不同浓度O_3对黄檗幼苗叶片生理特征的影响

以4年生黄檗(Phellodendron amurense Rupr.)幼苗为实验材料,利用开顶箱模拟法,开展了4个不同浓度O340(对照)、80、120和200 nmol·mol-1的气体熏蒸实验,测定并分析了黄檗叶片光合色素含量、净光合速率、膜脂质过氧化产物和抗氧化酶活性等生理指标的变化规律。结果表明:与CK相比,随着O3浓度增加和处理时间延长,黄檗幼苗叶片叶绿素总含量、类胡萝卜素含量和净光合速率显著下降(P0.05),最大降幅分别为52.76%、44.57%和54.67%;叶片离子渗漏率、丙二醛含量显著增加(P0.05),最大增幅分别为101.49%和70.21%。当O3浓度为80 nmol·mol-1时,叶片最大光化学效率(Fv/Fm)无显著变化,而O3浓度为120和200 nmol·mol-1时,Fv/Fm显著下降(P0.05),表明黄檗幼苗的光合系统功能在超高浓度臭氧下受到了显著伤害。叶片抗氧化超氧化物歧化酶活性在开始处理后的前14 d升高,随后下降,最大降幅为17.22%。O3浓度为120和200nmol·mol-1时,超氧阴离子(O2-·)含量显著升高(P0.05);O3浓度为80 nmol·mol-1时,O2-·含量在处理7 d与CK相比显著增加,随后无显著差异。表明黄檗幼苗在生长初期对中等高浓度(80 nmol·mol-1)臭氧熏蒸具有一定的适应和耐受性是因其具有较强抗氧化保护能力,更高浓度的O3或更长时间的熏蒸对叶片光合系统产生破坏。     

毛竹对大气臭氧胁迫的生理响应变化

以毛竹1年生盆栽苗为材料,运用开顶式气室(OTCs)模拟环境背景大气O3浓度(AA,40～45 nL·L-1)和高O3浓度(EO,92～106 nL·L-1)情景,分析毛竹叶片光合生理、脂质过氧化及抗氧化酶等主要生理指标的变化,为气候变化背景下的竹林培育应对策略提供理论依据.结果显示:(1)EO较AA在同一处理时间的毛竹叶片O3通量均显著升高,且二处理的叶片O3通量均随着处理时间的延长呈升高趋势.(2) EO较AA的光合速率(Pn)、气孔导度(Gs)和可溶性糖含量均显著下降,且叶片叶绿素(ChD含量、胞间CO2浓度(Ci)显著下降的时间点分别出现在EO处理的60 d和92 d,可溶性蛋白在处理60 d后显著升高;随处理时间的延长,EO的叶片Pn、Ci、Chl含量均呈下降趋势,可溶性糖和可溶性蛋白含量呈先升高后降低的趋势;Pn下降由气孔限制因素引起.(3)超氧自由基(O2)含量、丙二醛(MDA)含量、相对电导率分别在处理29 d、60 d、60 d后均显著升高,且随着处理时间的延长呈升高趋势.(4)超氧化物歧化酶(SOD)活性在高浓度O3处理60 d时显著升高,后显著下降,而POD活性均显著升高,且SOD和POD活性均随着处理时间呈先升高后降低的趋势.研究表明,毛竹对大气高O3胁迫存在着短时间的主动生理生化适应,但长期高O3胁迫会对毛竹造成严重的过氧化伤害,从而影响毛竹的正常生长.     

黄条金刚竹对高浓度CO2和O3及其复合作用的生理响应

以黄条金刚竹为试材,环境背景大气为对照,应用开顶式气室(OTCs)熏蒸法模拟大气高浓度CO2(700μmol.mol-1)、O3(100nmol.mol-1)及其复合作用情景,分析叶片光合色素、膜脂过氧化及抗氧化酶等的变化规律。结果显示:(1)与对照相比,高浓度O3处理103d的黄条金刚竹叶片叶绿素a、叶绿素b、总叶绿素含量和叶绿素a/b、叶绿素/类胡萝卜素及SOD、POD、CAT活性均显著下降,而超氧阴离子和丙二醛含量、相对电导率、APX活性均显著升高,类胡萝卜素含量变化不明显。(2)与对照相比,同期高浓度CO2处理的叶片叶绿素a、叶绿素b、总叶绿素含量和叶绿素/类胡萝卜素均显著升高,而叶绿素a/b和超氧阴离子、丙二醛含量及SOD、POD、CAT、APX活性显著降低,相对电导率和类胡萝卜素含量变化不明显。(3)高浓度O3和CO2复合作用下,除叶绿素a/b和CAT活性显著下降外,其余测定指标均与对照无明显变化。研究表明:高浓度O3使黄条金刚竹叶片活性氧产生速率提高,抗氧化酶活性和光合色素含量降低,膜脂过氧化程度加剧,膜结构破坏,表现出严重的伤害效应;而高浓度CO2能降低叶片活性氧产生速率,减轻膜脂过氧化程度,提高光合色素含量,表现出保护效应;高浓度O3和CO2复合处理能使叶片维持比高浓度O3处理更高的光合色素含量和抗氧化酶活性,即高浓度CO2能在一定程度上有效地缓解高浓度O3对黄条金刚竹所造成的生理伤害。     

施氮对镉胁迫下山杨幼苗叶片氮磷钾吸收及镉积累量的影响

为探究山杨在重金属镉胁迫与增施外源氮复合处理条件下的生长及元素积累差异,该研究以山杨幼苗为材料,通过盆栽实验研究在重金属镉(Cd)胁迫下增施外源氮(N)对其形态、叶绿素、淀粉和可溶性糖含量、叶片中全氮(N)、全磷(P)、全钾(K)以及Cd含量的影响。结果表明：(1)单独Cd处理显著抑制山杨幼苗的叶片宽度和茎粗;施加外源N能够缓解Cd胁迫对植株的毒害,其叶长、叶宽和茎粗较单独Cd处理显著增加。(2)单独Cd处理后山杨叶片的叶绿素含量和淀粉含量均比对照显著下降,而其可溶性糖含量显著增加;与单独Cd处理相比,Cd+N复合处理后山杨的叶绿素含量显著增加,淀粉含量却显著下降,而可溶性糖含量略有降低。(3)与对照相比,单独Cd处理后,山杨叶片全N含量显著下降,全P含量显著增加;单独N和Cd+N复合处理后叶片全N含量均显著增加,全P含量均显著下降;全K含量在各处理下均无显著差异。(4)在Cd胁迫下山杨幼苗叶片的Cd含量比对照极显著增加,且Cd+N复合处理后叶片的Cd含量约是单独Cd处理的2倍。研究认为,增施氮素处理可显著提高山杨幼苗对镉胁迫环境的适应能力以及叶片对镉的富集能力。     

钙对烟草叶片热激忍耐和活性氧代谢的影响

热胁迫导致烟草叶片细胞膜系统显著受损,表现为SOD活性降低和MDA含量明显升高,叶片叶绿素含量下降,活性氧增加。10mmol/LcaCl2溶液处理烟草幼苗后,能有效降低热胁迫下叶片细胞膜透性,维持较高的SOD和CAT等抗氧化酶活性,减缓O2^-形成和膜脂过氧化反应。研究结果表明,CaCl2处理提高了烟草叶片膜稳定性和膜保护酶活性,有利于保护细胞膜结构,降低高温对烟草幼苗的伤害。钙离子螯合剂EGTA能在一定程度上降低烟草叶片的抗热性。     

土壤水分和氮素有效性对刺槐幼苗叶片活性氧产生和清除的影响

采用施加氮肥和人工控水的方法,以一年生刺槐幼苗为材料进行盆栽实验,探讨提高土壤氮素含量对不同土壤水分条件下刺槐幼苗叶片中活性氧产生和清除的影响。结果表明:(1)相同氮素水平下,降低土壤水分含量引起刺槐生物量和叶片光合色素含量降低,而过氧化氢(H2O2)含量升高;抗氧化酶系统中的超氧化物歧化酶(SOD)和过氧化物酶(POD)活性和过氧化氢酶(CAT)活性不同程度降低;抗氧化剂中抗坏血酸(ASA)含量和还原型谷胱甘肽(GSH)含量均有所提高;MDA含量逐渐升高,而同期细胞膜相对电导率显著升高。(2)相同水分条件下,提高土壤氮素水平显著提高了刺槐幼苗叶片光合色素含量,同时也一定程度提高了总生物量,显著降低了H2O2含量;SOD、POD和CAT活性不同程度升高;ASA含量和GSH含量则表现出不同程度下降;相对电导率显著降低同时MDA含量一定程度降低。因此,增加土壤氮素有效性可显著提高刺槐幼苗叶片光合色素含量,显著抑制活性氧的产生,一定程度提高总生物量和抗氧化酶活性,降低膜脂过氧化程度,从而有利于缓解干旱引起的伤害。     

模拟大气硫酸铵污染对香樟幼苗生长及光合特性的影响

通过室内盆栽实验研究了大气颗粒污染物硫酸铵对香樟幼苗生长及光合特性的影响。结果表明,香樟幼苗叶片涂抹硫酸铵处理对植物生长无显著影响;低浓度硫酸铵(2 g·L^-1)提高了叶片叶绿素含量,而高浓度(4 g·L^-1)却降低了叶片叶绿素含量;与对照相比,低浓度处理的香樟叶片净光合速率、气孔导度、胞间二氧化碳浓度与蒸腾速率无显著差异;高浓度处理的香樟叶片净光合速率与蒸腾速率高于对照,而气孔导度与胞间二氧化碳浓度与对照无显著差异。机理分析表明,硫酸铵颗粒物主要通过影响叶片气孔导度来影响植物光合特性。     

高浓度O_3对银杏凋落叶化学组成的影响

北半球近地大气O3浓度在最近几十年里持续升高,对陆地生态系统产生深远的影响。本文选取5年生银杏(Ginkgo biloba)为研究对象,利用开顶式气室(OTCs)开展了3个不同浓度的O3(自然对照浓度约40 nmol·mol-1、处理浓度80和120 nmol·mol-1)熏蒸试验,持续熏蒸2个生长季(2012—2013年)后,于2013年11年月初收集其凋落叶,测定并分析不同处理下银杏凋落叶化学成分的变化。结果表明:与对照相比,高浓度O3处理组银杏凋落叶的N、K含量均显著升高,总酚含量以及C/N、木质素/N降低;而C、P、木质素含量以及C/P并未呈现出显著差异。可溶性糖、缩合单宁含量在两高浓度O3处理下变化趋势并不一致:与对照相比,可溶性糖含量仅在120 nmol·mol-1处理下表现出显著差异,降低38%;缩合单宁含量在80 nmol·mol-1处理下显著升高(343%),而在120 nmol·mol-1处理下无显著变化。木本植物凋落物的化学组成在O3熏蒸下会发生一定变化,这种变化可能会对树木凋落物在O3污染地区的分解及区域气候变化下森林生态系统的物质循环,特别是碳循环产生重要影响。     

大气CO2与O3浓度升高对毛竹叶片膜脂过氧化和抗氧化系统的影响

以盆栽一年生毛竹为材料,利用开顶式气室模拟大气O3和CO2浓度升高情景,分析毛竹叶片细胞膜脂过氧化、抗氧化酶(SOD、CAT、POD和APX)活性和渗透调节物质(可溶性糖和可溶性蛋白)含量的响应规律,为气候变化背景下的竹林适应性管理提供理论支撑.结果表明:(1)短期(30 d)高浓度O3(92～106 nL-L-1)胁迫能刺激毛竹叶片抗氧化酶活性和渗透调节物质含量显著提高,清除活性氧能力增强,并未出现细胞膜脂过氧化现象,即毛竹对短期高浓度O3具有较强的适应性;但长期(90 d)O3胁迫条件下,毛竹叶片抗氧化酶活性显著降低,叶片细胞膜脂过氧化程度加剧,膜结构破坏,发生严重的氧化伤害.(2)短期高浓度CO2(685～730 μmol·mol-1)处理总体上对毛竹叶片细胞膜脂过氧化和抗氧化酶系统活性影响并不明显;而长期高浓度CO2处理能一定程度上增强毛竹的抗氧化能力和渗透调节功能,减轻氧化损伤,体现了对毛竹的保护效应.(3)高浓度O3和CO2复合作用下,相对于单一高浓度O3胁迫,毛竹叶片能够维持较高的抗氧化酶活性和渗透调节物质含量,有效地调节活性氧产生与清除间的平衡,细胞膜脂过氧化程度变化不明显,说明高浓度CO2可在一定程度上缓解高浓度O3对毛竹所造成的生理伤害.     

外源抗坏血酸对臭氧胁迫下水稻叶片膜保护系统的影响

在田间原位条件下,运用OTCs（open top chamber）装置研究了外源抗坏血酸（exogenous ascorbate acid,ExAsA）对臭氧（O3）胁迫下水稻（Oryza Sativa L.）叶片膜保护系统的影响.研究发现,O3胁迫下的水稻叶片经过ExAsA处理后叶绿素a含量显著升高,而叶绿素b含量变化不明显;相对于对照,经ExAsA处理后的水稻叶片过氧化氢（H2O2）和丙二醛（MDA）含量及相对电导率（REC）均降低,超氧化物岐化酶（SOD）和抗坏血酸过氧化物酶（APX）活性明显提高,抗氧化剂类胡萝卜素（Carotene）含量升高.这表明,ExAsA改善了O3胁迫下水稻叶片的抗氧化系统功能,减少了叶片中活性氧（activity oxygen species,AOS）的积累,抑制了脂质过氧化（lipid peroxidation,LP）,延迟了O3对水稻叶片的老化作用,提高了水稻叶片对O3危害的抗性.     

Effects of ozone exposure on sub-tropical evergreen Cinnamomum camphora seedlings grown in different nitrogen loads

The present study aims in investigating the individual and combined effects of ozone (O₃) exposure and nitrogen (N) load on the growth and photosynthetic characters of Cinnamomum camphora seedlings, a dominant evergreen broadleaf tree species in sub-tropical regions. The seedlings were supplied with N as NH₄NO₃ solution at 0, 30 and 60 kg ha⁻¹ year⁻¹ (simplified as N0, N30, N60, respectively) and were exposed to ambient O₃ concentration (AA) or elevated [O₃] (E-O₃, AA +60 ppb) for one growth season. E-O₃ induced significant negative effects on foliar photosynthesis, including lower photosynthetic rate, reduced carboxylation efficiency, quantum yield of PSII and photosynthetic pigment contents, despite no effect on growth. In contrast, N load acted as fertilization effects. Medium N (N30) increased photosynthetic pigments and stem-base diameter growth relative to N0, whereas high N load (N60) significantly enhanced the growth, photosynthetic pigments, and dark and light action of photosynthesis of C. camphora seedlings. No significant interactive effects of O₃ and N load on the growth, net photosynthetic rate and pigment contents of the seedlings were found, suggesting that N supply to the soil at ≤60 kg ha⁻¹ year⁻¹ does not significantly change the sensitivity of C. camphora to ozone.     

Improving key enzyme activities and quality of rice under various methods of zinc application

Zinc (Zn) is an important micronutrient for the physiology of plants. It is poorly available to the plants in soil solution. A pot experiment was conducted to evaluate effectiveness of various Zn application methods on key enzyme activities and protein content of two contrasting rice genotypes viz., PD16 (Zn efficient) and NDR359 (Zn inefficient). The treatments were, control (0 mg Zn kg⁻¹ soil), soil application (5 mg Zn kg⁻¹ soil), foliar application (0.5 % ZnSO₄ + 0.25 % lime at 30, 60 and 90 days after transplanting), soil (5 mg Zn kg⁻¹ soil) + foliar application of 0.5 % ZnSO₄ + 0.25 % lime at 30, 60 and 90 days after transplanting. Among all the methods tested soil+foliar application of Zn fertilizers was found most effective in increasing superoxide dismutase (SOD) and carbonic anhydrase (CA) activities as well as chlorophyll and protein content in both the rice varieties. NDR359, showed higher enzyme activities and more chlorophyll content in leaves than PD16, when Zn was applied either through foliar spray alone or in soil along with foliar application. Regarding the protein content in grains, PD16 showed higher protein content than NDR359, thus showed better translocation of Zn from leaves to grains.     

Foliar fertilizers for the management of phoma leaf spot on coffee seedlings

The growing demand for alternative strategies for plant disease management has sought a reduction in the use of fungicides via the employment of resistance inducers and foliar fertilizers, among others. Therefore, this study aimed at evaluating the following foliar fertilizers for the management of phoma leaf spot: a foliar fertilizer based on macro‐ and micronutrients (Fmm: 10% N, 4% S, B, 5% Fe and 5% Zn); one based on cobalt and molybdenum (Fcm: 2% Co and 3% Mo); manganese phosphite (FMn: 30% P₂O₅ and 9% Mn); and the FMn+Fmm, Fcm+Fmm and FMn+Fcm+Fmm associations compared to a boscalid fungicide and a control with no treatment. The disease severity, the chlorophyll a and b contents, the net photosynthetic rate (LPR), the phenylalanine ammonia lyase (PAL) activity and the lignin content in leaves were assessment. Based on the severity, the area under the disease severity progress curve (AUSPC) and the efficiency of disease control in relation to untreated plants were calculated. The experimental design was in randomized blocks with eight treatments and three replications. Treatments FMn+Fmm and Fcm were the most effective in reducing the AUPSC in comparison with the control and promoted an increased activity of PAL. FMn was the treatment that promoted the highest increase in the LPR. There were no effects of the treatments on the lignin content compared to the control.     

Oligogalacturonides stimulate antioxidant system in alfalfa roots

Alfalfa (Medicago sativa L.) roots were treated with 50 and 100 μg cm⁻³ of oligogalacturonide (OGA) solutions with a degree of polymerization between 7 and 15. Changes in the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) as well as ascorbate (ASC) content were determined in crude extract of alfalfa roots after 30, 60 and 120 min of treatment. An increase in the SOD activity was observed in roots treated with 50 and 100 μg cm⁻³ OGA, which could be related to its O₂ ^·− scavenging function. As concern H₂O₂ scavenging, CAT activity was increased in the first 30 min by both OGA concentrations, while POX was a key enzyme at higher OGA concentration and treatment duration. ASC content firstly increased upon exposure to high OGA concentration, and then decreased after longer treatment while low OGA concentration had no effect on ASC content.     

Effects of nitrogen and phosphorus imbalance on photosynthetic traits of poplar Oxford clone under ozone pollution

Ozone (O₃) pollution and the availability of nitrogen (N) and phosphorus (P) in the soil both affect plant photosynthesis and chlorophyll (Chl) content, but the interaction of O₃ and nutrition is unclear. We postulated that the nutritional condition changes plant photosynthetic responses to O₃. An O₃-sensitive poplar clone (Oxford) was subject to two N levels (N0, 0 kg N ha^??1; N80, 80 kg N ha^??1), two P levels (P0, 0 kg P ha^??1; P80, 80 kg P ha^??1) and three levels of O₃ exposure (ambient concentration, AA; 1.5?×?AA; 2.0?×?AA) over a growing season in an O₃ free air controlled exposure (FACE) facility. The daily change of leaf gas exchange and dark respiration (R_d) were investigated at mid-summer (August). Chl a fluorescence was measured three times in July, August and September. At the end of the growing season, Chl content was measured. It was found that Chl content, the maximum quantum yield (F_v/F_m), Chl a fluorescence performance index (PI) and gas exchange were negatively affected by elevated O₃. Phosphorus may mitigate the O₃-induced reduction of the ratio of photosynthesis to stomatal conductance, while it exacerbated the O₃-induced loss of F_v/F_m. Nitrogen alleviated negative effects of O₃ on F_v/F_m and PI in July. Ozone-induced loss of net photosynthetic rate was mitigated by N in medium O₃ exposure (1.5?×?AA). However, such a mitigation effect was not observed in the higher O₃ level (2.0?×?AA). Nitrogen addition exacerbated O₃-induced increase of R_d suggesting an increased respiratory carbon loss in the presence of O₃ and N. This may result in a further reduction of the net carbon gain for poplars exposed to O₃.     

Responses of gas exchange,cellular membrane integrity,and antioxidant enzymes activities of salinity-stressed winter wheat to ozone pollution

A sand-culture experiment was conducted in open-top chambers which were constructed in a greenhouse to investigate the responses of salt-stressed wheat (Triticum aestivum L.) to O₃. Plant seeding of JN17 (a popular winter wheat cultivar) was grown in saltless (−S) and saline (+S, 100 mM NaCl) conditions combined with charcoal-filtered air (CF, < 5 ppb O₃) and elevated O₃ (+O₃, 80 ± 5 ppb, 8 h day⁻¹) for 30 d. O₃ significantly reduced net photosynthetic rate (PN), stomatal conductance, chlorophyll contents and plant biomass in -S treatment, but no considerable differences were noted in those parameters between +O₃+S and CF+S treatments. O₃-induced loss in cellular membrane integrity was significant in -S plants, but not in +S plants evidenced by significant elevations being measured in electrolyte leakage (EL) and malondialdehyde (MDA) content in -S plants, but not in +S plants. Both O₃ and salinity increased proline content and stimulated antioxidant enzymes activities. Soluble protein increased by salinity but decreased by O₃. Abscisic acid (ABA) was significantly elevated by O₃ in -S plants but not in +S plants. The results of this study suggested that the specificity of different agricultural environments should be considered in order to develop reliable prediction models on O₃ damage to wheat plants.     

Maturation in larch : I. Effect of age on shoot growth, foliar characteristics, and DNA methylation

The time course of maturation in eastern larch (Larix laricina [Du Roi] K. Koch) was examined by grafting scions from trees of different ages onto 2-year-old root stock and following scion development for several years. Height, diameter, foliar chlorophyll content, and rooting ability of scion-derived cuttings all varied linearly as a function of log₁₀ age. Chlorophyll content (milligrams per gram of dry weight) increased while height, diameter, and ability to root decreased with age (P < 0.01). The tendency toward orthotropic growth and branch formation per centimeter of main stem decreased abruptly between age 1 and 5 years (P < 0.01). Total chlorophyll content of both long and short shoot foliage increased by 30 to 50% with increasing age, but the chlorophyll a/b ratio did not change. Also, juvenile long shoot needles were significantly longer than mature (P < 0.01). Surprisingly, the juvenile scions produced more total strobili over two successive years, but the mature scions produced a significantly higher proportion of male strobili (P < 0.001 year 1; P < 0.02 year 2). The age-related changes in foliar traits were not associated with changes in DNA methylation between juvenile and mature scions. Using HPLC, we found that 20% of foliar DNA cytosine residues were methylated in both scion types.     

Retardation of Senescence in Red Clover Leaf Discs by a New Antiozonant, N-[2-(2-Oxo-1-imidazolidinyl)ethyl]-N'-phenylurea

Dark-induced senescence in leaf discs from O₃-sensitive red clover trifoliates (Trifolium pratense L. cv. `Pennscott') was markedly retarded by treatment with N-[2-(2-oxo-1-imidazolidinyl)ethyl-N′-phenylurea (EDU). EDU also protects against acute and chronic foliar O₃ injury when sprayed on intact leaves or supplied to the plants through soil application. Senescence retardation was measured by time-dependent analyses of chlorophyll, protein, and RNA in discs floated on aqueous EDU solutions ranging from 0 to 500 micrograms per milliliter EDU. Chlorophyll degradation, total protein, and nucleic acids were followed over 10-day test periods.     

Free Radical Scavengers and Photosynthetic Pigments in Pinus Cembra L. Needles as Affected by Ozone Exposure

The goal of this study was the characterization of the antioxidative protection system of current and 1-year-old needles of a cembran pine (Pinus cembra L.) and its possible responses to elevated concentrations of atmospheric O₃. Twigs of a mature cembran pine at the alpine timberline (1950 m a.s.l.) were exposed in climate-controlled twig chambers for 91 d to charcoal-filtered air (CF), ambient air O₃ concentration (A), and two-fold ambient air O₃ concentration (2A). Additionally, a chamberless control group (AA) was used to examine chamber effects. At the end of the fumigation period the contents of free radical scavengers and photosynthetic pigments were measured in the needles. Independent from O₃ exposure, total ascorbate and -tocopherol contents were higher in 1-year-old needles compared to the current flush while the opposite was found for glutathione. The amounts of pigments and antioxidants in P. cembra needles were comparable to those in other conifers growing at high-elevation sites. The only hint toward O₃ induced changes in the composition of antioxidants was an increase in the glutathione redox state toward more oxidation in 1-year-old needles upon exposure to A or AA conditions, but not upon 2A exposure. Chlorophyll and carotenoid contents were not affected by O₃ neither in current- nor in previous-year needles. The de-epoxidation state of the xanthophyll cycle pigments, however, was significantly increased in 1-year-old needles under A and AA compared to the CF control, but not under 2A. Hence, Pinus cembra, which is well adapted to the extreme environment of the timberline ecotone, exhibited only marginal biochemical changes in response to elevated O₃.