AgNO<Subscript>3</Subscript> prevents the occurrence of hyperhydricity in <Emphasis Type="Italic">Dianthus chinensis</Emphasis> L. by enhancing water loss and antioxidant capacity

Hyperhydricity symptoms are common and significant during the in vitro culture of Dianthus chinensis L. and greatly affect the micropropagation and regeneration of cultured plantlets. However, effective measures for preventing such abnormalities have not been developed for this species. Silver nitrate (AgNO₃) has been shown to revert hyperhydric plantlets to a normal state. Nevertheless, the effect of AgNO₃ on the prevention of hyperhydricity and the underlying mechanisms remain unclear. In the present study, 98.7% of the Dianthus chinensis L. plantlets cultured in a hyperhydricity induction medium (HIM) developed symptoms of hyperhydricity; however, hyperhydricity symptoms were inhibited to different degrees when D. chinensis L. plantlets were cultured in HIM supplemented with various concentrations of AgNO₃. In particular, approximately 97% of the D. chinensis L. plantlets grew normally and did not show any symptoms of hyperhydricity when cultured in HIM supplemented with 30 μmol L^?1 AgNO₃. Compared with the plantlets cultured in HIM alone, the plantlets cultured in HIM containing AgNO₃ displayed dramatic decreases in water content, ethylene content, and reactive oxygen species (ROS) production (particularly regarding H₂O₂ accumulation in guard cells) and showed increased antioxidant enzyme activity, stoma aperture, and water loss. These changes not only prevented excess water from accumulating in the tissues of plantlets but also improved the antioxidant capacity of plantlets, ultimately resulting in the prevention of hyperhydricity.     

Cell ultrastructure and chlorophyll pigments in hyperhydric and non-hyperhydric <Emphasis Type="Italic">Beta vulgaris</Emphasis> var. Conditiva plantlets,treated with deuterium depleted water

Hyperhydricity can cause significant loss in the in vitro propagated plantlets. In order to predict and control its occurrence, a better understanding of the structural aspects and physiological features of hyperhydric plantlets is required. In this study, the ultrastructural and physiological changes associated with hyperhydric red beet plantlets were investigated. Our objective was to establish a correlation between the ultrastructural aspects of Beta vulgaris var. Conditiva leaflets and hypocotyls and the content of chlorophyll pigments extracted in N,N-dimethylformamide (DMF) of two type of plantlets: hyperhydric from a basal culture medium Murashige and Skoog (JAMA 15:473–497, 1962) prepared with distilled water (DW—155 ppm Deuterium) and non-hyperhydric, cultivated on identical medium where distilled water was replaced with deuterium depleted water (DDW- 25 ppm Deuterium) as a method of preventing hyperhydricity. Cell ultrastructure in hyperhydricity, both from the leaves, but especially from hypocotyls, showed denatured chloroplasts in a myxoplasm mass formed by the damage of the tonoplast and the mixing of the cytoplasm with the vacuolar juice. The nuclei were picnotic, presenting paranucleolar corpuscles. The amount of assimilating pigments was significantly reduced in the plantlets grown on medium prepared with DW as compared to the normal, non-hyperhydric ones from medium prepared with DDW. Both evaluations showed that, in red beet, DDW also prevents the appearance of hyperhydricity.     

Hyperhydricity in micropropagated carnation shoots: the role of oxidative stress

The physiology of hyperhydricity in relation to oxidative stress, mineral nutrients, antioxidant enzymes and ethylene has been studied in three micropropagated carnation cultivars under experimentally induced hyperhydricity. A marked increase in Fe content in comparison with normal tissues was observed in the hyperhydric tissues from the three cultivars. The levels of ethylene, solute leakage and malondialdehyde content were also significantly higher in the hyperhydric tissues. In relation to the time course of H₂O₂ production measured by fluorescence quenching, a similar trend could be observed for the three cultivars, with a clear increase in the generation of hydrogen peroxide in hyperhydric tissues. The activities of all the antioxidative enzymes studied, except lipoxygenase, were higher in the hyperhydric shoots. Phenylalanine ammonia-lyase (PAL) showed a significant decrease in activity in the hyperhydric tissues in comparison with the controls for the three cultivars. Soluble guaiacol peroxidase had a strong increase in activity in hyperhydric shoots of the three cultivars. These results provide, for the first time, direct evidence of H₂O₂ generation in hyperhydric tissues, characterize the response of the antioxidant system to an oxidative stress during hyperhydricity in carnation leaves and point to the accumulation of toxic forms of oxygen as the inducer of some of the abnormalities observed.     

Proteomic and Antioxidant Analysis Elucidates the Underlying Mechanism of Tolerance to Hyperhydricity Stress in In Vitro Shoot Cultures of <Emphasis Type="Italic">Dianthus caryophyllus</Emphasis>

Hyperhydric disorders occur frequently in plant tissues cultured in vitro and cause several morphological and physiological abnormalities. However, a systematic defense response is triggered by hyperhydric conditions. The accumulation of reactive oxygen species (ROS), activities of antioxidant enzymes and their immunoblots, and the proteome-level changes in normal versus hyperhydric shoots of carnation (Dianthus caryophyllus) cultured in vitro were investigated. Total proteins were also extracted from the shoot and analyzed by two-dimensional electrophoresis. Among a total of 700 spots detected, only 40 had significant changes in abundance in the hyperhydric compared to the normal shoots, which were further identified by a mass spectrometer (MALDI-TOF MS). Most of them were involved in photosynthesis, RNA processing, and general metabolisms, while the rest were involved in secondary metabolic processes. These identified proteins in carnation shoots may provide novel evidences for stress tolerance against hyperhydricity.     

A Study on Comparative Morphology of Normal and Hyperhydric Stems and leaves from Sophora japonica Plantlets Cultured in Vitro

A large numar of plantlets were obtained from cotyledon explants of Sophom japonica L. cultured in vitro. They could be classified into 3 kinds according to their morphological characteristics, viz. the normal plantlets, the hyperhydric planfiets,and the intermediate state between the two or the sub-hyperhydric type. The free water content was more than 79% in the hyperhydric shoots,and 70% in the sub-hyperhydric shoots,while less than 50% in the normal shoots. The surface anatomy of normal, sub-hyperhydric and hyperhydric stems and leaves of the plantlets were compared by scanning electron microscopy. The surface structure of the normal plantlets was similar to those found in field-grown plants,but great change occurred in that of hyperhydric and the sub-hyperhydric plantlets. The stems and leaves surface of the hyperhydric and sub-hyperhydric plantlets appeared to be uneven, wrinkled, brittle and translucent and besides the leaves were thick, curled with a reduced surface area. There was little or no epicuticular wax on the surface of epidermal cells which had irregular shapes and patterns. All leaves were amphistomatic and the stomatal density, size and degree of opening were obviously bigger in the sub-hyperhydric and hyperhydric leaves than in the normal ones. Normal stomata had kidney-shaped guard cells and resembled closely those found in the feild-grown plants, whereas abnormal stomata had deformed guard cells. All of the morphological characteristics mentioned above indicated that the sub-hyperhydric and hyperhydric shoots bended to lose their water easily and resulted in desiccation, which might be one of the major causes of failure to transfer sub-hyperhydric and hyperhydric plantlets to soil.     

Analysis of ultrastructure and reactive oxygen species of hyperhydric garlic (Allium sativum L.) shoots

Hyperhydricity is a physiological disorder frequently affecting shoots propagated in vitro. Since it negatively affects shoot multiplication vigor, and impedes the successful transfer of micropropagated plants to in vivo conditions, hyperhydricity is a major problem in plant tissue culture. In commercial plant micropropagation, there are reports of up to 60% of cultured shoots or plantlets which demonstrate hyperhydricity, which reflects the pervasiveness of this problem. The phenomenon has been correlated to water availability, microelements, and/or hormonal imbalance in the tissue culture. In this study, the ultrastructure and the characteristics of reactive oxygen species between hyperhydric and normal shoots of garlic were studied. We observed that in some cells of hyperhydric tissues, the intranuclear inclusion was separated, the mitochondrion was swollen and its intracristae had splits, the organelles were compressed against the cell wall, and the chloroplasts and intergranal thylakoids were also compressed. Additionally, the content of chlorophyll and soluble protein in hyperhydric shoots decreased significantly. For instance, chlorophyll a decreased 43.61%, chlorophyll b decreased 49.29%, chlorophyll a+b decreased 48.10%, and soluble protein dropped 47.36%. In contrast, the O₂ generation rate and H₂O₂ level increased 45.36% and 63.98%, respectively, obviously higher than the normal shoots. Lipoxygenase activity and malondialdehyde content in the hyperhydric shoots increased significantly, while the electrolyte leakage rose, indicating a serious membrane lipid peroxidatic reaction. Superoxide dismutase, peroxidase, catalase, glutathione peroxidase, and ascorbate peroxidase activities in hyperhydric tissue were all significantly higher than in normal leaf tissue. The antioxidant metabolism demostrated a close connection between hyperhydricity and reactivated oxygen species.     

槐树试管正常苗与超度含水态苗茎叶的比较形态学研究

槐树(Sophora japonica L.)子叶经培养获得了大量的试管苗,依其形态正常与否可将其分为正常苗、超度含水态苗和介于二者之间的过度含水态苗。其自由水含量明显不同,正常苗低于50％,超度含水态苗高于79％,而过度含水态苗约为70％。以扫描电镜对其茎、叶的形态学结构进行了比较研究,结果表明:正常苗茎、叶表皮结构基本类似于实生苗,而过度及超度含水态苗的茎、叶表皮层结构变异较大。主要表现在其表面凹凸不平,表皮层外稀有或无蜡质存在,表皮细胞形状及排列不规则,叶片近、远轴两面气孔器密度、大小及开度均较正常苗显著增大;保卫细胞形态、结构异常。上述特征,均显示出槐树试管过度及超度含水态苗易失水干化,这可能是其在移栽过程中难以成活的主要原因之一。     

Red and Blue Light Emitting Diodes (LEDs) Participate in Mitigation of Hyperhydricity in In Vitro-Grown Carnation Genotypes (<Emphasis Type="Italic">Dianthus Caryophyllus</Emphasis>)

The present study was to determine the factors that can reduce hyperhydricity in in vitro-propagated carnation genotypes. The carnation genotypes (Green Beauty, Purple Beauty, and Inca Magic) were grown in vitro under normal and hyperhydric conditions in white fluorescent light (FL) in which half of the hyperhydric plants were grown in red and blue LEDs (light emitting diodes). It was observed that hyperhydricity leads to oxidative stress in terms of TBARS (thiobarbituric acid reactive substances) content, whereas stress was alleviated by R (red) and B (blue) LEDs. The multiprotein complex proteins such as ATPase (RCI?+?LHC1) PSII-core dimer, PSII-monomer/ATPs synthase, and PSII-monomer/cyt b₆f had decreased levels in hyperhydric conditions grown in white FL; however, the expression level of these photosynthetic proteins was retained in hyperhydric plants grown in R and B LEDs. Moreover, the immunoblots of two photosynthetic proteins (PsaA and PsbA) and stress-responsive proteins such as superoxide dismutase, ascorbate peroxidase, and catalase showed recovery of hyperhydricity in carnation genotypes grown in R and B LEDs. Our present study signifies that red (R) and blue light (B) LEDs reduced the hyperhydricity to control levels by maintaining the composition of thylakoid proteins and antioxidative defense mechanisms in carnation genotypes.     

Sub-cellular location of H2O2, peroxidases and pectin epitopes in control and hyperhydric shoots of carnation

In carnation shoots (Dianthus caryophyllus cv. Killer), hyperhydricity was induced in in vitro culture using a low agar concentration. Using transmission electron microscopy, cytochemical techniques and immunolocation of JIM5 and JIM7 pectin epitopes, we followed the sub-cellular modifications of cell walls in relation to peroxidase activity and hydrogen peroxide accumulation during hyperhydricity induction. Peroxidase activity revealed a significant induction of the stomatal and epidermal cells as well as of the intercellular spaces of hyperhydric leaves. Similarly, hydrogen peroxide accumulated in the epidermal cell walls and the intercellular spaces of hyperhydric leaves. Immunolocation of an epitope recognised by the JIM5 antibody revealed the main unesterified nature of the cell walls. Such an epitope was located in the epidermal cell walls as well as in the corners of cell junctions in control leaves. However, hyperhydric leaves showed a total reduction of JIM5 labelling in the corners of cell junctions and a significant reduction of the intercellular spaces and the middle lamella. Highly-methylsterified pectin, recognised by the JIM7 antibody, was present to a slight extent in cell walls in control and hyperhydric leaves. We propose that the altered anatomy observed in hyperhydric carnation leaves could be regulated by the concomitant actions of pectin methyl esterases and free radicals, modifying the structure of the pectin and polysaccharides of the cell walls.     

Effects of culture medium composition and PEG on hyperhydricity in Dendrobium officinale

Hyperhydricity is a physiological disorder during plant tissue culture that seriously affects regeneration and micropropagation. In this study, Dendrobium officinale plantlets were cultured on solid Murashige and Skoog (MS) medium supplemented with plant growth regulators and various concentrations of sucrose, agar, and polyethylene glycol (PEG)-6000 to explore the effect of osmotic stress on hyperhydricity. The results show that low concentrations of sucrose or agar, as well as PEG-6000 at various concentrations, significantly increase the hyperhydric rate of D. officinale, whereas high concentrations of sucrose or agar did not. Furthermore, high concentrations of PEG-6000 significantly increase total water content, free-water content, relative electrical conductivity, and peroxidase (POD) activity of D. officinale plantlets, whereas they significantly decrease bound-water content, proline content, soluble protein content, soluble sugar content, and superoxide dismutase (SOD) activity. These results indicate that PEG-6000 disrupts the antioxidant system and water metabolism in D. officinale plantlets, as well as increases cell membrane permeability, which might be the key factors for the occurrence of hyperhydricity in this species.

     

蓝莓玻璃化试管苗的显微结构及生理生化特性变化

研究了蓝莓试管苗玻璃化的显微结构、超微结构以及生理生化特性的影响。与正常试管苗相比,蓝莓玻璃化苗的茎、叶显微结构发生了明显的改变：叶片表皮细胞松散、不连续;气孔结构难以辨认;叶片增厚;缺少栅栏组织,海绵组织细胞间隙变大,部分细胞解体;茎的维管组织发育不良;亚显微结构观察发现,玻璃化苗叶肉细胞体积增大,细胞壁变薄;部分细胞缺少细胞核及线粒体;叶绿体数目减少,类囊体解体,缺乏淀粉体。玻璃化试管苗的生理生化特性也发生了显著的改变：玻璃化苗组织含水量显著增加;叶绿素、可溶性糖及可溶性蛋白含量显著降低;O2- 产生速率、H2O2积累量、MDA含量及相对电导率显著升高;活性氧清除酶系中POD活性显著升高,SOD和CAT活性显著降低;PAL活性下降。蓝莓玻璃化苗的形态结构异常,水分及物质代谢紊乱,活性氧清除能力降低,表明玻璃化与氧化胁迫相关。     

Regulation of plant water loss by manipulating the expression of phospholipase Dα

Phospholipase D (PLD) has been implicated in various processes, including signal transduction, membrane trafficking, and membrane degradation. Multiple forms of PLD with distinct biochemical properties have been described in the cell. In Arabidopsis, PLDalpha and PLDgamma, but not PLDbeta, were detected in guard cells, and antisense suppression resulted in a specific loss of PLDalpha. The abrogation of PLDalpha rendered plants less sensitive to abscisic acid and impaired stomatal closure induced by water deficits. PLDalpha-depleted plants exhibited accelerated transpirational water loss and a decreased ability to tolerate drought stress. Overexpression of PLDalpha enhanced the leaf's sensitivity to abscisic acid. These findings provide molecular and physiological evidence that PLDalpha plays a crucial role in regulating stomatal movement and plant-water status.     

In the light of stomatal opening: new insights into 'the Watergate'

Stomata can be regarded as hydraulically driven valves in the leaf surface, which open to allow CO2 uptake and close to prevent excessive loss of water. Movement of these 'Watergates' is regulated by environmental conditions, such as light, CO2 and humidity. Guard cells can sense environmental conditions and function as motor cells within the stomatal complex. Stomatal movement results from the transport of K+ salts across the guard cell membranes. In this review, we discuss the biophysical principles and mechanisms of stomatal movement and relate these to ion transport at the plasma membrane and vacuolar membrane. Studies with isolated guard cells, combined with recordings on single guard cells in intact plants, revealed that light stimulates stomatal opening via blue light-specific and photosynthetic-active radiation-dependent pathways. In addition, guard cells sense changes in air humidity and the water status of distant tissues via the stress hormone abscisic acid (ABA). Guard cells thus provide an excellent system to study cross-talk, as multiple signaling pathways induce both short- and long-term responses in these sensory cells.     

Reactive oxygen species signaling and stomatal movement in plant responses to drought stress and pathogen attack

Stomata, the pores formed by a pair of guard cells, are the main gateways for water transpiration and photosynthetic CO_2 exchange, as well as pathogen invasion in land plants. Guard cell movement is regulated by a combination of environmental factors, including water status, light, CO_2 levels and pathogen attack, as well as endogenous signals, such as abscisic acid and apoplastic reactive oxygen species(ROS). Under abiotic and bioticstress conditions, extracellular ROS are mainly produced by plasma membrane-localized NADPH oxidases, whereas intracellular ROS are produced in multiple organelles. These ROS form a sophisticated cellular signaling network, with the accumulation of apoplastic ROS an early hallmark of stomatal movement. Here, we review recent progress in understanding the molecular mechanisms of the ROS signaling network,primarily during drought stress and pathogen attack. We summarize the roles of apoplastic ROS in regulating stomatal movement, ABA and CO_2 signaling, and immunity responses.Finally, we discuss ROS accumulation and communication between organelles and cells. This information provides a conceptual framework for understanding how ROS signaling is integrated with various signaling pathways during plant responses to abiotic and biotic stress stimuli.     

Lipid peroxidation and antioxidant enzyme activities of Euphorbia millii hyperhydric shoots

A large number of micropropagated Euphorbia millii shoots from temporary immersion bioreactor showed thick broad leaves that were translucent, wrinkled and/or curled and brittle, symptoms of hyperhydricity. The environment inside bioreactor normally used in plant micropropagation is characterised by high relative humidity, poor gaseous exchange between the internal atmosphere of the bioreactor and its surrounding environment, and the accumulation of ethylene, conditions that may induce physiological disorders. A comparison of hyperhydric shoots (HS) with normal plants shows marked increase in malondialdehyde (MDA) content in HS plants. MDA, a decomposition product of polyunsaturated fatty acids hydroperoxides, has been utilized very often as a suitable biomarker for lipid peroxidation, which is an effect of oxidative damage. This hypothesis is also confirmed by the higher lipoxygenase (LOX) activity in HS plants. The potential role of antioxidant enzymes in protecting hyperhydric shoots from oxidative injury was examined by analyzing enzyme activities and isozyme profiles of hyperhydric and non-hyperhydric leaves of E. millii. Superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activity were significantly higher in hyperhydric tissue as compared to non-hyperhydric normal leaf tissue. After native polyacrylamide gel electrophoresis (PAGE) analysis, seven SOD isoenzymes were detected and the increase in SOD activity observed in hyperhydric tissue seemed to be mainly due to Mn-SOD and Cu/Zn-SOD. The activity of ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) was proportionally increased in HS tissue compared to normal leaves indicating a crucial role in eliminating toxic H₂O₂ from plant cells. The depletion of GSH and total glutathione in spite of higher GR activities observed in HS tissue indicates that mechanism of antioxidant defense was by enhanced oxidation of GSH to GSSG by DHAR yielding ascorbate (AA). The antioxidant metabolism has been shown to be important in determining the ability of plants to survive in hyperhydric stress and the up regulation of these enzymes would help to reduce the build up of ROS.     

Shoot multiplication kinetics and hyperhydric status of regenerated shoots of gladiolus in agar-solidified and matrix-supported liquid cultures

In vitro shoot regeneration of gladiolus in three different culture systems, viz., semi-solid agar (AS), membrane raft (MR), and duroplast foam liquid (DF) cultures was evaluated following the kinetics of shoot multiplication and hyperhydricity at optimized growth regulator combinations. Compared to the AS system, matrix-supported liquid cultures enhanced shoot multiplication. The peak of shoot multiplication rate was attained at 18 days of incubation in the MR and DF systems, whereas the maximum rate in the AS system was attained at 21 days. An early decline in acceleration trend was observed in liquid cultures than the AS culture. The hyperhydric status of the regenerated shoots in the different culture systems was assessed in terms of stomatal attributes and antioxidative status. Stomatal behavior appeared to be normal in the AS and MR systems. However, structural anomaly of stomata such as large, round shaped guard cells with damage in bordering regions of stomatal pores was pronounced in the DF system along with a relatively higher K⁺ ion concentration than in the AS and MR systems. Antioxidative status of regenerated shoots was comparable in the AS and MR systems, while a higher incidence of oxidative damages of lipid membrane as evidenced from malondialdehyde and ascorbate content was observed in the DF system. Higher oxidative stress in the DF system was also apparent by elevated activities of superoxide dismutase, ascorbate peroxidase, and catalase. Among the three culture systems, liquid culture with MR resulted in maximum shoot multiplication with little or no symptoms of hyperhydricity. Shoots in the DF system were more prone to hyperhydricity than those in the AS and MR systems. The use of matrix support such as membrane raft as an interface between liquid medium and propagating tissue could be an effective means for rapid and efficient mass propagation with little or no symptoms of hyperhydricity.     

The stomata frontline of plant interaction with the environment-perspectives from hormone regulation

Plants have evolved elaborate mechanisms to perceive and integrate signals from various environmental conditions.On leaf surface,stomata formed by pairs of guard cells mediate gas exchange,water transp...     

Gain of function of stomatal movements in rooting <Emphasis Type="Italic">Vitis vinifera</Emphasis> L. plants: regulation by H<Subscript>2</Subscript>O<Subscript>2</Subscript> is independent of ABA before the protruding of roots

Reactive oxygen species (ROS), namely superoxide radical (O₂ ⁻) and hydrogen peroxide (H₂O₂) are generated when plant tissues endure a variety of environmental stresses, including light stress. The extremely short life times of ROS makes the study of their production in planta very difficult. The use of ROS-specific tracer dyes, 3-3′ diaminobenzidine and nitroblue tetrazolium, together with high-resolution imaging provides the opportunity to identify sites of photooxidative stress response by ROS accumulation. This technique was applied to grapevine during the first 7 days after transfer from in vitro to ex vitro under an irradiance 4-fold higher than in vitro. ROS accumulation was detected in the first days of analysis, which gradually decreased to levels comparable to greenhouse leaves. O₂ ⁻ was uniformly distributed while H₂O₂ accumulated preferentially in veins, wounds and stomatal guard and surrounding cells. To evaluate the role of H₂O₂ in stomatal functioning and its crosstalk with abscisic acid (ABA) we focused on the percentage of coloured structures, stomatal aperture and ABA concentration. We propose that the high H₂O₂ level triggered by increased light is responsible for the activation of a signalling pathway over stomatal cells, in a process apparently irrespective of ABA regulation prior to root protrusion. This could explain the gain of function of a low yet consistent percentage of stomatal cells, essential for plant survival during the ontogenic period in analysis.     

Interactive effects of ozone and drought stress on plants: A review北大核心CSCD

Ground-level ozone (O3) and drought are two key factors limiting plant growth. O3 can enter into the plant tissue through the stomata, then causing the formation of reactive oxygen species (ROS) which inspires programmed cell death. Drought usually induces the accumulation of ROS due to damage to antioxidant systems of plants. The effects of two kinds of stress on plants are similar due to the accumulation of ROS, resulting in reduced photosynthesis rate and physiological metabolism, eventually decreased plant growth and biomass. Nevertheless, O3 and drought interacts synergistically to accumulate detrimental effects or antagonistically to reduce harmful effects. Actually, it is complex interactive process between O3 and drought. On the one hand, O3 triggers stomatal sluggishness or even dysfunction, which exacerbates water transpiration of leaves, water loss from plants and further O3 phytotoxicity. On the other hand, drought induces stomatal closure, and thus protecting plants against the O3 influx and evaporation of water. However, prolonged drought could limit the uptake of CO2 and thus result in reduced plant growth. The response of plants to both O3 and drought not only depends on the occurring sequence and duration of any factor but also rely on the difference in physiological metabolism of the plant itself. The interactive effects of O3 and drought on stomatal characteristics, photosynthetic carbon mechanism, antioxidant response and growth development are reviewed in this paper and the aspects to be further studied are also suggested.     

Interactive effects of ozone and drought stress on plants: A review

Ground-level ozone (O₃) and drought are two key factors limiting plant growth. O₃ can enter into the plant tissue through the stomata, then causing the formation of reactive oxygen species (ROS) which inspires programmed cell death. Drought usually induces the accumulation of ROS due to damage to antioxidant systems of plants. The effects of two kinds of stress on plants are similar due to the accumulation of ROS, resulting in reduced photosynthesis rate and physiological metabolism, eventually decreased plant growth and biomass. Nevertheless, O₃ and drought interacts synergistically to accumulate detrimental effects or antagonistically to reduce harmful effects. Actually, it is complex interactive process between O₃ and drought. On the one hand, O₃ triggers stomatal sluggishness or even dysfunction, which exacerbates water transpiration of leaves, water loss from plants and further O₃ phytotoxicity. On the other hand, drought induces stomatal closure, and thus protecting plants against the O₃ influx and evaporation of water. However, prolonged drought could limit the uptake of CO₂ and thus result in reduced plant growth. The response of plants to both O₃ and drought not only depends on the occurring sequence and duration of any factor but also rely on the difference in physiological metabolism of the plant itself. The interactive effects of O₃ and drought on stomatal characteristics, photosynthetic carbon mechanism, antioxidant response and growth development are reviewed in this paper and the aspects to be further studied are also suggested.