Contribution of deep soil layers to the transpiration of a temperate deciduous forest: Implications for the modelling of productivity.

Climate change is imposing drier atmospheric and edaphic conditions on temperate forests. Here, we investigated how deep soil (down to 300 cm) water extraction contributed to the provision of water in the Fontainebleau-Barbeau temperate oak forest over two years, including the 2018 record drought. Deep water provision was key to sustain canopy transpiration during drought, with layers below 150 cm contributing up to 60% of the transpired water in August 2018, despite their very low density of fine roots. We further showed that soil databases used to parameterize ecosystem models largely underestimated the amount of water extractable from the soil by trees, due to a considerable underestimation of the tree rooting depth. The consensus database established for France gave an estimate of 207 mm for the soil water holding capacity (SWHC) at Fontainebleau-Barbeau, when our estimate based on the analysis of soil water content measurements was 1.9 times as high, reaching 390 ± 17 mm. Running the CASTANEA forest model with the database-derived SWHC yielded a 185 gC m-2 y-1 average underestimation of annual gross primary productivity under current climate, reaching up to 687 ± 117 gC m-2 y-1 under climate change scenario RCP8.5. It is likely that the strong underestimation of SWHC that we show at our site is not a special case, and concerns a large number of forest sites. Thus, we argue for a generalisation of deep soil water content measurements in forests, in order to improve the estimation of SWHC and the simulation of the forest carbon cycle in the current context of climate change.

Polygenic risk scores predict diabetes complications and their response to intensive blood pressure and glucose control.

AIMS/HYPOTHESIS: Type 2 diabetes increases the risk of cardiovascular and renal complications, but early risk prediction could lead to timely intervention and better outcomes. Genetic information can be used to enable early detection of risk. METHODS: We developed a multi-polygenic risk score (multiPRS) that combines ten weighted PRSs (10 wPRS) composed of 598 SNPs associated with main risk factors and outcomes of type 2 diabetes, derived from summary statistics data of genome-wide association studies. The 10 wPRS, first principal component of ethnicity, sex, age at onset and diabetes duration were included into one logistic regression model to predict micro- and macrovascular outcomes in 4098 participants in the ADVANCE study and 17,604 individuals with type 2 diabetes in the UK Biobank study. RESULTS: The model showed a similar predictive performance for cardiovascular and renal complications in different cohorts. It identified the top 30% of ADVANCE participants with a mean of 3.1-fold increased risk of major micro- and macrovascular events (p = 6.3 × 10-21 and p = 9.6 × 10-31, respectively) and a 4.4-fold (p = 6.8 × 10-33) higher risk of cardiovascular death. While in ADVANCE overall, combined intensive blood pressure and glucose control decreased cardiovascular death by 24%, the model identified a high-risk group in whom it decreased the mortality rate by 47%, and a low-risk group in whom it had no discernible effect. High-risk individuals had the greatest absolute risk reduction with a number needed to treat of 12 to prevent one cardiovascular death over 5 years. CONCLUSIONS/INTERPRETATION: This novel multiPRS model stratified individuals with type 2 diabetes according to risk of complications and helped to target earlier those who would receive greater benefit from intensive therapy.

A Variant in the Nicotinic Acetylcholine Receptor Alpha 3 Subunit Gene Is Associated With Hypertension Risks in Hypogonadic Patients.

Ephb6 gene knockout causes hypertension in castrated mice. EPHB6 controls catecholamine secretion by adrenal gland chromaffin cells (AGCCs) in a testosterone-dependent way. Nicotinic acetylcholine receptor (nAChR) is a ligand-gated Ca2+/Na+ channel, and its opening is the first signaling event leading to catecholamine secretion by AGCCs. There is a possibility that nAChR might be involved in EPHB6 signaling, and thus sequence variants of its subunit genes are associated with hypertension risks. CHRNA3 is the major subunit of nAChR used in human and mouse AGCCs. We conducted a human genetic study to assess the association of CHRNA3 variants with hypertension risks in hypogonadic males. The study cohort included 1,500 hypogonadic Chinese males with (750 patients) or without (750 patients) hypertension. The result revealed that SNV rs3743076 in the fourth intron of CHRNA3 was significantly associated with hypertension risks in the hypogonadic males. We further showed that EPHB6 physically interacted with CHRNA3 in AGCCs, providing a molecular basis for nAChR being in the EPHB6 signaling pathway.

EPHA4 regulates vascular smooth muscle cell contractility and is a sex-specific hypertension risk gene in individuals with type 2 diabetes.

OBJECTIVE: We investigated the association of genetic variants of EPHA4, a receptor tyrosine kinase, with hypertension, and its role in vascular smooth muscle cell (VSMC) contractility. METHODS: Data from two human genetic studies, ADVANCE and HCHS/SOL, were analyzed for association of EPHA4 single nucleotide variants (SNVs) with hypertension risks. The effect of EPHA4 signalling on mouse VSMC contractility was assessed. RESULTS: We identified a SNV (rs75843691 hg19 chr2:g.222395371 C>G), located in the third intron of EPHA4 gene, being significantly associated with hypertension in human female patients (P value = 8.3 × 10, below the Bonferroni-corrected critical P value) but not male patients with type 2 diabetes from the ADVANCE clinical trial. We found that EPHA4 was expressed in VSMCs and its stimulation by anti-EPHA4 antibody led to reduced VSMC contractility. Estrogen enhanced the contractility-lowering effect of EPHA4 stimulation. Conversely, siRNA knockdown of Epha4 expression in VSMCs resulted in increased contractility of VSMCs from female mice but not from male mice. CONCLUSION: EPHA4 appears to be a sex-specific hypertension risk gene in type 2 diabetic patients. Forward EPHA4 signalling reduces VSMC contractility, and estrogen is a modifier of this effect. The effect of EPHA4 on VSMCs contractility explains the association of EPHA4 gene with hypertension risks in female patients.

Uromodulin in a Pathway Between Decreased Renal Urate Excretion and Albuminuria.

BACKGROUND: The mechanism explaining the inverse association between renal urate and albumin excretion remains unclear. First, we evaluated the impact of candidate variants in the main urate transporter genes (i.e., SLC2A9, SLC22A12, ABCG2) on the association between fractional excretion of uric acid (FEUA) and urinary albumin/creatinine ratio (uACR). Second, we examined uromodulin and sodium excretion as mediators of the association between FEUA and uACR. METHODS: We performed cross-sectional analysis of 737 French Canadians from the CARTaGENE cohort, a random sample of the Quebec population aged 40-69 years (a total of 20,004 individuals). Individuals with available genotyping and urinary data were obtained from a sub-study including gender-matched pairs with high and low Framingham Risk Score and vascular rigidity index. We further excluded individuals with an estimated glomerular filtration rate <60 ml/min/1.73 m2, glycosuria, and use of confounding medication. A spot urine sample was analyzed. Genotyping was performed using the Illumina Omni2.5-8 BeadChips. Genetic variants were analyzed using an additive model. RESULTS: Final analyses included 593 individuals (45.5% of men; mean age 54.3 ± 8.6). We observed an antagonistic interaction between rs13129697 variant of the SLC2A9 gene and FEUA tertiles on uACR (P = 0.002). Using the mediation analysis, uromodulin explained 32%, fractional excretion of sodium (FENa) 44%, and uromodulin together with FENa explained 70% of the inverse relationship between FEUA and uACR. Bootstrapping process confirmed the role of both mediators. CONCLUSIONS: Our data suggest that the association of albuminuria with decreased renal urate excretion may be modified by the transporter SLC2A9, and mediated by uromodulin and sodium handling.

Analysis of the association of EPHB6, EFNB1 and EFNB3 variants with hypertension risks in males with hypogonadism.

Several members of the EPH kinase family and their ligands are involved in blood pressure regulation, and such regulation is often sex- or sex hormone-dependent, based on animal and human genetic studies. EPHB6 gene knockout (KO) in mice leads to hypertension in castrated males but not in un-manipulated KO males or females. To assess whether this finding in mice is relevant to human hypertension, we conducted a human genetic study for the association of EPHB6 and its two ligands, EFNB1 and EFNB3, with hypertension in hypogonadic patients. Seven hundred and fifty hypertensive and 750 normotensive Han Chinese patients, all of whom were hypogonadic, were genotyped for single nucleotide polymorphisms (SNPs) within the regions of the genes, plus an additional 50 kb 5' of the genes for EPHB6, EFNB1 and EFNB3. An imputed insertion/deletion polymorphism, rs35530071, was found to be associated with hypertension at p-values below the Bonferroni-corrected significance level of 0.0024. This marker is located 5' upstream of the EFNB3 gene start site. Previous animal studies showed that while male EFNB3 gene knockout mice were normotensive, castration of these mice resulted in hypertension, corroborating the results of the human genetic study. Considering the significant associations of EFNB3 SNPs with hypertension in hypogonadic males and supporting evidence from castrated EFNB3 KO mice, we conclude that loss-of-function variants of molecules in the EPHB6 signaling pathway in the presence of testosterone are protective against hypertension in humans.

Evidence from single nucleotide polymorphism analyses of ADVANCE study demonstrates EFNB3 as a hypertension risk gene.

EPH kinases and their ligands, ephrins (EFNs), have vital and diverse biological functions. We recently reported that Efnb3 gene deletion results in hypertension in female but not male mice. These data suggest that EFNB3 regulates blood pressure in a sex- and sex hormone-dependent way. In the present study, we conducted a human genetic study to assess the association of EFNB3 single nucleotide polymorphisms with human hypertension risks, using 3,448 patients with type 2 diabetes from the ADVANCE study (Action in Diabetes and Vascular Disease: Peterax and Diamicron MR Controlled Evaluation). We have observed significant association between 2 SNPs in the 3' untranslated region or within the adjacent region just 3' of the EFNB3 gene with hypertension, corroborating our findings from the mouse model. Thus, our investigation has shown that EFNB3 is a hypertension risk gene in certain individuals.

PROX1 gene CC genotype as a major determinant of early onset of type 2 diabetes in slavic study participants from Action in Diabetes and Vascular Disease: Preterax and Diamicron MR Controlled Evaluation study.

BACKGROUND: The prevalence of diabetic nephropathy varies according to ethnicity. Environmental as well as genetic factors contribute to the heterogeneity in the presentation of diabetic nephropathy. Our objective was to evaluate this heterogeneity within the Caucasian population. METHODS: The geo-ethnic origin of the 3409 genotyped Caucasian type 2 diabetes (T2D) patients of Action in Diabetes and Vascular Disease: Preterax and Diamicron MR Controlled Evaluation was determined using principal component analysis. Genome-wide association studies analyses of age of onset of T2D were performed for geo-ethnic groups separately and combined. RESULTS: The first principal component separated the Caucasian study participants into Slavic and Celtic ethnic origins. Age of onset of diabetes was significantly lower in Slavic patients (P = 7.3 × 10), whereas the prevalence of hypertension (P = 4.9 × 10) and albuminuria (5.1 × 10) were significantly higher. Age of onset of T2D and albuminuria appear to have an important genetic component as the values of these traits were also different between Slavic and Celtic individuals living in the same countries. Common and geo-ethnic-specific loci were found to be associated to age of onset of diabetes. Among the latter, the PROX1/PROX1-AS1 genes (rs340841) had the highest impact. Single-nucleotide polymorphism rs340841 CC genotype was associated with a 4.4 year earlier onset of T2D in Slavic patients living or not in countries with predominant Slavic populations. CONCLUSION: These results reveal the presence of distinct genetic architectures between Caucasian ethnic groups that likely have clinical relevance, among them PROX1 gene is a strong candidate of early onset of diabetes with variations depending on ethnicity.

Environmental control of carbon allocation matters for modelling forest growth.

We aimed to evaluate the importance of modulations of within-tree carbon (C) allocation by water and low-temperature stress for the prediction of annual forest growth with a process-based model. A new C allocation scheme was implemented in the CASTANEA model that accounts for lagged and direct environmental controls of C allocation. Different approaches (static vs dynamic) to modelling C allocation were then compared in a model-data fusion procedure, using satellite-derived leaf production estimates and biometric measurements at c. 104 sites. The modelling of the environmental control of C allocation significantly improved the ability of CASTANEA to predict the spatial and year-to-year variability of aboveground forest growth along regional gradients. A significant effect of the previous year's water stress on the C allocation to leaves and wood was reported. Our results also are consistent with a prominent role of the environmental modulation of sink demand in the wood growth of the studied species. Data available at large scales can inform forest models about the processes driving annual and seasonal C allocation. Our results call for a greater consideration of C allocation drivers, especially sink-demand fluctuations, for the simulations of current and future forest productivity with process-based models.

Reduced blood pressure after smooth muscle EFNB2 deletion and the potential association of EFNB2 mutation with human hypertension risk.

Ephrin B2 (EFNB2) is a ligand for erythropoietin-producing hepatocellular kinases (EPH), the largest family of receptor tyrosine kinases. It has critical functions in many biological systems, but is not known to regulate blood pressure. We generated mice with a smooth muscle cell (SMC)-specific deletion of EFNB2 and investigated its roles in blood pressure regulation and vascular SMC (VSMC) contractility. Male Efnb2 knockout (KO) mice presented reduced blood pressure, whereas female KO mice had no such reduction. Both forward signaling from EFNB2 to EPHs and reverse signaling from EPHs to EFNB2 were involved in regulating VSMC contractility, with EPHB4 serving as a critical molecule for forward signaling, based on crosslinking studies. We also found that a region from aa 313 to aa 331 in the intracellular tail of EFNB2 was essential for reverse signaling regulating VSMC contractility, based on deletion mutation studies. In a human genetic study, we identified five SNPs in the 3' region of the EFNB2 gene, which were in linkage disequilibrium and were significantly associated with hypertension for male but not female subjects, consistent with our findings in mice. The coding (minor) alleles of these five SNPs were protective in males. We have thus discovered a previously unknown blood pressure-lowering mechanism mediated by EFNB2 and identified EFNB2 as a gene associated with hypertension risk in humans.

Leaf concentrate compared with skimmed milk as nutritional supplementation for HIV-infected children: a randomized controlled trial in Burundi.

OBJECTIVE: The effectiveness of leaf concentrate powder (LCP) as a nutritional supplement was established in trials conducted among adolescent girls and pregnant women in India. Here we evaluate LCP, compared with skimmed milk powder (SMP), as a supplement for antiretroviral-naïve children living with HIV in a sub-Saharan African country. DESIGN: Randomized controlled, two-arm, 6-month trial comparing effects of isoproteic (5 g) LCP (10 g daily) and SMP (15 g daily) on HIV-1 viral load, CD4+ cell count/percentage, weight/height-for-age, general blood parameters, diarrhoea, respiratory and HIV-related opportunistic infections. SETTING: Bujumbura and Kirundo, Burundi. SUBJECTS: Eighty-three HIV-positive, antiretroviral-naïve children aged 5-14 years: median (range) CD4+ count, 716 (361-1690) cells/mm3; log10 HIV-1 viral load, 4·39 (1·79-6·00). RESULTS: LCP was equivalent to SMP in relation to HIV-specific blood parameters and did not demonstrate superiority over SMP in relation to Hb. Three children in each arm (LCP, 7·1 % (3/42); SMP, 7·3 % (3/41)) proceeded to antiretroviral therapy because their CD4+ counts fell below 350 cells/mm3. Children in the LCP group reported higher levels of appetite and overall health at 6 months. There were no differences in clinical events or any other outcome measures. LCP was less palatable than SMP to the children in this population, but there were few negative perceptions of appearance, texture and taste. CONCLUSIONS: LCP appears to be equivalent to SMP as a nutritional supplement in this population, despite slightly lower palatability. In relation to viral load and CD4+ count, equivalence may indicate no effect in either group. Effectiveness relative to no supplementation remains to be determined.

Assessing the effects of management on forest growth across France: insights from a new functional-structural model.

BACKGROUND AND AIMS: The structure of a forest stand, i.e. the distribution of tree size features, has strong effects on its functioning. The management of the structure is therefore an important tool in mitigating the impact of predicted changes in climate on forests, especially with respect to drought. Here, a new functional-structural model is presented and is used to assess the effects of management on forest functioning at a national scale. METHODS: The stand process-based model (PBM) CASTANEA was coupled to a stand structure module (SSM) based on empirical tree-to-tree competition rules. The calibration of the SSM was based on a thorough analysis of intersite and interannual variability of competition asymmetry. The coupled CASTANEA-SSM model was evaluated across France using forest inventory data, and used to compare the effect of contrasted silvicultural practices on simulated stand carbon fluxes and growth. KEY RESULTS: The asymmetry of competition varied consistently with stand productivity at both spatial and temporal scales. The modelling of the competition rules enabled efficient prediction of changes in stand structure within the CASTANEA PBM. The coupled model predicted an increase in net primary productivity (NPP) with management intensity, resulting in higher growth. This positive effect of management was found to vary at a national scale across France: the highest increases in NPP were attained in forests facing moderate to high water stress; however, the absolute effect of management on simulated stand growth remained moderate to low because stand thinning involved changes in carbon allocation at the tree scale. CONCLUSIONS: This modelling approach helps to identify the areas where management efforts should be concentrated in order to mitigate near-future drought impact on national forest productivity. Around a quarter of the French temperate oak and beech forests are currently in zones of high vulnerability, where management could thus mitigate the influence of climate change on forest yield.

Transcriptional divergence plays a role in the rewiring of protein interaction networks after gene duplication.

Gene duplication plays a key role in the evolution of protein-protein interaction (PPI) networks. After a gene duplication event, paralogous proteins may diverge through the gain and loss of PPIs. This divergence can be explained by two non-exclusive mechanisms. First, mutations may accumulate in the coding sequences of the paralogs and affect their protein sequences, which can modify, for instance, their binding interfaces and thus their interaction specificity. Second, mutations may accumulate in the non-coding region of the genes and affect their regulatory sequences. The resulting changes in expression profiles can lead to paralogous proteins being differentially expressed and occurring in the cell with different sets of potential interaction partners. These changes could also alter splicing regulation and lead to the inclusion or exclusion of alternative exons. The evolutionary role of these regulatory mechanisms remains largely unexplored. We use bioinformatics analyses of existing PPI data and proteome-wide PPI screening to show that the divergence of transcriptional regulation between paralogs plays a significant role in determining their PPI specificity. Because many gene duplication events are followed by rapid changes in transcriptional regulation, our results suggest that PPI networks may be rewired by gene duplication, without the need for protein to diverge in their binding specificities. This article is part of a Special Issue entitled: From protein structures to clinical applications.

Climate change impacts on tree ranges: model intercomparison facilitates understanding and quantification of uncertainty.

Model-based projections of shifts in tree species range due to climate change are becoming an important decision support tool for forest management. However, poorly evaluated sources of uncertainty require more scrutiny before relying heavily on models for decision-making. We evaluated uncertainty arising from differences in model formulations of tree response to climate change based on a rigorous intercomparison of projections of tree distributions in France. We compared eight models ranging from niche-based to process-based models. On average, models project large range contractions of temperate tree species in lowlands due to climate change. There was substantial disagreement between models for temperate broadleaf deciduous tree species, but differences in the capacity of models to account for rising CO(2) impacts explained much of the disagreement. There was good quantitative agreement among models concerning the range contractions for Scots pine. For the dominant Mediterranean tree species, Holm oak, all models foresee substantial range expansion.

Impact of carbohydrate supply on stem growth, wood and respired CO2 delta13C: assessment by experimental girdling.

The present study examines the impact of the C source (reserves vs current assimilates) on tree C isotope signals and stem growth, using experimental girdling to stop the supply of C from leaves to stem. Two-year-old sessile oaks (Quercus petraea) were girdled at three different phenological periods during the leafy period: during early wood growth (Girdling Period 1), during late wood growth (Girdling Period 2) and just after growth cessation (Girdling Period 3). The measured variables included stem respiration rates, stem radial increment, delta(13)C of respired CO(2) and contents of starch and water-soluble fraction in stems (below the girdle) and leaves. Girdling stopped growth, even early in the growing season, leading to a decrease in stem CO(2) efflux (CO(2R)). Shift in substrate use from recently fixed carbohydrate to reserves (i.e., starch) induced (13)C enrichment of CO(2) respired by stem. However, change in substrate type was insufficient to explain alone all the observed CO(2R) delta(13)C variations, especially at the period corresponding to large growth rate of control trees. The below-girdle mass balance suggested that, during girdling periods, stem C was invested in metabolic pathways other than respiration and stem growth. After Girdling Period 1, the girdle healed and the effects of girdling on stem respiration were reversed. Stem growth restarted and total radial increment was similar to the control one, indicating that growth can be delayed when a stress event occurs early in the growth period. Concerning tree ring, seasonal shift in substrate use from reserves (i.e., starch) to recently fixed carbohydrate is sufficient to explain the observed (13)C depletion of tree ring during the early wood growth. However, the inter-tree intra-ring delta(13)C variability needs to be resolved in order to improve the interpretation of intra-seasonal ring signals in terms of climatic or ecophysiological information. This study highlighted, via carbohydrate availability effects, the importance of the characterization of stem metabolic pathways for a complete understanding of the delta(13)C signals.

Leaf and twig delta13C during growth in relation to biochemical composition and respired CO2.

In deciduous trees, the delta(13)C values of leaves are known to diverge during growth from those of woody organs. The main purpose of this study is to determine whether the divergence in delta(13)C between leaves and current-year twigs of Fagus sylvatica (L.) is influenced by changes (i) in the relative contents of organic matter fractions and (ii) in the delta(13)C of respired CO(2). The delta(13)C values of bulk matter, extractive-free matter, lignin, holocellulose, starch, soluble sugars, water-soluble fraction and respired CO(2), as well as their relative contents in bulk matter were determined. The delta(13)C values of biochemical fractions and respired CO(2) showed very similar temporal variations for both leaves and twigs. Variations in bulk matter delta(13)C during growth were, therefore, poorly explained by changes in biochemical composition or in respiratory fractionation and were attributed to the transition from (13)C-enriched reserves (mainly starch) to (13)C-depleted new photoassimilates. The divergence between leaves and twigs was related to higher values of soluble sugar delta(13)C in twigs. However, the difference between lignin and holocellulose delta(13)C varied during growth. This phenomenon was attributed to the delay between holocellulose and lignin deposition. These results may have implications for analysis of organic matter delta(13)C in trees and forest ecosystems.

Biochemical composition is not the main factor influencing variability in carbon isotope composition of tree rings.

From June to December, we determined the effects of variations in biochemical composition on delta(13)C of tree rings of 2-year-old oaks (Quercus petraea (Matt.) Liebl.) growing under semi-natural conditions, and the dependence of these effects of water stress during the growth season. Percent abundance, carbon concentration and delta(13)C were measured in holocellulose, lignin, extractive-free wood, starch and a water-soluble fraction. Relative concentrations of lignin and holocellulose in the extractive-free wood varied little during the season or in response to water stress, indicating that these compounds could not quantitatively explain the variations in whole-wood delta(13)C. Among all sampled tree rings, the relative concentration of each structural compound (holocellulose and lignin) accounted for less than 5% of the delta(13)C variability in whole wood. Variations in holocellulose and extractive-free wood delta(13)C between tree rings were almost identical (r > 0.95), whereas variations in lignin delta(13)C were less well correlated to these compounds. Whole-wood delta(13)C had a slightly altered isotopic signal compared with that of the structural compounds because of the presence of the extractive component. These results showed that variations in lignin delta(13)C and lignin concentration have little influence on extractive-free wood delta(13)C and whole-wood delta(13)C. Rather, holocellulose influences delta(13)C the most. Thus, we confirmed that, for climatic reconstruction from tree rings, removal of extractives by soxhlet is generally sufficient and sometimes unnecessary. Our findings also indicate that, in the case of rapid and severe water stress, the structural component did not accurately record the associated increase in delta(13)C because of dilution with previously formed organic matter and cessation of trunk growth. The effect of drought on carbon isotope ratios was more pronounced in the extractive compounds, making them good water stress indicators but only on a scale of days to months.

Interpreting the decrease in leaf photosynthesis during flowering in mango.

Little is known about the effect of flowering on leaf photosynthesis. To understand why net photosynthesis (A(net)) is lower in Mangifera indica L. leaves close to inflorescences than in leaves on vegetative shoots, we measured nitrogen and carbohydrate concentrations, chlorophyll a fluorescence and gas exchange in recently matured leaves on vegetative terminals and on floral terminals of 4-year-old trees. We used models to estimate photosynthetic electron fluxes and mesophyll conductance (g(m)). Lower A(net) in leaves close to developing inflorescences was attributable to substantial decreases in stomatal conductance and g(m), and also in photosynthetic capacity as indicated by the decrease in the light-saturated rate of photosynthetic electron transport (J(max)). The decrease in J(max) was the result of decreases in the amount of foliar nitrogen per unit leaf area, and may have been triggered by a decrease in sink activity as indicated by the increase in the hexose:sucrose ratio. Parameters measured on leaves close to panicles bearing set fruits were generally intermediate between those measured on leaves on vegetative shoots and on leaves close to inflorescences, suggesting that the changes in A(net) associated with flowering are reversible.