Alternative Electron Sinks Research Articles

Alternative electron sinks in chloroplasts and mitochondria of halophytes as a safety valve for controlling ROS production during salinity.

Electron flow through the electron transport chain (ETC) is essential for oxidative phosphorylation in mitochondria and photosynthesis in chloroplasts. Electron fluxes depend on environmental parameters, e.g., ionic and osmotic conditions and endogenous factors, and this may cause severe imbalances. Plants have evolved alternative sinks to balance the reductive load on the electron transport chains in order to avoid overreduction, generation of reactive oxygen species (ROS), and to cope with environmental stresses. These sinks act primarily as valves for electron drainage and secondarily as regulators of tolerance-related metabolism, utilizing the excess reductive energy. High salinity is an environmental stressor that stimulates the generation of ROS and oxidative stress, which affects growth and development by disrupting the redox homeostasis of plants. While glycophytic plants are sensitive to high salinity, halophytic plants tolerate, grow, and reproduce at high salinity. Various studies have examined the ETC systems of glycophytic plants, however, information about the state and regulation of ETCs in halophytes under non-saline and saline conditions is scarce. This review focuses on alternative electron sinks in chloroplasts and mitochondria of halophytic plants. In cases where information on halophytes is lacking, we examined the available knowledge on the relationship between alternative sinks and gradual salinity resilience of glycophytes. To this end, transcriptional responses of involved components of photosynthetic and respiratory ETCs were compared between the glycophyte Arabidopsis thaliana and the halophyte Schrenkiella parvula, and the time-courses of these transcripts were examined in A. thaliana. The observed regulatory patterns are discussed in the context of reactive molecular species formation in halophytes and glycophytes.

Anthocyanins act as a sugar-buffer and an alternative electron sink in response to starch depletion during leaf senescence: a case study on a typical anthocyanic tree species, Acer japonicum.

We hypothesized that anthocyanins act as a sugar-buffer and an alternative electron sink during leaf senescence to prevent sugar-mediated early senescence and photoinhibition. To elucidate the anthocyanin role, we monitored seasonal changes in photosynthetic traits, sugar, starch and N contents, pigment composition, and gene expression profiles in leaves exposed to substantially different light conditions within a canopy of an adult tree of fullmoon maple (Acer japonicum). Enhancement of starch amylolysis accompanied with cessation of starch synthesis occurred in the same manner independent of light conditions. Leaf sugar contents increased, but reached upper limits in the late stage of leaf senescence, even though leaf anthocyanins further increased after complete depletion of starch. Sun-exposed leaves maintained higher energy consumption via electron flow than shade-grown leaves during leaf N resorption. Thus, anthocyanins accumulated in sun-exposed leaves might have a regulative role as a sugar buffer, retarding leaf senescence, and an indirect photoprotective role as an alternative sink for electron consumption to compensate declines in other metabolic processes such as starch and protein synthesis. In this context, anthocyanins might be key substrates protecting both outer-canopy leaves (against photoinhibition) and inner-canopy leaves (via shading by outer-canopy leaves) from high light stress during N resorption.

Extra O2 evolution reveals an O2-independent alternative electron sink in photosynthesis of marine diatoms.

Following the principle of oxygenic photosynthesis, electron transport in the thylakoid membranes (i.e., light reaction) generates ATP and NADPH from light energy, which is subsequently utilized for CO2 fixation in the Calvin-Benson-Bassham cycle (i.e., dark reaction). However, light and dark reactions could discord when an alternative electron flow occurs with a rate comparable to the linear electron flow. Here, we quantitatively monitored O2 and total dissolved inorganic carbon (DIC) during photosynthesis in the pennate diatom Phaeodactylum tricornutum, and found that evolved O2 was larger than the consumption of DIC, which was consistent with 14CO2 measurements in literature. In our measurements, the stoichiometry of O2 evolution to DIC consumption was always around 1.5 during photosynthesis at different DIC concentrations. The same stoichiometry was observed in the cells grown under different CO2 concentrations and nitrogen sources except for the nitrogen-starved cells showing O2 evolution 2.5 times larger than DIC consumption. An inhibitor to nitrogen assimilation did not affect the extra O2 evolution. Further, the same physiological phenomenon was observed in the centric diatom Thalassiosira pseudonana. Based on the present dataset, we propose that the marine diatoms possess the metabolic pathway(s) functioning as the O2-independent electron sink under steady state photosynthesis that reaches nearly half of electron flux of the Calvin-Benson-Bassham cycle.

Photobioelectrocatalysis of Intact Photosynthetic Bacteria Exposed to Dinitrophenol

AbstractThe outstanding metabolic versatility of purple non‐sulphur bacteria makes these organisms an ideal candidate for developing photobioelectrochemical systems applicable in contaminated environments. Here, the effects of 2,4 dinitrophenol, a common contaminant, on purple bacteria photobioelectrocatalysis were investigated. The aromatic contaminant clearly affects current generation, with an enhanced photocurrent obtained at low dinitrophenol concentrations (0.5–1 μM), while higher values (up to 100 μM) resulted in a gradual decrease of photocurrent. The obtained electrochemical evidence, coupled to spectroscopic studies, allowed verifying the viability of the bacteria cells after exposure to dinitrophenol, and that no alteration of the photosynthetic apparatus was obtained. The results indicate that high dinitrophenol concentrations divert electrons from the extracellular electron pathway to an alternative electron sink. The present results open the door to the possible use of intact bacteria‐based photoelectrodes to develop technologies for sustainable biosensors with simultaneous environmental remediation.

Carbon and Nitrogen Allocation between the Sink and Source Leaf Tissue in Response to the Excess Excitation Energy Conditions.

Plants are inevitably exposed to extreme climatic conditions that lead to a disturbed balance between the amount of absorbed energy and their ability to process it. Variegated leaves with photosynthetically active green leaf tissue (GL) and photosynthetically inactive white leaf tissue (WL) are an excellent model system to study source-sink interactions within the same leaf under the same microenvironmental conditions. We demonstrated that under excess excitation energy (EEE) conditions (high irradiance and lower temperature), regulated metabolic reprogramming in both leaf tissues allowed an increased consumption of reducing equivalents, as evidenced by preserved maximum efficiency of photosystem II (ФPSII) at the end of the experiment. GL of the EEE-treated plants employed two strategies: (i) the accumulation of flavonoid glycosides, especially cyanidin glycosides, as an alternative electron sink, and (ii) cell wall stiffening by cellulose, pectin, and lignin accumulation. On the other hand, WL increased the amount of free amino acids, mainly arginine, asparagine, branched-chain and aromatic amino acids, as well as kaempferol and quercetin glycosides. Thus, WL acts as an important energy escape valve that is required in order to maintain the successful performance of the GL sectors under EEE conditions. Finally, this role could be an adaptive value of variegation, as no consistent conclusions about its ecological benefits have been proposed so far.

Symposium review: Effective nutritional strategies to mitigate enteric methane in dairy cattle.

Intensive research in the past decade has resulted in a better understanding of factors driving enteric methane (CH4) emissions in ruminants. Meta-analyses of large databases, developed through the GLOBAL NETWORK project, have identified successful strategies for mitigation of CH4 emissions. Methane inhibitors, alternative electron sinks, vegetable oils and oilseeds, and tanniferous forages are among the recommended strategies for mitigating CH4 emissions from dairy and beef cattle and small ruminants. These strategies were also effective in decreasing CH4 emissions yield and intensity. However, a higher inclusion rate of oils may negatively affect feed intake, rumen function, and animal performance, specifically milk components in dairy cows. In the case of nitrates (electron sinks), concerns with animal health may be impeding their adoption in practice, and potential emission trade-offs have to be considered. Tannins and tanniferous forages may have a negative effect on nutrient digestibility, and more research is needed to confirm their effects on overall animal performance in long-term experiments with high-producing animals. A meta-analysis of studies with dairy cows fed the CH4 inhibitor 3-nitrooxypropanol (3-NOP) at the Pennsylvania State University showed (1) a consistent 28 to 32% decrease in daily CH4 emissions or emissions yield and intensity; (2) no effect on dry matter intake, milk production, body weight, or body weight change, and a slight increase in milk fat concentration and yield (0.19 percentage units and 90 g/d, respectively); 3-NOP also appears to increase milk urea nitrogen concentration; (3) an exponential decrease in the mitigation effect of the inhibitor with increasing its dose (from 40 to 200 mg/kg of feed dry matter, corresponding to 3-NOP intake of 1 to 4.8 g/cow per day); and (4) a potential decrease in the efficacy of 3-NOP over time, which needs to be further investigated in long-term, full-lactation or multiple-lactation studies. The red macroalga Asparagopsis taxiformis has a strong CH4 mitigation effect, but studies are needed to determine its feasibility, long-term efficacy, and effects on animal production and health. We concluded that widespread adoption of mitigation strategies with proven effectiveness by the livestock industries will depend on cost, government policies and incentives, and willingness of consumers to pay a higher price for animal products with decreased carbon footprint.

The Dynamic Changes of Alternative Electron Flows upon Transition from Low to High Light in the Fern Cyrtomium fortune and the Gymnosperm Nageia nagi

In photosynthetic organisms except angiosperms, an alternative electron sink that is mediated by flavodiiron proteins (FLVs) plays the major role in preventing PSI photoinhibition while cyclic electron flow (CEF) is also essential for normal growth under fluctuating light. However, the dynamic changes of FLVs and CEF has not yet been well clarified. In this study, we measured the P700 signal, chlorophyll fluorescence, and electrochromic shift spectra in the fern Cyrtomium fortune and the gymnosperm Nageia nagi. We found that both species could not build up a sufficient proton gradient (∆pH) within the first 30 s after light abruptly increased. During this period, FLVs-dependent alternative electron flow was functional to avoid PSI over-reduction. This functional time of FLVs was much longer than previously thought. By comparison, CEF was highly activated within the first 10 s after transition from low to high light, which favored energy balancing rather than the regulation of a PSI redox state. When FLVs were inactivated during steady-state photosynthesis, CEF was re-activated to favor photoprotection and to sustain photosynthesis. These results provide new insight into how FLVs and CEF interact to regulate photosynthesis in non-angiosperms.

Exogenous melatonin strongly affects dynamic photosynthesis and enhances water-water cycle in tobacco.

Melatonin (MT), an important phytohormone synthesized naturally, was recently used to improve plant resistance against abiotic and biotic stresses. However, the effects of exogenous melatonin on photosynthetic performances have not yet been well clarified. We found that spraying of exogenous melatonin (100 μM) to leaves slightly affected the steady state values of CO2 assimilation rate (AN), stomatal conductance (gs) and mesophyll conductance (gm) under high light in tobacco leaves. However, this exogenous melatonin strongly delayed the induction kinetics of gs and gm, leading to the slower induction speed of AN. During photosynthetic induction, AN is mainly limited by biochemistry in the absence of exogenous melatonin, but by CO2 diffusion conductance in the presence of exogenous melatonin. Therefore, exogenous melatonin can aggravate photosynthetic carbon loss during photosynthetic induction and should be used with care for crop plants grown under natural fluctuating light. Within the first 10 min after transition from low to high light, photosynthetic electron transport rates (ETR) for AN and photorespiration were suppressed in the presence of exogenous melatonin. Meanwhile, an important alternative electron sink, namely water-water cycle, was enhanced to dissipate excess light energy. These results indicate that exogenous melatonin upregulates water-water cycle to facilitate photoprotection. Taking together, this study is the first to demonstrate that exogenous melatonin inhibits dynamic photosynthesis and improves photoprotection in higher plants.

One substrate, many fates: different ways of methanol utilization in the acetogen Acetobacterium woodii.

Acetogenic bacteria such as Acetobacterium woodii use the Wood-Ljungdahl pathway (WLP) for fixation of CO2 and energy conservation. This pathway enables conversion of diverse substrates to the main product of acetogenesis, acetate. Methyl group containing substrates such as methanol or methylated compounds, derived from pectin, are abundant in the environment and a source for CO2 . Methyl groups enter the WLP at the level of methyltetrahydrofolic acid (methyl-THF). For methyl transfer from methanol to THF a substrate-specific methyltransferase system is required. In this study, we used genetic methods to identify mtaBC2A (Awo_c22760-Awo_c22740) as the methanol-specific methyltransferase system of A. woodii. After methyl transfer, methyl-THF serves as carbon and/or electron source and the respiratory Rnf complex is required for redox homeostasis if methanol+CO2 is the substrate. Resting cells fed with methanol+CO2 , indeed converted methanol to acetate in a 4:3 stoichiometry. When methanol was fed in combination with other electron sources such as H2 +CO2 or CO, methanol was converted Rnf-independently and the methyl group was condensed with CO to build acetate. When fed in combination with alternative electron sinks such as caffeate methanol was oxidized only and resulting electrons were used for non-acetogenic growth. These different pathways for the conversion of methyl-group containing substrates enable acetogens to adapt to various ecological niches and to syntrophic communities.

Involvement of poly(ADP-ribose)ylation (PARylation) in the regulation of antioxidant defense system in Arabidopsis thaliana under salt stress

Posttranslational modifications (PTM) are one of the first responses of plants to environmental stress and involve changing the location and activity of proteins in the cell. Addition of poly(ADP-ribose) (PAR) to proteins, poly(ADP-ribose)ylation (PARylation), is a posttranslational modification resulting from the binding of ADP-ribose from NAD+ to target proteins by PAR polymerases (PARP). PARylation is involved in many physiological events in plants including abiotic and biotic stress response. The aim of this work was to understand involvement of PARylation in inducing enzymatic antioxidant defence and alternative electron sinks in response to salinity. For this purpose A. thaliana plants were treated with 3-aminobenzamide (3-AB), which is a PARP inhibitor, in the presence of 100 mM NaCl. The 3-AB treatment induced plant fresh weight under control and salinity conditions. Moreover, 100 mM NaCl + 3-AB treated plants had lower lipid peroxidation when compared to 100 mM NaCl group indicating mitigation of oxidative stress. This oxidative stress mitigation was achieved by significantly induced superoxide dismutase (SOD) activity and transcriptional activation of genes related to ROS scavenging such as MSD1, CAT1, APX1, GR1. On the other hand, transcriptional regulation of mitochondrial alternative oxidase (AOX) pathway was also induced with 3-AB treatment (AOX1a and AOX1d) under salt stress indicating that ROS avoidance mechanisms are also activated along with ROS scavenging. However, in contrast to AOX, chloroplastic plastid terminal oxidase pathway was not induced with 3-AB.

Physiological Roles of Flavodiiron Proteins and Photorespiration in the Liverwort Marchantia polymorpha.

Against the potential risk in oxygenic photosynthesis, that is, the generation of reactive oxygen species, photosynthetic electron transport needs to be regulated in response to environmental fluctuations. One of the most important regulations is keeping the reaction center chlorophyll (P700) of photosystem I in its oxidized form in excess light conditions. The oxidation of P700 is supported by dissipating excess electrons safely to O2, and we previously found that the molecular mechanism of the alternative electron sink is changed from flavodiiron proteins (FLV) to photorespiration in the evolutionary history from cyanobacteria to plants. However, the overall picture of the regulation of photosynthetic electron transport is still not clear in bryophytes, the evolutionary intermediates. Here, we investigated the physiological roles of FLV and photorespiration for P700 oxidation in the liverwort Marchantia polymorpha by using the mutants deficient in FLV (flv1) at different O2 partial pressures. The effective quantum yield of photosystem II significantly decreased at 2kPa O2 in flv1, indicating that photorespiration functions as the electron sink. Nevertheless, it was clear from the phenotype of flv1 that FLV was dominant for P700 oxidation in M. polymorpha. These data suggested that photorespiration has yet not replaced FLV in functioning for P700 oxidation in the basal land plant probably because of the lower contribution to lumen acidification, compared with FLV, as reflected in the results of electrochromic shift analysis.

Photosynthetic Linear Electron Flow Drives CO2 Assimilation in Maize Leaves.

Photosynthetic organisms commonly develop the strategy to keep the reaction center chlorophyll of photosystem I, P700, oxidized for preventing the generation of reactive oxygen species in excess light conditions. In photosynthesis of C4 plants, CO2 concentration is kept at higher levels around ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) by the cooperation of the mesophyll and bundle sheath cells, which enables them to assimilate CO2 at higher rates to survive under drought stress. However, the regulatory mechanism of photosynthetic electron transport for P700 oxidation is still poorly understood in C4 plants. Here, we assessed gas exchange, chlorophyll fluorescence, electrochromic shift, and near infrared absorbance in intact leaves of maize (a NADP-malic enzyme C4 subtype species) in comparison with mustard, a C3 plant. Instead of the alternative electron sink due to photorespiration in the C3 plant, photosynthetic linear electron flow was strongly suppressed between photosystems I and II, dependent on the difference of proton concentration across the thylakoid membrane (ΔpH) in response to the suppression of CO2 assimilation in maize. Linear relationships among CO2 assimilation rate, linear electron flow, P700 oxidation, ΔpH, and the oxidation rate of ferredoxin suggested that the increase of ΔpH for P700 oxidation was caused by the regulation of proton conductance of chloroplast ATP synthase but not by promoting cyclic electron flow. At the scale of intact leaves, the ratio of PSI to PSII was estimated almost 1:1 in both C3 and C4 plants. Overall, the photosynthetic electron transport was regulated for P700 oxidation in maize through the same strategies as in C3 plants only except for the capacity of photorespiration despite the structural and metabolic differences in photosynthesis between C3 and C4 plants.

Melatonin mitigates UV-B stress via regulating oxidative stress response, cellular redox and alternative electron sinks in Arabidopsis thaliana

Melatonin plays an active role in neutralizing free radicals, especially by triggering the defense system and certain enzymes that work under stress in both mammals and plant systems. Exposure to ultraviolet (UV–B) stress can be deadly for plants since UV-B can induce production of reactive oxygen species and damage nucleic acids. In the present study, to uncover the possible alleviative role of melatonin against UV-B stress, Arabidopsis thaliana plants were treated with melatonin (10 μM) and were exposed to UV-B stress for 90 min and 180 min (46 and 92 kJ m−2 d−1). Plants treated with melatonin had lower lipid peroxidation levels and higher Fv/Fm values at both time points. UV-B stress-induced activities of superoxide dismutase (SOD), glutathione reductase (GR) and ascorbate peroxidase (APX), but no additional induction was observed in melatonin treated groups. Moreover, melatonin differentially regulated the expression of glutathione peroxidase 2 (GPX2) and GPX7 genes under UV-B stress. Melatonin treatment did not have any effect on glutathione biosynthesis and catabolism genes. However, expression of alternative oxidase 1a (AOX1a) and AOX1d were lower in UV-B + melatonin treated plants when compared to only UV-B treated plants, which indicates lower oxidative load in mitochondria.

Alkalinity, and Not the Oxidation State of the Organic Substrate, Is the Key Factor in Domestic Wastewater Treatment by Mixed Cultures of Purple Phototrophic Bacteria

Domestic wastewater treatment by purple phototrophic bacteria (PPB) is based on the assimilative uptake of organics and nutrients into the bacterial biomass. Thereby, it strongly depends on the carbon/nutrients ratio of the wastewater. The physiological COD/N/P ratio for PPB growth in domestic wastewater makes the addition of an external organic carbon source necessary in order to allow for an efficient process. However, PPB need a source of alkalinity (as CO2) to grow on reduced organics that serves as an electron acceptor since biohydrogen production (an alternative electron sink) is inhibited by ammonium. A preliminary experiment showed that high nutrients-loading wastewater was limited by CO2 imbalance, leading to poor removal efficiencies. Subsequently, the effect of the oxidation state of the organics added as external organic carbon sources to PPB reactors treating low nutrients-loading domestic wastewater has been analyzed. Three organics were used as additives to PPB development in four consecutive batches: acetate (more oxidized), ethanol and butyrate (more reduced). The PPB population was settled and the general performance under the three situations, in terms of organics, N and P assimilation, and growth kinetics was not significantly different irrespective of the external organic carbon source. The reactors were dominated by PPB, though reduced organics allowed for dominance of Rhodopseudomonas palustris, whereas oxidized organics caused co-dominance of R. palustris and Rhodobacter capsulatus. Thereby, alkalinity (as bicarbonate), and not the oxidation state of the organics, is the key parameter for the efficient treatment of domestic wastewater by PPB.

Chlorophyll-depleted wheat mutants are disturbed in photosynthetic electron flow regulation but can retain an acclimation ability to a fluctuating light regime

The search for more productive crops is exploring accessions with reduced chlorophyll accumulation in leaves, which would promote plant growth mainly because of a higher light transmittance throughout the canopy. Under continuous light, chlorophyll-depleted chlorina mutants of wheat can reach yields similar to WT; however, their performance under fluctuating light could possibly be lowered by a disturbed photosynthetic electron transport.Six chlorina mutants of Triticum aestivum (ANBW4A, ANBW4B, ANK32A) or T. durum (ANDW7A, ANDW7B, ANDW8A) were compared to WT genotypes under continuous and fluctuating light regimes, the latter obtained through a randomized pattern of light intensity changes. After two weeks of plant acclimation under either regimes, light energy management was thoroughly analysed for four weeks.All chlorina mutants showed a defective ability to regulate the electron poise during a fast rise in irradiance. Different extent of deregulation depended on mutation and genomic background, durum wheat being more severely impaired than bread wheat. However, a great acclimative capacity to a fluctuating light regime unexpectedly still characterized the chlorina wheat plants, with the only exception of the mutants carrying the cn-B1b mutated locus (ANDW7B and ANBW4B). Under fluctuating light, all other chlorina mutants developed indeed an ability to improve their control of electron transport against the recurrent reducing bursts caused by lightflecks. Compensatory responses included the regulation of energy distribution between photosystems, the use of alternative electron sinks, and a structural reorganization of the thylakoid system. However, in no case a reduced chlorophyll content led to more productive plants. Among 23 measured photosynthetic parameters, especially the non-regulatory energy dissipation in photosystem II - Y(NO) –, when probed during a light rise, can be proposed in automated phenotyping experiments as a straightforward index to be used for chlorina wheat screening with respect to potential productivity.

Chlororespiration Serves as Photoprotection for the Photo-Inactivated Oxygen-Evolving Complex in Zostera marina, a Marine Angiosperm.

As an alternative electron sink, chlororespiration, comprising the NAD(P)H dehydrogenase complex and plastid terminal plastoquinone oxidase, may play a significant role in sustaining the redox equilibrium between stroma and thylakoid membrane. This study identified a distinct role for chlororespiration in the marine angiosperm Zostera marina, whose oxygen-evolving complex (OEC) is prone to photo-inactivation as a result of its inherent susceptibility to excess irradiation. The strong connectivity between OEC peripheral proteins and key chlororespiratory enzymes, as demonstrated in the interaction network of differentially expressed genes, suggested that the recovery of photo-inactivated OEC was connected with chlororespiration. Chlorophyll fluorescence, transcriptome and Western blot data verified a new physiological role for chlororespiration to function as photoprotection and generate a proton gradient across the thylakoid membrane for the recovery of photo-inactivated OEC. Chlororespiration was only activated in darkness following excess irradiation exposure, which might be related to electron deficiency in the electron transport chain because of the continuous impairment of the OEC. The activation of chlororespiration in Z. marina was prone to proactivity, which was also supported by the further activation of the oxidative pentose-phosphate pathway synthesizing NADPH to meet the demand of chlororespiration during darkness. This phenomenon is distinct from the common assumption that chlororespiration is prone to consuming redundant reducing power during the short transition phase from light to dark.

The water-water cycle is more effective in regulating redox state of photosystem I under fluctuating light than cyclic electron transport

Photosynthetic electron flux from water via photosystem II (PSII) and PSI to oxygen (water-water cycle) may act as an alternative electron sink under fluctuating light in angiosperms. We measured the P700 redox kinetics and electrochromic shift signal under fluctuating light in 11 Camellia species and tobacco leaves. Upon dark-to-light transition, these Camellia species showed rapid re-oxidation of P700. However, this rapid re-oxidation of P700 was not observed when measured under anaerobic conditions, as was in experiment with tobacco performed under aerobic conditions. Therefore, photo-reduction of O2 mediated by water-water cycle was functional in these Camellia species but not in tobacco. Within the first 10 s after transition from low to high light, PSI was highly oxidized in these Camellia species but was over-reduced in tobacco leaves. Furthermore, such rapid oxidation of PSI in these Camellia species was independent of the formation of trans-thylakoid proton gradient (ΔpH). These results indicated that in addition to ΔpH-dependent photosynthetic control, the water-water cycle can protect PSI against photoinhibition under fluctuating light in these Camellia species. We here propose that the water-water cycle is an overlooked strategy for photosynthetic regulation under fluctuating light in angiosperms.

Metabolic Hydrogen Flows in Rumen Fermentation: Principles and Possibilities of Interventions.

Rumen fermentation affects ruminants productivity and the environmental impact of ruminant production. The release to the atmosphere of methane produced in the rumen is a loss of energy and a cause of climate change, and the profile of volatile fatty acids produced in the rumen affects the post-absorptive metabolism of the host animal. Rumen fermentation is shaped by intracellular and intercellular flows of metabolic hydrogen centered on the production, interspecies transfer, and incorporation of dihydrogen into competing pathways. Factors that affect the growth of methanogens and the rate of feed fermentation impact dihydrogen concentration in the rumen, which in turn controls the balance between pathways that produce and incorporate metabolic hydrogen, determining methane production and the profile of volatile fatty acids. A basic kinetic model of competition for dihydrogen is presented, and possibilities for intervention to redirect metabolic hydrogen from methanogenesis toward alternative useful electron sinks are discussed. The flows of metabolic hydrogen toward nutritionally beneficial sinks could be enhanced by adding to the rumen fermentation electron acceptors or direct fed microbials. It is proposed to screen hydrogenotrophs for dihydrogen thresholds and affinities, as well as identifying and studying microorganisms that produce and utilize intercellular electron carriers other than dihydrogen. These approaches can allow identifying potential microbial additives to compete with methanogens for metabolic hydrogen. The combination of adequate microbial additives or electron acceptors with inhibitors of methanogenesis can be effective approaches to decrease methane production and simultaneously redirect metabolic hydrogen toward end products of fermentation with a nutritional value for the host animal. The design of strategies to redirect metabolic hydrogen from methane to other sinks should be based on knowledge of the physicochemical control of rumen fermentation pathways. The application of new –omics techniques together with classical biochemistry methods and mechanistic modeling can lead to exciting developments in the understanding and manipulation of the flows of metabolic hydrogen in rumen fermentation.

Photorespiration is the major alternative electron sink under high light in alpine evergreen sclerophyllous Rhododendron species

Owing to the high leaf mass per area, alpine evergreen sclerophyllous Rhododendron have low values of mesophyll conductance (gm). The resulting low chloroplast CO2 concentration aggravates photorespiration, which requires a higher ATP/NADPH ratio. However, the significance of photorespiration and underlying mechanisms of energy balance in these species are little known. In this study, eight alpine evergreen sclerophyllous Rhododendron species grown in a common garden were tested for their gm, electron flow to photorespiration, and energy balancing. Under saturating light, gm was the most limiting factor for net photosynthesis (AN) in all species, and the species differences in AN were primarily driven by gm rather than stomatal conductance. The total electron flow through photosystem II (ETRII) nearly equaled the electron transport required for Rubisco carboxylation and oxygenation. Furthermore, blocking electron flow to photosystem I with appropriate inhibitors showed that electron flow to plastic terminal oxidase was not observed. As a result, these studied species showed little alternative electron flow mediated by water-water cycle or plastic terminal oxidase. By comparison, the ratio of electron transport consumed by photorespiration to ETRII (JPR/ETRII), ranging from 43%∼55%, was negatively correlated to AN and gm. Furthermore, the increased ATP production required by enhanced photorespiration was regulated by cyclic electron flow around photosystem I. These results indicate that photorespiration is the major electron sink for dissipation of excess excitation energy in the alpine evergreen sclerophyllous Rhododendron species. The coordination of gm, photorespiration and cyclic electron flow is important for sustaining leaf photosynthesis.

Photoprotective strategies against drought are depending on the elevation provenance in Phacelia secunda

The central Chilean Andes are located in a Mediterranean-type climate zone, characterized by dry summers and high irradiance. This creates a contrasting elevational gradient because higher elevations get more solid precipitation and lower temperatures, resulting in higher soil humidity along the growing season compared with severe drought at lower elevations. Therefore, species with wide elevational distributions, such as Phacelia secunda, must have developed specific adaptations to cope with contrasting severity of drought stress-induced photoinhibition at different elevations. We hypothesize that P. secunda from lower elevation, is more tolerant to drought stress-induced photo-damage than plants from high elevation. This higher tolerance will be associated to a higher diversity of photoprotective strategies in plants that naturally suffers severe drought every growing season. To test this hypothesis, plants from 2700 and 3600 m in the central Chilean Andes were grown under the common garden and then subjected to water restriction. We measured stress indicators, photochemistry of PSII and PSI and estimate alternative electron sinks. Drought affected P. secunda photosynthetic performance differentially depending on the elevation of provenance. Plants from lower elevation exhibited higher drought tolerance than higher elevation ones. This was related to higher levels of heat dissipation and alternative electron sinks exhibited by plants from lower elevation under drought stress. We concluded that plants naturally subjected to recurrent drought are better adapted to respond to drought stress using additional photochemical photoprotective mechanisms and confirm the role of alternative electron sinks ameliorating photodamage.