Two negative cis-regulatory regions involved in fruit-specific promoter activity from watermelon (Citrullus vulgaris S.)

A 1.8 kb 5′-flanking region of the large subunit of ADP-glucose pyrophosphorylase, isolated from watermelon (Citrullus vulgaris S.), has fruit-specific promoter activity in transgenic tomato plants. Two negative regulatory regions, from –986 to –959 and from –472 to –424, were identified in this promoter region by fine deletion analyses. Removal of both regions led to constitutive expression in epidermal cells. Gain-of-function experiments showed that these two regions were sufficient to inhibit RFP (red fluorescent protein) expression in transformed epidermal cells when fused to the cauliflower mosaic virus (CaMV) 35S minimal promoter. Gel mobility shift experiments demonstrated the presence of leaf nuclear factors that interact with these two elements. A TCCAAAA motif was identified in these two regions, as well as one in the reverse orientation, which was confirmed to be a novel specific cis-element. A quantitative β-glucuronidase (GUS) activity assay of stable transgenic tomato plants showed that the activities of chimeric promoters harbouring only one of the two cis-elements, or both, were ∼10-fold higher in fruits than in leaves. These data confirm that the TCCAAAA motif functions as a fruit-specific element by inhibiting gene expression in leaves

An empirical study of normative dissociation in musical and non-musical everyday life experiences

Dissociative experiences involving music have received little research attention outside the field of ethnomusicology. This paper examines the psychological characteristics of normative dissociation (detachment) across musical and non-musical experiences in ‘real world’, everyday settings. It draws upon a subset of data arising from an empirical project designed to compare transformative shifts of consciousness, with and without music in daily life, and the ways in which use of music may facilitate the processes of dissociation and absorption. Twenty participants kept unstructured diaries for two weeks, recording free descriptions of involving experiences of any kind as soon as possible after their occurrence. All descriptions were subsequently subjected to Interpretative Phenomenological Analysis (IPA). Results suggest that dissociative experiences are a familiar occurrence in everyday life. Diary entries highlight an established practice of actively sought detachment from self, surroundings or activity, suggesting that, together with absorption, the processes of derealization (altered perception of surroundings) and depersonalization (detachment from self) constitute common means of selfregulation in daily life. Music emerges as a particularly versatile facilitator of dissociative experience because of its semantic ambiguity, portability, and the variety of ways in which it may mediate perception, so facilitating an altered relationship to self and environmen

Chemical PARP Inhibition Enhances Growth of Arabidopsis and Reduces Anthocyanin Accumulation and the Activation of Stress Protective Mechanisms

Poly-ADP-ribose polymerase (PARP) post-translationally modifies proteins through the addition of ADP-ribose polymers, yet its role in modulating plant development and stress responses is only poorly understood. The experiments presented here address some of the gaps in our understanding of its role in stress tolerance and thereby provide new insights into tolerance mechanisms and growth. Using a combination of chemical and genetic approaches, this study characterized phenotypes associated with PARP inhibition at the physiological level. Molecular analyses including gene expression analysis, measurement of primary metabolites and redox metabolites were used to understand the underlying processes. The analysis revealed that PARP inhibition represses anthocyanin and ascorbate accumulation under stress conditions. The reduction in defense is correlated with enhanced biomass production. Even in unstressed conditions protective genes and molecules are repressed by PARP inhibition. The reduced anthocyanin production was shown to be based on the repression of transcription of key regulatory and biosynthesis genes. PARP is a key factor for understanding growth and stress responses of plants. PARP inhibition allows plants to reduce protection such as anthocyanin, ascorbate or Non-Photochemical-Quenching whilst maintaining high energy levels likely enabling the observed enhancement of biomass production under stress, opening interesting perspectives for increasing crop productivity

The influence of a short-term mindfulness meditation intervention on emotion and visual attention

It has been suggested that mindfulness meditation (MM) improves psychological wellbeing via the focusing and broadening of attention. Whilst studies show that short term MM interventions can improve focused attention there is little evidence to support the broadening of attention. The current study investigated the influence of a short-term MM intervention on emotion and the scope of visual attention. Seventy participants completed a global-local processing task separated into three blocks of trials, with a ten minute break between each one. During the breaks a MM group engaged in a breath counting task and a control group engaged in a task of their choosing. Response times to global and local targets and a measure of self-reported emotional affect were recorded for each block. Mindfulness had no impact on attention however both positive and negative affect decreased for the MM group across the course of the experiment. The results suggest that MM can reduce the focus on negative (and positive) thoughts, indicating possible changes to focused attention, yet a short-term intervention is not sufficient to broaden attentio

Exploiting multisite gateway and pENFRUIT plasmid collection for fruit genetic engineering

El pdf es la versión post-print.MultiSite Gateway cloning techniques based on homologous recombination facilitate the combinatorial assembly of basic genetic pieces (i.e., promoters, CDS, and terminators) into gene expression or gene silencing cassettes. pENFRUIT is a collection of MultiSite Triple Gateway Entry vectors dedicated to genetic engineering in fruits. It comprises a number of fruit-operating promoters as well as C-terminal tags adapted to the Gateway standard. In this way, flanking regulatory/labeling sequences can be easily Gateway-assembled with a given gene of interest for its ectopic expression or silencing in fruits. The resulting gene constructs can be analyzed in stable transgenic plants or in transient expression assays, the latter allowing fast testing of the increasing number of combinations arising from MultiSite methodology. A detailed description of the use of MultiSite cloning methodology for the assembly of pENFRUIT elements is presented.This work was supported by the Spanish Ministry of Science and Education (Ramón y Cajal programme BIO2005-01015 and BIO2008-03434 projects), the European Commission (programme I3P and EU-SOL project) and Fundación Genoma España (ESP-SOL project).Peer reviewe