Isoprene (2-methyl-1,3-butadiene) emission varies diurnally in different species. In poplar (Populus spp.), it has recently been shown that the gene encoding the synthesizing enzyme for isoprene, isoprene synthase (ISPS), displays diurnal variation in expression. Working on shoot cultures of Grey poplar (Populus 3 canescens) placed under a different light regime in phytochambers, we showed that these variations in PcISPS gene expression, measured by quantitative real-time polymerase chain reaction, are not only due to day-night changes, but also are linked to an internal circadian clock. Measurement of additional selected isoprenoid genes revealed that phytoene synthase (carotenoid pathway) displays similar fluctuations, whereas 1-deoxy-D-xylulose 5-phosphate reductoisomerase, possibly the first committed enzyme of the 1-deoxy-D-xylulose 5-phosphate pathway, only shows light regulation. On the protein level, it appeared that PcISPS activity and protein content became reduced under constant darkness, whereas under constant light, activity and protein content of this enzyme were kept high. In contrast, isoprene emission rates under continuous irradiation displayed circadian changes as is the case for gene expression of PcISPS. Furthermore, binding assays with Arabidopsis (Arabidopsis thaliana) late elongated hypocotyl, a transcription factor of Arabidopsis involved in circadian regulation, clearly revealed the presence of circadian-determining regulatory elements in the promoter region of PcISPS.

Isoprene is the most abundant volatile compound emitted by vegetation. It influences air chemistry and is part of plant defense against abiotic stresses. However, whether isoprene influences biotic interactions between plants and other organisms has not been investigated to date. Here we show a new effect of isoprene, namely its influence on interactions between plants and insects. Herbivory induces the release of plant volatiles that attract the herbivore's enemies, such as parasitic wasps, as a kind of bodyguard. We used transgenic isoprene-emitting Arabidopsis plants in behavioral, chemical, and electrophysiological studies to investigate the effects of isoprene on ecological interactions in 2 tritrophic systems. We demonstrate that isoprene is perceived by the chemoreceptors of the parasitic wasp Diadegma semiclausum and interferes with the attraction of this parasitic wasp to volatiles from herbivore-infested plants. We verified this repellent effect on D. semiclausum female wasps by adding external isoprene to the volatile blend of wild-type plants. In contrast, the antennae of the parasitic wasp Cotesia rubecula do not perceive isoprene and the behavior of this wasp was not altered by isoprene emission. In addition, the performance of the 2 examined lepidopteran herbivores (Pieris rapae and Plutella xylostella) was not affected by isoprene emission. Therefore, attraction of parasitic wasps to host-infested herbaceous plants in the neighborhood of high isoprene emitters, such as poplar or willow, may be hampered by the isoprene emission that repels plant bodyguards.Arabidopsis ͉ isoprene emission ͉ plant-insect interactions ͉ tritrophic interactions ͉ parasitoid P lants interact with their abiotic and biotic environments through volatile organic compounds, of which terpenes [isoprene (C 5 ), mono-(C 10 ), sesqui-(C 15 ), and homoterpenes (C 11 , C 16 )] form the most prominent group (1, 2). Global emission of the highly reactive hemiterpene isoprene is estimated to be 440-660 Tg carbon per year (3), which amounts to 44% of the overall nonmethane volatiles released from ecosystems. Some plants, mainly tree species (1), may even emit up to 15% of photosynthetically fixed carbon back to the atmosphere as isoprene (4). The compound may help the photosynthetic apparatus to recover from brief, high-temperature episodes (5). This effect was recently demonstrated with transgenic isoprene nonemitting poplar leaves in which gene expression of isoprene synthase (PcISPS) was knocked down (6). Isoprene is thought to act physically by stabilizing the thylakoid membranes at high temperatures (5) or by quenching reactive oxygen species, such as ozone, which can lead to membrane damage (7).Isoprene emission was first observed 51 years ago (8) and has been widely studied by atmospheric chemists and plant physiologists (5, 9). To our knowledge, a potential effect of isoprene on interactions between the emitting plants and other biota remains unexplored to date. Attack by insect herbivores results in the biosynthesis of a pla...

The volatile hemiterpene isoprene is emitted from plants and can affect atmospheric chemistry. Although recent studies indicate that isoprene can enhance thermotolerance or quench oxidative stress, the underlying physiological mechanisms are largely unknown. In this work, Arabidopsis (Arabidopsis thaliana), a natural nonemitter of isoprene and the model plant for functional plant analyses, has been constitutively transformed with the isoprene synthase gene (PcISPS) from Grey poplar (Populus x canescens). Overexpression of poplar ISPS in Arabidopsis resulted in isoprene-emitting rosettes that showed transiently enhanced growth rates compared to the wild type under moderate thermal stress. The findings that highest growth rates, higher dimethylallyl diphosphate levels, and enzyme activity were detected in young plants during their vegetative growth phase indicate that enhanced growth of transgenic plants under moderate thermal stress is due to introduced PcISPS. Dynamic gasexchange studies applying transient cycles of heat stress to the wild type demonstrate clearly that the prime physiological role of isoprene formation in Arabidopsis is not to protect net assimilation from damage against thermal stress, but may instead be to retain the growth potential or coordinated vegetative development of the plant. Hence, this study demonstrates the enormous potential but also the pitfalls of transgenic Arabidopsis (or other nonnatural isoprenoid emitters) in studying isoprene biosynthesis and its biological function(s).

In the present study, we combined transient temperature and light stress (sunfleck) and comparably analyzed photosynthetic gas exchange in Grey poplar which has been genetically modified in isoprene emission capacity. Overall, we demonstrate that for poplar leaves the ability to emit isoprene is crucial to maintain photosynthesis when exposed to sunflecks. Net CO2 assimilation and electron transport rates were strongly impaired in sunfleck-treated non-isoprene emitting poplars. Similar impairment was not detected when the leaves were exposed to high light (lightflecks) only. Within 10 h non-isoprene emitting poplars recovered from sunfleck-related impairment as indicated by chlorophyll fluorescence and microarray analysis. Unstressed leaves of non-isoprene emitting poplars had higher ascorbate contents, but also higher contents of malondialdehyde than wild-type. Microarray analyses revealed lipid and chlorophyll degradation processes in the non-isoprene emitting poplars. Thus, there is evidence for an adjustment of the antioxidative system in the non-isoprene emitting poplars even under normal growth conditions.

Plants interact with their environment through a wide variety of biogenic volatile organic compounds (BVOCs), with isoprenoids ( identical with terpenes), i.e. isoprene, mono- and sesquiterpenes, playing an important role. Isoprene, a hemiterpene, is the simplest isoprenoid compound mainly emitted by tree species like poplars, oaks and willows. Woody plants alone comprise 75% of the global isoprene emitted to the atmosphere. Due to its significant influence on atmospheric chemistry, research has been focused on this C5 compound, with poplar being the most prominent model system. Recent studies indicate that isoprene can enhance thermotolerance or quench oxidative stress, while also interfering with the attraction of herbivores and parasitoids to plants. In this paper, we report on biosynthesis, regulation and function of isoprene and other stress-induced volatile isoprenoids in poplar, and discuss the future scientific challenges in this genus with respect to the importance of plant volatiles in high-density poplar biomass plantations.