Biochemical properties of isoprene synthase in poplar (Populus × canescens)

Schnitzler, Jörg‐Peter; Zimmer, Ina; Bachl, Anette; Arend, Matthias; Fromm, Jörg; Fischbach, Robert J.

doi:10.1007/s00425-005-0022-1

Cited by 84 publications

(79 citation statements)

References 40 publications

(66 reference statements)

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“…While DMAPP is found both in the cytosol (from MVA pathway flux) and the chloroplast (from MEP pathway flux), IspS is localized in the chloroplast (Wildermuth and Fall, 1996;Schnitzler et al, 2005;Vickers et al, 2010), so it can only use DMAPP from the chloroplastic pool. Leaf isoprene emission is directly correlated with extractible enzyme activity (Monson et al, 1992) as well as with the amount of IspS in the leaf (Vickers et al, 2010), and IspS levels do not change rapidly in response to changing environmental conditions (Vickers et al, 2011), suggesting that the enzyme itself is not under direct regulation and isoprene production is largely driven by the availability of DMAPP in the chloroplast.…”

Section: A Rapid Mechanism For Balancing Availability Of Carbon For Imentioning

confidence: 99%

Dynamic Balancing of Isoprene Carbon Sources Reflects Photosynthetic and Photorespiratory Responses to Temperature Stress

et al. 2014

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The volatile gas isoprene is emitted in teragrams per annum quantities from the terrestrial biosphere and exerts a large effect on atmospheric chemistry. Isoprene is made primarily from recently fixed photosynthate; however, alternate carbon sources play an important role, particularly when photosynthate is limiting. We examined the relative contribution of these alternate carbon sources under changes in light and temperature, the two environmental conditions that have the strongest influence over isoprene emission. Using a novel real-time analytical approach that allowed us to examine dynamic changes in carbon sources, we observed that relative contributions do not change as a function of light intensity. We found that the classical uncoupling of isoprene emission from net photosynthesis at elevated leaf temperatures is associated with an increased contribution of alternate carbon. We also observed a rapid compensatory response where alternate carbon sources compensated for transient decreases in recently fixed carbon during thermal ramping, thereby maintaining overall increases in isoprene production rates at high temperatures. Photorespiration is known to contribute to the decline in net photosynthesis at high leaf temperatures. A reduction in the temperature at which the contribution of alternate carbon sources increased was observed under photorespiratory conditions, while photosynthetic conditions increased this temperature. Feeding [2- CO 2 , supporting the possibility that photorespiration can provide an alternate source of carbon for isoprene synthesis. Our observations have important implications for establishing improved mechanistic predictions of isoprene emissions and primary carbon metabolism, particularly under the predicted increases in future global temperatures.

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Section: A Rapid Mechanism For Balancing Availability Of Carbon For Imentioning

confidence: 99%

Dynamic Balancing of Isoprene Carbon Sources Reflects Photosynthetic and Photorespiratory Responses to Temperature Stress

et al. 2014

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“…Protein transfer was achieved using a Millipore semidry electroblot system following the manufacturer's instructions. Immunoassaying of ISPS was performed according to Schnitzler et al (2005). Alkaline phosphatase-conjugated secondary anti-rabbit antibody (SigmaAldrich) was detected with addition of 5-bromo-4-chloroindolyl-3-phosphate and nitro blue tetrazolium (10 mg of 5-bromo-4-chloroindolyl-3-phosphate and 5 mg of nitro blue tetrazolium in 20 mL of 1 M Tris-HCl, pH 8.8, 1 mM MgCl 2 ).…”

Section: Quantification Of Transcript Levels By Quantitative Reverse mentioning

confidence: 99%

Arabidopsis, a Model to Study Biological Functions of Isoprene Emission?

Loivamäki¹,

Gilmer²,

Fischbach³

et al. 2007

Plant Physiol.

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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).

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“…Quantification of ISPS protein was performed according to Schnitzler et al (2005) using purified polyclonal anti-PcISPS IgG generated against N-terminal 6x-His-tagged PcISPS (Miller et al, 2001). For use as second antibody in ELISA, anti-PcISPS IgG was conjugated with horseradish peroxidase by BioGenes.…”

Section: Quantification Of Pcisps Protein With Elisamentioning

confidence: 99%

Circadian Rhythms of Isoprene Biosynthesis in Grey Poplar Leaves

et al. 2006

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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.

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Biochemical properties of isoprene synthase in poplar (Populus × canescens)

Cited by 84 publications

References 40 publications

Dynamic Balancing of Isoprene Carbon Sources Reflects Photosynthetic and Photorespiratory Responses to Temperature Stress

Dynamic Balancing of Isoprene Carbon Sources Reflects Photosynthetic and Photorespiratory Responses to Temperature Stress

Arabidopsis, a Model to Study Biological Functions of Isoprene Emission?

Circadian Rhythms of Isoprene Biosynthesis in Grey Poplar Leaves

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