No abstract

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.

No abstract

Although the effects of high-frequency electromagnetic fields on biological systems have been studied frequently, unequivocal results have rarely been obtained, primarily because suitably controlled experiments could not be performed. In the present work, tomato plants were exposed to a homogeneous and isotropic field (900 MHz) using a mode stirred reverberation chamber, and the stress-related transcripts (calmodulin, protease inhibitor and chloroplast mRNA-binding protein) were assayed by real-time quantitative PCR. Exposure to an electromagnetic field induced a biphasic response, in which the levels of all three transcripts increased four-to six-fold 15 min after the end of electromagnetic stimulation, dropped to close to initial levels by 30 min, and then increased again at 60 min. We deliberately focused on the very early molecular responses to high-frequency electromagnetic fields in order to minimize secondary effects.

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. Radiolabelled 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 baker's yeast demonstrate that RhSUC2 encodes a sucrose/proton co-transporter with a Km 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.

Using an especially-designed facility, the Mode Stirred Reverberation Chamber, we exposed tomato plants (Lycopersicon esculentum Mill. VFN8) to low level (900 MHz, 5 V m(-1)) electromagnetic fields for a short period (10 min) and measured changes in abundance of three specific mRNA soon after exposure. Within minutes of electromagnetic stimulation, stress-related mRNA (calmodulin, calcium-dependent protein kinase and proteinase inhibitor) accumulated in a rapid, large and 3-phase manner typical of an environmental stress response. Accumulation of these transcripts into the polysomal RNA also took place (indicating that the encoded proteins were translated) but was delayed (indicating that newly-synthesized mRNA was not immediately recruited into polysomes). Transcript accumulation was maximal at normal Ca(2+) levels and was depressed at higher Ca(2+), especially for those encoding calcium-binding proteins. Removal of Ca(2+) (by addition of chelating agents or Ca(2+) channel blocker) led to total suppression of mRNA accumulation. Finally, 30 min after the electromagnetic treatment, ATP concentration and adenylate energy charge were transiently decreased, while transcript accumulation was totally prevented by application of the uncoupling reagent, CCCP. These responses occur very soon after exposure, strongly suggesting that they are the direct consequence of application of radio-frequency fields and their similarities to wound responses strongly suggests that this radiation is perceived by plants as an injurious stimulus.

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.

Exposing all of a wild-type tomato plant to electromagnetic radiation evoked rapid and substantial accumulation of basic leucine-zipper transcription factor (bZIP) mRNA in the terminal leaf (#4) with kinetics very similar to that seen in response to wounding, while in the abscisic acid (ABA) mutant (Sitiens), the response was more rapid, but transient. Submitting just the oldest leaf (#1) of a wild-type plant to irradiation evoked bZIP mRNA accumulation both locally in the exposed leaf and systemically in the unexposed (distant) leaf #4, although systemic accumulation was delayed somewhat. Accumulation of Pin2 mRNA was less than bZIP in both the exposed and distant leaves in wild type, but there was no delay in the systemic response. In Sitiens, bZIP mRNA accumulation was far less than in wild type in both local and distant leaves, while Pin2 mRNA accumulation was stronger in the exposed leaf, but totally prevented in the systemic leaf. In the jasmonic acid (JA) mutant (JL-5) and in wild-type plants treated with the ABA biosynthesis inhibitor, naproxen, responses were similar to those in the ABA mutant, while treatment of the exposed leaf with calcium antagonists totally abolished both local and systemic increases in bZIP transcript accumulation.