Responses to Drought Stress in Poplar: What Do We Know and What Can We Learn?

Poplar (Populus spp.) is a high-value crop for wood and biomass production and a model organism for tree physiology and genomics. The early release, in 2006, of the complete genome sequence of P. trichocarpa was followed by a wealth of studies that significantly enriched our knowledge of complex pathways inherent to woody plants, such as lignin biosynthesis and secondary cell wall deposition. Recently, in the attempt to cope with the challenges posed by ongoing climate change, fundamental studies and breeding programs with poplar have gradually shifted their focus to address the responses to abiotic stresses, particularly drought. Taking advantage from a set of modern genomic and phenotyping tools, these studies are now shedding light on important processes, including embolism formation (the entry and expansion of air bubbles in the xylem) and repair, the impact of drought stress on biomass yield and quality, and the long-term effects of drought events. In this review, we summarize the status of the research on the molecular bases of the responses to drought in poplar. We highlight how this knowledge can be exploited to select more tolerant genotypes and how it can be translated to other tree species to improve our understanding of forest dynamics under rapidly changing environmental conditions.

Thylakoid proteome variation of Eutrema salsugineum in response to drought and salinity combined stress.

It is well known that plant responses to stress involve different events occurring at different places of the cell/leaf and at different time scales in relation with the plant development. In fact, the organelles proteomes include a wide range of proteins that could include a wide range of proteins showing a considerable change in cellular functions and metabolism process. On this basis, a comparative proteomics analysis and fluorescence induction measurements were performed to investigate the photosynthetic performance and the relative thylakoid proteome variation in Eutrema salsugineum cultivated under salt stress (200 mM NaCl), water deficit stress (PEG) and combined treatment (PEG + NaCl) as a hyperosmotic stress. The obtained results showed a significant decrease of plant growth under drought stress conditions, with the appearance of some toxicity symptoms, especially in plants subjected to combined treatment. Application of salt or water stress alone showed no apparent change in the chlorophyll a fluorescence transients, primary photochemistry (fluorescence kinetics of the O-J phase), the PQ pool state (J-I phase changes), (Fv/Fm) and (Fk/Fj) ratios. However, a considerable decrease of all these parameters was observed under severe osmotic stress (PEG + NaCl). The thylakoid proteome analysis revealed 58 proteins showing a significant variation in their abundance between treatments (up or down regulation). The combined treatment (PEG + NaCl) induced a decrease in the expression of the whole PSII core subunit (D1, D2, CP43, CP47, PsbE and PsbH), whereas the OEC subunits proteins remained constant. An increase in the amount of PsaD, PsaE, PsaF, PsaH, PsaK and PsaN was detected under drought stress (PEG5%). No significant change in the accumulation of Cyt b6 and Cyt f was observed. Some regulated proteins involved in cellular redox homeostasis were detected (glutamine synthetase, phosphoglycerate kinase, transketolase), and showed a significant decrease under the combined treatment. Some oxidative stress related proteins were significantly up-regulated under salt or drought stress and could play a crucial role in the PSI photoprotection and the control of ROS production level.

Photosynthetic performance of quinoa (Chenopodium quinoa Willd.) after exposure to a gradual drought stress followed by a recovery period.

Drought is an abiotic scourge, one of the major environmental stress factors that adversely affect plant growth and photosynthesis machinery through a disruption of cell organelles, arrangement thylakoid membranes and the electron transport chain. Herein, we probed the effect of drought stress on photosynthetic performance of Chenopodium quinoa Willd. Beforehand, plants were subjected to water deficit (as 15% Field Capacity, FC) for one (D-1W) or two weeks (D-2W), and were then re-watered at 95% FC for 2 weeks. Light and electron microscopy analysis of leaves showed no apparent changes in mesophyll cell organization and chloroplast ultrastructure after one week of drought stress, while a swelling of thylakoids and starch accumulation were observed after the prolonged drought (D-2W). The latter induced a decrease in both PSI and PSII quantum yields which was accompanied by an increase in F0 (minimum fluorescence) and a decline in Fm (maximum fluorescence). Drought stress influenced the fluorescence transients, where the major changes at the OJIP prompt FI level were detected in the OJ and IP phases. Prolonged drought induced a decrease in chl a fluorescence at IP phase which was readjusted and established back after re-watering and even more an increase was observed after 2 weeks of recovery. The maximum quantum yield of primary photochemistry (φPo) was unaffected by the different drought stress regimes. Drought induced an increase in the ABS/RC and DI0/RC ratios which was concurrent to a stable φPo (maximum quantum yield of PSII primary photochemistry). A substantial decrease in PI(ABS) was detected especially, during severe drought stress (D-2W) suggesting a drop in the PSII efficiency and the level of electron transport through the plastoquinone pool (PQ pool) towards oxidized PSI RCs (P700+). The immunoblot analysis of the main PSII proteins revealed considerable changes in the D1, D2, CP47, OEC, PsbQ and LHCII proteins under drought. These changes depend on the stress duration and recovery period. The main message of this investigation is the elevated recovery capacities of PSII and PSI photochemical activities after re-watering.

Phytoremediation of a Highly Arsenic Polluted Site, Using Pteris vittata L. and Arbuscular Mycorrhizal Fungi.

Phytoremediation is a promising green technique for the restoration of a polluted environment, but there is often a gap between lab and field experiments. The fern, Pteris vittata L., can tolerate a high soil arsenic concentration and rapidly accumulate the metalloid in its fronds. Arbuscular mycorrhizal fungi (AMF) are mutualistic fungi that form a symbiosis with most land plants' roots, improve their growth, and induce stress tolerance. This paper reports the results obtained using P. vittata inoculated with AMF, to extract Arsenic (As) from an industrial site highly contaminated also by other pollutants. Two experiments have been performed. In the first one, AMF colonized ferns were grown for two years under controlled conditions in soil coming from the metallurgic site. Positive effects on plant health and As phytoextraction and accumulation were detected. Then, considering these results, we performed a three year in situ experiment in the industrial site, to assess the remediation of As at two different depths. Our results show that the colonization of P. vittata with AMF improved the remediation process of As with a significant impact on the depth 0-0.2 m.

Tomato responses to Funneliformis mosseae during the early stages of arbuscular mycorrhizal symbiosis.

The concept of symbiosis can be described as a continuum of interactions between organisms ranging from mutualism to parasitism that can also change over time. Arbuscular mycorrhizal fungi (AMF) are among the most important obligate plant symbionts. Once the symbiosis is well established, mycorrhizal plants are more tolerant to biotic or abiotic stresses, so the AMF relationship with the host plant is generally described as mutualistic. However, little is known about AMF effects on the plant during the early stages of root colonization. The aim of this work was to assess the type of interaction (mutualistic or parasitic) between the arbuscular mycorrhizal (AM) fungus Funelliformis mosseae and Solanum lycopersicum cv. Rio Grande plants, at 7, 14, 21, and 28 days after inoculation (DAI), considering that in the adopted experimental design (one plant per pot), the seedling was the only carbon source for fungus development in the absence of common mycorrhizal networks with other plants. At each harvest, mycorrhizal colonization, shoot and root weights, morphometric parameters, and photosynthetic efficiency were evaluated. The presence of the AM fungus in the tomato root system was observed starting from the 14th DAI, and its level increased over time. Few effects of the fungus presence on the considered parameters were observed, and no stress symptoms ever appeared; so, we can state that the fungus behaved as a mutualistic symbiont during the early stages of plant growth. Moreover, a trend towards a positive effect on plant growth was observed at 28 DAI in mycorrhizal plants.

Light Weight, Easy Formable and Non-ToxicPolymer-Based Composites for Hard X-rayShielding: A Theoretical and Experimental Study.

Composite lightweight materials for X-ray shielding applications were studied anddeveloped with the goal of replacing traditional screens made of lead and steel, with innovativematerials with similar shielding properties, but lighter, more easily formed and workable, with lowerimpact on the environment and reduced toxicity for human health. New epoxy-based compositesadditivated with barium sulfate and bismuth oxide were designed through simulations performedwith software based on Geant4. Then, they were prepared and characterized using differenttechniques starting from digital radiography in order to test the radiopacity of the composites,in comparison with traditional materials. The lower environmental impact and toxicity of theseinnovative screens were quantified by Life Cycle Assessment (LCA) calculation based on the ecoinventdatabase, within the openLCA framework. Optimized mixtures are (i) 20% epoxy/60% bismuthoxide/20% barite, which guarantees the best performance in X-ray shielding, largely overcomingsteel, but higher in costs and a weight reduction of circa 60%; (ii) 20% epoxy/40% bismuth oxide/40%barite which has slightly lower performances in shielding, but it is lighter and cheaper than thefirst one and (iii) the 20% epoxy/20% bismuth oxide/60% barite which is the cheapest material, stillmaintaining the X-ray shielding of steel. Depending on the cost/efficiency request of the specificapplication (industrial ra.

Hybrid inorganic (nonporous silica)/organic (alginate) core-shell platform for targeting a cisplatin-based Pt(IV) anticancer prodrug.

Nonporous silica nanoparticles with an external shell containing the 3­aminopropyl arm (SiNP) were further decorated with alginic acid (SiNP-ALG) as a potential biocompatible delivery system for Pt antitumor agents. Such particles were coupled with the prodrug (OC­6­44)­acetato(ß­alaninato)diamminedichloridoplatinum(IV), 1, through the formation of amide bonds between the pendant carboxylate groups on SiNP-ALG and the free amino group of the complex. Cytosol extracted from tumor cells was able to quickly and efficiently reduce the Pt(IV) prodrug, and produces the active metabolite cisplatin. SiNP-ALG-Pt conjugate was more active than both cisplatin and 1, due to its more efficient cell uptake, whereas the SiNP-ALG unplatinated nanoparticles were deprived of any nonspecific toxicity.

Comparative analysis of salt stress, duration and intensity, on the chloroplast ultrastructure and photosynthetic apparatus in Thellungiella salsuginea.

Salinity is one of the most important abiotic stress affecting plant growth and productivity worldwide. Photosynthesis, together with cell growth, is among the primary process affected by salinity. Here, we report the effects of salt stress on photosynthesis in the model halophyte Thellungiella salsuginea. Plants were grown in hydroponic system and then treated for 2 weeks with different NaCl concentrations (0, 100, 200 and 400 mM). Leaf analysis using both photonic and transmission electron microscopes showed some changes in mesophyll cell organization, including shape and dimension. Under high NaCl concentration (400 mM) a swelling of thylakoids and starch accumulation was also observed. The obtained results also showed a change in the photosynthetic efficiency of both photosystems (PSI and PSII), depending on both NaCl concentrations and duration of the stress treatment. Under moderate salinity (100 and 200 mM NaCl) no significant variation was observed in PSI and PSII yield parameters. Chlorophyll a fluorescence transient showed some variations in OJ, JI and IP phases under salt stress depending also on NaCl levels and the duration of stress. Under high salinity PSII donor side was affected as well as quantum yield of PSI which also showed a donor side limitation. A significant decrease on quantum yields Y(I) and Y(II) under high salt treatment (400 mM NaCl) for prolonged period of time (15 days) was observed. The decrease of these parameters was quantitatively compensated by a corresponding increase of energy thermal dissipation Y(NPQ) in photosystem II and a increase in the Y(ND) in PSI. Analysis of derived parameters from the OJIP transient curve revealed that ABS/RC decreased under NaCl treatment by reason of the increase in size of antenna of active reaction centers. An increase in the performance index PI(ABS), a slight decrease in the rate of DIO/RC, TRO/RC and the level of electron transport per PSII RC (ETO/RC) were observed during the first days of salt stress treatment reflecting a high PSII efficiency.

Arbuscular mycorrhizal fungi and plant growth-promoting pseudomonads improve yield, quality and nutritional value of tomato: a field study.

The aim of this work was to assess the effects of plant-beneficial microorganisms (two Pseudomonas strains and a mixed mycorrhizal inoculum, alone or in combination) on the quality of tomato fruits of plants grown in the field and subjected to reduced fertilization. Pseudomonas strain 19Fv1T was newly characterized during this study. The size and quality of the fruits (concentration of sugars, organic acids and vitamin C) were assessed. The microorganisms positively affected the flower and fruit production and the concentrations of sugars and vitamins in the tomato fruits. In particular, the most important effect induced by arbuscular mycorrhizal (AM) fungi was an improvement of citric acid concentration, while bacteria positively modulated sugar production and the sweetness of the tomatoes. The novelty of the present work is the application of soil microorganisms in the field, in a real industrial tomato farm. This approach provided direct information about the application of inocula, allowed the reduction of chemical inputs and positively influenced tomato quality.

Palladium uptake by Pisum sativum: partitioning and effects on growth and reproduction.

Environmental palladium levels are increasing because of anthropogenic activities. The considerable mobility of the metal, due to solubilisation phenomena, and its known bioavailability may indicate interactions with higher organisms. The aim of the study was to determine the Pd uptake and distribution in the various organs of the higher plant Pisum sativum and the metal-induced effects on its growth and reproduction. P. sativum was grown in vermiculite with a modified Hoagland's solution of nutrients in the presence of Pd at concentrations ranging 0.10-25 mg/L. After 8-10 weeks in a controlled environment room, plants were harvested and dissected to isolate the roots, stems, leaves, pods and peas. The samples were analysed for Pd content using AAS and SEM-EDX. P. sativum absorbed Pd, supplied as K2PdCl4, beginning at seed germination and continuing throughout its life. Minimal doses (0.10-1.0 mg Pd/L) severely inhibited pea reproductive processes while showing a peculiar hormetic effect on root development. Pd concentrations ≥1 mg/L induced developmental delay, with late growth resumption, increased leaf biomass (up to 25%) and a 15-20% reduction of root mass. Unsuccessful repeated blossoming efforts led to misshapen pods and no seed production. Photosynthesis was also disrupted. The absorbed Pd (ca. 0.5 % of the supplied metal) was primarily fixed in the root, specifically in the cortex, reaching concentrations up to 200 µg/g. The metal moved through the stem (up to 1 µg/g) to the leaves (2 µg/g) and pods (0.3 µg/g). The presence of Pd in the pea fruits, together with established evidence of environmental Pd accumulation and bioavailability, suggests possible contamination of food plants and propagation in the food chain and must be the cause for concern.

AM fungi and PGP pseudomonads increase flowering, fruit production, and vitamin content in strawberry grown at low nitrogen and phosphorus levels.

There is increasing interest in the quality of crops because of the implications concerning health, economic revenue, and food quality. Here we tested if inoculation with a mixture of arbuscular mycorrhizal fungi (AMF) and/or two strains of plant growth-promoting bacteria (PGPB), in conditions of reduced chemical inputs, affects the quality and yield of strawberry fruits. Fruit quality was measured by concentrations of soluble sugars, various organic acids, and two vitamins (ascorbic and folic acid). Co-inoculation with the AMF and each of the two PGPB resulted in increased flower and fruit production, larger fruit size, and higher concentrations of sugars and ascorbic and folic acid in comparison with fruits of uninoculated plants. These results provide further evidence that rhizospheric microorganisms affect fruit crop quality and show that they do so even under conditions of reduced chemical fertilization and can thus be exploited for sustainable agriculture.

Arbuscular mycorrhizal fungi and plant growth-promoting pseudomonads increases anthocyanin concentration in strawberry fruits (Fragaria x ananassa var. Selva) in conditions of reduced fertilization.

Anthocyanins are a group of common phenolic compounds in plants. They are mainly detected in flowers and fruits, are believed to play different important roles such as in the attraction of animals and seed dispersal, and also in the increase of the antioxidant response in tissues directly or indirectly affected by biotic or abiotic stress factors. As a major group of secondary metabolites in plants commonly consumed as food, they are of importance in both the food industry and human nutrition. It is known that arbuscular mycorrhizal (AM) fungi can influence the plant secondary metabolic pathways such as the synthesis of essential oils in aromatic plants, of secondary metabolites in roots, and increase flavonoid concentration. Plant Growth-Promoting Bacteria (PGPB) are able to increase plant growth, improving plant nutrition and supporting plant development under natural or stressed conditions. Various studies confirmed that a number of bacterial species living on and inside the root system are beneficial for plant growth, yield and crop quality. In this work it is shown that inoculation with AM fungi and/or with selected and tested Pseudomonas strains, under conditions of reduced fertilization, increases anthocyanin concentration in the fruits of strawberry.

Effects of Pseudomonas putida S1Pf1Rif against chrysanthemum yellows phytoplasma infection.

Phytoplasmas cause damage on a number of plant species leading to relevant economical loss. Up to now, strategies to limit their spread led to only partial success. In this context, the use of plant-beneficial bacteria to control phytoplasmas has never been explored. The aim of this work was to assess the effect of Pseudomonas putida S1Pf1Rif against chrysanthemum yellows phytoplasma (CYP) infection of daisy. Plant biomass, root architecture, symptom severity, phytoplasma titer, and viability were evaluated in inoculated and control plants. CYP reduced plant growth and root development. Although the phytoplasma titer in young apical leaves was not affected by inoculation with S1Pf1Rif, the pseudomonad improved plant growth of CYP-infected plants. Whereas CYP titer increased over time in uninoculated plants, its viability decreased, regardless of the presence of P. putida S1Pf1Rif. Finally, phytoplasma cells in fully developed leaves of CYP-infected plants inoculated with S1Pf1Rif often appeared degenerated. Overall, our results indicate that P. putida S1Pf1Rif is able to alleviate the disease, although it does not affect the presence of viable phytoplasmas in young, developing leaves of the infected plants.