Stimulation of PGP Bacteria on the Development of Seeds, Plants and Cuttings of the Obligate Halophyte Arthrocaulon (Arthrocnemum) macrostachyum (Moric.) Piirainen & G. Kadereit.

The Earth is undergoing alterations at a high speed, which causes problems such as environmental pollution and difficulty in food production. This is where halophytes are interesting, due to their high potential in different fields, such as remediation of the environment and agriculture. For this reason, it is necessary to deepen the knowledge of the development of halophytes and how plant growth-promoting bacteria (PGP) can play a fundamental role in this process. Therefore, in this work were tested the effects of five PGP bacteria on its rhizosphere and other endophytic bacteria at different concentrations of NaCl on seed germination, plant growth (0 and 171 mM) and cutting growth (0 mM) of Arthrocaulon macrostachyum. The growth promotion in this strict halophyte is highlighted due to the presence of PGP bacteria and the fact that no salt is needed. Thus, without salt, the bacterial strains Kocuria polaris Hv16, Pseudarthrobacter psychrotolerans C58, and Rahnella aceris RTE9 enhanced the biomass production by more than 60% in both stems and roots. Furthermore, germination was encouraged by more than 30% in the presence of both R. aceris RTE9 and K. polaris Hv16 at 171 mM NaCl; the latter also had a biocontrol effect on the fungi that grew on the seeds. Additionally, for the first time in cuttings of this perennial species, the root biomass was improved thanks to the consortium of K. polaris Hv16 and P. psychrotolerans C58. Finally, this study demonstrates the potential of PGPs for optimising the development of halophytes, either for environmental or agronomic purposes.

Salinity Modulates Juncus acutus L. Tolerance to Diesel Fuel Pollution.

Soil contamination with petroleum-derived substances such as diesel fuel has become a major environmental threat. Phytoremediation is one of the most studied ecofriendly low-cost solutions nowadays and halophytes species has been proved to have potential as bio-tools for this purpose. The extent to which salinity influences diesel tolerance in halophytes requires investigation. A greenhouse experiment was designed to assess the effect of NaCl supply (0 and 85 mM NaCl) on the growth and photosynthetic physiology of Juncus acutus plants exposed to 0, 1 and 2.5% diesel fuel. Relative growth rate, water content and chlorophyll a derived parameters were measured in plants exposed to the different NaCl and diesel fuel combinations. Our results indicated that NaCl supplementation worsened the effects of diesel toxicity on growth, as diesel fuel at 2.5% reduced relative growth rate by 25% in the absence of NaCl but 80% in plants treated with NaCl. Nevertheless, this species grown at 0 mM NaCl showed a high tolerance to diesel fuel soil presence in RGR but also in chlorophyll fluorescence parameters that did not significantly decrease at 1% diesel fuel concentration in absence of NaCl. Therefore, this study remarked on the importance of knowing the tolerance threshold to abiotic factors in order to determine the bioremediation capacity of a species for a specific soil or area. In addition, it showed that NaCl presence even in halophytes does not always have a positive effect on plant physiology and it depends on the pollutant nature.

Sarcocornia fruticosa photosynthetic response to short-term extreme temperature events in combination with optimal and sub-optimal salinity concentrations.

Climate change would increase frequency and intensity of extreme events as heat and cold waves. There is a lack of studies that consider the co-occurrence of these waves with other abiotic factors relevant on a climate change scenario as salinity. Therefore, it could be interesting to improve our knowledge about the effects that this co-occurrence could have in different species due to the species specific effect of the photosynthesis tolerance to extreme temperatures. A controlled condition experiment was performed using the salt marsh species Sarcocornia perrnis with eight experimental blocks combining temperature ranges (40-28/22-15/13-5 °C) and salinity concentration on the growth solution (171/1050 mM NaCl). After 3 days of treatment, gas exchange, chlorophyll a fluorescence, pigment profile and water state measurement were applied. Photosynthetic machinery function of this perennial species decreased on for both high and low temperature range. Nevertheless, at 13-5 °C the effect of the salinity was mainly due to diffusion limitations more than to damage on the photosystems. At 40-28 °C, in presence of optimal salinity S. fruticosa was not altered overall. However, high temperatures in combination with high salinity reduced the photosynthetic capacity mainly by reducing the efficiency of the electron transport chain.

Impact of short-term extreme temperature events on physiological performance of Salicornia ramosissima J. Woods under optimal and sub-optimal saline conditions.

Increasing extreme temperature climatic events could exert an important effect on plant photosynthetic performance, which could be modulated by the co-occurrence with other environmental factors, such as salinity, in estuarine ecosystems. Therefore, a mesocosm experiment was designed to assess the impact of temperature events for three days (13/5 °C, 25/13 °C and 40/28 °C) in combination with two NaCl concentrations (171 and 1050 mM NaCl) on the physiological performance of Salicornia ramosissima. Extreme temperature events had a negative impact on S. ramosissima photosynthetic efficiency, this effect being more marked with cold wave at both salinities, compared with heat wave, even in presence of NaCl excess. This differential thermotolerance in the photosynthetic apparatus was ascribed to the greater integrity and functioning of its photosynthetic pathway at high temperature, as indicated by constant gs, Vc,max values at optimal salinity and the higher values of those parameters and gm recorded in combination with NaCl excess. Moreover, S. ramosissima was able to upregulate the energy sink capacity of its photochemical apparatus at elevated temperature and salinity by a greater energy excess dissipation capacity. This could have contributed to reducing the risk of oxidative stress, along with the recorded higher capacity for antioxidant enzyme activity modulation under these conditions.

Investigating the physiological mechanisms underlying Salicornia ramosissima response to atmospheric CO2 enrichment under coexistence of prolonged soil flooding and saline excess.

A 45-days long climatic chamber experiment was design to evaluate the effect of 400 and 700 ppm atmospheric CO2 treatments with and without soil water logging in combination with 171 and 510 mM NaCl in the halophyte Salicornia ramosissima. In order to ascertain the possible synergetic impact of these factors associate to climatic change in this plant species physiological and growth responses. Our results indicated that elevated atmospheric CO2 concentration improved plant physiological performance under suboptimal root-flooding and saline conditions plants. Thus, this positive impact was mainly ascribed to an enhancement of energy transport efficiency, as indicated the greater PG, N and Sm values, and the maintaining of carbon assimilation capacity due to the higher net photosynthetic rate (AN) and water use efficiency (iWUE). This could contribute to reduce the risk of oxidative stress owing to the accumulation of reactive oxygen species (ROS). Moreover, plants grown at 700 ppm had a greater capacity to cope with flooding and salinity synergistic impact by a greater efficiency in the modulation in enzyme antioxidant machinery and by the accumulation of osmoprotective compounds and saturated fatty acids in its tissues. These responses indicate that atmospheric CO2 enrichment would contribute to preserve the development of Salicornia ramosissima against the ongoing process of increment of soil stressful conditions linked with climatic change.

Combined effect of Cr-toxicity and temperature rise on physiological and biochemical responses of Atriplex halimus L.

An experiment was conducted to evaluate the combined effect of temperature (26 and 30 °C) and Cr toxicity (0, 100 and 1000 µM Cr) on growth, photosynthesis, water content, Cr and nutrients uptake and translocation. The role of antioxidative enzyme towards stresses tolerance was also investigated. Results showed that the maximum relative growth rate and leaf area per plant of Atriplex halimus L. were recorded at 100 µM Cr and 26 °C. However, presence of Cr reduced net photosynthetic and stomatal conductance rates. Overall, temperature rise enhanced the toxic effect of Cr by reducing growth and photosynthesis and inducing antioxidant enzymes activities. Furthermore, temperature rise increased nutrient uptake, as well as nutrient translocation to aboveground tissues; while it diminished Cr translocation. Finally, roots were the main sink for Cr accumulation in A. halimus. At 1000 µM Cr, root Cr concentrations reached 7.2 and 9.1 mg g-1 at 26 and 30 °C, respectively; while shoot Cr concentrations were 0.45 and 0.44 mg g-1 (26 and 30 °C, respectively). The high Cr-accumulation in roots suggests that A. halimus presents a great potential for phytoremediation, especially phytostabilisation of Cr contaminated soils.

Disentangling the effect of atmospheric CO2 enrichment on the halophyte Salicornia ramosissima J. Woods physiological performance under optimal and suboptimal saline conditions.

A mesocosm experiment was designed to assess the effect of atmospheric CO2 increment on the salinity tolerance of the C3 halophyte Salicornia ramosissima. Thus, the combined effect of 400 ppm and 700 ppm CO2 at 0, 171 and 510 mM NaCl on plants growth, gas exchange, chlorophyll fluorescence parameters, pigments profiles, antioxidative enzyme activities and water relations was studied. Our results highlighted a positive effect of atmospheric CO2 increment on plant physiological performance under suboptimal salinity concentration (510 mM NaCl). Thus, we recorded higher net photosynthetic rate (AN) values under saline conditions and 700 ppm CO2, being this effect mainly mediated by a reduction of mesophyll (gm) and biochemical limitation imposed to salt excess. In addition, rising atmospheric CO2 led to a better plant water balance, linked with a reduction of stomatal conductante (gs) and an overall increment of osmotic potential (Ѱo) with NaCl concentration increment. In spite of these positive effects, there were no significant biomass variations between any treatments. Being this fact ascribed by the investment of the higher energy fixed for salinity stress defence mechanisms, which allowed plants to maintain more active the photochemical machinery even at high salinities, reducing the risk of ROS production, as indicated an improvement of the electron flux and a rise of the energy dissipation. Finally, the positive effect of the CO2 was also supported by the modulation of pigments profiles (mainly zeaxhantin and violaxhantin) concentrations and anti-oxidative stress enzymes, such as superoxide dismutase (SOD) and ascorbate peroxidase (APx).

Physiological and biochemical mechanisms preventing Cd-toxicity in the hyperaccumulator Atriplex halimus L.

The xero-halophyte Atriplex halimus L., recently described as Cd-hyperaccumulator, was examined to determine Cd toxicity threshold and the physiological mechanisms involved in Cd tolerance. An experiment was conducted to investigate the effect of cadmium from 0 to 1350 µM on chlorophyll fluorescence parameters, gas exchange, photosynthetic pigment concentrations and antioxidative enzyme activities of A. halimus. Cadmium, calcium, iron, manganese, magnesium, potassium, phosphorous, sodium and zinc concentrations were also analyzed. Plants of A. halimus were not able to survive at 1350 µM Cd and the upper tolerance limit was recorded at 650 µM Cd; although chlorosis was observed from 200 µM Cd. Cadmium accumulation increased with increase in Cd supply, reaching maxima of 0.77 and 4.65 mg g(-1) dry weight in shoots and roots, respectively, at 650 µM Cd. Dry mass, shoot length, specific leaf area, relative growth rate, net photosynthetic rate, stomatal conductance, pigments contents and chlorophyll fluorescence were significantly reduced by increasing Cd concentration. However, the activities of superoxide dismutase (SOD; EC1.15.1.1), catalase (CAT; EC1.11.1.6) and guaiacol peroxidase (GPx; EC1.11.1.7) were significantly induced by Cd. Exposures to Cd caused also a significant decrease in P contents in roots, Mg and Mn contents in shoots and Fe and K contents in roots and shoots and had no effect on Ca, Na and Zn contents. The tolerance of A. halimus to Cd stress might be related with its capacity to avoid the translocation of great amounts of Cd in its aboveground tissues and higher activities of enzymatic antioxidants in the leaf.