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  1. Cory, Jenny (Ed.)
    Abstract Nitrogen (N) is a key nutrient required by all living organisms for growth and development, but is a limiting resource for many organisms. Organisms that feed on material with low N content, such as wood, might be particularly prone to N limitation. In this study, we investigated the degree to which the xylophagous larvae of the stag beetle Ceruchus piceus (Weber) use associations with N-fixing bacteria to acquire N. We paired acetylene reduction assays by cavity ring-down absorption spectroscopy (ARACAS) with 15N2 incubations to characterize rates of N fixation within C. piceus. Not only did we detect significant N fixation activity within C. piceus larvae, but we calculated a rate that was substantially higher than most previous reports for N fixation in insects. While taking these measurements, we discovered that N fixation within C. piceus can decline rapidly in a lab setting. Consequently, our results demonstrate that previous studies, which commonly keep insects in the lab for long periods of time prior to and during measurement, may have systematically under-reported rates of N fixation in insects. This suggests that within-insect N fixation may contribute more to insect nutrition and ecosystem-scale N budgets than previously thought. 
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    Free, publicly-accessible full text available July 7, 2024
  2. Abstract

    Nitrogen (N)‐fixing trees are thought to break a basic rule of leaf economics: higher leaf N concentrations do not translate into higher rates of carbon assimilation. Understanding how leaf N affects photosynthesis and water use efficiency (WUE) in this ecologically important group is critical.

    We grew six N‐fixing and four non‐fixing tree species for 4–5 years at four fertilization treatments in field experiments in temperate and tropical regions to assess how functional type (N fixer vs. non‐fixer) and N limitation affected leaf N and how leaf N affected light‐saturated photosynthesis (Asat), stomatal conductance (gsw) and WUE (WUEiand δ13C).

    Asat, WUEiand δ13C, but notgsw, increased with higher leaf N. Surprisingly, N‐fixing and non‐fixing trees displayed similar scaling between leaf N and these physiological variables, and this finding was supported by reanalysis of a global dataset. N fixers generally had higher leaf N than non‐fixers, even when non‐fixers were not N‐limited at the leaf level. Leaf‐level N limitation did not alter the relationship ofAsat,gsw, WUEiand δ13C with leaf N, although it did affect the photosynthetic N use efficiency. Higher WUE was associated with higher productivity, whereas higherAsatwas not.

    Synthesis: The ecological success of N‐fixing trees depends on the effect of leaf N on carbon gain and water loss. Using a field fertilization experiment and reanalysis of a global dataset, we show that high leaf‐level photosynthesis and WUE in N fixers stems from their higher average leaf N, rather than a difference between N fixers and non‐fixers in the scaling of photosynthesis and WUE with leaf N. By clarifying the mechanism by which N fixers achieve and benefit from high WUE, our results further the understanding of global N fixer distributions.

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  3. Abstract

    Symbiotic nitrogen fixation (SNF) is a key ecological process whose impact depends on the strategy of SNF regulation—the degree to which rates of SNF change in response to limitation by N versus other resources. SNF that is obligate or exhibits incomplete downregulation can result in excess N fixation, whereas a facultative SNF strategy does not. We hypothesized that tree‐based SNF strategies differed by latitude (tropical vs. temperate) and symbiotic type (actinorhizal vs. rhizobial). Specifically, we expected tropical rhizobial symbioses to display strongly facultative SNF as an explanation of their success in low‐latitude forests. In this study we used15N isotope dilution field experiments in New York, Oregon, and Hawaii to determine SNF strategies in six N‐fixing tree symbioses. Nitrogen fertilization with +10 and +15 g N m−2 year−1for 4–5 years alleviated N limitation in all taxa, paving the way to determine SNF strategies. Contrary to our hypothesis, all six of the symbioses we studied sustained SNF even at high N.Robinia pseudoacacia(temperate rhizobial) fixed 91% of its N (%Ndfa) in controls, compared to 64% and 59% in the +10 and +15 g N m−2 year−1treatments. ForAlnus rubra(temperate actinorhizal), %Ndfawas 95%, 70%, and 60%. For the tropical species, %Ndfawas 86%, 80%, and 82% forGliricidia sepium(rhizobial); 79%, 69%, and 67% forCasuarina equisetifolia(actinorhizal); 91%, 42%, and 67% forAcacia koa(rhizobial); and 60%, 51%, and 19% forMorella faya(actinorhizal). Fertilization with phosphorus did not stimulate tree growth or SNF. These results suggest that the latitudinal abundance distribution of N‐fixing trees is not caused by a shift in SNF strategy. They also help explain the excess N in many forests where N fixers are common.

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  4. Abstract

    Symbiotic nitrogen fixation (SNF) by higher plants and their bacterial symbionts is a globally important input of nitrogen. Our understanding of the mechanisms that control SNF and the time‐scales over which they operate has been constrained by the limitations of the existing methods for measuring SNF. One method, Acetylene Reduction Assays by Cavity ring‐down laser Absorption Spectroscopy (ARACAS), seems promising, as it is highly sensitive and gives rapid, continuous, repeatable and real‐time measurements of nitrogenase activity. ARACAS has been used to study nitrogen fixation in lichens, mosses and asymbiotic bacteria, but adapting it to higher plants poses challenges because acetylene and ethylene can influence plant function.

    Here, we report modifications to ARACAS that allow it to be used on higher plants in an environmentally controlled incubation chamber. The modifications include lower concentrations of acetylene (2%) and ethylene and concurrent measurements of whole‐chamber CO2exchange, H2O exchange and nitrogenase activity, linking nitrogenase activity to whole‐plant rates of photosynthesis and respiration.

    After propagating the error terms from all sources, we establish the following parameters of the method: (a) The detection limit of our method was 2–3 ppbv C2H4per hour, although it rose substantially when we used tank‐derived acetylene, which has much higher ethylene contamination; (b) Repeated measures at a frequency of 3 days or longer did not diminish nitrogenase activity or photosynthesis, although daily measurements diminished nitrogenase activity; (c) This method can detect changes at time‐scales as short as seconds; (d) Continuous measurement of nitrogenase activity is maintained above 90% of the maximum rate for 7.0 ± 1.3 (M ± SD) hours.

    This method has the potential to improve our understanding of the controls over SNF, and therefore, how SNF and global nitrogen and carbon cycling are likely to be affected by global change.

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