Environmental and Experimental Botany 60 (2007) 159–169 Roles of hydroxyl radical generating/scavenging mechanisms in pseudo polluted dew in reducing the foliar CO 2 assimilation rate and biomass production of Japanese red pine (Pinus densiflora Sieb. et Zucc.) seedlings Nobutake Nakatani a, , Sachiko Akane a , Masaaki Chiwa a,b , Tsuyoshi Kobayashi c , Hiroshi Sakugawa a a Graduate School of Biosphere Science, Hiroshima University, 1-7-1 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8521, Japan b Department of Forest and Forest Products Sciences, Faculty of Agriculture, Graduate School of Kyushu University, 394 Tsubakuro, Sasaguri 811-2415, Japan c Faculty of Agriculture, Kagawa University, 2393 Ikenobe, Miki-cho, Kita-gun, Kagawa 761-0795, Japan Received 8 September 2006; accepted 1 October 2006 Abstract Hydroxyl radical ( OH) is one of the most highly reactive of all active oxygen species. Aqueous-phase OH is photochemically generated in polluted dew droplets on needle surfaces of Japanese red pine (Pinus densiflora Sieb. et Zucc., an evergreen coniferous tree) in declining pine forests. In former studies, we examined the effects of OH-generating mist solutions, which simulate polluted dew droplets, on needle ecophysiological traits of pine seedlings. Two types of solution with different OH-generating sources, i.e. the photo-Fenton reaction (H 2 O 2 -Fe- oxalate) and photolysis of HONO and NO 2 - [N(III)], had contrasting functional effects on needle gas exchange characteristics even though they had similar OH photoformation rates. In the present study, we investigated the effects of OH-generating mist solutions containing mixed-sources of OH (photo-Fenton and N(III)) on needle gas exchange and the biomass production of potted Japanese red pine seedlings. The N(III) and H 2 O 2 concentrations of the OH-generating mist solutions were regulated to the same concentration (0, 25, 50, 75 or 100 M, pH 5.2–5.3). Treated needles with the lowest photoformation rate and scavenging rate constant of OH had the smallest CO 2 assimilation rate (A max ) and needle conductance (g n ) of all treated pine seedlings. This suggests that the photoformation rate and scavenging rate constant do not always explain the ecophysiological disorders of pine needles subjected to OH-generating wet deposition. On the other hand, the calculated steady-state concentration of OH in the mist solutions was significantly negatively correlated with A max . In addition, pine seedlings with reduced A max showed suppressed biomass (dry weight) production. These results suggest that the OH concentration in the mist solutions is the decisive factor in explaining the negative effects of pseudo dew droplets not only on needle but also plant productivity. We propose that the deposition of polluted dew with chemical components and systems that result in a high OH concentration is one of the causes of growth decline of Japanese red pine. © 2006 Elsevier B.V. All rights reserved. Keywords: Hydroxyl radical; Mist treatment; Photosynthesis; Steady-state concentration; Tree decline; Wet acidic deposition 1. Introduction Active oxygen species (AOS), such as superoxide anion rad- ical (O 2 •- ), hydrogen peroxide (H 2 O 2 ), singlet oxygen ( 1 O 2 ) and hydroxyl radical ( OH), are produced during plant metabolic processes and are normally kept at a low concentration by vari- ous enzymatic and chemical scavenging mechanisms (Asada, 1999; Foyer and Noctor, 2000; Apel and Hirt, 2004). How- Corresponding author. Tel.: +81 82 424 6504; fax: +81 82 424 6504. E-mail address: nnakatan@hiroshima-u.ac.jp (N. Nakatani). ever, when these scavenging mechanisms are disturbed and/or the production of AOS exceeds the capacity to reduce them, AOS accumulate in plant cells. For example, environmental fac- tors that induce stressful conditions such as drought (Egert and Tevini, 2002), cold (Bowers, 1994) and salinity (Gueta-Dahan et al., 1997) promote the accumulation of AOS in plant cells. In addition to these factors, gaseous atmospheric pollutants also induce excess AOS production in plant cells (Morita and Tanaka, 2002). Air pollutants such as sulfur dioxide (SO 2 ) and ozone (O 3 ) are taken into the leaves through the stomata and dissolved in cellular fluid (Kondo and Saji, 1992). In both enzy- matic and non-enzymatic processes, AOS are then produced 0098-8472/$ – see front matter © 2006 Elsevier B.V. All rights reserved. doi:10.1016/j.envexpbot.2006.10.006