Differential physiological responses to environmental change promote woody shrub expansion

Direct and indirect effects of warming are increasingly modifying the carbon-rich vegetation and soils of the Arctic tundra, with important implications for the terrestrial carbon cycle. Understanding the biological and environmental influences on the pr

Differential physiological responses to environmental change promote woody shrub expansion

© The Author(s), 2013. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Ecology and Evolution 3 (2013): 1149–1162, doi:10.1002/ece3.525.Direct and indirect effects of warming are increasingly modifying the carbon-rich vegetation and soils of the Arctic tundra, with important implications for the terrestrial carbon cycle. Understanding the biological and environmental influences on the processes that regulate foliar carbon cycling in tundra species is essential for predicting the future terrestrial carbon balance in this region. To determine the effect of climate change impacts on gas exchange in tundra, we quantified foliar photosynthesis (Anet), respiration in the dark and light (RD and RL, determined using the Kok method), photorespiration (PR), carbon gain efficiency (CGE, the ratio of photosynthetic CO2 uptake to total CO2 exchange of photosynthesis, PR, and respiration), and leaf traits of three dominant species – Betula nana, a woody shrub; Eriophorum vaginatum, a graminoid; and Rubus chamaemorus, a forb – grown under long-term warming and fertilization treatments since 1989 at Toolik Lake, Alaska. Under warming, B. nana exhibited the highest rates of Anet and strongest light inhibition of respiration, increasing CGE nearly 50% compared with leaves grown in ambient conditions, which corresponded to a 52% increase in relative abundance. Gas exchange did not shift under fertilization in B. nana despite increases in leaf N and P and near-complete dominance at the community scale, suggesting a morphological rather than physiological response. Rubus chamaemorus, exhibited minimal shifts in foliar gas exchange, and responded similarly to B. nana under treatment conditions. By contrast, E. vaginatum, did not significantly alter its gas exchange physiology under treatments and exhibited dramatic decreases in relative cover (warming: −19.7%; fertilization: −79.7%; warming with fertilization: −91.1%). Our findings suggest a foliar physiological advantage in the woody shrub B. nana that is further mediated by warming and increased soil nutrient availability, which may facilitate shrub expansion and in turn alter the terrestrial carbon cycle in future tundra environments.This study was supported by the National Science Foundation #0732664; Australian Research Council DP0986823; and Marsden Fund of the Royal Society of New Zealand

Ion antiport accelerates photosynthetic acclimation in fluctuating light environments

Many photosynthetic organisms globally, including crops, forests and algae, must grow in environments where the availability of light energy fluctuates dramatically. How photosynthesis maintains high efficiency despite such fluctuations in its energy source remains poorly understood. Here we show that Arabidopsis thaliana ​K+ efflux antiporter (​KEA3) is critical for high photosynthetic efficiency under fluctuating light. On a shift from dark to low light, or high to low light, ​kea3 mutants show prolonged dissipation of absorbed light energy as heat. ​KEA3 localizes to the thylakoid membrane, and allows proton efflux from the thylakoid lumen by proton/potassium antiport. ​KEA3’s activity accelerates the downregulation of pH-dependent energy dissipation after transitions to low light, leading to faster recovery of high photosystem II quantum efficiency and increased ​CO2 assimilation. Our results reveal a mechanism that increases the efficiency of photosynthesis under fluctuating light. [EN]This project was funded by the Carnegie Institution for Science, by ERDF-cofinanced grants from the Ministry of Economy and Competitiveness (BIO2012-33655) and Junta de Andalucia (CVI-7558) to K.V., the Natural Sciences and Engineering Research Council of Canada (NSERC) PGS-D3 scholarship to L.P. and Deutsche Forschungsgemeinschaft grants (JA 665/10-1 and GRK 1525 to P.J.; AR 808/1-1 to U.A.).Peer reviewe

Connecting active to passive fluorescence with photosynthesis: a method for evaluating remote sensing measurements of Chl fluorescence

Recent advances in the retrieval of Chl fluorescence from space using passive methods (solar-induced Chl fluorescence, SIF) promise improved mapping of plant photosynthesis globally. However, unresolved issues related to the spatial, spectral, and temporal dynamics of vegetation fluorescence complicate our ability to interpret SIF measurements. We developed an instrument to measure leaf-level gas exchange simultaneously with pulse-amplitude modulation (PAM) and spectrally resolved fluorescence over the same field of view – allowing us to investigate the relationships between active and passive fluorescence with photosynthesis. Strongly correlated, slope-dependent relationships were observed between measured spectra across all wavelengths (Fλ, 670–850 nm) and PAM fluorescence parameters under a range of actinic light intensities (steady-state fluorescence yields, Ft) and saturation pulses (maximal fluorescence yields, Fm). Our results suggest that this method can accurately reproduce the full Chl emission spectra – capturing the spectral dynamics associated with changes in the yields of fluorescence, photochemical (ΦPSII), and nonphotochemical quenching (NPQ). We discuss how this method may establish a link between photosynthetic capacity and the mechanistic drivers of wavelength-specific fluorescence emission during changes in environmental conditions (light, temperature, humidity). Our emphasis is on future research directions linking spectral fluorescence to photosynthesis, ΦPSII, and NPQ

Rab7 Associates with Early Endosomes to Mediate Sorting and Transport of Semliki Forest Virus to Late Endosomes

Semliki forest virus (SFV) is internalized by clathrin-mediated endocytosis, and transported via early endosomes to late endosomes and lysosomes. The intracellular pathway taken by individual fluorescently labeled SFV particles was followed using immunofluorescence in untransfected cells, and by video-enhanced, triple-color fluorescence microscopy in live cells transfected with GFP- and RFP-tagged Rab5, Rab7, Rab4, and Arf1. The viruses progressed from Rab5-positive early endosomes to a population of early endosomes (about 10% of total) that contained both Rab5 and Rab7. SFV were sequestered in the Rab7 domains, and they were sorted away from the early endosomes when these domains detached as separate transport carriers devoid of Rab5, Rab4, EEA1, Arf1, and transferrin. The process was independent of Arf1 and the acidic pH in early endosomes. Nocodazole treatment showed that the release of transport carriers was assisted by microtubules. Expression of constitutively inactive Rab7T22N resulted in accumulation of SFV in early endosomes. We concluded that Rab7 is recruited to early endosomes, where it forms distinct domains that mediate cargo sorting as well as the formation of late-endosome-targeted transport vesicles

The Role of Alternative Oxidase (AOX) in Plant Stress: do Plants Increase the Activity of AOX in Response to Nutrient Stress Under Field Conditions?

RATIONALE: Recent studies indicate that plants can partition electron transport through alternative oxidase (AOX) and cytochrome c oxidase (COX) in response to environmental cues, thus modulating respiratory efficiency. The ¹⁸O discrimination method necessary for measuring electron partitioning in vivo, however, has been restricted to laboratory settings due to equipment constraints. Since plants grown in more natural and variable environments may not respond as predicted by laboratory experiments, I developed a new field-compatible analytical method and then applied it to three ecophysiological studies. METHODS: To address these needs, I developed a field-compatible method in which plant tissue was incubated in 12 mL septum-capped vials for 0.5 – 3 h, after which the incubation air was transferred to 3.7 mL storage vials for subsequent measurement by mass spectrometry. I also developed mathematical tools to correct for unavoidable contamination, and to detect and address curvature in the data – whether intrinsic to the respiration or due to contamination, – and to extend the usable dynamic range of the mass spectrometer. These methods were used to investigate respiratory responses (1) in canopy trees growing across a soil nutrient gradient at the Franz Josef chronosequence, New Zealand; (2) in a nutrient manipulation experiment of Griselinea littoralis; and (3) in a long-term nutrient-, temperature-, and light manipulation at Toolik, Alaska, USA. Leaf dry matter content, specific leaf area, nitrogen (N), phosphorus (P), sugars, starch, and AOX/COX protein concentrations were also measured as explanatory variables. (Leaf Cu and Fe were measured at the Franz Josef chronosequence.) RESULTS: Discrimination values computed using my methods replicated previously reported results over a range of 10 – 31‰, with precision generally better than ±0.5‰, thus demonstrating its validity as tool for measuring respiratory electron partitioning. Foliar respiration declined with site age across the soil chronosequence, increasing with leaf N levels, r² = 0.8, but electron partitioning declined with increasing N/P, r² = 0.23. AP activity was positively correlated with leaf P, Cu, and starch, r² = 0.71. In younger soils, however, declines in respiration were attributed to declines in cytochrome pathway (CP) activity, whereas across the older sites respiration declined due to a reduction in AOX pathway (AP) activity. The Griselinia nutrient-manipulation experiment partially confirmed these results: AOX protein levels were highest in N-deficient plants rather than in plants deficient in both N and P. AP activity was very low in all leaves, however, possibly due to low illumination. In support of this claim, leaves that had developed in the sun had higher AOX/COX protein ratios than those that had developed in the greenhouse. In Griselinia roots, CP activity declined by more than half in response to nutrient deficiency, whereas AP activity was unchanged. At the Arctic site, only one species changed electron partitioning in response to nutrient addition. Betula nana, the most successful adapter to improved mineral nutrition, doubled leaf CP activity without changing AP activity. Species grown in full sun at that site also had higher AOX/COX protein ratios than those that grew in enclosures. CONCLUSIONS: This is the first study of how engagement of terminal respiratory oxidases in plants responds to multiple nutrient deficiencies, both in nature and in a controlled environment. I have uncovered some intriguing relationships, including the possible importance of N/P to electron partitioning, as well as a role for Cu. The results also suggest that electron partitioning is sensitive to plant energy balance, as suggested by the low AP activities and low AOX/COX protein ratios in shaded plants. Perhaps most significantly, the AP and CP appeared to act independently of each other, rather than through a concerted “partitioning” process. In addition to their own scientific merit, these results illustrate the value of using the new field-compatible method to conduct ecophysiological investigations of plant respiratory electron partitioning on a much large scale, and under more realistic conditions, than has been previously possible

Modulation of respiratory metabolism in response to nutrient changes along a soil chronosequence

Laboratory studies indicate that plant respiratory efficiency may decrease in response to low nutrient availability due to increased partitioning of electrons to the energy-wasteful alternative oxidase (AOX); however, field confirmation of this hypothesisWe are especially grateful to the Department of Conservation at Franz Josef for providing access to the sites.This work was supported by a grant from the Marsden Fund of the Royal Society of New Zealand as well as scholarship grants for A.K. from Education New Zealand and the University of Canterbury, with additional partial funding from the Natural Environment Research Council (NERC) in the UK (NE/D01168X/1 and NE/F002149/1) and the Australian Research Council (ARC FT0991448 and DP0986823)

Respiratory flexibility and efficiency are affected by simulated global change in Arctic plants

Laboratory studies indicate that, in response to environmental conditions, plants modulate respiratory electron partitioning between the 'energy-wasteful' alternative pathway (AP) and the 'energy-conserving' cytochrome pathway (CP). Field data, however, are scarce. Here we investigate how 20-yr field manipulations simulating global change affected electron partitioning in Alaskan Arctic tundra species. We sampled leaves from three dominant tundra species - Betula nana, Eriophorum vaginatum and Rubus chamaemorus - that had been strongly affected by manipulations of soil nutrients, light availability, and warming. We measured foliar dark respiration, in-vivo electron partitioning and alternative oxidase/cytochrome c oxidase concentrations in addition to leaf traits and mitochondrial ultrastructure. Changes in leaf traits and ultrastructure were similar across species. Respiration at 20°C (R20) was reduced 15% in all three species grown at elevated temperature, suggesting thermal acclimation of respiration. In Betula, the species with the largest growth response to added nutrients, CP activity increased from 9.4 ± 0.8 to 16.6 ± 1.6 nmol O2 g-1 DM s-1 whereas AP activity was unchanged. The ability of Betula to selectively increase CP activity in response to the environment may contribute to its overall ecological success by increasing respiratory energy efficiency, and thus retaining more carbon for growth

Plant-mediated partner discrimination in ectomycorrhizal mutualisms

Although ectomycorrhizal fungi have well-recognized effects on ecological processes ranging from plant community dynamics to carbon cycling rates, it is unclear if plants are able to actively influence the structure of these fungal communities. To address this knowledge gap, we performed two complementary experiments to determine (1) whether ectomycorrhizal plants can discriminate among potential fungal partners, and (2) to what extent the plants might reward better mutualists. In experiment 1, split-root Larix occidentalis seedlings were inoculated with spores from three Suillus species (S. clintonianus, S. grisellus, and S. spectabilis). In experiment 2, we manipulated the symbiotic quality of Suillus brevipes isolates on split-root Pinus muricata seedlings by changing the nitrogen resources available, and used carbon-13 labeling to track host investment in fungi. In experiment 1, we found that hosts can discriminate in multi-species settings. The split-root seedlings inhibited colonization by S. spectabilis whenever another fungus was available, despite similar benefits from all three fungi. In experiment 2, we found that roots and fungi with greater nitrogen supplies received more plant carbon. Our results suggest that plants may be able to regulate this symbiosis at a relatively fine scale, and that this regulation can be integrated across spatially separated portions of a root system

A field-compatible method for measuring alternative respiratory pathway activities in vivo using stable O2 isotopes

Plants can alter rates of electron transport through the alternative oxidase (AOX) pathway in response to environmental cues, thus modulating respiratory efficiency, but the 18O discrimination method necessary for measuring electron partitioning in vivo has been restricted to laboratory settings. To overcome this limitation, we developed a field-compatible analytical method. Series of plant tissue subsamples were incubated in 12 mL septum-capped vials for 0.5-4 h before aliquots of incubation air were injected into 3.7 mL evacuated storage vials. Vials were stored for up to 10 months before analysis by mass spectrometry. Measurements were corrected for unavoidable contamination. Additional mathematical tools were developed for detecting and addressing non-linearity (whether intrinsic or due to contamination) in the data used to estimate discrimination values. Initial contamination in the storage vials was 0.03 ± 0.01 atm; storing the gas samples at -17 °C eliminated further contamination effects over 10 months. Discrimination values obtained using our offline incubation and computation method replicated previously reported results over a range of 10-31‰, with precision generally better than ±0.5‰. Our method enables large-scale investigations of plant alternative respiration along natural environmental gradients under field conditions