Alain Vian

Regulation of RhSUC2, a sucrose transporter, is correlated with the light control of bud burst in Rosa sp.

In roses, light is a central environmental factor controlling bud break and involves a stimulation of sugar metabolism. Very little is known about the role of sucrose transporters in the bud break process and its regulation by light. In this study, we show that sugar promotes rose bud break and that bud break is accompanied by an import of sucrose. Radio-labelled sucrose accumulation is higher in buds exposed to light than to darkness and involves an active component. Several sucrose transporter (RhSUC1, 2, 3 and 4) transcripts are expressed in rose tissues, but RhSUC2 transcript level is the only one induced in buds exposed to light after removing the apical dominance. RhSUC2 is preferentially expressed in bursting buds and stems. Functional analyses in bakers yeast demonstrate that RhSUC2 encodes a sucrose/proton co-transporter with a K(m) value of 2.99 mm at pH 4.5 and shows typical features of sucrose symporters. We therefore propose that bud break photocontrol partly depends upon the modulation of sucrose import into buds by RhSUC2.

Plant Responses to High Frequency Electromagnetic Fields

High frequency nonionizing electromagnetic fields (HF-EMF) that are increasingly present in the environment constitute a genuine environmental stimulus able to evoke specific responses in plants that share many similarities with those observed after a stressful treatment. Plants constitute an outstanding model to study such interactions since their architecture (high surface area to volume ratio) optimizes their interaction with the environment. In the present review, after identifying the main exposure devices (transverse and gigahertz electromagnetic cells, wave guide, and mode stirred reverberating chamber) and general physics laws that govern EMF interactions with plants, we illustrate some of the observed responses after exposure to HF-EMF at the cellular, molecular, and whole plant scale. Indeed, numerous metabolic activities (reactive oxygen species metabolism, α- and β-amylase, Krebs cycle, pentose phosphate pathway, chlorophyll content, terpene emission, etc.) are modified, gene expression altered (calmodulin, calcium-dependent protein kinase, and proteinase inhibitor), and growth reduced (stem elongation and dry weight) after low power (i.e., nonthermal) HF-EMF exposure. These changes occur not only in the tissues directly exposed but also systemically in distant tissues. While the long-term impact of these metabolic changes remains largely unknown, we propose to consider nonionizing HF-EMF radiation as a noninjurious, genuine environmental factor that readily evokes changes in plant metabolism.

Cytokinins Are Initial Targets of Light in the Control of Bud Outgrowth

Light perception by buds triggers a rapid cytokinin signal that controls major downstream regulators, including sugars, auxin, and strigolactones, of bud outgrowth. Bud outgrowth is controlled by environmental and endogenous factors. Through the use of the photosynthesis inhibitor norflurazon and of masking experiments, evidence is given here that light acts mainly as a morphogenic signal in the triggering of bud outgrowth and that initial steps in the light signaling pathway involve cytokinins (CKs). Indeed, in rose (Rosa hybrida), inhibition of bud outgrowth by darkness is suppressed solely by the application of CKs. In contrast, application of sugars has a limited effect. Exposure of plants to white light (WL) induces a rapid (after 3–6 h of WL exposure) up-regulation of CK synthesis (RhIPT3 and RhIPT5), of CK activation (RhLOG8), and of CK putative transporter RhPUP5 genes and to the repression of the CK degradation RhCKX1 gene in the node. This leads to the accumulation of CKs in the node within 6 h and in the bud at 24 h and to the triggering of bud outgrowth. Molecular analysis of genes involved in major mechanisms of bud outgrowth (strigolactone signaling [RwMAX2], metabolism and transport of auxin [RhPIN1, RhYUC1, and RhTAR1], regulation of sugar sink strength [RhVI, RhSUSY, RhSUC2, and RhSWEET10], and cell division and expansion [RhEXP and RhPCNA]) reveal that, when supplied in darkness, CKs up-regulate their expression as rapidly and as intensely as WL. Additionally, up-regulation of CKs by WL promotes xylem flux toward the bud, as evidenced by Methylene Blue accumulation in the bud after CK treatment in the dark. Altogether, these results suggest that CKs are initial components of the light signaling pathway that controls the initiation of bud outgrowth.

Mode Stirred Reverberation Chamber (MSRC): a large and efficient tool to lead high frequency bioelectromagnetic in vitro experimentation

An important aspect of the studies undertaken in bioelectromagnetism relates to the choice of exposure facility. The characteristics of a real electromagnetic environment are far more complex compared to the one plane wave irradiation set-up used in the majority of bioelectromagnetic studies. Moreover, biological requirements should represent the starting point in the design of an in vitro exposure system. Indeed it is important to avoid altering the electromagnetic properties of the exposure system in the presence of biological equipments. Related to these two essential points, this article contributes to show the advantages of a Mode Stirred Reverberation Chamber (MSRC) to guarantee a controlled electromagnetic environment around biological materials for in vitro experimentation. An example of irradiation of in vitro human skin cells cultures is considered to illustrate this paper. In order to show that the biological conditions and the physical requirements for in vitro experiments are checked, two aspects are described. Firstly we achieved the characterization of the electromagnetic fleld generated around the biological system (both equipments and cultures). Secondly the analysis of the Speciflc Absorption Rate (SAR) inside the biological medium was evaluated both numerically and experimentally. Initially,

Light and nitrogen nutrition regulate apical control in Rosa hybrida L.

Apical control is defined as the inhibition of basal axillary bud outgrowth by an upper actively growing axillary axis, whose regulation is poorly understood yet differs markedly from the better-known apical dominance. We studied the regulation of apical control by environmental factors in decapitated Rosa hybrida in order to remove the apical hormonal influence and nutrient sink. In this plant model, all the buds along the main axis have a similar morphology and are able to burst in vitro. We concentrated on the involvement of light intensity and nitrate nutrition on bud break and axillary bud elongation in the primary axis pruned above the fifth leaf of each rose bush. We observed that apical control took place in low light (92 μmol m(-2)s(-1)), where only the 2-apical buds grew out, both in low (0.25 mM) and high (12.25 mM) nitrate. In contrast, in high light (453 μmol m(-2)s(-1)), the apical control only operates in low nitrate while all the buds along the stem grew out when the plant was supplied with a high level of nitrate. We found a decreasing photosynthetic activity from the top to the base of the plant concomitant with a light gradient along the stem. The quantity of sucrose, fructose, glucose and starch are higher in high light conditions in leaves and stem. The expression of the sucrose transporter RhSUC2 was higher in internodes and buds in this lighting condition, suggesting an increased capacity for sucrose transport. We propose that light intensity and nitrogen availability both contribute to the establishment of apical control.

Low-amplitude, high-frequency electromagnetic field exposure causes delayed and reduced growth in Rosa hybrida

It is now accepted that plants perceive high-frequency electromagnetic field (HF-EMF). We wondered if the HF-EMF signal is integrated further in planta as a chain of reactions leading to a modification of plant growth. We exposed whole small ligneous plants (rose bush) whose growth could be studied for several weeks. We performed exposures at two different development stages (rooted cuttings bearing an axillary bud and 5-leaf stage plants), using two high frequency (900MHz) field amplitudes (5 and 200Vm(-1)). We achieved a tight control on the experimental conditions using a state-of-the-art stimulation device (Mode Stirred Reverberation Chamber) and specialized culture-chambers. After the exposure, we followed the shoot growth for over a one-month period. We observed no growth modification whatsoever exposure was performed on the 5-leaf stage plants. When the exposure was performed on the rooted cuttings, no growth modification was observed on Axis I (produced from the elongation of the axillary bud). Likewise, no significant modification was noted on Axis II produced at the base of Axis I, that came from pre-formed secondary axillary buds. In contrast, Axis II produced at the top of Axis I, that came from post-formed secondary buds consistently displayed a delayed and significant reduced growth (45%). The measurements of plant energy uptake from HF-EMF in this exposure condition (SAR of 7.2 10(-4)Wkg(-1)) indicated that this biological response is likely not due to thermal effect. These results suggest that exposure to electromagnetic field only affected development of post-formed organs.

Signalomics: Diversity and Methods of Analysis of Systemic Signals in Plants

We provide a brief definition and history of signals, pointing out how differences in body plan between plants and animals require fundamentally different signaling mechanisms, and then list the diversity of chemical and physical signals along with their pathways of transmission, providing details on molecular signals and focusing on the phloem and xylem as being the main conduits for (rapid) systemic signaling. The two major electrical (action potentials and variation potentials) as well as hydraulic signals are then described. The latter part of the chapter deals with methods of analysis of molecular signals, including accessing the phloem and identifying the array of gene products transported therein. A description is provided of the modern methods used in metabolomics and phenotyping to analyze the metabolic consequences of signal action. Conventional techniques for analyzing electrical and hydraulic signals and their ionic components using electrodes are then furnished. Finally we describe novel techniques developed recently in the animal field using fluorescence to monitor real-time changes in membrane potential, which could be adapted for plants to open up new vistas in our understanding of electrical signals in plants.

Asparagine and sugars are both required to sustain secondary axis elongation after bud outgrowth in Rosa hybrida

Nitrogen is required for optimal plant growth, especially in young organs such as secondary axes (axes II) after axillary bud outgrowth. Several studies have shown an increase of nitrogen concentration in xylem sap concomitantly with bud outgrowth, but the relation between nitrogen, sugars and plant hormones in axis II still remains unclear. We investigated in Rosa hybrida the involvement of nitrogen nutrition in axis II elongation in relation with sugars and cytokinins using 15N-labeled nitrate and sugars, amino acids and cytokinin quantifications. Besides, we measured the effect of the exogenous supply of these compounds on axis II elongation using in vitro excised bud culture. We demonstrated that nitrogen in the axis II comes mainly from new root uptake after decapitation. Asparagine, which concentration increases in sap exudates and tissues during axis II elongation, was the sole amino acid able to sustain an efficient elongation in vitro when supplied in combination with sucrose.

Intercellular Communication in Plants: Evidence for an EMF-Generated Signal that Evokes Local and Systemic Transcriptional Responses in Tomato

Exposing the oldest leaf of a tomato plant to a short (10 min), low-amplitude (5 V.m−1), high-frequency (900 MHz) electromagnetic field evoked a rapid (15 min) and systemic accumulation of the stress-related transcript LebZIP-1 in the exposed leaf and in the distant, terminal leaf that is protected from EMF radiation. The accumulation was prevented by calcium counteracting drugs both locally and systemically. It was also prevented, but only in the distant tissue, in the ABA tomato mutant Sitiens or in wild-type tomato grown in the presence of the ABA synthesis inhibitor naproxen.