Quantum dot and superparamagnetic nanoparticle interaction with pathogenic fungi: internalization and toxicity profile.

For several years now, nanoscaled materials have been implemented in biotechnological applications related to animal (in particular human) cells and related pathologies. However, the use of nanomaterials in plant biology is far less widespread, although their application in this field could lead to the future development of plant biotechnology applications. For any practical use, it is crucial to elucidate the relationship between the nanomaterials and the target cells. In this work we have evaluated the behavior of two types of nanomaterials, quantum dots and superparamagnetic nanoparticles, on Fusarium oxysporum, a fungal species that infects an enormous range of crops causing important economic losses and is also an opportunistic human pathogen. Our results indicated that both nanomaterials rapidly interacted with the fungal hypha labeling the presence of the pathogenic fungus, although they showed differential behavior with respect to internalization. Thus, whereas magnetic nanoparticles appeared to be on the cell surface, quantum dots were significantly taken up by the fungal hyphae showing their potential for the development of novel control approaches of F. oxysporum and related pathogenic fungi following appropriate functionalization. In addition, the fungal germination and growth, accumulation of ROS, indicative of cell stress, and fungal viability have been evaluated at different nanomaterial concentrations showing the low toxicity of both types of nanomaterials to the fungus. This work represents the first study on the behavior of quantum dots and superparamagnetic particles on fungal cells, and constitutes the first and essential step to address the feasibility of new nanotechnology-based systems for early detection and eventual control of pathogenic fungi.

CdSe/ZnS quantum dots trigger DNA repair and antioxidant enzyme systems in Medicago sativa cells in suspension culture.

BACKGROUND: Nanoparticles appear to be promising devices for application in the agriculture and food industries, but information regarding the response of plants to contact with nano-devices is scarce. Toxic effects may be imposed depending on the type and concentration of nanoparticle as well as time of exposure. A number of mechanisms may underlie the ability of nanoparticles to cause genotoxicity, besides the activation of ROS scavenging mechanisms. In a previous study, we showed that plant cells accumulate 3-Mercaptopropanoic acid-CdSe/ZnS quantum dots (MPA-CdSe/ZnS QD) in their cytosol and nucleus and increased production of ROS in a dose dependent manner when exposed to QD and that a concentration of 10 nM should be cyto-compatible. RESULTS: When Medicago sativa cells were exposed to 10, 50 and 100 nM MPA-CdSe/ZnS QD a correspondent increase in the activity of Superoxide dismutase, Catalase and Glutathione reductase was registered. Different versions of the COMET assay were used to assess the genotoxicity of MPA-CdSe/ZnS QD. The number of DNA single and double strand breaks increased with increasing concentrations of MPA-CdSe/ZnS QD. At the highest concentrations, tested purine bases were more oxidized than the pyrimidine ones. The transcription of the DNA repair enzymes Formamidopyrimidine DNA glycosylase, Tyrosyl-DNA phosphodiesterase I and DNA Topoisomerase I was up-regulated in the presence of increasing concentrations of MPA-CdSe/ZnS QD. CONCLUSIONS: Concentrations as low as 10 nM MPA-CdSe/ZnS Quantum Dots are cytotoxic and genotoxic to plant cells, although not lethal. This sets a limit for the concentrations to be used when practical applications using nanodevices of this type on plants are being considered. This work describes for the first time the genotoxic effect of Quantum Dots in plant cells and demonstrates that both the DNA repair genes (Tdp1ß, Top1ß and Fpg) and the ROS scavenging mechanisms are activated when MPA-CdSe/ZnS QD contact M. sativa cells.

The impact of CdSe/ZnS Quantum Dots in cells of Medicago sativa in suspension culture.

BACKGROUND: Nanotechnology has the potential to provide agriculture with new tools that may be used in the rapid detection and molecular treatment of diseases and enhancement of plant ability to absorb nutrients, among others. Data on nanoparticle toxicity in plants is largely heterogeneous with a diversity of physicochemical parameters reported, which difficult generalizations. Here a cell biology approach was used to evaluate the impact of Quantum Dots (QDs) nanocrystals on plant cells, including their effect on cell growth, cell viability, oxidative stress and ROS accumulation, besides their cytomobility. RESULTS: A plant cell suspension culture of Medicago sativa was settled for the assessment of the impact of the addition of mercaptopropanoic acid coated CdSe/ZnS QDs. Cell growth was significantly reduced when 100 mM of mercaptopropanoic acid -QDs was added during the exponential growth phase, with less than 50% of the cells viable 72 hours after mercaptopropanoic acid -QDs addition. They were up taken by Medicago sativa cells and accumulated in the cytoplasm and nucleus as revealed by optical thin confocal imaging. As part of the cellular response to internalization, Medicago sativa cells were found to increase the production of Reactive Oxygen Species (ROS) in a dose and time dependent manner. Using the fluorescent dye H2DCFDA it was observable that mercaptopropanoic acid-QDs concentrations between 5-180 nM led to a progressive and linear increase of ROS accumulation. CONCLUSIONS: Our results showed that the extent of mercaptopropanoic acid coated CdSe/ZnS QDs cytotoxicity in plant cells is dependent upon a number of factors including QDs properties, dose and the environmental conditions of administration and that, for Medicago sativa cells, a safe range of 1-5 nM should not be exceeded for biological applications.