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Main conclusion: Ambient concentrations of atmospheric nitrogen dioxide (NO2) inhibit the binding of PIF4 to promoter regions of auxin pathway genes to suppress hypocotyl elongation in Arabidopsis. Ambient concentrations (10–50 ppb) of atmospheric nitrogen dioxide (NO2) positively regulate plant growth to the extent that organ size and shoot biomass can nearly double in various species, including Arabidopsis thaliana (Arabidopsis). However, the precise molecular mechanism underlying NO2-mediated processes in plants, and the involvement of specific molecules in these processes, remain unknown. We measured hypocotyl elongation and the transcript levels of PIF4, encoding a bHLH transcription factor, and its target genes in wild-type (WT) and various pif mutants grown in the presence or absence of 50 ppb NO2. Chromatin immunoprecipitation assays were performed to quantify binding of PIF4 to the promoter regions of its target genes. NO2 suppressed hypocotyl elongation in WT plants, but not in the pifq or pif4 mutants. NO2 suppressed the expression of target genes of PIF4, but did not affect the transcript level of the PIF4 gene itself or the level of PIF4 protein. NO2 inhibited the binding of PIF4 to the promoter regions of two of its target genes, SAUR46 and SAUR67. In conclusion, NO2 inhibits the binding of PIF4 to the promoter regions of genes involved in the auxin pathway to suppress hypocotyl elongation in Arabidopsis. Consequently, PIF4 emerges as a pivotal participant in this regulatory process. This study has further clarified the intricate regulatory mechanisms governing plant responses to environmental pollutants, thereby advancing our understanding of how plants adapt to changing atmospheric conditions. [ABSTRACT FROM AUTHOR]

To gain more insight into the physiological function of nitrogen dioxide ( NO2), we investigated the effects of exogenous NO2 on growth in Arabidopsis thaliana., Plants were grown in air without NO2 for 1 wk after sowing and then grown for 1-4 wk in air with (designated treated plants) or without (control plants) NO2. Plants were irrigated semiweekly with a nutrient solution containing 19.7 mM nitrate and 10.3 mM ammonium., Five-week-old plants treated with 50 ppb NO2 showed a ≤ 2.8-fold increase in biomass relative to controls. Treated plants also showed early flowering. The magnitude of the effects of NO2 on leaf expansion, cell proliferation and enlargement was greater in developing than in maturing leaves. Leaf areas were 1.3-8.4 times larger on treated plants than corresponding leaves on control plants. The NO2-induced increase in leaf size was largely attributable to cell proliferation in developing leaves, but was attributable to both cell proliferation and enlargement in maturing leaves. The expression of different sets of genes for cell proliferation and/or enlargement was induced by NO2, but depended on the leaf developmental stage., Collectively, these results indicated that NO2 regulates organ growth by controlling cell proliferation and enlargement. [ABSTRACT FROM AUTHOR]

Assimilatory nitrite reductase (aNiR) reduces nitrite to an ammonium ion and has siroheme and a [Fe4S4] cluster as prosthetic groups. A reaction mechanism for Nii3, an aNiR from tobacco, is proposed based on high resolution X-ray structures and UV-Vis (ultraviolet-visible) microspectroscopy of Nii3-ligand complexes. Analysis of UV-Vis spectral changes in Nii3 crystals with increasing X-ray exposure showed prosthetic group reductions. In Nii3-NO [ABSTRACT FROM AUTHOR]

Tobacco expresses four isomers of assimilatory nitrite reductase (aNiR), leaf-type (Nii1 and Nii3), and root-type (Nii2 and Nii4). The high-resolution crystal structures of Nii3 and Nii4, determined at 1.25 and 2.3 Å resolutions, respectively, revealed that both proteins had very similar structures. The Nii3 structure provided detailed geometries for the [4Fe-4S] cluster and the siroheme prosthetic groups. We have generated two types of Nii3 variants: one set focuses on residue Met175 (Nii3-M175G, Nii3-M175E, and Nii3-M175K), a residue that is located on the substrate entrance pathway; the second set targets residue Gln448 (Nii3-Q448K), a residue near the prosthetic groups. Comparison of the structures and kinetics of the Nii3 wild-type (Nii3-WT) and the Met175 variants showed that the hydrophobic side-chain of Met175 facilitated enzyme efficiency ( kcat/ Km). The Nii4-WT has Lys449 at the equivalent position of Gln448 in Nii3-WT. The enzyme activity assay revealed that the turnover number ( kcat) and Michaelis constant ( Km) of Nii4-WT were lower than those of Nii3-WT. However, the kcat/ Km of Nii4-WT was about 1.4 times higher than that of Nii3-WT. A comparison of the kinetics of the Nii3-Q448K and Nii4-K449Q variants revealed that the change in kcat/ Km was brought about by the difference in Residue 448 (defined as Gln448 in Nii3 and Lys449 in Nii4). By combining detailed crystal structures with enzyme kinetics, we have proposed that Nii3 is the low-affinity and Nii4 is the high-affinity aNiR. [ABSTRACT FROM AUTHOR]

Production of novel mutants with a high ability to mitigate pollutants is important for phytoremediation. We investigated the use of ion beam irradiation to produce mutants of Ficus pumila L. with an improved ability to mitigate atmospheric nitrogen dioxide (NO2). More than 25,000 shoot explants were irradiated with an ion beam (12C5+, 12C6+, or 4He2+), from which 263 independent plant lines were obtained. The plants were analyzed for NO2 uptake by fumigation with 1 ppm 15N-labeled NO2 for 8 h in light, followed by mass spectrometric analysis. The mean NO2 uptake values of each of the 263 lines differed over a 110-fold range. Propagation was attempted using cuttings from 44 lines showing the greatest NO2 uptake; in total, 15 lines were propagated. Two of the 15 lines showed a mean NO2 uptake 1.7- to 1.8-fold greater than that of the wild-type. This increase in NO2 uptake was heritable in both lines; their progenies showed a significantly greater ability to take up and assimilate NO2 than did the wild-type. RAPD analysis demonstrated DNA variation between the progeny plants and the wild type, suggesting that the progeny were true mutants. These mutants of F. pumila may prove useful in mitigating atmospheric NO2. [ABSTRACT FROM AUTHOR]

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As reported previously, atmospheric nitrogen dioxide (NO2) at an ambient level increased plant size and the contents of cell constituents. We investigated this effect of atmospheric NO2 on decontamination of cadmium (Cd) by kenaf (Hibiscus cannabinus). Seventeen-day-old seedlings of kenaf were grown in air either with NO2 or without NO2. (Plants were exposed to 100 ± 50 ppb NO2 for 10 d under irrigation of 0.1% Hyponex supplemented with 20 μM CdCl2.) Plants were then harvested and the biomass of stems, leaves, and roots, as well as the content of Cd in the organs, was determined. The stem and root biomass per plant were 1.25-1.27-fold greater in +NO2 plants than in -NO2 plants. The Cd content per stem was more than 30% greater in +NO2 plants than in -NO2 plants. [ABSTRACT FROM AUTHOR]

• We report the unexpected novel finding that exogenously supplied atmospheric NO2 at an ambient concentration is a plant vitalization signal to double shoot size and the contents of cell constituents. • When seedlings of Nicotiana plumbaginifolia were grown for 10 wk under natural light and irrigation with 10 m m KNO3 in air containing (+NO2 plants) or not containing (–NO2 plants) 15NO2 (150 ± 50 ppb), shoot biomass, total leaf area, and contents per shoot of carbon (C), nitrogen (N), sulphur (S), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), free amino acids and crude proteins were all approximately 2 times greater in +NO2 plants than in –NO2 plants. • In mass spectrometric analysis of the 15N/14N ratio, it was found that NO2-derived N (NO2-N) comprised < 3% of total plant N, indicating that the contribution of NO2-N to total N was very minor. • It thus seems very likely that the primary role of NO2 is as a multifunctional signal to stimulate plant growth, nutrient uptake and metabolism. [ABSTRACT FROM AUTHOR]

All plants examined to date possess non-symbiotic hemoglobin whose physiological role remains unclear. The present study explored the catalytic function of three representative classes of the plant hemoglobin from Arabidopsis thaliana: AtGLB1, AtGLB2, and AtGLB3. Purified recombinant proteins of these hemoglobins displayed hydrogen peroxide-dependent oxidation of several peroxidase substrates that was sensitive to cyanide, revealing intrinsic peroxidase-like activity. In the presence of nitrite and hydrogen peroxide, AtGLB1 was the most efficient at mediating tyrosine nitration of its own and other proteins via the formation of reactive nitrogen species as a result of nitrite oxidation. AtGLB1 mRNA significantly accumulated in Arabidopsis seedlings exposed to nitrite, supporting the physiological relevance of its function to nitrite and nitrite-derived reactive nitrogen species. [Copyright &y& Elsevier]

S–Nitrosoglutathione (GSNO), an adduct of nitric oxide (NO) with glutathione, is known as a biological NO reservoir. Heterologous expression in Escherichia coli of a cDNA encoding a glutathione-dependent formaldehyde dehydrogenase of Arabidopsis thaliana showed that the recombinant protein reduces GSNO. The identity of the cDNA was further confirmed by functional complementation of the hypersensitivity to GSNO of a yeast mutant with impaired GSNO metabolism. This is the first demonstration of a plant GSNO reductase, suggesting that plants possess the enzymatic pathway that modulates the bioactivity and toxicity of NO. [Copyright &y& Elsevier]

Summary A 12.5-kb DNA fragment with junction regions between the transgene and genomic DNA was cloned from a transgenic tobacco cell line obtained by microprojectile bombardment of plasmid pCaMVNEO. Nucleotide sequence analysis of the fragment (DDBJ accession no. D84238) showed that it carried a 7.7-kb core sequence (concatemer of a complete pCaMVNEO and a partial pCaMVNEO) and two identical 1.3-kb junction sequences that flanked both the 5′ and 3′ ends of the core sequence and had inverted orientations. These sequences had topoisomerase II (Topo II) cleavage sites and adenine and thimine-rich sequences known to be specific to nuclear scaffold-attachment regions (SARs). An in vitro binding assay showed that a 507-bp fragment (designated TJ1) from the 1.3-kb sequence had the ability to bind to nuclear scaffold preparations of cultured tobacco cells, confirmation that the 1.3-kb sequence is an SAR. Insertion of TJ1at the 5′ and 3′ sides of the expression cassette for the npt II gene increased transformant yields 5- to 10-fold and the NPT II enzyme activity per copy of the gene 5-fold. TJ1enhances the integration or expression of the transgene, or both. Clearly, TJ1is very useful for producing transgenic plants. This is the first report on an SAR-like sequence that is located in the transgene locus and enhances transformation efficiency in eukaryotic cells. The possible role of TJ1-SAR in the molecular evolution of plant genome is discussed. [ABSTRACT FROM AUTHOR]

Particle bombardment has proved to be useful for the transformation of plants. We have previously reported successful transient expression of the β-glucuronidase (GUS) gene in cultured plant cells and tissues and the stable transformation of various plants using a pneumatic particle gun. In this chapter, we describe transient expression of the GUS gene in Arabidopsis thaliana leaves and roots using the pneumatic particle gun. [ABSTRACT FROM AUTHOR]

Summary: The emission of N[sub 2] and N[sub 2]O from intact transgenic tobacco (clone 271) expressing antisense nitrite reductase (NiR) mRNA, and wild-type plants grown aseptically, on NO[sub 3][sup –], NO[sub 2][sup –] or NH[sub 4][sup +] -containing medium was investigated. [sup 15]N contents of gas sampled from gas-sealed pots, in which the plants were grown on [sup 15]N-containing medium, were analyzed by gas chromato- graphy and mass spectrometry (GC–MS). No emission of N[sub 2] was detected in either of the gas samples from plant clone 271 or the wild-type grown on NO[sub 3][sup –]-containing medium. N[sub 2]O emission from clone 271 grown on NO[sub 3][sup –]-containing medium was detected, but not from the wild-type plants. The N[sub 2]O emission rate of clone 271 was 106 ng N[sub 2]O mg[sup –1] incorporated N week[sup –1] and the N[sub 2]O emission was inhibited by tungstate (a nitrate reductase inhibitor). No emission of N[sub 2]O was found from clone 271 or wild-type plants grown on medium containing NH[sub 4][sup +]. Emission of N[sub 2]O also was detected from clone 271 grown on NO[sub 2][sup –]-containing medium and its emission rate increased with increasing NO[sub 2][sup –] levels in plants. We speculate that NO[sub 3][sup –] is reduced to NO[sub 2][sup –] and that a part of NO[sub 2][sup –] is metabolized to N[sub 2]O in clone 271. [ABSTRACT FROM AUTHOR]

Suspension-cultured cells (A-18 line) of the liverwort Marchanta polymorpha were bombarded by a pneumatic particle gun with plasmid pCH harbouring the hygromycin phosphotransferase (HPT) gene ( hpt) under the control of the cauliflower mosaic virus (CaMV) 35 S promoter and the nopaline synthase polyadenylation region. Nine weeks after bombardments, 128 hygromycin-resistant calluses were obtained from an approximate total of 7×10 cells. Ten cell lines chosen randomly were analysed further. Southern blot analysis showed that all of the ten lines contain the hpt gene in the genome, demonstrating that these lines are transformants. An HPT enzyme activity assay confirmed the expression of the gene in all of the transformant lines. [ABSTRACT FROM AUTHOR]

Immature pollen from Nicotiana rustica was bombarded with gold particles coated with plasmid DNA encoding neomycin phosphotransferase II (NPTII) and β-glucuronidase (GUS) genes which, respectively, are under the control of the cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (NOS) terminator in the plasmid. Kanamycin-resistant pollen embryoids were selected from the bombarded pollen cells and two independent lines of transgenic plants were regenerated. Enzyme assays showed that one has both NPTII and GUS activities and the other only weak NPTII activity. Southern blot analyses indicated that the former has a DNA fragment corresponding to the intact expression cassettes for both genes in its genome; whereas the latter lacks intact expression cassettes for both genes and has only the intact nptII coding sequence in its genome. The transgenic plants of both lines have 24 chromosomes, confirming haploidy, and they are infertile. These results indicate that transgenic haploid plants can be produced directly by the bombardment-mediated transformation of immature pollen. [ABSTRACT FROM AUTHOR]

Assimilation of nitrogen dioxide in response to fumigation with 15N-labelled nitrogen dioxide was studied in 217 plant taxa. The taxa included 50 wild herbaceous plants collected from roadsides (42 genera, 15 families), 60 cultivated herbaceous plants (55 genera, 30 families) and 107 cultivated woody plants (74 genera, 45 families). Two parameters, the ‘NO2-N content’, or NO2-derived reduced nitrogen content in fumigated plant leaves (mg N g–1 dry weight), and the ‘NO2-utilization index’, or percentage of the NO2-derived reduced nitrogen in the total reduced nitrogen, were determined. The NO2-N content differed 657-fold between the highest (Eucalyptus viminalis; 6·57) and lowest (Tillandsia ionantha and T. caput-medusae; 0·01) values in the 217 taxa; 62-fold in a family (Theaceae) and 26-fold in a species (Solidago altissima). Nine species had NO2-utilization indices greater than 10%, of which Magnolia kobus, Eucalyptus viminalis, Populus nigra, Nicotiana tabacum and Erechtites hieracifolia had NO2-N contents > 4·9. These plants can be considered ‘NO2-philic’ because in them NO2-nitrogen has an important function(s). The Compositae and Myrtaceae had high values for both parameters, whereas the monocots and gymnosperms had low ones. These findings suggest that the metabolic pathway of NO2-nitrogen differs among plant species. The information presented here will be useful for creating a novel vegetation technology to reduce the atmospheric concentration of nitrogen dioxide. [ABSTRACT FROM AUTHOR]

Stable transformation of suspension-cultured cells of tobacco with plasmid DNA (pCaMV-NEO) harbouring the neomycin phosphotransferase II ( npt-II) gene was achieved using a previously described gas-pressure-driven particle acceleration device. The cells were bombarded with DNA-coated gold particles accelerated by the device, and callus containing the introduced gene was selected in the presence of geneticin disulphate. The geneticin-resistant callus exhibited npt-II enzyme activity, and Southern analysis confirmed the stable integration of the foreign gene into the tobacco genome. Transformants were obtained at a rate of more than 1.9 × 10. [ABSTRACT FROM AUTHOR]

Cultured cells of Lavandula vera were entrapped with photosensitive synthetic resin prepolymers (PVA-SbQ). PVA-SbQ-entrapped cells grew well inside gel matrices and synthesized de novo blue pigments in the presence of l-cysteine as an inducer. The entrapped cells were superior to calcium alginate-entrapped cells judging from cell growth and total pigment productivity. Release of the pigments, which were almost insoluble in water, from the gels was markedly enhanced by the increase in hydrophilicity of the cell-entrapping gels. The entrapped cells could be used repeatedly for the pigment production. [ABSTRACT FROM AUTHOR]

It was found that plasmid DNA (YEp13) can be introduced into intact yeast cells by electric field pulses. The frequency of transformation by this electroinjection method depend upon the initial electric field intensity, the capacitance of the electric discharge capacitor, and the number of pulses applied. A maximum number of transformants (90±20/μgDNA) was obtained by three successive pulses with an initial intensity of 5 KV/cm and with a capacitance of 1 μF. The present electroinjection method is simple, and transformants can be obtained within 2 to 3 days after transformation treatment. [ABSTRACT FROM AUTHOR]

Gold particles coated with β-glucuronidase (GUS) mRNA with a 5′ cap structure that had been synthesized in vitro were introduced, by use of a pneumatic particle gun, into pollen grains of lily ( Lilium longiflorum), freesia ( Freesia refracta) and tulip ( Tulipa gesneriana). A fluorometric assay for the GUS activity indicated that in vitro synthesized GUS mRNA introduced into these pollen cells by particle bombardment was successfully expressed. GUS activity in extracts of the bombarded lily pollen became detectable fluorometrically within 30 min after bombardment, peaked at 6 h, then gradually decreased. This activity changed as a function of the developmental stage of the pollen cell of lily. [ABSTRACT FROM AUTHOR]

A number of A/T-rich sequences and a CATGCAT/A sequence are contained in the 5′-upstream regions of the genes encoding concanavalin A (Con A) and canavalin, two major seed storage proteins of Canavalia gladiata D.C. To study the role of these sequences in the seed-specific gene expression, we constructed 5′-deletion mutants and examined the transient expression of β-glucuronidase reporter gene by particle bombardment and the stable expression by Agrobacterium-mediated transformation of tobacco plants. Positive regulatory elements were located in the −894/−602 and −602/−74 regions of the Con A gene, and in the −428/−376, −281/−155 and −155/−50 regions of the canavalin gene. In addition, the results suggested that the A/T-rich sequences in the 5′-upstream region of the Con A gene play a role in transcriptional activation, but that those of the canavalin gene have little effect on the gene expression. The CATGCAT/A sequence was not sufficient by itself for high levels of expression of both the Con A and canavalin genes. The canavalin polypeptide amounted to about 1% of the total extractable protein in the transgenic tobacco seeds, but the Con A polypeptide was not detected in the extractable protein. [ABSTRACT FROM AUTHOR]

Successful transient expression of β-glucuronidase (GUS) in Arabidopsis thaliana leaves and roots and Brassica napus stems was obtained after gene delivery with a pneumatic particle gun driven by compressed air. Effects of the pneumatic pressure used to accelerate the particles (accelerating pressure; 85 to 200 kg/cm) and of preculture periods of plant tissues (0 to 6 days) on the efficiency of gene delivery were studied. In A. thaliana leaves, best results were obtained at 115 kg/cm of accelerating pressure and 3 days of preculture. In A. thaliana roots, the optimum was at 200 kg/cm of accelerating pressure and 3 days of preculture. These results indicate that both preculture period and accelerating pressure are vital factors that determine the efficiency of gene delivery by particle gun. [ABSTRACT FROM AUTHOR]

A plasmid pARK 22 harbouring the bar gene encoding phosphinothricin acetyltransferase (PAT) under the control of the cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (NOS) terminator was constructed and introduced into root sections of Arabidopsis thaliana using the pneumatic particle gun. The root sections that had been bombarded with this plasmid gave four to eight times higher yield of drug-resistant calluses than those sections bombarded with pCaMVNEO or pCH, which respectively contain the neomycin phosphotransferase and hygromycin phosphotransferase genes. Among a number of primary transformant (T) plants obtained from independent bialaphos-resistant calluses, three were studied by Southern blot hybridization and PAT enzyme activity analyses, confirming the stable integration of the foreign gene into the Arabidopsis genome and its expression in plants. The progeny analysis showed transmission of the foreign gene and its expression in up to the T generation. Some of the T progeny showed morphological abnormalities. Thus, the bar gene can be used effectively to allow selection of transgenic A. thalianna plants. [ABSTRACT FROM AUTHOR]

Transformed roots of Cucumis sativus were obtained from cotyledon tissues that had been bombarded with gold particles coated with plasmid pE7.4 using a pneumatic particle gun. This plasmid contains rolA, rolB, rolC genes and ORF 13 of the 7.4 kb Eco RI fragment of T-DNA of pRi 1724 isolated from Agrobacterium rhizogenes MAF 03-01724. The nature of the tissue and the composition of the culture media used greatly influenced the recovery of transformed roots. The transgenic nature of the derived roots was confirmed by the vigorous. highly-branched growth seen on a phytohormone-free medium. The stable integration of rol genes into the cucumber genome was confirmed by Southern blot analysis. [ABSTRACT FROM AUTHOR]

Tobacco (Nicotiana tabacum L. cv. Xanthi XHFD8) possesses four nitrite reductase (NiR) genes: nii1, nii2, nii3, and nii4. Their differential expression in leaves and roots was investigated by quantitative competitive RT-PCR using gene-specific primer pairs. These results appear to contradict existing views on the expression of these NiR genes: (i) the mRNA of each of the four NiR genes was distinguishable both in leaves and roots; (ii) nitrate treatment increased nii1 and nii3 mRNA in leaves and roots by at least 4-fold (at least 5-fold in nii2 and nii4 mRNA); and (iii) the steady-state levels of nii1 and nii3 mRNA were almost the same in leaves (6–7×105 and about 3×106 copies μg−1 of total RNA before and after nitrate treatment, respectively) and in roots (3–4×104 and 3–6×105 copies μg−1 of total RNA before and after nitrate treatment, respectively). Very similar relationships were obtained for the steady-state levels of nii2 and nii4 mRNA in roots (2–4×105 and 8×106 copies μg−1 of total RNA before and after nitrate treatment, respectively), and in leaves (5–9×104 and 4×105 copies μg−1 of total RNA before and after nitrate treatment, respectively). These results demonstrate that nii1 and nii3 transcripts are a dominating, but not exclusive, NiR mRNA in leaves, and the same is true for nii2 and nii4 transcripts in roots. [ABSTRACT FROM PUBLISHER]

Levels of NO-3, NO-2 and NH+4 ions in leaves of six plant species were determined by capillary electrophoresis. Levels of NO-3 ions differed by a factor of more than 350 in the six species. NO-2 and NH+4 ions were detected in all species examined but at lower and more similar respective levels than NO-3 ions. [ABSTRACT FROM AUTHOR]

Successful transient expression of genes (luciferase and β-glucuronidase) in cultured tobacco cells and adventitious shoots of egg-plant hypocotyls can be obtained by use of a simple biolistic device. This device is driven by controlled gas pressure from a cylinder of nitrogen gas. It has the advantages of being free of explosive heat and of avoiding cell damage caused by expanding gas, due to the 'self-sealing' effect of the projectile. [ABSTRACT FROM AUTHOR]

Exposing Arabidopsis thaliana (Arabidopsis) seedlings fed with soil nitrogen to 10–50 ppb nitrogen dioxide (NO2) for several weeks stimulated the uptake of major elements, photosynthesis, and cellular metabolisms to more than double the biomass of shoot, total leaf area and contents of N, C P, K, S, Ca and Mg per shoot relative to non-exposed control seedlings. The 15N/14N ratio analysis by mass spectrometry revealed that N derived from NO2 (NO2-N) comprised < 5% of the total plant N, showing that the contribution of NO2-N as N source was minor. Moreover, histological analysis showed that leaf size and biomass were increased upon NO2 treatment, and that these increases were attributable to leaf age-dependent enhancement of cell proliferation and enlargement. Thus, NO2 may act as a plant growth signal rather than an N source. Exposure of Arabidopsis leaves to 40 ppm NO2 induced virtually exclusive nitration of PsbO and PsbP proteins (a high concentration of NO2 was used). The PMF analysis identified the ninth tyrosine residue of PsbO1 (9Tyr) as a nitration site. 9Tyr of PsbO1 was exclusively nitrated after incubation of the thylakoid membranes with a buffer containing NO2 and NO2− or a buffer containing NO2− alone. Nitration was catalyzed by illumination and repressed by photosystem II (PSII) electron transport inhibitors, and decreased oxygen evolution. Thus, protein tyrosine nitration altered (downregulated) the physiological function of cellular proteins of Arabidopsis leaves. This indicates that NO2-induced protein tyrosine nitration may stimulate plant growth. We hypothesized that atmospheric NO2 at ambient concentrations may induce tyrosine nitration of PYR/PYL/RCAR receptors in Arabidopsis leaves, followed by degradation of PYR/PYL/RCAR, upregulation of target of rapamycin (TOR) regulatory complexes, and stimulation of plant growth. [ABSTRACT FROM AUTHOR]