Maria Luce Bartucca

Iron deficiency in barley plants: phytosiderophore release, iron translocation, and DNA methylation

All living organisms require iron (Fe) to carry out many crucial metabolic pathways. Despite its high concentrations in the geosphere, Fe bio-availability to plant roots can be very scarce. To cope with Fe shortage, plants can activate different strategies. For these reasons, we investigated Fe deficient Hordeum vulgare L. plants by monitoring growth, phytosiderophores (PS) release, iron content, and translocation, and DNA methylation, with respect to Fe sufficient ones. Reductions of plant growth, roots to shoots Fe translocation, and increases in PS release were found. Experiments on DNA methylation highlighted significant differences between fully and hemy-methylated sequences in Fe deficient plants, with respect to Fe sufficient plants. Eleven DNA bands differently methylated were found in starved plants. Of these, five sequences showed significant alignment to barley genes encoding for a glucosyltransferase, a putative acyl carrier protein, a peroxidase, a β-glucosidase and a transcription factor containing a Homeodomin. A resupply experiment was carried out on starved barley re-fed at 13 days after sowing (DAS), and it showed that plants did not recover after Fe addition. In fact, Fe absorption and root to shoot translocation capacities were impaired. In addition, resupplied barley showed DNA methylation/demethylation patterns very similar to that of barley grown in Fe deprivation. This last finding is very encouraging because it indicates as these variations/modifications could be transmitted to progenies.

Italian ryegrass for the phytoremediation of solutions polluted with terbuthylazine

The phytoextraction capacity of Italian ryegrass (Lolium multiflorum L.) to remove terbuthylazine (TBA) from aqueous solution has been assessed using a plant-based biotest (RHIZOtest). Three TBA concentrations (0.5, 1.0 and 2.0 mg L(-1)) were chosen to evaluate the tolerance capacity of the ryegrass. Even though the treatments negatively affected plants, they were able to remove up to 30-40% of TBA. In addition, some enzymatic activities involved in the response to TBA-induced stress were determined. Glutathione S-transferase (GST) has been activated with a TBA-dose dependent trend; ascorbate peroxidase (APX) activities have been induced within the first hours after the treatments, followed by decreases or disappearance in plants exposed to two higher dosages. In conclusion, this case-study highlights that the combination of ryegrass and RHIZOtest resulted to be effective in the remediation of aqueous solutions polluted by TBA.

Nitrate removal from polluted water by using a vegetated floating system

Nitrate (NO3(-)) water pollution is one of the most prevailing and relevant ecological issues. For instance, the wide presence of this pollutant in the environment is dramatically altering the quality of superficial and underground waters. Therefore, we set up a floating bed vegetated with a terrestrial herbaceous species (Italian ryegrass) with the aim to remediate hydroponic solutions polluted with NO3(-). The floating bed allowed the plants to grow and achieve an adequate development. Ryegrass was not affected by the treatments. On the contrary, plant biomass production and total nitrogen content (N-K) increased proportionally to the amount of NO3(-) applied. Regarding to the water cleaning experiments, the vegetated floating beds permitted to remove almost completely all the NO3(-) added from the hydroponic solutions with an initial concentration of 50, 100 and 150 mg L(-1). Furthermore, the calculation of the bioconcentration factor (BCF) indicated this species as successfully applicable for the remediation of solutions polluted by NO3(-). In conclusion, the results highlight that the combination of ryegrass and the floating bed system resulted to be effective in the remediation of aqueous solutions polluted by NO3(-).

Use of two grasses for the phytoremediation of aqueous solutions polluted with terbuthylazine

ABSTRACT The capacity of two grasses, tall fescue (Festuca arundinacea) and orchardgrass (Dactylis glomerata), to remove terbuthylazine (TBA) from polluted solutions has been assessed in hydroponic cultures. Different TBA concentrations (0.06, 0.31, 0.62, and 1.24 mg/L) were chosen to test the capacity of the two grasses to resist the chemical. Aerial biomass, effective concentrations (to cause reductions of 10, 50, and 90% of plant aerial biomass) and chlorophylls contents of orchardgrass were found to be more affected. Tall fescue was found to be more capable of removing the TBA from the growth media. Furthermore, enzymes involved both in the herbicide detoxification and in the response to herbicide-induced oxidative stress were investigated. Glutathione S-transferase (GST, EC. 2.5.1.18) and ascorbate peroxidase (APX, EC. 1.11.1.11) of tall fescue were found to be unaffected by the chemical. GST and APX levels of orchardgrass were decreased by the treatment. These negative modulations exerted by the TBA on the enzyme of orchardgrass explained its lower capacity to cope with the negative effects of the TBA.

Effect of three safeners on sulfur assimilation and iron deficiency response in barley (Hordeum vulgare) plants

BACKGROUND Safeners are agrochemicals used in agriculture to protect crops from herbicide injuries. They act by stimulating herbicide metabolism. As graminaceous plants, to cope with iron (Fe) deficiency, activate sulfur (S) metabolism and release huge amounts of Fe-chelating compounds, or phytosiderophores (PSs), we investigated, in barley plants (Hordeum vulgare, L.) grown in Fe deficiency, the effects of three safeners on two enzymes of S assimilation, cysteine (Cys) and glutathione (GSH), and PS release. Finally, we monitored the root Fe content in plants treated with the most effective safener. RESULTS Generally, all the safeners activated S metabolism and increased Cys and GSH contents. In addition, the safened plants excreted higher levels of PSs. Given that mefenpyr-diethyl (Mef) was the most effective in causing these effects, we assessed the Fe concentration in Mef-treated barley and found higher Fe levels than those in untreated plants. CONCLUSION The three safeners, in different ways but specifically, activated S reductive metabolism and regulated Cys and GSH contents, PS release rate and Fe content (Mef-treated barley). The results of this research provide new indications of the biochemical and physiological mechanisms involved in the safening action.

Interference of three herbicides on iron acquisition in maize plants

The use of herbicides to control weed species could lead to environmental threats due to their persistence and accumulation in the ecosystems and cultivated fields. Nonetheless, the effect of these compounds on plant mineral nutrition in crops has been barely investigated. This study aimed at ascertaining the effect of three herbicides (S-metolachlor, metribuzin and terbuthylazine) on the capacity of maize to acquire iron (Fe). Interferences on plant growth and reductions on the Fe contents were found in the plants treated. Furthermore, root cell viability and functionality losses were ascertained following the treatments, which, in turn, decreased the amount of phytosiderophores (PSs) released by the roots. An investigation carried out in greater depth on root apices of treated plants using an FE-SEM (Scanning Electron Microscope) coupled with EDX (Energy Dispersive X-ray) indicated that the reductions on Fe content started in this part of the roots. Lastly, decreases were found also in copper (Cu+2), zinc (Zn+2) and manganese (Mn+2) content in root apices.

The treatment of duckweed with a plant biostimulant or a safener improves the plant capacity to clean water polluted by terbuthylazine.

Water pollution is becoming alarming since thousands contaminants are dispersed in the aquatic environments, and agricultural practices, for the massive use of pesticides, are contributing to exacerbating this problem. In this context, a research aimed at investigating the ability of duckweed (Lemna minor), a free-floating aquatic species widespread throughout the world, to remediate water polluted with five different concentrations of a herbicide - terbuthylazine (TBA) - was carried out. In addition, duckweed was treated with a plant biostimulant and a safener with the aim of increasing the plants capacity to tolerate and remove the TBA from the water. The results evidenced that the herbicide affected the duckweed already at the lower concentrations, reducing its capacity to proliferate and the area of its fronds. On the contrary, when the TBA treatments were performed in combination with the biostimulant or the safener the average area of the fronds was affected of lesser extents, compared to the plants treated with the herbicide only. Antioxidant enzymes, namely ascorbate peroxidases (APX) and catalases (CAT), were investigated and it was found that the biostimulated and safened duckweed showed increased activities of these enzymes, compared to the plants treated with TBA only. At last, some phytofiltration experiments were planned. The biostimulated and safened duckweed removed more TBA from polluted water than the plants treated with the herbicide alone. In conclusion, this research showed that duckweed is suitable for cleaning water polluted with TBA and this potential can be successfully improved by treating the species with a biostimulant or a safener.