil N fluxes and, in turn, tree N nutrition is not well-studied. In the present study, we quantified N input and output fluxes via dry matter input, throughfall, and soil leachates. Furthermore, we investigated the effects of mass insect herbivory on tree N acquisition (i.e., organic and inorganic 15N net uptake capacity of fine roots) as well as N pools in fine roots and needles in a Scots pine (Pinus sylvestris L.) forest over an entire vegetation period. Plots were either infested by the nun moth (Lymantria monacha L.) or served as controls. Our results show an increased N input by insect feces, litter, and throughfall at the infested plots compared to controls, as well as increased leaching of nitrate. However, the additional N input into the soil did not increase, but reduce inorganic and organic net N uptake capacity of Scots pine roots. N pools in the fine roots and needles of infested trees showed an accumulation of total N, amino acid-N, protein-N, and structural N in the roots and the remaining needles as a compensatory response triggered by defoliation. Thus, although soil N availability was increased via surplus N input, trees did not respond with an increased N acquisition, but rather invested resources into defense by accumulation of amino acid-N and protein-N as a survival strategy.