Changes induced by lead in root system architecture of Arabidopsis seedlings are mediated by PDR2-LPR1/2 phosphate dependent way.

As sessile organisms, plants respond to changing environments modulating their genetic expression, metabolism and postembryonic developmental program (PDP) to adapt. Among environmental stressor, lead (Pb) is one of the most hazardous pollutants which limits crop productivity. Here, we describe in detail the effects of a wide range of concentrations of Pb on growth and development and a possible convergence with phosphate (Pi) starvation response. We found that the response to Pb presents a biphasic curve dose response in biomass accumulation: below 400 µM show a stimulatory effect meanwhile at Pb doses up to 600 µM effects are inhibitory. We found that +Pb (800 µM) modifies root system architecture (RSA) and induces acidification media, according to in silico ion interaction, in the growing medium Pb and Pi coprecipitate and plants grow in both Pi deficiency and Pb stress at the same time, however in spite of seedlings are under Pi starvation AtPT2 expression are Pb downregulated indicating that in addition to Pi starvation stress, Pb regulates physiological responses in root system. Using the mutants stop1, lpr1/2 and lpi3, which are affected in Pi starvation response, we found that changes in RSA by +Pb is genetically regulated and there are shared pathways with Pi starvation response mediated by PDR2-LPR1/2 and LPI3 pathways since lpr1/2 and lpi3 mutants are insensitive to +Pb and Pi starvation. Taking together, these results indicate that similar changes in RSA induced by independent environmental stimuli +Pb and Pi starvation are due to similar mediated response by PDR2-LPR1/2 pathway.

Expression analysis of the Arabidopsis thaliana AtSpen2 gene, and its relationship with other plant genes encoding Spen proteins

Abstract Proteins of the Split ends (Spen) family are characterized by an N-terminal domain, with one or more RNA recognition motifs and a SPOC domain. In Arabidopsis thaliana, the Spen protein FPA is involved in the control of flowering time as a component of an autonomous pathway independent of photoperiod. The A. thaliana genome encodes another gene for a putative Spen protein at the locus At4g12640, herein named AtSpen2. Bioinformatics analysis of the AtSPEN2 SPOC domain revealed low sequence similarity with the FPA SPOC domain, which was markedly lower than that found in other Spen proteins from unrelated plant species. To provide experimental information about the function of AtSpen2, A. thaliana plants were transformed with gene constructs of its promoter region with uidA::gfp reporter genes; the expression was observed in vascular tissues of leaves and roots, as well as in ovules and developing embryos. There was absence of a notable phenotype in knockout and overexpressing lines, suggesting that its function in plants might be specific to certain endogenous or environmental conditions. Our results suggest that the function of Atspen2 diverged from that of fpa due in part to their different transcription expression pattern and divergence of the regulatory SPOC domain.

Plant γH2AX foci are required for proper DNA DSB repair responses and colocalize with E2F factors.

Cellular responses to DNA double-strand breaks (DSBs) are linked in mammals and yeasts to the phosphorylated histones H2AX (γH2AX) repair foci which are multiproteic nuclear complexes responsible for DSB sensing and signalling. However, neither the components of these foci nor their role are yet known in plants. In this paper, we describe the effects of γH2AX deficiency in Arabidopsis thaliana plants challenged with DSBs in terms of genotoxic sensitivity and E2F-mediated transcriptional responses. We further establish the existence, restrictive to the G1/S transition, of specific DSB-induced foci containing tobacco E2F transcription factors, in both A. thaliana roots and BY-2 tobacco cells. These E2F foci partially colocalize with γH2AX foci while their formation is ataxia telangiectasia mutated (ATM)-dependent, requires the E2F transactivation domain with its retinoblastoma-binding site and is optimal in the presence of functional H2AXs. Overall, our results unveil a new interplay between plant H2AX and E2F transcriptional activators during the DSB response.

The xipotl mutant of Arabidopsis reveals a critical role for phospholipid metabolism in root system development and epidermal cell integrity.

Phosphocholine (PCho) is an essential metabolite for plant development because it is the precursor for the biosynthesis of phosphatidylcholine, which is the major lipid component in plant cell membranes. The main step in PCho biosynthesis in Arabidopsis thaliana is the triple, sequential N-methylation of phosphoethanolamine, catalyzed by S-adenosyl-l-methionine:phosphoethanolamine N-methyltransferase (PEAMT). In screenings performed to isolate Arabidopsis mutants with altered root system architecture, a T-DNA mutagenized line showing remarkable alterations in root development was isolated. At the seedling stage, the mutant phenotype is characterized by a short primary root, a high number of lateral roots, and short epidermal cells with aberrant morphology. Genetic and biochemical characterization of this mutant showed that the T-DNA was inserted at the At3g18000 locus (XIPOTL1), which encodes PEAMT (XIPOTL1). Further analyses revealed that inhibition of PCho biosynthesis in xpl1 mutants not only alters several root developmental traits but also induces cell death in root epidermal cells. Epidermal cell death could be reversed by phosphatidic acid treatment. Taken together, our results suggest that molecules produced downstream of the PCho biosynthesis pathway play key roles in root development and act as signals for cell integrity.