Erwin Dreyer

Mesophyll diffusion conductance to CO2: An unappreciated central player in photosynthesis

Mesophyll diffusion conductance to CO(2) is a key photosynthetic trait that has been studied intensively in the past years. The intention of the present review is to update knowledge of g(m), and highlight the important unknown and controversial aspects that require future work. The photosynthetic limitation imposed by mesophyll conductance is large, and under certain conditions can be the most significant photosynthetic limitation. New evidence shows that anatomical traits, such as cell wall thickness and chloroplast distribution are amongst the stronger determinants of mesophyll conductance, although rapid variations in response to environmental changes might be regulated by other factors such as aquaporin conductance. Gaps in knowledge that should be research priorities for the near future include: how different is mesophyll conductance among phylogenetically distant groups and how has it evolved? Can mesophyll conductance be uncoupled from regulation of the water path? What are the main drivers of mesophyll conductance? The need for mechanistic and phenomenological models of mesophyll conductance and its incorporation in process-based photosynthesis models is also highlighted.

Gradual Soil Water Depletion Results in Reversible Changes of Gene Expression, Protein Profiles, Ecophysiology, and Growth Performance in Populus euphratica , a Poplar Growing in Arid Regions

The responses of Populus euphratica Oliv. plants to soil water deficit were assessed by analyzing gene expression, protein profiles, and several plant performance criteria to understand the acclimation of plants to soil water deficit. Young, vegetatively propagated plants originating from an arid, saline field site were submitted to a gradually increasing water deficit for 4 weeks in a greenhouse and were allowed to recover for 10 d after full reirrigation. Time-dependent changes and intensity of the perturbations induced in shoot and root growth, xylem anatomy, gas exchange, and water status were recorded. The expression profiles of approximately 6,340 genes and of proteins and metabolites (pigments, soluble carbohydrates, and oxidative compounds) were also recorded in mature leaves and in roots (gene expression only) at four stress levels and after recovery. Drought successively induced shoot growth cessation, stomatal closure, moderate increases in oxidative stress-related compounds, loss of CO2 assimilation, and root growth reduction. These effects were almost fully reversible, indicating that acclimation was dominant over injury. The physiological responses were paralleled by fully reversible transcriptional changes, including only 1.5% of the genes on the array. Protein profiles displayed greater changes than transcript levels. Among the identified proteins for which expressed sequence tags were present on the array, no correlation was found between transcript and protein abundance. Acclimation to water deficit involves the regulation of different networks of genes in roots and shoots. Such diverse requirements for protecting and maintaining the function of different plant organs may render plant engineering or breeding toward improved drought tolerance more complex than previously anticipated.

Gene expression and metabolite profiling of Populus euphratica growing in the Negev desert

BackgroundPlants growing in their natural habitat represent a valuable resource for elucidating mechanisms of acclimation to environmental constraints. Populus euphratica is a salt-tolerant tree species growing in saline semi-arid areas. To identify genes involved in abiotic stress responses under natural conditions we constructed several normalized and subtracted cDNA libraries from control, stress-exposed and desert-grown P. euphratica trees. In addition, we identified several metabolites in desert-grown P. euphratica trees.ResultsAbout 14,000 expressed sequence tag (EST) sequences were obtained with a good representation of genes putatively involved in resistance and tolerance to salt and other abiotic stresses. A P. euphratica DNA microarray with a uni-gene set of ESTs representing approximately 6,340 different genes was constructed. The microarray was used to study gene expression in adult P. euphratica trees growing in the desert canyon of Ein Avdat in Israel. In parallel, 22 selected metabolites were profiled in the same trees.ConclusionOf the obtained ESTs, 98% were found in the sequenced P. trichocarpa genome and 74% in other Populus EST collections. This implies that the P. euphratica genome does not contain different genes per se, but that regulation of gene expression might be different and that P. euphratica expresses a different set of genes that contribute to adaptation to saline growth conditions. Also, all of the five measured amino acids show increased levels in trees growing in the more saline soil.

Is mesophyll conductance to CO2 in leaves of three Eucalyptus species sensitive to short-term changes of irradiance under ambient as well as low O2?

Mesophyll conductance to CO2 (g m) limits the diffusion of CO2 to the sites of carboxylation, and may respond rapidly (within minutes) to abiotic factors. Using three Eucalyptus species, we tested the rapid response of g m to irradiance under 21% and 1% O2. We used simultaneous measurements of leaf gas exchange and discrimination against 13CO2 with a tuneable diode laser absorption spectrometer. Measurements under 1% O2 were used to limit uncertainties due to 13C-12C fractionation occurring during photorespiration. Switching irradiance from 600 to 200µmolm-2s-1 led to a ≈60% decrease of g m within minutes in all species under both 21% O2 and 1% O2. The g m response to irradiance is unlikely to be a computation artefact since using different values for the parameters of the discrimination model changed the absolute values of g m but did not affect the relative response to irradiance. Simulations showed that possible rapid changes of any parameter were unable to explain the observed variations of g m with irradiance, except for13C-12C fractionation during carboxylation (b), which, in turn, is dependent on the fraction of leaf C assimilated by phospho-enol pyruvate carboxylase (PEPc) (β). g m apparently increased by ≈30% when O2 was switched from 21% to 1% O2. Again, possible changes of β with O2 could explain this apparent g m response to O2. Nevertheless, large irradiance or O2-induced changes in β would be required to fully explain the observed changes in g m, reinforcing the hypothesis that g m is responsive to irradiance and possibly also to O2.

Performance of a new dynamic model for predicting diurnal time courses of stomatal conductance at the leaf level

Under natural conditions, plants are subjected to continuous changes of irradiance that drive variations of stomatal conductance to water vapour (g(s)). We propose a dynamic model to predict the temporal response of g(s) at the leaf level using an asymmetric sigmoid function with a unique parameter describing time constants for increasing and decreasing g(s). The model parameters were adjusted to observed data using Approximate Bayesian Computation. We tested the model performance for (1) instant changes of irradiance; or (2) continuous and controlled variations of irradiance simulating diurnal time courses. Compared with the two mostly used steady-state models, our dynamic model described daily time courses of g(s) with a higher accuracy. In particular, it was able to describe the hysteresis of g(s) responses to increasing/decreasing irradiance and the resulting rapid variations of intrinsic water-use efficiency. Compared to the mechanistic model of temporal responses of g(s) by Kirschbaum, Gross & Pearcy, for which time constants were estimated with a large variance, our model estimated time constants with a higher precision. It is expected to improve predictions of water loss and water-use efficiency in higher scale models by using a small number of parameters.

Do trees use reserve or newly assimilated carbon for their defense reactions? A 13C labeling approach with young Scots pines inoculated with a bark-beetle-associated fungus (Ophiostoma brunneo ciliatum)

Three-year-old saplings of Pinus sylvestris L. were labeled with 13CO2 prior to inoculating the trunk with Ophiostoma brunneo ciliatum, a blue-staining fungus usually associated to Ips sexdentatus. During incubation, half the trees were submitted to a severe drought that decreased photosynthesis and natural 13C content in non-labeled saplings. A large 13C-excess was obtained in wood and phloem, especially in the fractions of soluble proteins, starch and soluble sugars of labeled saplings. Drought increased 13C-excess, due to reduced photosynthesis and smaller dilution of 13C by the addition of newly assimilated 12C. The induced-reaction zones in inoculated saplings displayed large total C (58 g 100 g−1) because of the accumulation of secondary metabolites. They also showed much larger 13C-excess than any other compartment: the contribution of stored C to the reaction zones was much higher than that of currently assimilated C. Moreover, drought lowered the contribution of the latter, as shown by the increase of 13C in the reaction zones. We conclude that stored C was readily mobilized for the construction of reaction tissues, and that the contribution of currently assimilated C was only minor.RésuméDe jeunes pins sylvestres (Pinus sylvestris L.) âgés de trois ans ont été marqués avec du 13CO2 puis inoculés dans le tronc avec Ophiostoma brunneo ciliatum, un champignon habituellement associé au scolyte Ips sexdentatus. Pendant l’incubation, la moitié des arbres a été soumise à une sécheresse sévère qui a fortement réduit la photosynthèse et l’abondance naturelle en 13C des individus non marqués. Un fort excès en 13C a été détecté dans le bois et le phloème ainsi que dans les protéines solubles, l’amidon et les sucres solubles des individus marqués. La sécheresse a amplifié cet excès, du fait d’une photosynthèse réduite et donc d’une moindre dilution du 13C par du 12C récemment assimilé. Les zones de réaction induite autour des points d’inoculation présentaient de fortes teneurs en C (58 g 100 g−1), du fait de l’accumulation massive de métabolites secondaires. Elles présentaient également un excès de 13C plus marqué que n’importe quel autre tissu : ces zones de réaction étaient donc essentiellement constituées à partir de C provenant des réserves avec une faible contribution de C récemment assimilé. De plus, la sécheresse a augmenté la contribution du C de réserve, comme le montre l’augmentation de l’excès de 13C dans les zones de réaction.

Impact of Erysiphe alphitoides on transpiration and photosynthesis in Quercus robur leaves

Oak powdery mildew, (Erysiphe alphitoides) causes one of the most common diseases of oaks. We assessed the impact of this pathogen on photosynthesis and water relations of infected leaves using greenhouse-grown oak seedlings. Transpiration of seedlings infected by oak powdery mildew was also investigated. Altogether, E. alphitoides had a low impact on host gas exchange whether at the leaf or whole plant scale. Maximal stomatal conductance of infected leaves was reduced by 20–30% compared to healthy controls. Severely infected seedlings did not experience any detectable change of whole plant transpiration. The reduction in net CO2 assimilation, An, was less than proportional to the fraction of leaf area infected. Powdery mildew reduced both the maximal light-driven electron flux (Jmax) and the apparent maximal carboxylation velocity (Vcmax) although Vcmax was slightly more impacted than Jmax. No compensation for the infection occurred in healthy leaves of partly infected seedlings as the reduced photosynthesis in the infected leaves was not paralleled by increased An levels in the healthy leaves of the seedlings. However, E. alphitoides had a strong impact on the leaf life-span of infected leaves. It is concluded that the moderate effect of E. alphitoides on oak might be related to the small impact on net CO2 assimilation rates and on tree transpiration; nevertheless, the severe reduction in leaf life-span of heavily infected leaves may lead to decreased carbon uptake over the growth season.

Response to canopy opening does not act as a filter to Fagus sylvatica and Acer sp. advance regeneration in a mixed temperate forest

Abstract• In mixed-species forest stands, large losses in tree species diversity often occur during the regeneration phase. In a former coppice-with-standards, we investigated whether the limiting stage in the recruitment process of advance regeneration is the immediate seedling response to canopy release. Experimental canopy gaps were opened and the survival and growth of advance seedlings (Fagus sylvatica, Acer pseudoplatanus, Acer campestre, Acer platanoides) growing in the gaps or under closed canopy were monitored for three years.• All species responded positively and rapidly to canopy release. Survival was not affected by gap opening. Diameter increment after gap opening was similar across species, and height increment was greater for Acer platanoides and for Acer pseudoplatanus. Post-release diameter and height growth were mainly determined by pre-release seedling size. Competition from neighbouring seedlings did not affect growth in the three years following canopy opening.• In the recruitment process of F. sylvatica and Acer sp. advance regeneration, the recovery from canopy release did not appear as a limiting step that would filter against some species. Pre-release size was the main factor accounting for post-release growth and is probably a major determinant of long-term seedling dominance.Résumé• Dans les peuplements mélangés, une forte réduction de la diversité spécifique a lieu au cours de la phase de régénération. Nous avons examiné dans un ancien taillis-sous-futaie dans quelle mesure la réponse immédiate des semis à l’ouverture du couvert adulte pouvait limiter le recrutement de semis préexistants. Des trouées ont été ouvertes dans le couvert, et la survie et la croissance de semis préexistants (Fagus sylvatica, Acer pseudoplatanus, Acer campestre, Acer platanoides), situés dans les trouées ou bien sous couvert fermé, ont été suivies pendant trois années.• Les quatre espèces ont répondu positivement et rapidement à l’ouverture du couvert. La survie n’a pas été affectée par l’ouverture. Pour les semis dans les trouées, l’accroissement en diamètre était similaire pour les quatre espèces et l’accroissement en hauteur était plus important pour Acer platanoides et Acer pseudoplatanus. La croissance en hauteur et en diamètre après ouverture étaient principalement déterminées par les dimensions des semis avant ouverture. La compétition exercée par les semis voisins n’a pas affecté significativement la croissance dans les trois années qui ont suivi l’ouverture.• La période d’acclimatation des semis aux nouvelles conditions après ouverture de la canopée n’est pas apparue comme un stade limitant le recrutement des semis préexistants de Fagus sylvatica, Acer sp., ni comme un filtre vis-à-vis de l’une des quatre espèces. Les dimensions initiales des semis constituaient le facteur principal expliquant la croissance après ouverture, et probablement un facteur explicatif important pour la dominance future des semis vis-à-vis de leurs voisins.

Vulnerability to embolism differs in roots and shoots and among three Mediterranean conifers: consequences for stomatal regulation of water loss?

We investigated the potential links between stomatal control of transpiration and the risk of embolism in root and shoot xylem of seedlings of three Mediterranean conifers (Cupressus sempervirens, Pinus halepensis and P. nigra) grown in a greenhouse under semi-controlled conditions. We measured the intrinsic vulnerability to embolism in roots and current year shoots by the air injection method. Root and shoot segments were subjected to increasing pressures, and the induced loss of hydraulic conductivity recorded. The three species displayed very different vulnerabilities in shoots, with P. nigra being much more vulnerable than P. halepensis and C. sempervirens. Roots were distinctly more vulnerable than shoots in C. sempervirens and P. halepensis (50% loss of conductivity induced at 3.0 MPa and 1.7 MPa higher xylem water potential in roots vs shoots). In P. nigra, no significant difference of vulnerability between shoots and roots was found. Seedlings were subjected to soil drought, and stomatal conductance, twig hydraulic conductivity and needle water potential were measured. The water potential resulting in almost complete stomatal closure (90%) was very close to the threshold water potential inducing loss of conductivity (10%) in twigs in P nigra, resulting in a very narrow safety margin between stomatal closure and embolism induction. The safety margin was larger in P. halepensis and greatest in C. sempervirens. Unexpectedly, this water potential threshold produced a 30–50% loss of conductivity in 3–5 mm diameter roots, depending on the species. The implications of this finding are discussed.

Stomatal conductance and root-to-shoot signalling in chestnut saplings exposed to Phytophthora cinnamomi or partial soil drying

The effects of root infection by Phytophthora cinnamomi on stomatal conductance in Castanea sativa L. saplings were investigated to determine the potential role of root-derived chemical signals. A split-root experiment was carried out, in which inoculation of the pathogen or drought was applied to the root systems in either one or both compartments. At the end of the experiment plant sap extracts were collected and their effects on stomatal conductance were determined by leaf bioassay. Inoculation or drought imposed in both compartments resulted in decreases in stomatal conductance (gs), transpiration rate, soil-to-leaf specific hydraulic conductance, leaf water potential, xylem [ABA] and root biomass, but not in the ratio of root-to-leaf mass in inoculated plants. Conversely, only gs and xylem [ABA] were affected in plants inoculated or droughted in one compartment, and no changes were detectable in leaf water potential and soil-to-leaf specific hydraulic conductance. The leaf bioassay showed that gs in chestnut was sensitive to ABA but not to Phytophthora elicitins. Stomatal conductance was reduced by some sap extracts, both from control and inoculated plants. Our results suggest the involvement of different signals, chemical and hydraulic, in regulating stomatal conductance of chestnut at different stages of stress.