José N. Semedo

Long-term elevated air [CO2 ] strengthens photosynthetic functioning and mitigates the impact of supra-optimal temperatures in tropical Coffea arabica and C. canephora species.

The tropical coffee crop has been predicted to be threatened by future climate changes and global warming. However, the real biological effects of such changes remain unknown. Therefore, this work aims to link the physiological and biochemical responses of photosynthesis to elevated air [CO2 ] and temperature in cultivated genotypes of Coffea arabica L. (cv. Icatu and IPR108) and Coffea canephora cv. Conilon CL153. Plants were grown for ca. 10 months at 25/20°C (day/night) and 380 or 700 μl CO2 l(-1) and then subjected to temperature increase (0.5°C day(-1) ) to 42/34°C. Leaf impacts related to stomatal traits, gas exchanges, C isotope composition, fluorescence parameters, thylakoid electron transport and enzyme activities were assessed at 25/20, 31/25, 37/30 and 42/34°C. The results showed that (1) both species were remarkably heat tolerant up to 37/30°C, but at 42/34°C a threshold for irreversible nonstomatal deleterious effects was reached. Impairments were greater in C. arabica (especially in Icatu) and under normal [CO2 ]. Photosystems and thylakoid electron transport were shown to be quite heat tolerant, contrasting to the enzymes related to energy metabolism, including RuBisCO, which were the most sensitive components. (2) Significant stomatal trait modifications were promoted almost exclusively by temperature and were species dependent. Elevated [CO2 ], (3) strongly mitigated the impact of temperature on both species, particularly at 42/34°C, modifying the response to supra-optimal temperatures, (4) promoted higher water-use efficiency under moderately higher temperature (31/25°C) and (5) did not provoke photosynthetic downregulation. Instead, enhancements in [CO2 ] strengthened photosynthetic photochemical efficiency, energy use and biochemical functioning at all temperatures. Our novel findings demonstrate a relevant heat resilience of coffee species and that elevated [CO2 ] remarkably mitigated the impact of heat on coffee physiology, therefore playing a key role in this crop sustainability under future climate change scenarios.

Sustained Photosynthetic Performance of Coffea spp. under Long-Term Enhanced [CO2]

Coffee is one of the world’s most traded agricultural products. Modeling studies have predicted that climate change will have a strong impact on the suitability of current cultivation areas, but these studies have not anticipated possible mitigating effects of the elevated atmospheric [CO2] because no information exists for the coffee plant. Potted plants from two genotypes of Coffea arabica and one of C. canephora were grown under controlled conditions of irradiance (800 μmol m-2 s-1), RH (75%) and 380 or 700 μL CO2 L-1 for 1 year, without water, nutrient or root development restrictions. In all genotypes, the high [CO2] treatment promoted opposite trends for stomatal density and size, which decreased and increased, respectively. Regardless of the genotype or the growth [CO2], the net rate of CO2 assimilation increased (34-49%) when measured at 700 than at 380 μL CO2 L-1. This result, together with the almost unchanged stomatal conductance, led to an instantaneous water use efficiency increase. The results also showed a reinforcement of photosynthetic (and respiratory) components, namely thylakoid electron transport and the activities of RuBisCo, ribulose 5-phosphate kinase, malate dehydrogenase and pyruvate kinase, what may have contributed to the enhancements in the maximum rates of electron transport, carboxylation and photosynthetic capacity under elevated [CO2], although these responses were genotype dependent. The photosystem II efficiency, energy driven to photochemical events, non-structural carbohydrates, photosynthetic pigment and membrane permeability did not respond to [CO2] supply. Some alterations in total fatty acid content and the unsaturation level of the chloroplast membranes were noted but, apparently, did not affect photosynthetic functioning. Despite some differences among the genotypes, no clear species-dependent responses to elevated [CO2] were observed. Overall, as no apparent sign of photosynthetic down-regulation was found, our data suggest that Coffea spp. plants may successfully cope with high [CO2] under the present experimental conditions.

Drought effect on photosynthetic activity, osmolyte accumulation and membrane integrity of two Cicer arietinum genotypes

Drought was induced in chickpea (Cicer arietinum L.) genotypes (ChK 3226 and ILC 3279) differing in yield capacity. Water stress (S1, RWC around 55–50%; S2, RWC ≤ 40%) drastically reduced stomatal conductance (gs) and net photosynthetic rate (PN) in both genotypes. ILC 3279 showed greater photosynthetic capacity (Amax) decreases. Maximum PSII photochemical efficiency (Fv/Fm), photochemical quenching (qP), total chlorophylls (Chls) and carotenoids (Cars) content showed stability in both genotypes under stress, but in S2 ILC 3279 presented an increase in basal fluorescence (F0) and a greater reduction in estimation of quantum yield of linear electron transport (Φe) than ChK 3226. Membrane damage evaluated by electrolyte leakage occurred earlier and was greater in ILC 3279. It also presented a decrease of total fatty acids (TFA) along drought, while in ChK 3226 greater amounts of TFA were observed in S1. In rehydration, PN of S1 plants completely recovered (ILC 3279) or remained slightly below control (ChK 3226). As regards S2 plants, ILC 3279 showed stronger PN and gs reductions than ChK 3226, despite both genotypes totally recovered Amax and chlorophyll (Chl) a fluorescence. ChK 3226 recovered more efficiently from membrane damage. Under control conditions, greater amounts of most of the studied soluble metabolites occurred in ChK 3226 plants. Malate and citrate decreased with water stress (S2) in both genotypes. Sucrose and pinitol (that had a higher concentration than sucrose in both genotypes) increased in ILC 3279 (S1 and S2), and decreased in ChK 3226 (S2). In ILC 3279 proline and asparagine followed similar patterns. Genotypes showed a similar shoot dry mass (DM) in control plants, but root DM was higher in ChK 3226. Drought reduced root and shoot DM in ChK 3226 already under S1, while in ILC 3279 root DM was unaffected by drought and shoot biomass decreased only in S2. Root/shoot ratio was always higher in ChK 3226 but tended to decrease under stress, while the opposite was observed in ILC 3279. No pods were obtained from control plants of both genotypes, or droughted ILC 3279 plants. ChK 3226 produced pods under S1 (higher yield) and S2. Under stress conditions, ChK 3226 was less affected in photosynthetic activity and membrane integrity, showing a better tolerance to drought. This agrees with the better yield of this genotype under water stress. Distinct strategies seem to underlie the different physiological responses of the two genotypes to water deficit. In spite of its significant solutes accumulation, ILC 3279 was more affected in photosynthetic activity and membrane integrity during water stress than ChK 3226, which showed better yield under drought. A relation could not be established between solutes accumulation of ILC 3279 and yield.

Is salt stress tolerance in Casuarina glauca Sieb. ex Spreng. associated with its nitrogen-fixing root-nodule symbiosis? An analysis at the photosynthetic level

Casuarina glauca is an actinorhizal tree which establishes root-nodule symbiosis with N2-fixing Frankia bacteria. This plant is commonly found in saline zones and is widely used to remediate marginal soils and prevent desertification. The nature of its ability to survive in extreme environments and the extent of Frankia contribution to stress tolerance remain unknown. Thus, we evaluated the ability of C. glauca to cope with salt stress and the influence of the symbiosis on this trait. To this end, we analysed the impact of salt on plant growth, mineral contents, water relations, photosynthetic-related parameters and non-structural sugars in nodulated vs. non-nodulated plants. Although the effects on photosynthesis and stomatal conductance started to become measurable in the presence of 200 mM NaCl, photochemical (e.g., photosynthetic electron flow) and biochemical (e.g., activity of photosynthetic enzymes) parameters were only strongly impaired when NaCl levels reached 600 mM. These results indicate the maintenance of high tissue hydration under salt stress, probably associated with enhanced osmotic potential. Furthermore, the maintenance of photosynthetic assimilation potential (A(max)), together with the increase in the quantum yield of down-regulated energy dissipation of PSII (Y(NPQ)), suggested a down-regulation of photosynthesis instead of photo-damaging effects. A comparison of the impact of increasing NaCl levels on the activities of photosynthetic (RubisCO and ribulose-5 phosphate kinase) and respiratory (pyruvate kinase and NADH-dependent malate dehydrogenase) enzymes vs. photosynthetic electron flow and fluorescence parameters, revealed that biochemical impairments are more limiting than photochemical damage. Altogether, these results indicate that, under controlled conditions, C. glauca tolerates high NaCl levels and that this capacity is linked to photosynthetic adjustments.

Plant growth-promoting Burkholderia species isolated from annual ryegrass in Portuguese soils

To search for culturable Burkholderia species associated with annual ryegrass in soils from natural pastures in Portugal, with plant growth‐promoting effects.

Antioxidative ability and membrane integrity in salt-induced responses of Casuarina glauca Sieber ex Spreng. in symbiosis with N2-fixing Frankia Thr or supplemented with mineral nitrogen

The actinorhizal tree Casuarina glauca tolerates extreme environmental conditions, such as high salinity. This species is also able to establish a root-nodule symbiosis with N2-fixing bacteria of the genus Frankia. Recent studies have shown that C. glauca tolerance to high salt concentrations is innate and linked to photosynthetic adjustments. In this study we have examined the impact of increasing NaCl concentrations (200, 400 and 600mM) on membrane integrity as well as on the control of oxidative stress in branchlets of symbiotic (NOD+) and non-symbiotic (KNO3+) C. glauca. Membrane selectivity was maintained in both plant groups at 200mM NaCl, accompanied by an increase in the activity of antioxidative enzymes (superoxide dismutase, ascorbate peroxidase, glutathione reductase and catalase). Regarding cellular membrane lipid composition, linolenic acid (C18:3) showed a significant decline at 200mM NaCl in both NOD+ and KNO3+ plants. In addition, total fatty acids (TFA) and C18:2 also decreased in NOD+ plants at this salt concentration, resulting in malondialdehyde (MDA) production. Such initial impact at 200mM NaCl is probably due to the fact that NOD+ plants are subjected to a double stress, i.e., salinity and low nitrogen availability. At 400mM NaCl a strong reduction of TFA and C18:3 levels was observed in both plant groups. This was accompanied by a decrease in the unsaturation degree of membrane lipids in NOD+. However, in both NOD+ and KNO3+ lipid modifications were not reflected by membrane leakage at 200 or 400mM, suggesting acclimation mechanisms at the membrane level. The fact that membrane selectivity was impaired only at 600mM NaCl in both groups of plants points to a high tolerance of C. glauca to salt stress independently of the symbiotic relation with Frankia.

Colonization and beneficial effects on annual ryegrass by mixed inoculation with plant growth promoting bacteria

Multi-strain inoculants have increased potential to accomplish a diversity of plant needs, mainly attributed to its multi-functionality. This work evaluated the ability of a mixture of three bacteria to colonize and induce a beneficial response on the pasture crop annual ryegrass. Pseudomonas G1Dc10 and Paenibacillus G3Ac9 were previously isolated from annual ryegrass and were selected for their ability to perform multiple functions related to plant growth promotion. Sphingomonas azotifigens DSMZ 18530T was included due to nitrogen fixing ability. The effects of the bacterial mixture were assessed in gnotobiotic plant inoculation assays and compared with single and dual inoculation treatments. Triple inoculation with 3×108 bacteria significantly increased plant dry weight and leaf pigments, indicating improved photosynthetic performance. Plant lipid biosynthesis was enhanced by 65%, mainly due to the rise of linolenic acid, an omega-3 fatty acid with high dietary value. Electrolyte leakage, an indicator of plant membrane stability under stress, was decreased pointing to a beneficial effect by inoculation. Plants physiological condition was more favoured by triple inoculation than by single, although benefits on biomass were only evident relative to non-inoculated plants. The colonization behaviour and coexistence in plant tissues were assessed using FISH and GFP-labelling, combined with confocal microscopy and a cultivation-based approach for quantification. The three strains occupied the same sites, localizing preferentially along root hairs and in stem epidermis. Endophytic colonization was observed as bacteria entered root and stem inner tissues. This study reveals the potential of this mixture of strains for biofertilization, contributing to improve crop productivity and nutritional value.

Sequential zinc and iron biofortification of bread-wheat grains: from controlled to uncontrolled environments

Abstract. The development of knowledge on bread wheat (Triticum aestivum L.) biofortification in zinc (Zn) and iron (Fe), related to its potential agronomical use and the nutritional and technological implications, is becoming important to strategies for improving human nutrition. In this context, we studied the accumulation of Zn and Fe in grains, considering potential uptake and translocation kinetics, photoassimilate production and deposition, and related yields, in grains of cv. Roxo produced under controlled-environment conditions and used thereafter in field trials. The metabolic plasticity of this wheat genotype grown under controlled-environment conditions allowed a 10- and 4-fold enhancement in accumulation of Zn and Fe in the grains after nutrient supplementation with a 5-fold concentrated Hoagland solution (5S), after two generations. Moreover, when these seeds were sown under field conditions and the resulting plants supplemented with or without Zn and Fe, the accumulation of these nutrients decreased within the next two generations. Such field seeds obtained without further Zn and Fe supplementation (with nitrogen only; F3(S) and F4(S)) maintained enhanced levels of Zn (∼400%) and Fe (40–50%) compared with the initial seeds. If Zn and Fe supplement was given to the plants germinated from F2(5S), the subsequent F3(5S) and F4(5S) seeds maintained the Zn increase (∼400%), whereas a further enhancement was observed for Fe, to 75% and 89%, respectively. Toxic limits were not reached for photosynthetic functioning. Even under the highest Zn and Fe supplement dose given to the F3(5S) plants, there was only a slight effect on photosystem II photochemical performance; in fact, enhanced net photosynthesis values were observed. In conclusion, within this experimental design, Zn and Fe biofortification can be obtained without toxicity effects on photosynthetic performance and with negligible modifications to grain texture and nutritional value (protein quality and contents as well as fatty acids).

Salt Stress Tolerance in Casuarina glauca and Its Relation with Nitrogen-Fixing Frankia Bacteria

Salinity is one of the most widespread abiotic stresses. It is estimated that salt stress will cause the loss of more than 50 % of arable land by the year 2050. A promising solution for the recovery of saline soils encompasses the use of actinorhizal plants, a group of perennial dicotyledonous angiosperms highly resilient to extreme environmental conditions. These plants are also able to establish a root nodule symbiosis with N2-fixing actinobacteria of the genus Frankia . Casuarina glauca , the model actinorhizal species, tolerates NaCl concentrations above seawater levels. Such ability seems to be innate and independent of the symbiotic relationship with N2-fixing Frankia. In this work, we present a mini review of the basic mechanisms underlying salt tolerance in C. glauca focusing on the impact of salt on the photosynthesis, redox status, and membrane integrity.

Coffee Responses to Drought, Warming and High [CO 2 ] in a Context of Future Climate Change Scenarios

Climate variability strongly determines agricultural productivity, further causing important economic and social impacts. In a context of global climate changes, the continuous enhancement of agricultural production in the coming years is a major challenge for plant science research. Coffee, one of the most important agricultural commodities worldwide, is grown in more than 80 countries in the tropical region. Several estimates point to a strong reduction on both coffee yields and suitable areas in a near future, mostly related to predicted rising temperature, but also due to changes in intra- and inter-annual rainfall amounts and distributions. Nonetheless, recent findings from our team has shown that the coffee plant is more resilient that usually accepted, and that the negative impacts of rising temperature, at physiological and biochemical levels, were strongly mitigated by enhanced air [CO2], which is considered one of the promoting agents of temperature rise. Also, the identification of ecophysiological and molecular traits that can promote plant acclimation to warming, in particular those related to the C-assimilation pathway, would foster the selection of more adapted/tolerant genotypes. In this context, this work aims at envisage leaf physiological responses in Coffea spp. subjected to supra-optimal temperatures, increased [CO2], and water shortage conditions, contributing to this crop sustainability.