Interactive effects of ozone and drought stress on pigments and activities of antioxidative enzymes in Pinus halepensis

The aim of this study was to investigate the interactive effects of ozone (O₃) and drought on pigments and antioxidant enzymes of Aleppo pine (Pinus halepensis). Two‐year‐old seedlings were exposed in open‐top chambers to charcoal‐filtered air or non‐filtered air plus an additional 40 nL L^?1 of ozone. After 20 months of O₃ exposure, a subset of plants was subjected to drought stress by withholding water supply for 11 d. Ozone induced higher guaiacol peroxidase, catalase and KCN‐resistant superoxide dismutase (SOD) activities in young needles, while drought stress increased glutathione reductase and CuZnSOD. One‐year‐old needles showed lower capacity to activate these enzymes in response to stress. Both ozone and drought activated the xanthophyll cycle pool and reduced chlorophyll contents in both current and 1‐year‐old needles. The combined effects of ozone and drought decreased antioxidant enzyme activities and the capacity of recovering after re‐watering. Similarly, interactive effects of O₃ and drought reduced xanthophyll‐mediated photoprotection capacity in 1‐year‐old needles but induced a higher conversion of the cycle in current‐year needles. These results showed that ozone modified the Aleppo pine response to drought stress, suggesting that this pollutant might be reducing the ability of this species to withstand other environmental stresses.     

Drought tolerance of Periploca sepium during seed germination: antioxidant defense and compatible solutes accumulation

Periploca sepium Bunge is a native and widespread shrub on the Loess Plateau, an arid and semi-arid region in China. To understand the adaptability of its seed germination to dry environments, we investigated the germination rate, water relations, lipid peroxidation, antioxidant capacity and accumulation of major organic solutes during seed germination under water deficit conditions. Results showed that seeds pre-treated by hydration–dehydration or ?0.9 MPa PEG germinated faster than control seeds, indicating strong resistance of P. sepium to drought condition. The re-dried seeds showed higher proline, total free amino acids (TFAA) and soluble proteins (SP) contents than control dry seeds, indicating the maintenance of physiological advancement when dehydrated. Osmotic stress made seed germination stay on the plateau phase (phase II). However, germinating seeds moved into phase III immediately once transferred into distilled water. Large increases in SP and soluble sugars (SS) of both re-dried and osmotic stressed seeds help themselves to resist drought stress. The re-hydrated seeds showed significantly higher levels of proline, TFAA, SP and SS than control seeds. The largely accumulated SS during osmotic stress declined sharply when transferred into distilled water. Our data demonstrate that P. sepium’s tolerance to drought stresses during germination is associated with enhanced activity of antioxidant enzymes and accumulation of some compatible solutes. Seed physiological advancement progressed slowly under low water conditions and it was maintained when seeds were air dried. This strategy ensures high and more rapid seed germination of P. sepium under drying and wetting conditions in drought-prone regions.     

Changes in antioxidant enzyme activities and gene expression in two muskmelon genotypes under progressive water stress

Responses of two muskmelon (Cucumis melo L.) genotypes (drought tolerant SC-15 and drought susceptible EC-564755) were analyzed at 0, 7, 14, and 21 d of progressive water stress. Although water deficit caused a significant decline in relative water content, the magnitude of reduction was lower in SC-15. Electrolyte leakage, hydrogen peroxide, and malonydialdehyde generation were higher in EC-564755, whereas accumulation of proline was higher in SC-15. Higher activities of antioxidant enzymes, such as catalase, superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, and glutathione reductase, and higher expression of the respective genes were recorded in SC-15 than in EC-564755. Expressions of DREB2C and DREB3 in SC-15 revealed a fluctuating pattern with down-regulation on days 7 and 21 of water stress, whereas up-regulation was observed on day 14. Concurrently, both genes in EC-564755 showed continuous down-regulation on days 7, 14, and 21 of water stress. Expressions of RD22 and dehydrin recorded on days 7, 14, and 21 were lower in SC-15. The cluster analysis showed that, these two genotypes had a clear distinction in physiological and biochemical properties and gene expressions under water stress and the genotype SC-15 had more efficient osmoprotectant mechanism than genotype EC-564755 under water deficit conditions.     

Alterations in photosynthetic pigments, protein and osmotic components in cotton genotypes subjected to short-term drought stress followed by recovery

In order to assess drought tolerance mechanism in cotton, short-term drought-induced biochemical responses were monitored in two cotton (Gossypium hirsutum L.) genotypes contrasting their tolerance to water deficit. The seeds of two genotypes, namely GM 090304 (moderately drought tolerant) and Ca/H 631 (drought sensitive), were sown in pots containing soil, sand and peat in the ratio of 1:1:1, and irrigated every alternate day up to 45 days after sowing when each genotype was subjected to a cycle of water stress by withholding irrigation for 7 days. The stress cycle was terminated by re-watering the stressed plants for 7 days. The leaf of the drought tolerant genotype (GM 090304) maintained higher relative water content under water stress than that of the drought sensitive genotype (Ca/H 631). The levels of biochemical components, such as chlorophylls, carotenoids, total protein, free proline, total free amino acids, sugars, starch and polyphenols, were measured during the stress as well as the recovery periods. The chlorophylls, carotenoids, protein and starch contents decreased in drought stressed plants as compared to control and tended to increase when the plants were recovered from stress. The degree of decrease in chlorophylls, carotenoids and protein contents under drought was higher in the sensitive genotype (Ca/H 631) as compared to the moderately tolerant genotype (GM 090304). However, proline, total free amino acids, total sugars, reducing sugars and polyphenol contents were increased in drought stressed plants and tended to decrease during the period of recovery. Drought-induced increases in total free amino acids, proline, sugars and polyphenols were significantly higher in the moderately tolerant genotype (GM 090304) than in the sensitive genotype (Ca/H 631). These results suggest that proline, sugars and polyphenols act as main compatible solutes in cotton in order to maintain osmotic balance, to protect cellular macromolecules, to detoxify the cells, and to scavenge free radicals under water stress condition.     

Changes in content of free,conjugated and bound polyamines and osmotic adjustment in adaptation of vetiver grass to water deficit

Osmotic adjustment and alteration of polyamines (PAs) have been suggested to play roles in plant adaptation to water deficit/drought stress. In this study, the changes in cell intactness, photosynthesis, compatible solutes and PAs [including putrescine (Put), spermidine (Spd) and spermine (Spm) each in free, conjugated and bound forms] were investigated in leaves of vetiver grass exposed to different intensity of water deficit stress and subsequent rewatering. The results showed that, when vetiver grass was exposed to the moderate (20% and 40% PEG-6000 solutions) and severe (60% PEG solution) water deficit for 6 days, the plant injury degree (expressed as the parameters of plant growth, cell membrane integrity, water relations and photosynthesis) increased and contents of free and conjugated Put decreased with the rise of PEG concentration. Under the moderate water deficit, the plants could survive by the reduced osmotic potential (ψ_s), increased free and conjugated Spd and Spm in leaves. After subsequent rewatering, the osmotic balance was re-established, most of the above investigated physiological parameters were fully or partly recovered to the control levels. However, it was not the case for the severely-stressed and rewatering plants. It indicates that, vetiver grass can cope well with the moderate water deficit/drought stress by using the strategies of osmotic adjustment and maintenance of total contents of free, conjugated and bound PAs in leaves.     

Photochemical responses and oxidative stress in two clones of Coffea canephora under water deficit conditions

The effects of water deficit on photochemical parameters and activities of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX), as well as, cellular damages were investigated in two clones of Coffea canephora differing in drought tolerance. After 6 days without irrigation, predawn leaf water potential fell to −3.0 MPa that was accompanied by the suppression of net photosynthesis in both clones. The variable to maximum chlorophyll fluorescence ratio remained unchanged regardless of the imposed treatments. Both clones showed a similar decline (about 25%) in the photochemical quenching coefficient, but only the drought-sensitive clone exhibited an enhancement (31%) of thermal deactivation under water deficit conditions. The quantum yield of electron transport decreased similarly in both genotypes. Under drought conditions, activities of SOD, CAT and APX increased to a greater extent in the drought-tolerant clone than in the drought-sensitive one. This seemed to be matched with higher protection against oxidative stress, as judged from the lower levels of lipid peroxidation and electrolyte leakage in the drought-tolerant clone. Thus, the ability to increase the antioxidant system activity in order to limit cellular damages might be an important attribute linked to the drought tolerance in C. canephora.     

Mycorrhization and phosphorus nutrition affect water relations and CAM induction by drought in seedlings of Clusia minor

Background and Aims

Crassulacean acid metabolism (CAM) is currently viewed as an adaptation to water deficit. In plants of Clusia minor, which grow mostly on acidic, P-deficient soils, CAM is induced by water deficit. The symbiosis between plants and mycorrhizal fungi alleviates the symptoms of P deficiency and may influence drought resistance. Therefore, the effect of P supply, modified by three different experimental treatments, on the induction of CAM by drought in C. minor was investigated to test the hypothesis that P deficiency will produce greater CAM activity and, in addition, that treatment will modify drought tolerance.

Methods

Seedlings were grown in forest soil sterilized and inoculated with Scutellospora fulgida (SF treatment), sterilized and supplemented with P (Ph treatment) or non-sterilized and containing native mycorrhizae (Nat treatment). Leaf turgor potential (ψ_T) was determined psychrometrically, and CAM activity as nocturnal acid accumulation (ΔH⁺) by titration of dawn and dusk leaf sap.

Key Results

Plant mass and P content were higher in SF and Ph than in Nat seedlings. After 21 d of water deficit, ψ_T increased in SF, decreased in Ph and remained unchanged in Nat, and, after 7 and 14 d of water deficit, ΔH⁺ in Nat was three times higher than at the beginning of drought, whereas in SF and Ph ΔH⁺ was lower than on day 0.

Conclusions

P deficiency in Nat seedlings was ameliorated by inoculation or P addition. The SF and Nat seedlings showed greater tolerance of drought than Ph. P deficiency promoted the induction of CAM by drought in Nat seedlings, whereas P fertilization and mycorrhization did not. Nocturnal acid accumulation was highly and negatively correlated with plant P and N contents, indicating that P and N deficiencies are promoters of CAM in droughted plants of C. minor.Key words: Clusia minor, crassulacean acid metabolism, CAM, mycorrhiza, drought, phosphorus deficiency, nitrogen–water relations     

杠柳幼苗对不同强度干旱胁迫的生长与生理响应

以黄土丘陵区常见灌木杠柳(Periploca sepium Bunge.)的两年生苗木为试验材料,模拟不同程度的土壤干旱环境,研究了土壤干旱对杠柳生长和生理生化特性的影响。结果表明,在3种土壤水分条件下杠柳的耗水和快速生长期均集中在6-8月,与黄土丘陵区雨热期重叠。在干旱胁迫下杠柳生长减慢,生物量累积减小,生物量优先向根系分配,根冠比显著增大。与适宜水分下相比,干旱胁迫下杠柳的水分利用效率随生物量与耗水量减小而显著升高,表明杠柳具有节约型水分利用对策。杠柳在干旱前期和中期丙二醛(MDA)含量下降、膜透性略有增加,干旱末期MDA含量和细胞膜透性与适宜水分相比显著升高。在中度干旱与严重干旱下,杠柳的超氧化物歧化酶与过氧化物保护酶活性持续上升直到试验末期才稍有下降。干旱胁迫对杠柳叶片渗透调节物质含量的影响显著,脯氨酸、可溶性糖和可溶性蛋白含量均随着干旱胁迫程度的加大而显著升高,且随着胁迫时间的延长,脯氨酸和可溶性糖含量一直保持上升趋势;可溶性蛋白含量在前期急剧上升,中、后期下降,但含量一直高于适宜水分处理。本研究表明,杠柳能够充分利用黄土丘陵区局部雨热资源优势、具备减少地上蒸发面积、增加地下生长、吸收深层土壤水源等御旱策略以及保持较高的抗氧化酶活性和渗透调节能力等生理学耐旱机制,本研究从生长、生理特征上揭示了杠柳在黄土高原植被自然恢复中普遍存在的原因。     

Physiological and biochemical changes in different drought-tolerant alfalfa (<Emphasis Type="Italic">Medicago sativa</Emphasis> L.) varieties under PEG-induced drought stress

Medicago sativa L. cv. Longzhong is a nutritious forage plant in dryland regions of the Loess Plateau with strong drought tolerance and broad adaptability. To understand the adaptation mechanism of alfalfa (M. sativa L. cv. Longzhong) to drought stress, growth, and physiological parameters including levels of chlorophyll content, osmotic adjustment, reactive oxygen species (ROS), and antioxidant enzymes and antioxidants were measured under simulated levels of drought (? 0.40, ? 0.80, ? 1.20, ? 1.60, and ? 2.00 MPa). The changes in M. sativa L. cv. Longzhong were compared with those of plants of M. sativa L. cv. Longdong control (Variety I) suited to moderate rainfall areas and M. sativa L. cv. Gannong No. 3 (Variety II) suited to irrigated areas. The results showed that root–shoot ratio, the chlorophyll (a + b) and osmolytes contents, the degree of lipid peroxidation and ROS production, and the levels of antioxidative enzymes and antioxidants increased significantly with increasing drought stress, whereas plant height, aboveground biomass, chlorophyll a/b ratio, leaf water potential (Ψ₁), and relative water content (RWC) decreased in response to drought. The Longzhong variety responded early to beginning drought stress (between 0 and ? 0.4 MPa) compared with the controls. Under drought stress (between ? 0.4 and ? 2.0 MPa), the Longzhong variety had significantly higher belowground biomass, root–shoot ratio, Ψ₁, RWC, catalase (CAT) activity and reduced glutathione content than those of Varieties I and II, but hydrogen peroxide and hydroxyl free radical (OH·) contents were significantly lower. Step regression analysis showed that OH·, CAT, malondialdehyde, superoxide anion-free radical (O ₂ ^·? ), and superoxide dismutase of Longzhong had the most marked response to drought stress. In conclusion, the stronger drought tolerance of the Longzhong variety might be due to its higher water-holding capacity, root–shoot ratio, and ability to coordinate enzymatic and non-enzymatic antioxidant systems, which coordinate the peroxidation and oxidative systems.     

Effect of ploidy levels on the activities of Δ1-pyrroline-5-carboxylate synthetase,superoxide dismutase and peroxidase in Cenchrus species grown under water stress

The effects of ploidy levels on the activities of Δ¹-pyrroline-5-carboxylate synthetase (P5CS; EC not assigned), superoxide dismutase (SOD; EC 1.15.1.1) and guaiacol peroxidase (POX; EC 1.11.1.7), as well as malondialdehyde (MDA) and proline contents were studied in two months old plants of Cenchrus species. The Cenchrus species represent three ploidy levels: diploid, tetraploid, hexaploid and two life spans: annual and perennial. Plants were subjected to water stress for 2, 4, 6 and 8 d by withholding water under glasshouse conditions. Although the levels of proline increased with the magnitude of water stress, the P5CS activity did not show a corresponding increase in all species. Peroxidase and superoxide dismutase activities showed an increase or steady state in the early phase of drought and then declined with the further increase in the magnitude of water stress, indicating differing behaviors of species towards drought tolerance. Under drought, diploid Cenchrus species had a higher POX activity, MDA accumulation and lower proline content than tetraploid species. Lower POX and higher P5CS activities and proline contents, however, were observed in hexaploid and tetraploid species. Taken together, our findings suggest that diploid species have a less efficient antioxidant system to scavenge reactive oxygen species than tetra and hexaploid Cenchrus. This may result in a corresponding variability in growth and persistence under natural grasslands. The study also paves the way for investigations on the molecular events associated with drought in Cenchrus species differing in ploidy and life span.     

Inoculation with arbuscular mycorrhizal fungi alleviates harmful effects of drought stress on damask rose

This study was conducted to examine the role of arbuscular mycorrhiza fungi (AMF) in alleviating the adverse effects of drought stress on damask rose (Rosa damascena Mill.) plants. Four levels of drought stress (100, 75, 50, and 25% FC) were examined on mycorrhizal and non-mycorrhizal plants in pots filled with sterilized soil. Our results showed that increasing drought stress level decreased all growth parameters, nutrient contents, gas exchange parameters, and water relations indicators. Under different levels of drought stress, mycorrhizal colonization significantly increased all studied parameters. P_n, g_s, and E of the mycorrhizal plants was higher than those of non-mycorrhizal plants under different levels of drought stress. The increase in those rates was proportional the level of the mycorrhizal colonization in the roots of these plants. Majority of growth, nutrition, water status and photosynthetic parameters had a great dependency on the mycorrhizal colonization under all levels of drought stress. The results obtained in this study provide a clear evidence that AMF colonization can enhance growth, flower quality and adaptation of rose plants under different drought stress levels, particularly at high level of drought stress via improving their water relations and photosynthetic status. It could be concluded that colonization with AMF could help plants to tolerate the harmful effects caused by drought stress in arid and semi-arid regions.     

干旱胁迫对发菜超微结构及抗性生理的影响

发菜是一种陆生固氮蓝藻,具有强烈的旱生生态适应性。对干旱胁迫条件下发菜超微结构和抗性生理进行了研究。结果表明：随着干旱胁迫加重,发菜细胞大小和细胞壁厚度变化不显著,胶质鞘趋于紧密,类囊体排列趋于紊乱,多角体变得模糊不清甚至消失,糖原颗粒数目减少,但结构颗粒数目没有明显变化。随着干旱胁迫的加重,发菜SOD、CAT活性呈先升高再下降趋势,SOD在含水量为120%时达到高峰;CAT活性在含水量为445%时达到高峰;MDA,氧自由基随着干旱胁迫加重,其含量呈上升趋势;H₂O₂含量随干旱胁迫加重呈先升高再下降趋势,在含水量为120%时达到峰值。干燥储存1年的发菜与恢复活性的发菜有明显差异性,其SOD、CAT活性较低、MDA、H₂O₂含量较少,但是氧自由基含量最高。研究结果对深入研究发菜的耐旱机理奠定了基础。     

Oxidative stress, protein carbonylation, proteolysis and antioxidative defense system as a model for depicting water deficit tolerance in Indica rice seedlings

Water deficit is an important constraint to rice (Oryza sativa L.) productivity. The present study was undertaken to investigate whether the level of oxidative stress, carbonylation of proteins, proteolysis and status of antioxidative defense could serve as a model to distinguish water deficit tolerant and sensitive rice cultivars. When 10-day-grown seedlings of two rice cultivars, Malviya-36 (drought-sensitive) and Brown Gora (drought-tolerant) were subjected to ?1.0 and ?2.1 MPa water deficit treatments for 24–72 h with polyethylene glycol 6000 in the medium, a greater decline in the growth of the seedlings and levels of leaf water potential, relative water content, Chl a, Chl b, carotenoids and greater increase in leaf water loss were observed in the sensitive cultivar than the tolerant. Under similar level of water deficit seedlings of sensitive cultivar showed higher level of superoxide anion generation, H₂O₂, lipid peroxidation and proteolysis in roots as well as shoots compared to the tolerant. Drought-tolerant cultivar had higher constitutive level of antioxidative enzymes superoxide dismutase and catalase and the activities of these two enzymes alongwith of guaiacol peroxidase showed greater increase in this cultivar under water deficit compared to the sensitive. A significant decline in the level of protein thiol and a higher increase in protein carbonyls content, also confirmed by protein gel blot analysis with an antibody against 2,4-dinitrophenylhydrazine was observed in the seedlings of drought sensitive cv. Malviya-36 compared to the tolerant cv. Brown Gora when subjected to similar level of water deficit. Seedlings of drought sensitive cultivar, under water deficit, showed higher proteolytic activity, higher number of in-gel activity stained proteolytic bands and higher expression of oxidized proteins in roots compared to the tolerant cultivar. Results suggest that poor capacity of antioxidative enzymes could be, at least partly, correlated with water deficit sensitivity of sensitive cultivar and that higher activity of antioxidative enzymes superoxide dismutase, catalase, guaiacol peroxidase, low proteolytic activity, lower level of protein carbonyls and protein thiolation could serve as a model to depict water deficit tolerance in Indica rice seedlings.     

内蒙古高原锦鸡儿属植物的形态和生理生态适应性

比较研究内蒙古高原锦鸡儿属(Caragana)中生种,旱生种和强旱生种的叶片形态结构、渗透调节、气孔调节和保护酶,目的是揭示锦鸡儿属不同类型植物的生态适应策略。中生种叶片平展,被稀疏绿色短柔毛;旱生种叶片平展或呈瓦状,被灰色柔毛;强旱生种叶片呈瓦状或卷成筒状,被直立或伏帖绢毛。叶片厚度强旱生种>旱生种>中生种,叶片面积、生物量和比叶面积(SLA)强旱生种<旱生种<中生种。叶片长宽比,强旱生种和旱生种大于中生种。这些形态结构导致保水能力强旱生种>旱生种>中生种,光能利用能力中生种>旱生种>强旱生种。渗透调节物质含量、细胞质离子浓度和细胞渗透势强旱生种>旱生种>中生种。渗透调节物质含量的差异主要表现在强旱生种可溶性糖和无机离子含量远高于旱生种,后者又远高于中生种。叶含水量、自由水含量、叶水势和气孔导度中生种>旱生种>强旱生种,束缚水含量、束缚水/自由水比值、POD和SOD活性正好相反,CAT活性旱生种>中生种>强旱生种。这些生理特性导致抗旱能力强旱生种>旱生种>中生种,但代谢速率正好相反。旱生种和中生种表现出较少的日水分亏缺,强旱生种水分亏缺从清晨到傍晚持续大幅增加。细胞膜相对透性和MDA含量强旱生种>旱生种>中生种。自由基含量表现为旱生种>中生种>强旱生种。这些数据说明虽然旱生种和强旱生种形成了多种特点来适应干旱环境,但仍然是不充分的。结论:(1)分布于半湿润至半干旱区的锦鸡儿属中生种依靠活跃的代谢、大量的水分消耗和快速生长使其在生物环境中取得竞争优势;生活在干旱地区和强干旱地区的旱生种和强旱生种依靠低代谢、节水和高抗旱性来抵抗苛刻的非生物环境。(2) 旱生种和强旱生种主要通过可溶性糖和无机离子的积累,调节细胞质渗透势,保持水分平衡,这是一种相对节省能量的适应对策。     

Organ-dependent responses of Periploca sepium to repeated dehydration and rehydration

Plant responses to water deficit occur in a complex framework of organ interactions, but few studies focus on the effect of drought stress on all organs in a whole-plant. The effects of repeated dehydration and rehydration (DH) on physiological and biochemical responses in various organs of Periploca sepium Bunge (P. sepium) were investigated. The leaf relative water content decreased significantly during drought, but recovered and showed an increase when compared to well-watered control plants. The malondialdehyde (MDA) content increased in mature and old leaves, but decreased in young leaves, new stems and fine roots during drought, indicating that the young and vigorous tissues of a whole-plant are protected preferentially from the oxidative stress. Among all organs, the fine roots showed the highest levels of proline, total free amino acids (TFAA) and Na⁺, while the leaves showed the highest levels of total soluble sugars (TSS), soluble proteins (SP), Ca²⁺ and Mg²⁺. The response to DH differed in different organs, both in magnitude and in the type of solutes involved. Drought stress increased the contents of proline, TFAA, TSS, SP and K⁺ in all organs of P. sepium plants, while the accumulation amounts were obviously different among the organs. The storage starch in stems and roots plays an important role in providing carbohydrates for growth. Changes in Na⁺, Ca²⁺ and Mg²⁺ under DH presented a high degree of organ specificity. Our data indicates that response strategies are different between different organs; therefore, evidence the needs to integrate all the information in order to better understand plant tolerance mechanisms.     

Effect of drought on pigments, osmotic adjustment and antioxidant enzymes in six woody plant species in karst habitats of southwestern China

Drought stress is one of the most important factors limiting the survival and growth of plants in the harsh karst habitats of southwestern China. Detailed knowledge about the ecophysiological responses of native plants with different growth forms to drought stress could contribute to the success of re-vegetation programs. Two shrubs, Pyracantha fortuneana and Rosa cymosa, and four trees, Broussonetia papyrifera, Cinnamomum bodinieri, Platycarya longipes and Pteroceltis tatarinowii, were randomly assigned to four drought treatments, i.e. well-watered, mild drought stress, moderate drought stress, and severe drought stress. Midday water potential, the maximum quantum efficiency of PSII photochemistry (F_v/F_m), pigments, osmotic solutes (soluble sugars and proline), cellular damages, and antioxidant enzymes (superoxide dismutase, catalase and peroxidase) were investigated. Drought stress significantly decreased pigments content, but increased the ratio of carotenoids to total chlorophylls in the studied species. After prolonged severe drought stress, the two shrubs exhibited higher F_v/F_m, less reductions of midday water potential, and lower increases of malondialdehyde content and ion leakage than the four trees. Prolonged severe drought stress largely decreased accumulations of osmotic solutes and activities of antioxidant enzymes in the four trees, but significantly increased proline content and superoxide dismutase activity in the two shrubs and peroxidase activity in P. fortuneana. The positive relationships were observed among activities of antioxidant enzymes, and between contents of osmotic solutes and activities of antioxidant enzymes. These findings suggested that the two shrubs had higher tolerance to severe drought stress than the four trees due to higher capacities of osmotic adjustment and antioxidant protection.