Marie-Andrée Esnault

Root uptake and phytotoxicity of nanosized molybdenum octahedral clusters

Here are examined the root uptake and phytotoxicity of octahedral hexamolybdenum clusters on rapeseed plants using the solid state compound Cs(2)Mo(6)Br(14) as cluster precursor. [Mo(6)Br(14)](2-) cluster units are nanosized entities offering a strong and stable emission in the near-infrared region with numerous applications in biotechnology. To investigate cluster toxicity on rapeseed plants, two different culture systems have been set up, using either a water-sorbing suspension of cluster aggregates or an ethanol-sorbing solution of dispersed nanosized clusters. Size, shape, surface area and state of clusters in both medium were analyzed by FE-SEM, BET and XPS. The potential contribution of cluster dissolution to phytotoxicity was evaluated by ICP-OES and toxicity analysis of Mo, Br and Cs. We showed that the clusters did not affect seed germination but greatly inhibited plant growth. This inhibition was much more important when plants were treated with nanosized entities than with microsized cluster aggregates. In addition, nanosized clusters affected the root morphology in a different manner than microsized cluster aggregates, as shown by FE-SEM observations. The root penetration of the clusters was followed by secondary ion mass spectroscopy with high spatial resolution (NanoSIMS) and was also found to be much more important for treatments with nanosized clusters.

Kinetic transcriptomic approach revealed metabolic pathways and genotoxic-related changes implied in the Arabidopsis response to ionising radiations.

Plants exposed to ionising radiation (IR) have to face direct and indirect (oxidative stress) deleterious effects whose intensity depends on the dose applied and led to differential genome regulation. Transcriptomic analyses were conducted with CATMA microarray technology on Arabidopsis thaliana plantlets, 2 and 26h after exposure to the IR doses 10Gy and 40Gy. 10Gy treatment seemed to enhance antioxidative compound biosynthetic pathways whereas the 40Gy dose up-regulated ROS-scavenging enzyme genes. Transcriptomic data also highlighted a differential regulation of chloroplast constituent genes depending on the IR dose, 10Gy stimulating and 40Gy down-regulating. This probable 40Gy decrease of photosynthesis could help for the limitation of ROS production and may be coupled with programmed cell death (PCD)/senescence phenomena. Comparisons with previous transcriptomic studies on plants exposed to a 100Gy dose revealed 60 dose-dependent up-regulated genes, including notably cell cycle checkpoints to allow DNA repairing phenomena. Furthermore, the alteration of some cellular structure related gene expression corroborated a probable mitotic arrest after 40Gy. Finally, numerous heat-shock protein and chaperonin genes, known to protect proteins against stress-dependent dysfunction, were up-regulated after IR exposure.

Application of proteomics to the assessment of the response to ionising radiation in Arabidopsis thaliana

Ionising radiation (IR) affects cellular and tissue function. However, the biological effects and interactions induced by IR are unclear. The aim of this study was to decipher the proteomic patterns that influence these pathways. The proteomes of Arabidopsis thaliana roots and rosettes were analysed in response to sub-lethal IR doses (0, 10, and 40 Gy). For each dose, the dynamic response was observed at different time points (2, 24 and 72 h). To quantitatively measure the effect of IR on the proteome, total proteins were extracted and subjected to 2-DE, and the changes in the 2-DE protein profiles were analysed. Statistical analysis revealed a total of 172 proteins (145 in leaves and 27 in roots) that were differentially expressed. These proteins were subsequently analysed by MALDI-TOF/TOF MS and comparative database analysis, and 144 (118 in leaves and 26 in roots) proteins were identified. The changes in the protein profile were quantitatively more significant for the 40 Gy dose than for the 10 Gy dose. In addition, specific functional groups of proteins were identified based on the consistency of the dose- and time-responses. The molecular mechanisms involved in the response to IR and a comparison of the observed responses are discussed.