Lower Mesophyll Conductance Research Articles

Halotolerant PGPB Staphylococcus sciuri ET101 protects photosynthesis through activation of redox dissipation pathways in Lycopersicon esculentum

Photosynthesis is known to be seriously affected by salt stress. The stress induced membrane damage leads to disrupted photosynthetic components causing imbalance between production and utilization of ATP/NADPH with generation of ROS leading to photoinhibition and photodamage. In the current study, role of halotolerant plant growth promoting bacteria (PGPB) Staphylococcus sciuri ET101 in protection of photosynthesis in tomato plants during salinity stress was evaluated by analysing changes in antioxidant defense and activation of redox dissipation pathways. Inoculation of S. sciuri ET101 significantly enhanced the growth of tomato plants with significantly higher photosynthetic rates (PN) under normal and salinity stress conditions. Further, increased membrane stability, soluble sugar accumulation and significant decrease in malondialdehyde (MDA) content in leaves of ET101 inoculated tomato plants under normal and salinity were observed along with increased expression of antioxidant genes for efficient ROS detoxification and suppression of oxidative damage. Additionally, salinity induced decrease in rate of photosynthesis (PN) due to lowered chloroplastic CO2 concentration (Cc) attributed by low mesophyll conductance (gm) in uninoculated plants was alleviated by ET101 inoculation showing significantly higher carboxylation rate (Vcmax), RuBP generation (Jmax) and increased photorespiration (PR). The genes involved in photorespiratory process, cyclic electron flow (CEF), and alternative oxidase (AOX) pathway of mitochondrial respiration were abundantly expressed in leaves of ET101 inoculated plants indicating their involvement in protecting photosynthesis from salt stress induced photoinhibition. Collectively, our results indicated that S. sciuri ET101 has the potential in protecting photosynthesis of tomato plants under salinity stress through activation of redox dissipation pathways.

Physiological and Transcriptomic Analyses Revealed the Implications of Abscisic Acid in Mediating the Rate-Limiting Step for Photosynthetic Carbon Dioxide Utilisation in Response to Vapour Pressure Deficit in Solanum Lycopersicum (Tomato).

The atmospheric vapour pressure deficit (VPD) has been demonstrated to be a significant environmental factor inducing plant water stress and affecting plant photosynthetic productivity. Despite this, the rate-limiting step for photosynthesis under varying VPD is still unclear. In the present study, tomato plants were cultivated under two contrasting VPD levels: high VPD (3–5 kPa) and low VPD (0.5–1.5 kPa). The effect of long-term acclimation on the short-term rapid VPD response was examined across VPD ranging from 0.5 to 4.5 kPa. Quantitative photosynthetic limitation analysis across the VPD range was performed by combining gas exchange and chlorophyll fluorescence. The potential role of abscisic acid (ABA) in mediating photosynthetic carbon dioxide (CO2) uptake across a series of VPD was evaluated by physiological and transcriptomic analyses. The rate-limiting step for photosynthetic CO2 utilisation varied with VPD elevation in tomato plants. Under low VPD conditions, stomatal and mesophyll conductance was sufficiently high for CO2 transport. With VPD elevation, plant water stress was gradually pronounced and triggered rapid ABA biosynthesis. The contribution of stomatal and mesophyll limitation to photosynthesis gradually increased with an increase in the VPD. Consequently, the low CO2 availability inside chloroplasts substantially constrained photosynthesis under high VPD conditions. The foliar ABA content was negatively correlated with stomatal and mesophyll conductance for CO2 diffusion. Transcriptomic and physiological analyses revealed that ABA was potentially involved in mediating water transport and photosynthetic CO2 uptake in response to VPD variation. The present study provided new insights into the underlying mechanism of photosynthetic depression under high VPD stress.

High leaf mass per area Oryza genotypes invest more leaf mass to cell wall and show a low mesophyll conductance.

The intraspecific variations of leaf structure and anatomy in rice leaves and their impacts on gas diffusion are still unknown. Researches about the tradeoff between structural compositions and intracellular chemical components within rice leaves are still lacking. The objectives of the present study were to investigate the varietal differences in leaf structure and leaf chemical compositions, and the tradeoff between leaf structural tissues and intracellular chemical components in rice leaves. Leaf structure, leaf anatomy, leaf chemical composition concentrations and gas exchange parameters were measured on eight Oryza sativa L. genotypes to investigate the intraspecific variations in leaf structure and leaf anatomy and their impacts on gas exchange parameters, and to study the tradeoff between leaf structural compositions (cell wall compounds) and intracellular chemical components (non-structural carbohydrates, nitrogen, chlorophyll). Leaf thickness increased with leaf mass per area (LMA), while leaf density did not correlate with LMA. Mesophyll cell surface area exposed to intercellular airspace (IAS) per leaf area, the surface area of chloroplasts exposed to IAS and cell wall thickness increased with LMA. Cell wall compounds accounted for 71.5 % of leaf dry mass, while mass-based nitrogen and chlorophyll concentrations decreased with LMA. Mesophyll conductance was negatively correlated with LMA and cell wall thickness. High LMA rice genotypes invest more leaf mass to cell wall and possess a low mesophyll conductance.

Waterlogging of Winter Crops at Early and Late Stages: Impacts on Leaf Physiology, Growth and Yield.

Waterlogging is expected to increase as a consequence of global climate change, constraining crop production in various parts of the world. This study assessed tolerance to 14-days of early- or late-stage waterlogging of the major winter crops wheat, barley, rapeseed and field pea. Aerenchyma formation in adventitious roots, leaf physiological parameters (net photosynthesis, stomatal and mesophyll conductances, chlorophyll fluorescence), shoot and root growth during and after waterlogging, and seed production were evaluated. Wheat produced adventitious roots with 20–22% of aerenchyma, photosynthesis was maintained during waterlogging, and seed production was 86 and 71% of controls for early- and late-waterlogging events. In barley and rapeseed, plants were less affected by early- than by late-waterlogging. Barley adventitious roots contained 19% aerenchyma, whereas rapeseed did not form aerenchyma. In barley, photosynthesis was reduced during early-waterlogging mainly by stomatal limitations, and by non-stomatal constraints (lower mesophyll conductance and damage to photosynthetic apparatus as revealed by chlorophyll fluorescence) during late-waterlogging. In rapeseed, photosynthesis was mostly reduced by non-stomatal limitations during early- and late-waterlogging, which also impacted shoot and root growth. Early-waterlogged plants of both barley and rapeseed were able to recover in growth upon drainage, and seed production reached ca. 79–85% of the controls, while late-waterlogged plants only attained 26–32% in seed production. Field pea showed no ability to develop root aerenchyma when waterlogged, and its photosynthesis (and stomatal and mesophyll conductances) was rapidly decreased by the stress. Consequently, waterlogging drastically reduced field pea seed production to 6% of controls both at early- and late-stages with plants being unable to resume growth upon drainage. In conclusion, wheat generates a set of adaptive responses to withstand 14 days of waterlogging, barley and rapeseed can still produce significant yield if transiently waterlogged during early plant stages but are more adversely impacted at the late stage, and field pea is not suitable for areas prone to waterlogging events of 14 days at either growth stage.

Mesophyll conductance in cotton bracts: anatomically determined internal CO2 diffusion constraints on photosynthesis.

Mesophyll conductance (gm) has been shown to affect photosynthetic capacity and thus the estimates of terrestrial carbon balance. While there have been some attempts to model gm at the leaf and larger scales, the potential contribution of gm to the photosynthesis of non-leaf green organs has not been studied. Here, we investigated the influence of gm on photosynthesis of cotton bracts and how it in turn is influenced by anatomical structures, by comparing leaf palisade and spongy mesophyll with bract tissue. Our results showed that photosynthetic capacity in bracts is much lower than in leaves, and that gm is a limiting factor for bract photosynthesis to a similar extent to stomatal conductance. Bract and the spongy tissue of leaves have lower mesophyll conductance than leaf palisade tissue due to the greater volume fraction of intercellular air spaces, smaller chloroplasts, lower surface area of mesophyll cells and chloroplasts exposed to leaf intercellular air spaces and, perhaps, lower membrane permeability. Comparing bracts with leaf spongy tissue, although bracts have a larger cell wall thickness, they have a similar gm estimated from anatomical characteristics, likely due to the cumulative compensatory effects of subtle differences in each subcellular component, especially chloroplast traits. These results provide the first evidence for anatomical constraints on gm and photosynthesis in non-leaf green organs.

Mesophyll conductance does not contribute to greater photosynthetic rate per unit nitrogen in temperate compared with tropical evergreen wet-forest tree leaves.

Globally, trees originating from high-rainfall tropical regions typically exhibit lower rates of light-saturated net CO2 assimilation (A) compared with those from high-rainfall temperate environments, when measured at a common temperature. One factor that has been suggested to contribute towards lower rates of A is lower mesophyll conductance. Using a combination of leaf gas exchange and carbon isotope discrimination measurements, we estimated mesophyll conductance (gm ) of several Australian tropical and temperate wet-forest trees, grown in a common environment. Maximum Rubisco carboxylation capacity, Vcmax , was obtained from CO2 response curves. gm and the drawdown of CO2 across the mesophyll were both relatively constant. Vcmax estimated on the basis of intercellular CO2 partial pressure, Ci , was equivalent to that estimated using chloroplastic CO2 partial pressure, Cc , using 'apparent' and 'true' Rubisco Michaelis-Menten constants, respectively Having ruled out gm as a possible factor in distorting variations in A between these tropical and temperate trees, attention now needs to be focused on obtaining more detailed information about Rubisco in these species.

A few enlarged chloroplasts are less efficient in photosynthesis than a large population of small chloroplasts in Arabidopsis thaliana

The photosynthetic, biochemical, and anatomical traits of accumulation and replication of chloroplasts (arc) mutants of Arabidopsis thaliana were investigated to study the effects of chloroplast size and number on photosynthesis. Chloroplasts were found to be significantly larger, and the chloroplast surface area exposed to intercellular air spaces (Sc) significantly lower in the mutants than in their wild-types. The decreased Sc and increase cytoplasm thickness in the mutants resulted in a lower mesophyll conductance (gm) and a consequently lower chloroplast CO2 concentration (Cc). There were no significant differences between the mutants and their wild-types in maximal carboxylation rate (Vcmax), maximal electron transport (Jcmax), and leaf soluble proteins. Leaf nitrogen (N) and Rubisco content were similar in both Wassilewskija (Ws) wild-type (Ws-WT) and the Ws mutant (arc 8), whereas they were slightly higher in Columbia (Col) wild-type (Col-WT) than the Col mutant (arc 12). The photosynthetic rate (A) and photosynthetic N use efficiency (PNUE) were significantly lower in the mutants than their wild-types. The mutants showed similar A/Cc responses as their wild-type counterparts, but A at given Cc was higher in Col and its mutant than in Ws and its mutant. From these results, we conclude that decreases in gm and Cc are crucial to the reduction in A in arc mutants.

Dissecting molecular and physiological response mechanisms to high solar radiation in cyanic and acyanic leaves: a case study on red and green basil.

Photosynthetic performance and the expression of genes involved in light signaling and the biosynthesis of isoprenoids and phenylpropanoids were analysed in green ('Tigullio', TIG) and red ('Red Rubin', RR) basil. The aim was to detect the physiological and molecular response mechanisms to high sunlight. The attenuation of blue-green light by epidermal anthocyanins was shown to evoke shade-avoidance responses with consequential effects on leaf morpho-anatomical traits and gas exchange performance. Red basil had a lower mesophyll conductance, partially compensated by the less effective control of stomatal movements, in comparison with TIG. Photosynthesis decreased more in TIG than in RR in high sunlight, because of larger stomatal limitations and the transient impairment of PSII photochemistry. The methylerythritol 4-phosphate pathway promoted above all the synthesis and de-epoxidation of violaxanthin-cycle pigments in TIG and of neoxanthin and lutein in RR. This enabled the green leaves to process the excess radiant energy effectively, and the red leaves to optimize light harvesting and photoprotection. The greater stomatal closure observed in TIG than in RR was due to enhanced abscisic acid (ABA) glucose ester deglucosylation and reduced ABA oxidation, rather than to superior de novo ABA synthesis. This study shows a strong competition between anthocyanin and flavonol biosynthesis, which occurs at the level of genes regulating the oxidation of the C2-C3 bond in the dihydro-flavonoid skeleton.

Extremely thick cell walls and low mesophyll conductance: welcome to the world of ancient living!

Mesophyll conductance is thought to be an important photosynthetic limitation in gymnosperms, but they currently constitute the most understudied plant group in regard to the extent to which photosynthesis and intrinsic water use efficiency are limited by mesophyll conductance. A comprehensive analysis of leaf gas exchange, photosynthetic limitations, mesophyll conductance (calculated by three methods previously used for across-species comparisons), and the underlying ultra-anatomical, morphological and chemical traits in 11 gymnosperm species varying in evolutionary history was performed to gain insight into the evolution of structural and physiological controls on photosynthesis at the lower return end of the leaf economics spectrum. Two primitive herbaceous species were included in order to provide greater evolutionary context. Low mesophyll conductance was the main limiting factor of photosynthesis in the majority of species. The strongest sources of limitation were extremely thick mesophyll cell walls, high chloroplast thickness and variation in chloroplast shape and size, and the low exposed surface area of chloroplasts per unit leaf area. In gymnosperms, the negative relationship between net assimilation per mass and leaf mass per area reflected an increased mesophyll cell wall thickness, whereas the easy-to-measure integrative trait of leaf mass per area failed to predict the underlying ultrastructural traits limiting mesophyll conductance.

The photosynthetic capacity in 35 ferns and fern allies: mesophyll CO2 diffusion as a key trait.

Ferns and fern allies have low photosynthetic rates compared with seed plants. Their photosynthesis is thought to be limited principally by physical CO2 diffusion from the atmosphere to chloroplasts. The aim of this study was to understand the reasons for low photosynthesis in species of ferns and fern allies (Lycopodiopsida and Polypodiopsida). We performed a comprehensive assessment of the foliar gas-exchange and mesophyll structural traits involved in photosynthetic function for 35 species of ferns and fern allies. Additionally, the leaf economics spectrum (the interrelationships between photosynthetic capacity and leaf/frond traits such as leaf dry mass per unit area or nitrogen content) was tested. Low mesophyll conductance to CO2 was the main cause for low photosynthesis in ferns and fern allies, which, in turn, was associated with thick cell walls and reduced chloroplast distribution towards intercellular mesophyll air spaces. Generally, the leaf economics spectrum in ferns follows a trend similar to that in seed plants. Nevertheless, ferns and allies had less nitrogen per unit DW than seed plants (i.e. the same slope but a different intercept) and lower photosynthesis rates per leaf mass area and per unit of nitrogen.

The arc mutants of Arabidopsis with fewer large chloroplasts have a lower mesophyll conductance.

Photosynthetic cells of most land plant lineages have numerous small chloroplasts even though most algae, and even the early diverging land plant group the hornworts, tend to have one or a few large chloroplasts. One constraint that small chloroplasts could improve is the resistance to CO2 diffusion from the atmosphere to the chloroplast stroma. We examined the mesophyll conductance (inverse of the diffusion resistance) of mutant Arabidopsis thaliana plants with one or only a few large chloroplasts per cell. The accumulation and replication of chloroplasts (arc) mutants of A. thaliana were studied by model fitting to gas exchange data and (13)CO2 discrimination during carbon fixation. The two methods generally agreed, but the value of the CO2 compensation point of Rubisco (Γ *) used in the model had a large impact on the estimated photosynthetic parameters, including mesophyll conductance. We found that having only a few large chloroplasts per cell resulted in a 25-50 % reduction in the mesophyll conductance at ambient CO2.

Ecophysiological significance of leaf traits in Cypripedium and Paphiopedilum

There is a close phylogenetic relationship between Paphiopedilum and Cypripedium, but these two genera diverge considerably in terms of their leaf traits. To understand the evolution and the ecophysiological significance of leaf traits, we investigated the leaf traits of three Paphiopedilum species and three Cypripedium species in southwestern China. Cypripedium tibeticum and C. flavum showed a significantly higher light-saturated photosynthetic rate (P(max) ), stomatal conductance (g(s) ), photosynthetic nitrogen utilization efficiency (PNUE) and specific leaf area (SLA), but lower ratio of leaf carbon to nitrogen content (C/N) and leaf construction cost (CC) than Paphiopedilum. These leaf traits of Cypripedium suggest its high resource use efficiency and high growth rate reflecting adaptation to a short growing period and abundant soil nutrients and water in alpine habitats. Conversely, the low P(max) , g(s) , PNUE, SLA and the ratio of chlorophyll a to chlorophyll b (Chl a/b), but high leaf nitrogen investment in light-harvesting component (P(L) ), CC and C/N in Paphiopedilum indicate its adaptation to a low light, nutrient-poor and limited soil water habitats in karst areas. As a sympatric species of Paphiopedilum, although C. lentiginosum retained the phylogenetic leaf traits of Cypripedium, such as high mass-based light-saturated photosynthetic rate (P(max-M) ), g(s) and PNUE, it had some similar leaf traits to those of Paphiopedilum, such as low mesophyll conductance (g(m) ) and Chl a/b, and high P(L) , which reflected an adaptation to the same habitat. Our results show that the evolution of the leaf traits of Paphiopedilum and Cypripedium are shaped by both phylogeny and environment.

Leaf succulence determines the interplay between carboxylase systems and light use during Crassulacean acid metabolism in Kalanchoë species*

The photosynthetic physiology of Crassulacean acid metabolism was investigated in two Kalanchoë species with differing leaf succulence. The magnitude of CAM was higher for the more succulent leaves of K. daigremontiana, compared to the less succulent leaves of K. pinnata. High succulence was related to low mesophyll conductance: K. pinnata was able to maximize diurnal carbon gain by the C(3) pathway, whereas increased succulence is associated with a higher commitment to the CAM cycle in K. daigremontiana. The Rubisco specificity factor, tau, determining selectivity for CO(2) over O(2), was similar for both species (approximately 88), and lower than that of Spinacea (approximately 95), but in contrast to C(4) plants, the Rubisco K(mCO(2)) (determined independently) was also lower in Kalanchoë spp. than in spinach. Differences in light use were related to the nature of the sink strength in each Phase of CAM, with PEPC activity resulting in low electron transport rates. Decarboxylation was marked by high, non-saturated rates of electron transport, with Rubisco activity and activation state increasing in both species during the course of the light period. The degree of succulence, and extent of CAM activity, was associated with a progressive inhibition of PSII photochemistry and potential Rubisco activity during the night in both species. Rubisco could be 'woken up' more rapidly in K. pinnata, whereas 45 min light acclimation was required for full recovery of electron transport and Rubisco activity in K. daigremontiana. Leaf morphology therefore seems to alter the expression of and dependence on CAM, but also the extent of co-regulation of carboxylase networks and light use capacity.

Photosynthesis and photoprotection responses to water stress in the wild-extinct plant Lysimachia minoricensis

Lysimachia minoricensis is an endemic species of the Balearic Islands that has become extinct in the wild, but persists in botanical gardens. Attempts of re-introducing the species into its natural habitat, which consisted in temporary dry streams, have failed. Low genetic variability has been reported for the garden individuals, suggesting that a reduced potential to adapt to environmental changes could be among the reasons for its extinction. In the present study, we particularly test whether photosynthesis and photoprotection responses of this species to water stress could help explaining the lack of success of this species in its natural habitat. Plants of L. minoricensis were grown in pots in a growth chamber. Soil water depletion was imposed over 20 days by stopping irrigation. Early stomata closure was observed in response to soil water depletion while leaves desiccated progressively. Although net photosynthesis was low in irrigated plants, due to a remarkably low mesophyll conductance to CO 2, substantial photosynthetic activity was kept at severe drought, where leaf relative water content was as low as 50%, suggesting that L. minoricensis is a very drought-tolerant species. In parallel with decreased photosynthesis, thermal dissipation of the excess light and photorespiration progressively increased. The former was linearly related to increased de-epoxidation of the xanthophylls cycle. Photoprotection was effective, as pre-dawn maximum photochemical efficiency was maintained higher than 0.75 through the entire experiment. Moreover, photosynthetic capacity was largely (80%) recovered only 24 h after re-watering. These results show that stomatal regulation, photosynthetic metabolism and photoprotection in L. minoricensis are well adapted to water stress, suggesting that additional factors may be responsible for its status as a wild-extinct plant.

Relationships between leaf conductance to CO2 diffusion and photosynthesis in micropropagated grapevine plants, before and after ex vitro acclimatization

In vitro-cultured plants typically show a low photosynthetic activity, which is considered detrimental to subsequent ex vitro acclimatization. Studies conducted so far have approached this problem by analysing the biochemical and photochemical aspects of photosynthesis, while very little attention has been paid to the role of leaf conductance to CO(2) diffusion, which often represents an important constraint to CO(2) assimilation in naturally grown plants. Mesophyll conductance, in particular, has never been determined in in vitro plants, and no information exists as to whether it represents a limitation to carbon assimilation during in vitro growth and subsequent ex vitro acclimatization. In this study, by means of simultaneous gas exchange and chlorophyll fluorescence measurements, the stomatal and mesophyll conductance to CO(2) diffusion were assessed in in vitro-cultured plants of the grapevine rootstock '41B' (Vitis vinifera 'Chasselas'xVitis berlandieri), prior to and after ex vitro acclimatization. Their impact on electron transport rate partitioning and on limitation of potential net assimilation rate was analysed. In vitro plants had a high stomatal conductance, 155 versus 50 mmol m(-2) s(-1) in acclimatized plants, which ensured a higher CO(2) concentration in the chloroplasts, and a 7% higher electron flow to the carbon reduction pathway. The high stomatal conductance was counterbalanced by a low mesophyll conductance, 43 versus 285 mmol m(-2) s(-1), which accounted for a 14.5% estimated relative limitation to photosynthesis against 2.1% estimated in acclimatized plants. It was concluded that mesophyll conductance represents an important limitation for in vitro plant photosynthesis, and that in acclimatization studies the correct comparison of photosynthetic activity between in vitro and acclimatized plants must take into account the contribution of both stomatal and mesophyll conductance.

The influence of water stress on leaf and stem photosynthesis in Spartium junceum L.

ABSTRACTStem and leaf photosynthetic responses to environmental parameters were studied in Spartium junceum L., a legume with chlorophyllous stems. Stem net photosynthesis (Pn) was consistently lower than leaf Pn. The low stem Pn was due to lower quantum yield, lower mesophyll conductance and lower CO2‐saturated Pn than that of leaf Pn. Stomatal limitations to leaf and stem Pn were similar (25%). Water stress caused a greater reduction in leaf Pn than that of stems. Leaf Pn was also reduced in water‐stressed plants following rehydration. The reduced leaf Pn was associated with a reduced photon saturated Pn rate and a reduced CO2 saturated Pn rate. Apparent quantum yield, mesophyll conductance and stomatal limitation of leaves were unaffected by water‐stress. Stem Pn following rehydration was not influenced by the water‐stress treatment. In general, leaf Pn was more responsive to environmental parameters and more sensitive to water stress than stem Pn. These data support the hypothesis that stem Pn has greater tolerance of water stress, but is limited to low Pn by biochemical means compared to leaves.

Estimation of Mesophyll Conductance to CO2 Flux by Three Different Methods

The resistance to diffusion of CO(2) from the intercellular airspaces within the leaf through the mesophyll to the sites of carboxylation during photosynthesis was measured using three different techniques. The three techniques include a method based on discrimination against the heavy stable isotope of carbon, (13)C, and two modeling methods. The methods rely upon different assumptions, but the estimates of mesophyll conductance were similar with all three methods. The mesophyll conductance of leaves from a number of species was about 1.4 times the stomatal conductance for CO(2) diffusion determined in unstressed plants at high light. The relatively low CO(2) partial pressure inside chloroplasts of plants with a low mesophyll conductance did not lead to enhanced O(2) sensitivity of photosynthesis because the low conductance caused a significant drop in the chloroplast CO(2) partial pressure upon switching to low O(2). We found no correlation between mesophyll conductance and the ratio of internal leaf area to leaf surface area and only a weak correlation between mesophyll conductance and the proportion of leaf volume occupied by air. Mesophyll conductance was independent of CO(2) and O(2) partial pressure during the measurement, indicating that a true physical parameter, independent of biochemical effects, was being measured. No evidence for CO(2)-accumulating mechanisms was found. Some plants, notably Citrus aurantium and Simmondsia chinensis, had very low conductances that limit the rate of photosynthesis these plants can attain at atmospheric CO(2) level.