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Indicates that all correspondence concerning 'The Plant Journal' paper titled 'Emission of nitrous oxide (N[sub 2]O) from transgenic tobacco expressing antisense NiR mRNA' should be addressed to Hiromichi Morikawa, Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima 739-8526, Japan.

Abstract: Atmospheric nitrogen dioxide (NO2) is an environmental oxidant that is removed through direct uptake by foliage, but plant responses to this highly reactive gas are not well understood at the molecular level. From NO2-exposed leaves of a woody azalea (Rhododendron mucronatum), we cloned two cDNAs (RmGLP1 and RmGLP2) for germin-like proteins (GLPs), a group of ubiquitous plant proteins that have been implicated in various plant physiological and developmental processes. Quantitative analysis of mRNA expression, together with immunoblotting data, showed that foliar exposure to NO2 caused a robust induction of these GLP-encoding genes. When produced in tobacco cell culture, recombinant RmGLP2 was secreted into the apoplast, where it exhibited superoxide dismutase activity. RmGLP1 and RmGLP2 represent the first examples of plant genes that are responsive to airborne NO2. These enzymes might have a potential role in extracellular defense mechanisms through attenuation of interactions between reactive nitrogen and oxygen species. [Copyright &y& Elsevier]

It is known that when plant leaves are exposed to exogenously applied nitrogen dioxide (NO 2 ), nitrogen derived from NO 2 is reduced to amino acid nitrogen. However, whether this is the sole metabolic fate of exogenously applied NO 2 is unclear. In this study, Arabidopsis leaves were exposed to 4 ppm 15 N-labeled NO 2 for 4 h in light, followed by capillary ion analysis and elemental analysis-mass spectrometry with an elemental analyzer connected directly to a mass spectrometer. We found that leaf cells exposed to 15 N-labeled NO 2 accumulated a large amount of 15 N-labeled nitrate. Neither 15 N-labeled nitrite nor endogenous nitrite was present in exposed leaves. It is likely that exogenously applied NO 2 is first converted to nitrite, and that nitrite is oxidized to nitrate in Arabidopsis leaf cells. The complete disappearance of nitrite derived from exogenously applied NO 2 and endogenous nitrite supports this mechanism.

To better understand the response of plants to atmospheric nitrogen dioxide (NO2), we investigated biomass accumulation in 3 accessions of Arabidopsis thaliana: C24, Columbia (Col-0), and Landsberg erecta (Ler). Plants were grown in NO2-free air for 1 week after sowing, followed by 3 (Col-0 and Ler) to 4 (C24) weeks in air with or without NO 2 (10 or 50 ppb). NO2 treatment increased the biomass of all 3 accessions to varying extents. Treatment with 10 ppb NO2 increased shoot biomass in C24, Col-0, and Ler by 3.2-, 1.4-, and 2.3-fold, respectively, compared with control. Treatment with 50 ppb gave similar increases, except in C24 (2.7-fold). The physiological, evolutionary, and genetic significance of these results are discussed below.

When stirred with powdered samarium and iodine in methanol at ice-cold temperature, ethyl 1,2,3-thiadiazole-4-carboxylate 5 underwent unusual reduction involving the dimeric ring enlargement with a sulfur addition, giving dimethyl 1,2,5-trithiepan-4,6-dicarboxylates 7a,b as a cis/trans-isomeric mixture in acceptable yield. The 1,2,3-thiadiazole ring of 5 proved to resist reduction by ordinary reducing agents, where the only choice in most cases was either recovery of the unchanged substrate or decomposition to an intractable mixture of unidentified products and tar., (Copyright 2004 The Royal Society of Chemistry)

Plants take up inorganic nitrogen and store it unchanged or convert it to organic forms. The nitrogen in such organic compounds is stoichiometrically recoverable by the Kjeldahl method. The sum of inorganic nitrogen and Kjeldahl nitrogen has long been known to equal the total nitrogen in plants. However, in our attempt to study the mechanism of nitrogen dioxide (NO(2)) metabolism, we unexpectedly discovered that about one-third of the total nitrogen derived from (15)N-labeled NO(2) taken up by Arabidopsis thaliana (L.) Heynh. plants was converted to neither inorganic nor Kjeldahl nitrogen, but instead to an as yet unknown nitrogen compound(s). We here refer to this nitrogen as unidentified nitrogen ( UN). The generality of the formation of UN across species, nitrogen sources and cultivation environments for plants has been shown as follows. Firstly, all of the other 11 plant species studied were found to form the UN in response to fumigation with (15)NO(2). Secondly, tobacco ( Nicotiana tabacum L.) plants fed with (15)N-nitrate appeared to form the UN. And lastly, the leaves of naturally fed vegetables, grass and roadside trees were found to possess the UN. In addition, the UN appeared to comprise a substantial proportion of total nitrogen in these plant species. Collectively, all of our present findings imply that there is a novel nitrogen mechanism for the formation of UN in plants. Based on the analyses of the exhaust gas and residue fractions of the Kjeldahl digestion of a plant sample containing the UN, probable candidates for compounds that bear the UN were deduced to be those containing the heat-labile nitrogen-oxygen functions and those recalcitrant to Kjeldahl digestion, including organic nitro and nitroso compounds. We propose UN-bearing compounds may provide a chemical basis for the mechanism of the reactive nitrogen species (RNS), and thus that cross-talk may occur between UN and RNS metabolisms in plants. A mechanism for the formation of UN-bearing compounds, in which RNS are involved as intermediates, is proposed. The important broad impact of this novel nitrogen metabolism, not only on the general physiology of plants, but also on plant substances as human and animal food, and on plants as an integral part of the global environment, is discussed.

The importance of nitric oxide (NO) as a signaling molecule to various plant physiological and pathophysiological processes is becoming increasingly evident. However, little is known about how plants protect themselves from nitrosative and oxidative damage mediated by NO and NO-derived reactive nitrogen species (RNS). Peroxynitrite, the product of the reaction between NO and superoxide anion, is considered to play a central role in RNS-induced cytotoxicity, as a result of its potent ability to oxidize diverse biomolecules. Employing heterologous expression in bacteria and yeast, we investigated peroxynitrite-scavenging activity in plants of 2-Cys peroxiredoxin (2CPRX), originally identified as a hydroperoxide-reducing peroxidase that is ubiquitously distributed among organisms. The putative mature form of a chloroplast-localized 2CPRX from Arabidopsis thaliana was overproduced in Escherichia coli as an amino-terminally hexahistidine-tagged fusion protein. The purified recombinant 2CPRX, which was catalytically active as peroxidase, efficiently prevented the peroxynitrite-induced oxidation of a sensitive compound. We also examined in vivo the ability of the Arabidopsis 2CPRX to complement the 2CPRX deficiency of a Saccharomyces cerevisiae mutant. Functional expression in the mutant strain of the Arabidopsis 2CPRX not only increased cellular tolerance to hydrogen peroxide, but also complemented the hypersensitive growth defect induced by nitrite-mediated cytotoxicity. The complemented cells significantly enhanced the capacity to reduce RNS-mediated oxidative damages. The results presented here demonstrate a new role of plant 2CPRX as a critical determinant of the resistance to RNS, and support the existence of a plant enzymatic basis for RNS metabolism.