Sustainable bioenergy for climate mitigation: developing drought-tolerant trees and grasses.

BACKGROUND AND AIMS: Bioenergy crops are central to climate mitigation strategies that utilize biogenic carbon, such as BECCS (bioenergy with carbon capture and storage), alongside the use of biomass for heat, power, liquid fuels and, in the future, biorefining to chemicals. Several promising lignocellulosic crops are emerging that have no food role - fast-growing trees and grasses - but are well suited as bioenergy feedstocks, including Populus, Salix, Arundo, Miscanthus, Panicum and Sorghum. SCOPE: These promising crops remain largely undomesticated and, until recently, have had limited germplasm resources. In order to avoid competition with food crops for land and nature conservation, it is likely that future bioenergy crops will be grown on marginal land that is not needed for food production and is of poor quality and subject to drought stress. Thus, here we define an ideotype for drought tolerance that will enable biomass production to be maintained in the face of moderate drought stress. This includes traits that can readily be measured in wide populations of several hundred unique genotypes for genome-wide association studies, alongside traits that are informative but can only easily be assessed in limited numbers or training populations that may be more suitable for genomic selection. Phenotyping, not genotyping, is now the major bottleneck for progress, since in all lignocellulosic crops studied extensive use has been made of next-generation sequencing such that several thousand markers are now available and populations are emerging that will enable rapid progress for drought-tolerance breeding. The emergence of novel technologies for targeted genotyping by sequencing are particularly welcome. Genome editing has already been demonstrated for Populus and offers significant potential for rapid deployment of drought-tolerant crops through manipulation of ABA receptors, as demonstrated in Arabidopsis, with other gene targets yet to be tested. CONCLUSIONS: Bioenergy is predicted to be the fastest-developing renewable energy over the coming decade and significant investment over the past decade has been made in developing genomic resources and in collecting wild germplasm from within the natural ranges of several tree and grass crops. Harnessing these resources for climate-resilient crops for the future remains a challenge but one that is likely to be successful.

Closing in on maximum yield of chlorophyll fluorescence using a single multiphase flash of sub-saturating intensity.

Estimation of the maximum chlorophyll fluorescence yield under illumination, or Fm', by traditional single saturation pulse (SP) methodology is prone to underestimation error because of rapid turnover within photosystem (PS) II. However, measurements of fluorescence yield during several single pulses of variable intensity describes the irradiance dependence of apparent Fm', from which estimates of Fm' at infinite irradiance can be derived. While such estimates have been shown to result in valid approximations of Fm', the need to apply several single pulses limits its applicability. We introduce a novel approach that determines the relationship between apparent Fm' and variable irradiance within a single ∼1 s multiphase flash (MPF). Through experiments and simulations, we demonstrate that the rate of variation in irradiance during an MPF is critical for achieving quasi-steady-state changes in the proportions of PSII acceptor side redox intermediates and the corresponding fluorescence yields, which are prerequisites for accurately estimating Fm' at infinite irradiance. The MPF methodology is discussed in the context of improving the accuracy of various parameters derived from chlorophyll fluorescence measurements, such as photochemical and non-photochemical quenchings and efficiencies. The importance of using MPF methodology for interpreting chlorophyll fluorescence, in particular for integrating fluorescence and gas exchange measurements, is emphasized.

Heterogeneous inhibition of photosynthesis over the leaf surface of rosa rubiginosa L. during water stress and abscisic acid treatment: induction of a metabolic component by limitation of CO(2) diffusion

The contribution of changes in stomatal conductance and metabolism in determining heterogeneous photosynthesis inhibition during dehydration and abscisic acid (ABA) feeding was investigated using detached leaves of Rosa rubiginosa L. The steady-state and maximal rates of electron transport under a transient high CO(2) concentration were monitored using chlorophyll fluorescence imaging. The decrease in electron transport rate induced by dehydration and ABA treatment almost reverted to the control rate under transient high CO(2) availability. Therefore, inhibition of photosynthesis was mainly mediated through stomatal closure. However, since reversion was not complete, a metabolic inhibition was also identified as a decrease in the maximal electron transport rate driven by carboxylation. Under dehydration or ABA feeding, as under low ambient CO(2) treatment, in 21% or 0.4% O(2), the lower the steady-state electron transport was, the lower was the maximal electron transport rate during transient high CO(2) availability. We conclude that low CO(2) availability reduced the capacity of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) to drive electron transport. The potential contribution of Rubisco deactivation mediated by stomatal closure is discussed.

Mapping intercellular CO2 mole fraction (Ci) in rosa rubiginosa leaves fed with abscisic acid by using chlorophyll fluorescence imaging. Significance Of ci estimated from leaf gas exchange

Imaging of photochemical yield of photosystem II (PSII) computed from leaf chlorophyll fluorescence images and gas-exchange measurements were performed on Rosa rubiginosa leaflets during abscisic acid (ABA) addition. In air ABA induced a decrease of both the net CO2 assimilation (An) and the stomatal water vapor conductance (gs). After ABA treatment, imaging in transient nonphotorespiratory conditions (0.1% O2) revealed a heterogeneous decrease of PSII photochemical yield. This decline was fully reversed by a transient high CO2 concentration (7400 mol mol-1) in the leaf atmosphere. It was concluded that ABA primarily affected An by decreasing the CO2 supply at ribulose-1,5-bisphosphate carboxylase/oxygenase. Therefore, the An versus intercellular mole fraction (Ci) relationship was assumed not to be affected by ABA, and images of Ci and gs were constructed from images of PSII photochemical yield under nonphotorespiratory conditions. The distribution of gs remained unimodal following ABA treatment. A comparison of calculations of Ci from images and gas exchange in ABA-treated leaves showed that the overestimation of Ci estimated from gas exchange was only partly due to heterogeneity. This overestimation was also attributed to the cuticular transpiration, which largely affects the calculation of the leaf conductance to CO2, when leaf conductance to water is low.

The consequences of chlorophyll deficiency for photosynthetic light use efficiency in a single nuclear gene mutation of cowpea.

The light harvesting and photosynthetic characteristics of a chlorophyll-deficient mutant of cowpea (Vigna unguilata), resulting from a single nuclear gene mutation, are examined. The 40% reduction in total chlorophyll content per leaf area in the mutant is associated with a 55% reduction in pigment-proteins of the light harvesting complex associated with Photosystem II (LHC II), and to a lesser extent (35%) in the light harvesting complex associated with Photosystem I (LHC I). No significant differences were found in the Photosystem I (PS I) and Photosystem II (PS II) contents per leaf area of the mutant compared to the wildtype parent. The decreases in the PS I and PS II antennae sizes in the mutant were not accompanied by any major changes in quantum efficiencies of PS I and PS II in leaves at non-saturating light levels for CO2 assimilation. Although the chlorophyll deficiency resulted in an 11% decrease in light absorption by mutant leaves, their maximum quantum yield and light saturated rate of CO2 assimilation were similar to those of wildtype leaves. Consequently, the large and different decreases in the antennae of PS II and PS I in the mutant are not associated with any loss of light use efficiency in photosynthesis.

Relationship between Photosynthetic Electron Transport and Stromal Enzyme Activity in Pea Leaves : Toward an Understanding of the Nature of Photosynthetic Control.

The responses of the quantum efficiencies of photosystem (PS) II and PSI measured in vivo simultaneously with estimations of the activities and activation states of NADP-malate dehydrogenase, chloroplast fructose-1,6-bisphosphatase, and ribulose-1,5-bisphosphate carboxylase were used to study the relationship between electron transport and carbon metabolism. The effects of varying irradiance and CO(2) partial pressure on the relationship between the quantum efficiencies of PSI and II, and the activity of these enzymes shows that the interrelationships vary according to the limitations placed on the system. The relationship between the quantum efficiencies of PSII and PSI was linear in most situations. In response to increasing irradiance, the activity of all three enzymes increased. In the case of NADP-malate dehydrogenase this increase was well correlated with the estimated flux of electrons through PSI and PSII. The other two enzymes showed a more complex relationship with the estimated flux of electrons through both photosystems. These relationships are consistent with the known interactions between these stromal enzymes and the thylakoids. The response to varying CO(2) partial pressure is more complex. The efficiencies of PSI and II declined with decreasing CO(2) partial pressure and the activity of each enzyme varied uniquely. However, there are clear correlations between the activities of the enzymes and the flux of electrons through the photosystems. In contrast to the data obtained under conditions of varying irradiance, there is clear evidence of photosynthetic control of electron transport when the CO(2) concentration is varied.

The relationship between CO2 assimilation and electron transport in leaves.

The inter-relationships between the quantum efficiencies of photosystems I (φI) and II (φII) and the quantum yield of CO2 fixation % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0dh9WrFfpC0xh9vqqj-hEeeu0xXdbba9frFj0-OqFf% ea0dXdd9vqaq-JfrVkFHe9pgea0dXdar-Jb9hs0dXdbPYxe9vr0-vr% 0-vqpWqaaeaabaGaciaacaqabeaadaqaaqaaaOqaaiabeA8aMnaaBa% aaleaacaWGdbGaam4tamaaBaaameaacaaIYaaaleqaaaqabaaaaa!3BD3!\[\phi _{CO_2 } \] were investigated in pea (Pisum sativum (L)) leaves with differing rates of photosynthesis using both photorespiratory and non-photorespiratory conditions, and in a leaf of Hedera helix (L) under photorespiratory conditions. The results indicate that under photorespiratory conditions the relationship between % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0dh9WrFfpC0xh9vqqj-hEeeu0xXdbba9frFj0-OqFf% ea0dXdd9vqaq-JfrVkFHe9pgea0dXdar-Jb9hs0dXdbPYxe9vr0-vr% 0-vqpWqaaeaabaGaciaacaqabeaadaqaaqaaaOqaaiabeA8aMnaaBa% aaleaacaWGdbGaam4tamaaBaaameaacaaIYaaaleqaaaqabaaaaa!3BD3!\[\phi _{CO_2 } \] and both φI and φII is non-linear and variable. The relationship between φI and φII under these circumstances remains predominantly linear. Under non-photorespiratory conditions, leaves with a low rate of photosynthesis due to sink limitation exhibit a non-linear relationship between φI and φII, though the relationship between φI and φII remains linear suggesting a close relationship between linear electron flow and CO2 fixation. Leaves irradiated at the CO2 compensation point also exhibit a non-linear relationship between φI and φII. These results suggest that for leaves in air linear electron flow is the predominant source of energy for metabolism. The role of cyclic electron transport is considered when the requirement for the products of linear electron transport is depressed.

The relationship between non-photochemical quenching of chlorophyll fluorescence and the rate of photosystem 2 photochemistry in leaves.

It has been suggested previously that non-photochemical quenching of chlorophyll fluorescence is associated with a decrease in the rate of photosystem 2 (PS 2) photochemistry. In this study analyses of fluorescence yield changes, induced by flashes in leaves exhibiting different amounts of non-photochemical quenching of fluorescence, are made to determine the effect of non-photochemical excitation energy quenching processes on the rate of PS 2 photochemistry. It is demonstrated that both the high-energy state and the more slowly relaxing components of non-photochemical quenching reduce the rate of PS 2 photochemistry. Flash dosage response curves for fluorescence yield show that non-photochemical quenching processes effectively decrease the relative effective absorption cross-section for PS 2 photochemistry. It is suggested that non-photochemical quenching processes exert an effect on the rate of PS 2 photochemistry by increasing the dissipation of excitation energy by non-radiative processes in the pigment matrices of PS 2, which consequently results in a decrease in the efficiency of delivery of excitation energy for PS 2 photochemistry.

Non-photochemical quenching of Fo in leaves is emission wavelength dependent: consequences for quenching analysis and its interpretation.

The effects of light-induced non-photochemical quenching on the minimal Fo, and variable Fv, fluorescence emissions at 690 and 730 nm in leaves were determined. Non-photochemical quenching of Fo, but not Fv, was found to be dependent upon the wavelength of emission, and was greater at 690 nm than at 730 nm. For emission at 730, compared to at 690 nm, approx. 30% of Fo was not affected by non-photochemical quenching processes in leaves of C3 plants; in maize leaves this was found to be approx. 50%. The data indicate that a substantial proportion of the pigments contributing to Fo emission at 730 nm are not quenched by light-induced, non-photochemical quenching processes and that there are large differences in the pigment matrices contributing to Fo and Fv emissions at 730 nm, compared to those at 690 nm. These findings have important implications for the accurate estimation and interpretation of non-photochemical quenching of fluorescence parameters and their use in the calculation of photochemical efficiencies in leaves. Measurements of fluorescence emissions at wavelengths above 700 nm are likely to give rise to significant errors when used for determinations of photochemical and non-photochemical quenching parameters.

Relationship between the Quantum Efficiencies of Photosystems I and II in Pea Leaves.

The irradiance dependence of the efficiencies of photosystems I and II were measured for two pea (Pisum sativum [L.]) varieties grown under cold conditions and one pea variety grown under warm conditions. The efficiencies of both photosystems declined with increasing irradiance for all plants, and the quantum efficiency of photosystem I electron transport was closely correlated with the quantum efficiency of photosystem II electron transport. In contrast to the consistent pattern shown by efficiency of the photosystems, the redox state of photosystem II (as estimated from the photochemical quenching coefficient of chlorophyll fluorescence) exhibited relationships with both irradiance and the reduction of P-700 that varied with growth environment and genotype. This variability is considered in the context of the modulation of photosystem II quantum efficiency by both photochemical and nonphotochemical quenching of excitation energy.

Effects of drought on primary photosynthetic processes of cotton leaves.

The effects of drought on Photosystem II (PSII) fluorescence and photosynthetic electron transport activities were analyzed in cotton. Water stress did not modify the amplitude of leaf variable fluorescence at room temperature in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) nor at 77 K. It is therefore concluded that photon collection, their distribution between the two photosystems, and PSII photochemistry are unaffected by the stress. In droughted leaves at room temperature under low exciting light, the transitory maximum (F(p)) and steady state (F(t)) fluorescence levels are increased; under high exciting light, F(p) level and the rise time from the initial level (F(o)) to F(p) are unchanged, whereas F(p) to F(t) decay time is increased. These results infer that the drought slows the rate of plastoquinone reoxidation. This conclusion agrees with a larger proportion of reduced primary PSII electron acceptor Q(A) measured at the steady state under low light. In thylakoids isolated from droughted leaves, PSII mediated electron flow was the same as in thylakoids from control leaves, whereas PSI mediated electron transport was inhibited. It is shown that water stress does not induce sensitization to photoinhibition in cotton.