Exploring the potential of Δ17O in CO2 for determining mesophyll conductance.

Mesophyll conductance to CO2 from the intercellular air space to the CO2-H2O exchange site has been estimated using δ18O measurements (gm18). However, the gm18 estimates are affected by the uncertainties in the δ18O of leaf water where the CO2-H2O exchange takes place and the degree of equilibration between CO2 and H2O. We show that measurements of Δ17O (i.e.Δ17O=δ17O-0.528×δ18O) can provide independent constraints on gm (gmΔ17) and that these gm estimates are less affected by fractionation processes during gas exchange. The gm calculations are applied to combined measurements of δ18O and Δ17O, and gas exchange in two C3 species, sunflower (Helianthus annuus L. cv. 'sunny') and ivy (Hedera hibernica L.), and the C4 species maize (Zea mays). The gm18 and gmΔ17 estimates agree within the combined errors (P-value, 0.876). Both approaches are associated with large errors when the isotopic composition in the intercellular air space becomes close to the CO2-H2O exchange site. Although variations in Δ17O are low, it can be measured with much higher precision compared with δ18O. Measuring gmΔ17 has a few advantages compared with gm18: (i) it is less sensitive to uncertainty in the isotopic composition of leaf water at the isotope exchange site and (ii) the relative change in the gm due to an assumed error in the equilibration fraction θeq is lower for gmΔ17 compared with gm18. Thus, using Δ17O can complement and improve the gm estimates in settings where the δ18O of leaf water varies strongly, affecting the δ18O (CO2) difference between the intercellular air space and the CO2-H2O exchange site.

A meta-analysis of responses of C3 plants to atmospheric CO2 : dose-response curves for 85 traits ranging from the molecular to the whole-plant level.

Generalised dose-response curves are essential to understand how plants acclimate to atmospheric CO2 . We carried out a meta-analysis of 630 experiments in which C3 plants were experimentally grown at different [CO2 ] under relatively benign conditions, and derived dose-response curves for 85 phenotypic traits. These curves were characterised by form, plasticity, consistency and reliability. Considered over a range of 200-1200 µmol mol-1 CO2 , some traits more than doubled (e.g. area-based photosynthesis; intrinsic water-use efficiency), whereas others more than halved (area-based transpiration). At current atmospheric [CO2 ], 64% of the total stimulation in biomass over the 200-1200 µmol mol-1 range has already been realised. We also mapped the trait responses of plants to [CO2 ] against those we have quantified before for light intensity. For most traits, CO2 and light responses were of similar direction. However, some traits (such as reproductive effort) only responded to light, others (such as plant height) only to [CO2 ], and some traits (such as area-based transpiration) responded in opposite directions. This synthesis provides a comprehensive picture of plant responses to [CO2 ] at different integration levels and offers the quantitative dose-response curves that can be used to improve global change simulation models.

Dividing the pie: A quantitative review on plant density responses.

Plant population density is an important variable in agronomy and forestry and offers an experimental way to better understand plant-plant competition. We made a meta-analysis of responses of even-aged mono-specific stands to population density by quantifying for 3 stand and 33 individual plant variables in 334 experiments how much both plant biomass and phenotypic traits change with a doubling in density. Increasing density increases standing crop per area, but decreases the mean size of its individuals, mostly through reduced tillering and branching. Among the phenotypic traits, stem diameter is negatively affected, but plant height remains remarkably similar, partly due to an increased stem length-to-mass ratio and partly by increased allocation to stems. The reduction in biomass is caused by a lower photosynthetic rate, mainly due to shading of part of the foliage. Total seed mass per plant is also strongly reduced, marginally by lower mass per seed, but mainly because of lower seed numbers. Plants generally have fewer shoot-born roots, but their overall rooting depth seems hardly affected. The phenotypic plasticity responses to high densities correlate strongly with those to low light, and less with those to low nutrients, suggesting that at high density, shading affects plants more than nutrient depletion.

Oxygen isotopes in tree rings are a good proxy for Amazon precipitation and El Nino-Southern Oscillation variability.

We present a unique proxy for the reconstruction of variation in precipitation over the Amazon: oxygen isotope ratios in annual rings in tropical cedar (Cedrela odorata). A century-long record from northern Bolivia shows that tree rings preserve the signal of oxygen isotopes in precipitation during the wet season, with weaker influences of temperature and vapor pressure. Tree ring δ(18)O correlates strongly with δ(18)O in precipitation from distant stations in the center and west of the basin, and with Andean ice core δ(18)O showing that the signal is coherent over large areas. The signal correlates most strongly with basin-wide precipitation and Amazon river discharge. We attribute the strength of this (negative) correlation mainly to the cumulative rainout processes of oxygen isotopes (Rayleigh distillation) in air parcels during westward transport across the basin. We further find a clear signature of the El Niño-Southern Oscillation (ENSO) in the record, with strong ENSO influences over recent decades, but weaker influence from 1925 to 1975 indicating decadal scale variation in the controls on the hydrological cycle. The record exhibits a significant increase in δ(18)O over the 20th century consistent with increases in Andean δ(18)O ice core and lake records, which we tentatively attribute to increased water vapor transport into the basin. Taking these data together, our record reveals a fresh path to diagnose and improve our understanding of variation and trends of the hydrological cycle of the world's largest river catchment.

Gap effects on leaf traits of tropical rainforest trees differing in juvenile light requirement.

The relationships of 16 leaf traits and their plasticity with the dependence of tree species on gaps for regeneration (gap association index; GAI) were examined in a Neotropical rainforest. Young saplings of 24 species with varying GAI were grown under a closed canopy, in a medium-sized and in a large gap, thus capturing the full range of plasticity with respect to canopy openness. Structural, biomechanical, chemical and photosynthetic traits were measured. At the chloroplast level, the chlorophyll a/b ratio and plasticity in this variable were not related to the GAI. However, plasticity in total carotenoids per unit chlorophyll was larger in shade-tolerant species. At the leaf level, leaf mass per unit area (LMA) decreased with the GAI under the closed canopy and in the medium gap, but did not significantly decrease with the GAI in the large gap. This was a reflection of the larger plasticity in LMA and leaf thickness of gap-dependent species. The well-known opposite trends in LMA for adaptation and acclimation to high irradiance in evergreen tropical trees were thus not invariably found. Although leaf strength was dependent on LMA and thickness, plasticity in this trait was not related to the GAI. Photosynthetic capacity expressed on each basis increased with the GAI, but the large plasticity in these traits was not clearly related to the GAI. Although gap-dependent species tended to have a greater plasticity overall, as evident from a principle component analysis, leaf traits of gap-dependent species are thus not invariably more phenotypically plastic.

Interaction of temperature and irradiance effects on photosynthetic acclimation in two accessions of Arabidopsis thaliana.

The effect of temperature and irradiance during growth on photosynthetic traits of two accessions of Arabidopsis thaliana was investigated. Plants were grown at 10 and 22 °C, and at 50 and 300 µmol photons m(-2) s(-1) in a factorial design. As known from other cold-tolerant herbaceous species, growth of Arabidopsis at low temperature resulted in increases in photosynthetic capacity per unit leaf area and chlorophyll. Growth at high irradiance had a similar effect. However, the growth temperature and irradiance showed interacting effects for several capacity-related variables. Temperature effects on the ratio between electron transport capacity and carboxylation capacity were also different in low compared to high irradiance grown Arabidopsis. The carboxylation capacity per unit Rubisco, a measure for the in vivo Rubisco activity, was low in low irradiance grown plants but there was no clear growth temperature effect. The limitation of photosynthesis by the utilization of triose-phosphate in high temperature grown plants was less when grown at low compared to high irradiance. Several of these traits contribute to reduced efficiency of the utilization of resources for photosynthesis of Arabidopsis at low irradiance. The two accessions from contrasting climates showed remarkably similar capabilities of developmental acclimation to the two environmental factors. Hence, no evidence was found for photosynthetic adaptation of the photosynthetic apparatus to specific climatic conditions.

Ontogenetic changes in leaf traits of tropical rainforest trees differing in juvenile light requirement.

Relationships between leaf traits and the gap dependence for regeneration, and ontogenetic changes therein, were investigated in juvenile and adult tropical rainforest tree species. The juveniles of the 17 species included in the study were grown in high light, similar to the exposed crowns of the adult trees. The traits were structural, biomechanical, chemical and photosynthetic. With increasing species gap dependence, leaf mass per area (LMA) decreased only slightly in juveniles and remained constant in adults, whereas punch strength together with tissue density decreased, and photosynthetic capacity and chlorophyll increased. Contrary to what has been mostly found in evergreen tropical rainforest, the trade-off between investment in longevity and in productivity was evident at an essentially constant LMA. Of the traits pertaining to the chloroplast level, photosynthetic capacity per unit chlorophyll increased with gap dependence, but the chlorophyll a/b ratio showed no relationship. Adults had a twofold higher LMA, but leaf strength was on average only about 50% larger. Leaf tissue density, and chlorophyll and leaf N per area were also higher, whereas chlorophyll and leaf N per unit dry mass were lower. Ranking of the species, relationships between traits and with the gap dependence of the species were similar for juveniles and adults. However, the magnitudes of most ontogenetic changes were not clearly related to a species' gap dependence. The adaptive value of the leaf traits for juveniles and adults is discussed.

Leaf scale quantification of the effect of photosynthetic gas exchange on Δ47 of CO2.

The clumped isotope composition (Δ47, the anomaly of the mass 47 isotopologue relative to the abundance expected from a random isotope distribution) of CO2 has been suggested as an additional tracer for gross CO2 fluxes. However, the effect of photosynthetic gas exchange on Δ47 has not been directly determined and two indirect/conceptual studies reported contradicting results. In this study, we quantify the effect of photosynthetic gas exchange on Δ47 of CO2 using leaf cuvette experiments with one C4 and two C3 plants. The experimental results are supported by calculations with a leaf cuvette model. Our results demonstrate the important roles of the Δ47 value of CO2 entering the leaf, kinetic fractionation as CO2 diffuses into, and out of the leaf and CO2-H2O isotope exchange with leaf water. We experimentally confirm the previously suggested dependence of Δ47 of CO2 in the air surrounding a leaf on the stomatal conductance and back-diffusion flux. Gas exchange can enrich or deplete the Δ47 of CO2 depending on the Δ47 of CO2 entering the leaf and the fraction of CO2 exchanged with leaf water and diffused back to the atmosphere, but under typical ambient conditions, it will lead to a decrease in Δ47.

Estimating mesophyll conductance to CO2: methodology, potential errors, and recommendations.

The three most commonly used methods for estimating mesophyll conductance (g(m)) are described. They are based on gas exchange measurements either (i) by themselves; (ii) in combination with chlorophyll fluorescence quenching analysis; or (iii) in combination with discrimination against (13)CO(2). To obtain reliable estimates of g(m), the highest possible accuracy of gas exchange is required, particularly when using small leaf chambers. While there may be problems in achieving a high accuracy with leaf chambers that clamp onto a leaf with gaskets, guidelines are provided for making necessary corrections that increase reliability. All methods also rely on models for the calculation of g(m) and are sensitive to variation in the values of the model parameters. The sensitivity to these factors and to measurement error is analysed and ways to obtain the most reliable g(m) values are discussed. Small leaf areas can best be measured using one of the fluorescence methods. When larger leaf areas can be measured in larger chambers, the online isotopic methods are preferred. Using the large CO(2) draw-down provided by big chambers, and the isotopic method, is particularly important when measuring leaves with high g(m) that have a small difference in [CO(2)] between the substomatal cavity and the site of carboxylation in the chloroplast (C(i)-C(c) gradient). However, equipment for the fluorescence methods is more easily accessible. Carbon isotope discrimination can also be measured in recently synthesized carbohydrates, which has its advantages under field conditions when large number of samples must be processed. The curve-fitting method that uses gas exchange measurements only is not preferred and should only be used when no alternative is available. Since all methods have their weaknesses, the use of two methods for the estimation of g(m), which are as independent as possible, is recommended.

Evolution of Increased Photosynthetic Capacity and Its Underlying Traits in Invasive Jacobaea vulgaris.

The evolution of increased competitive ability (EICA) hypothesis and the shifting defense hypothesis (SDH) predict that evolutionary changes occur in a suite of traits related to defense and growth in invasive plant species as result of the absence of specialist herbivores. We tested how this suite of traits changed due to the absence of specialist herbivores in multiple invasive regions that differ in climatic conditions with native and invasive Jacobaea vulgaris in a controlled environment. We hypothesized that invasive J. vulgaris in all invasive regions have i) a higher plant growth and underlying traits, such as photosynthetic capacity, ii) lower regrowth-related traits, such as carbohydrate storage, and iii) an increased plant qualitative defense, such as pyrrolizidine alkaloids (PAs). Our results show that invasive J. vulgaris genotypes have evolved a higher photosynthetic rate and total PA concentration but a lower investment in root carbohydrates, which supports the SDH hypothesis. All the traits changed consistently and significantly in the same direction in all four invasive regions, indicative of a parallel evolution. Climatic and soil variables did differ between ranges but explained only a very small part of the variation in trait values. The latter suggests that climate and soil changes were not the main selective forces on these traits.

Functional significance of shade-induced leaf senescence in dense canopies: an experimental test using transgenic tobacco.

Canopy photosynthesis models have predicted an optimal leaf area index (LAI; leaf area per unit surface area) and leaf nitrogen distribution at which whole-plant carbon gain per unit N is maximized. In this study we experimentally tested these models, using transgenic P(SAG12)-IPT tobacco (SAG; Nicotiana tabacum L.) plants with delayed leaf senescence and therefore a greater LAI and more uniform N distribution than the wild type (WT). In a competition experiment, the increased density of surrounding WT plants caused a greater reduction in dry mass of mature SAG target plants than in that of WT target plants, indicating negative effects of delayed leaf senescence on performance at high canopy density. Vegetative SAG plants achieved a lower calculated daily carbon gain than competing WT plants because the former retained leaves with a negative carbon gain in the shaded, lower part of the canopy. Sensitivity analyses showed that the carbon gain of SAG plants would increase if these lower leaves were shed and the N reallocated from these leaves were used to form additional leaf area at the canopy top. This strategy, which is adopted by the WT, is most advantageous because it results in the shading of competing neighbors.

(15)N in tree rings as a bio-indicator of changing nitrogen cycling in tropical forests: an evaluation at three sites using two sampling methods.

Anthropogenic nitrogen deposition is currently causing a more than twofold increase of reactive nitrogen input over large areas in the tropics. Elevated (15)N abundance (δ(15)N) in the growth rings of some tropical trees has been hypothesized to reflect an increased leaching of (15)N-depleted nitrate from the soil, following anthropogenic nitrogen deposition over the last decades. To find further evidence for altered nitrogen cycling in tropical forests, we measured long-term δ(15)N values in trees from Bolivia, Cameroon, and Thailand. We used two different sampling methods. In the first, wood samples were taken in a conventional way: from the pith to the bark across the stem of 28 large trees (the "radial" method). In the second, δ(15)N values were compared across a fixed diameter (the "fixed-diameter" method). We sampled 400 trees that differed widely in size, but measured δ(15)N in the stem around the same diameter (20 cm dbh) in all trees. As a result, the growth rings formed around this diameter differed in age and allowed a comparison of δ(15)N values over time with an explicit control for potential size-effects on δ(15)N values. We found a significant increase of tree-ring δ(15)N across the stem radius of large trees from Bolivia and Cameroon, but no change in tree-ring δ(15)N values over time was found in any of the study sites when controlling for tree size. This suggests that radial trends of δ(15)N values within trees reflect tree ontogeny (size development). However, for the trees from Cameroon and Thailand, a low statistical power in the fixed-diameter method prevents to conclude this with high certainty. For the trees from Bolivia, statistical power in the fixed-diameter method was high, showing that the temporal trend in tree-ring δ(15)N values in the radial method is primarily caused by tree ontogeny and unlikely by a change in nitrogen cycling. We therefore stress to account for tree size before tree-ring δ(15)N values can be properly interpreted.

Establishment of Plantago lanceolata L. and Plantago major L. among grass : II. Shade tolerance of seedlings and selection on time of germination.

Establishment of Plantago lanceolata and P. major ssp major among grass was studied in a field experiment in which survival and selection on date of seedling emergence and plant size was investigated in relation to the vegetation structure. P. major - in contrast to P. lanceolata - was not able to establish itself in grass because of its lower competitive ability caused by later germination, smaller seedling size, and shorter leaves. In both species there was selection for early germination. For P. lanceolata a significant correlation was found between the strength of selection and the light climate, determined by the structure of the grass sward. Plants that germinated early were at an advantage because they were larger, especially the leaves, when compared with plants that germinated late. It seems likely that selection was mainly by competition for light. Contrary to expectation P. major-seedlings had a higher shade tolerance than those of P. lanceolata. The performance of both species is discussed in relation to their different life strategies.

Establishment of Plantago lanceolata L. and Plantago major L. among grass : I. Significance of light for germination.

The influence of temperature, stratification, red/far-red ratio of the light (R/FR) and combinations of these factors on the germination of Plantago lanceolata and Plantago major ssp major have been investigated under controlled conditions. Only quantitative differences were found in the response of the species to the various environmental factors investigated except for germination in darkness. P. lanceolata germinated in darkness, the percentage depending on environmental factors and seed collection, whereas P. major seed swere found to have a virtually absolute light requirement. P. lanceolata germinated at lower temperatures and was less inhibited at a low R/FR than P. major. Pot experiments and a field experiment were carried out in order to study the influence of the presence of grass on germination and emergence. P. major was more inhibited than P. lanceolata when sown among grass and the presence of vegetation did not inhibit the germination of P. lanceolata in the field experiment. The latter could be ascribed to the dormancy alleviating effect of low winter temperatures to which the seeds had been subjected. The inhibition of emergence of P. major among grass could be explained by the light factor only, whereas in the case of P. lanceolata an additional factor must be involved.

Photosynthetic induction in saplings of three shade-tolerant tree species: comparing understorey and gap habitats in a French Guiana rain forest.

The photosynthetic induction response under constant and fluctuating light was examined in naturally occurring saplings (about 0.5-2 m in height) of three shade-tolerant tree species, Pourouma bicolor spp digitata, Dicorynia guianensis, and Vouacapoua americana, growing in bright gaps and in the shaded understorey in a Neotropical rain forest. Light availability to saplings was estimated by hemispherical photography. Photosynthetic induction was measured in the morning on leaves that had not yet experienced direct sunlight. In Dicorynia, the maximum net photosynthesis rate (A max) was similar between forest environments (ca 4 µmol m-2 s-1), whereas for the two other species, it was twice as high in gaps (ca 7.5) as in the understorey (ca 4.5). However, the time required to reach 90% of A max did not differ among species, and was short, 7-11 min. Biochemical induction was fast in leaves of Pourouma, as about 3 min were needed to reach 75% of maximum carboxylation capacity (V cmax); the two other species needed 4-5 min. When induction continued after reaching 75% of V cmax, stomatal conductance increased in Pourouma only (ca 80%), causing a further increase in its net photosynthesis rate. When fully induced leaves were shaded for 20 min, loss of induction was moderate in all species. However, gap saplings of Dicorynia had a rapid induction loss (ca 80%), which was mainly due to biochemical limitation as stomatal conductance decreased only slowly. When leaves were exposed to a series of lightflecks separated by short periods of low light, photosynthetic induction increased substantially and to a similar extent in all species. Although A max was much lower in old than in young leaves as measured in Dicorynia and Vouacapoua, variables of the dynamic response of photosynthesis to a change in light tended to be similar between young and old leaves. Old leaves, therefore, might remain important for whole-plant carbon gain, especially in understorey environments. The three shade-tolerant species show that, particularly in low light, they are capable of efficient sunfleck utilization.

Importance of the gradient in photosynthetically active radiation in a vegetation stand for leaf nitrogen allocation in two monocotyledons.

Carex acutiformis and Brachypodium pinnatum were grown with a uniform distribution of photosynthetic photon flux density (PFD) with height, and in a vertical PFD gradient similar to the PFD gradient in a leaf canopy. Distribution of organic leaf N and light-saturated rates of photosynthesis were determined. These parameters were also determined on plants growing in a natural vegetation stand. The effect of a PFD gradient was compared with the effect of a leaf canopy. In Brachypodium, plants growing in a vegetation stand had increasing leaf N with plant height. However, distribution of leaf N was not influenced by the PFD gradient treatment. The gradient of leaf N in plants growing in a leaf canopy was not due to differences within the long, mostly erect, leaves but to differences between leaves. In Carex, however, the PFD gradient caused a clear increase of leaf N with height in individual leaves and thus also in plants. The leaf N gradient was similar to that of plants growing in a leaf canopy. Leaf N distribution was not affected by nutrient availability in Carex. In most cases, photosynthesis was positively related to leaf N. Hence, lightsaturated rates of photosynthesis increased towards the top of the plants growing in leaf canopies in both species and, in Carex, also in the PFD gradient, thus contributing to increased N use efficiency for photosynthesis of the whole plant. It is concluded that in Carex the PFD gradient is the main environmental signal for leaf N allocation in response to shading in a leaf canopy, but one or more other signals must be involved in Brachypodium.

Analysis of differences in photosynthetic nitrogen use efficiency of alpine and lowland Poa species.

This study investigates factors determining variation in photosynthetic nitrogen use efficiency (φN) in seven slow- and fast-growing Poa species from altitudinally contrasting sites. The species and their environmental origin were (in order of increasing relative growth rate): two alpine (Poa fawcettiae and P. costiniana), one sub-alpine (P. alpina) and three temperate lowland perennials (P. pratensis, P. compressa and P. trivialis), as well as one temperate lowland annual (P. annua). Plants were grown hydroponically under identical conditions with free access to nutrients in a growth room. Photosynthesis per unit leaf area measured at growth irradiance (500 µmol m-2 s-1) was slightly higher in the slow-growing alpine species. At saturating light intensities, photosynthesis was considerably higher in the alpine species than in the lowland species. Carboxylation capacity and Rubisco content per unit leaf area were also greater in the alpine species. Despite variation between the species, the in vivo specific activity of Rubisco showed little relationship to relative growth rate or photosynthetic rate. Both at light saturation and at the growth irradiance, φN was lowest in the slow-growing alpine species P. fawcettiae, P. costiniana and P. alpina, and highest in the fast-growing P. compressa and P. annua. The proportion of leaf nitrogen that was allocated to photosynthetic capacity and the in vivo catalytic constant of Rubisco accounted for most of the variation in φN at light saturation. Minor variations in intercellular CO2 partial pressure also contributed to some extent to the variations in φN at light saturation. The low φN values at growth irradiance exhibited by the alpine species were additionally due to a lower percentage utilisation of their high photosynthetic capacity compared to the lowland species.

Midday depression of net photosynthesis in the tropical rainforest tree Eperua grandiflora: contributions of stomatal and internal conductances, respiration and Rubisco functioning.

High midday temperatures can depress net photosynthesis. We investigated possible mechanisms underlying this phenomenon in leaves of Eperua grandiflora (Aubl.) Benth. saplings. This tropical tree establishes in small gaps in the rainforest canopy where direct sunlight can raise midday temperatures markedly. We simulated this microclimate in a growth chamber by varying air temperature between 28 and 38 degrees C at constant vapor pressure. A decrease in stomatal conductance in response to an increase in leaf-to-air vapor pressure difference (deltaW) caused by an increase in leaf temperature (Tleaf) was the principal reason for the decrease in net photosynthesis between 28 and 33 degrees C. Net photosynthesis decreased further between 33 and 38 degrees C. Direct effects on mesophyll functioning and indirect effects through deltaW were of similar magnitude in this temperature range. Mitochondrial respiration during photosynthesis was insensitive to Tleaf over the investigated temperature range; it thus did not contribute to midday depression of net photosynthesis. Internal conductance for CO2 diffusion in the leaf, estimated by combined gas exchange and chlorophyll fluorescence measurements, decreased slightly with increasing Tleaf. However, the decrease in photosynthetic rate with increasing Tleaf was larger and thus the difference in CO2 partial pressure between the substomatal cavity and chloroplast was smaller, leading to the conclusion that this factor was not causally involved in midday depression. Carboxylation capacity inferred from the CO2 response of photosynthesis increased between 28 and 33 degrees C, but remained unchanged between 33 and 38 degrees C. Increased oxygenation of ribulose-1,5-bisphosphate relative to its carboxylation and the concomitant increase in photorespiration with increasing Tleaf were thus not compensated by an increase in carboxylation capacity over the higher temperature range. This was the principal reason for the negative effect of high midday temperatures on mesophyll functioning.

The effect of irradiance on the carbon balance and tissue characteristics of five herbaceous species differing in shade-tolerance.

The carbon balance is defined here as the partitioning of daily whole-plant gross CO2 assimilation (A) in C available for growth and C required for respiration (R). A scales positively with growth irradiance and there is evidence for an irradiance dependence of R as well. Here we ask if R as a fraction of A is also irradiance dependent, whether there are systematic differences in C-balance between shade-tolerant and shade-intolerant species, and what the causes could be. Growth, gas exchange, chemical composition and leaf structure were analyzed for two shade-tolerant and three shade-intolerant herbaceous species that were hydroponically grown in a growth room at five irradiances from 20 µmol m(-2) s(-1) (1.2 mol m(-2) day(-1)) to 500 µmol m(-2) s(-1) (30 mol m(-2) day(-1)). Growth analysis showed little difference between species in unit leaf rate (dry mass increase per unit leaf area) at low irradiance, but lower rates for the shade-tolerant species at high irradiance, mainly as a result of their lower light-saturated rate of photosynthesis. This resulted in lower relative growth rates in these conditions. Daily whole-plant R scaled with A in a very tight manner, giving a remarkably constant R/A ratio of around 0.3 for all but the lowest irradiance. Although some shade-intolerant species showed tendencies toward a higher R/A and inefficiencies in terms of carbon and nitrogen investment in their leaves, no conclusive evidence was found for systematic differences in C-balance between the shade-tolerant and intolerant species at the lowest irradiance. Leaf tissue of the shade-tolerant species was characterized by high dry matter percentages, C-concentration and construction costs, which could be associated with a better defense in shade environments where leaf longevity matters. We conclude that shade-intolerant species have a competitive advantage at high irradiance due to superior potential growth rates, but that shade-tolerance is not necessarily associated with a better C-balance at low irradiance. Under those conditions tolerance to other stresses is probably more important for the performance of shade-tolerant species.

The art of growing plants for experimental purposes: a practical guide for the plant biologist.

Every year thousands of experiments are conducted using plants grown under more-or-less controlled environmental conditions. The aim of many such experiments is to compare the phenotype of different species or genotypes in a specific environment, or to study plant performance under a range of suboptimal conditions. Our paper aims to bring together the minimum knowledge necessary for a plant biologist to set up such experiments and apply the environmental conditions that are appropriate to answer the questions of interest. We first focus on the basic choices that have to be made with regard to the experimental setup (e.g. where are the plants grown; what rooting medium; what pot size). Second, we present practical considerations concerning the number of plants that have to be analysed considering the variability in plant material and the required precision. Third, we discuss eight of the most important environmental factors for plant growth (light quantity, light quality, CO2, nutrients, air humidity, water, temperature and salinity); what critical issues should be taken into account to ensure proper growth conditions in controlled environments and which specific aspects need attention if plants are challenged with a certain a-biotic stress factor. Finally, we propose a simple checklist that could be used for tracking and reporting experimental conditions.