Effect of light irradiations on photosynthetic machinery and antioxidative enzymes during ex vitro acclimatization of Tylophora indica plantlets

Abstract

In the present communication, we studied the effect of light stress on pigment contents, net photosynthetic rate, electrolyte leakage, malondialdehyde concentration and various antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and ascorbate peroxidase (APX) during acclimatization of micropropagated Tylophora indica plantlets. Pigment (Chlorophyll a, b and carotenoids) contents in ex vitro formed leaves were found significantly higher compared to the in vitro formed ones. In vitro plantlets (day 0) exhibited a low photosynthetic activity, but with the emergence of new leaves a significant increase in net photosynthetic rates was observed. Changes in the activity of the antioxidant enzymes system were also observed during the critical days of acclimatization. Plantlets showed increased levels of SOD production, indicating its preventive mechanism of membrane oxidation and damage to biological molecules in high light (HL) acclimatized plantlets. The CAT activity increased at both low lights (LL) and HL during the whole period of acclimatization. Likewise, photoexposure of plantlets at LL and HL showed elevated activity of GR and APX against 0 day plantlets.     

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.     

The system modulating ROS content in germinating seeds of two Brazilian savanna tree species exposed to As and Zn

The effects of increasing arsenic (0, 10, 50, 100 mg L^?1) and zinc (0, 50, 80, 120, 200 mg L^?1) doses on germination and oxidative stress markers (H₂O₂, MDA, SOD, CAT, APX, and GR) were examined in two Brazilian savanna tree species (Anadenanthera peregrina and Myracrodruon urundeuva) commonly used to remediate contaminated soils. The deleterious effects of As and Zn on seed germination were due to As- and Zn-induced H₂O₂ accumulation and inhibition of APX and GR activities, which lead to oxidative damage by lipid peroxidation. SOD and CAT did not show any As- and Zn-induced inhibition of their activities as was seen with APX and GR. We investigated the close relationships between seed germination success under As and Zn stress in terms of GR and, especially, APX activities. Increased germination of A. peregrina seeds exposed to 50 mg L^?1 of Zn was related to increased APX activity, and germination in the presence of As (10 mg L^?1) was observed only in M. urundeuva seeds that demonstrated increased APX activity. All the treatments for both species in which germination decreased or was inhibited showed decreases in APX activity. A. peregrina seeds showed higher Zn-tolerance than M. urundeuva, while the reverse was observed with arsenic up to exposures of 10 mg L^?1.     

Enhanced multiplication and improved <Emphasis Type="Italic">ex vitro</Emphasis> acclimatization of <Emphasis Type="Italic">Decalepis arayalpathra</Emphasis>

The proposed work describes a protocol for high-frequency in vitro regeneration through nodal segments and shoot tips in Decalepsis arayalpathra, a critically endangered medicinal liana of the Western Ghats. Nodal segments were more responsive than shoot tips in terms of shoot proliferation. Murashige and Skoog’s (MS) basal medium supplemented with 5.0 μM 6-benzyladenine (BA) was optimum for shoot initiation through both the explants. Among different combinations of plant growth regulators and growth additive screened, MS medium added with 5.0 μM BA + 0.5 μM indole-3-acetic acid + 20.0 μM adenine sulphate effectuated the highest response: 11.8 shoots per nodal segment and 5.5 shoots per shoot tip with mean shoot length of 9.2 and 4.8 cm, respectively. Half-strength MS medium with 2.5 μM α-naphthalene acetic acid was optimum for in vitro root induction. The plantlets with the well developed shoot and root were acclimatized in Soilrite? with 92 % survival rate in the field conditions. During acclimatization, chlorophyll content, net photosynthetic rate, stomatal conductance, and transpiration rate were gradually changed in dependence of formation of new leaves. Further, the changes in activities of antioxidant enzymes, i.e., superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) as well as activity of carbonic anhydrase were also observed: a continuous rise in SOD activity, but a rise and fall in the activities of CAT, APX, and GR were also noticed. Maximum fresh mass (3.1 g plant^-1), dry mass (0.35 g plant^-1) of roots and 2-hydroxy-4-methoxybenzaldehyde content of 9.22 μg cm^-3(root extract) were recorded after 8 weeks of acclimatization.     

Responses to drought stress in two poplar species originating from different altitudes

Cuttings of Populus kangdingensis and Populus cathayana, originating from high and low altitudes in the eastern Himalaya, respectively, were examined during one growing season in a greenhouse to determine their responses to drought stress (soil moisture decreased from 100 to 55 or 25 % field capacity). Compared to control plants grown under 100 % field capacity, those poplars grown under 55 and 25 % field capacity possessed lower increases in height and stem diameter, and higher contents of soluble sugars, free proline, malondialdehyde (MDA) and hydrogen peroxide, and higher activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX) and glutathione reductase (GR). Compared with P. cathayana with greater leaf area, P. kangdingensis with greater root/shoot ratio exhibited lower MDA and H₂O₂ contents, higher soluble sugar and free proline contents, and higher activities of CAT, SOD, POD, APX and GR. These results suggested that P. kangdingensis was more drought tolerant than P. cathayana.     

Differential response to paraquat induced oxidative stress in two rice cultivars on antioxidants and chlorophyll a fluorescence

The responses of antioxidative system and photosystem II photochemistry of rice (Oryza sativa L.) to paraquat induced oxidative stress were investigated in a chilling-tolerant cultivar Xiangnuo no. 1, and a chilling-susceptible cultivar, IR-50. Electrolyte leakage and malondialdehyde (MDA) content of Xiangnuo no. 1 were little affected by paraquat, but they increased in IR-50. After paraquat treatment, superoxide dismutase (SOD) activity remained high in Xiangnuo no. 1, while it declined in IR-50. Activities of catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) declined with oxidative stress in both cultivars, but Xiangnuo no. 1 had higher GR activity than IR-50. Under paraquat induced oxidative stress, ascorbic acid (AsA) and reduced glutathione (GSH) concentrations remained high in Xiangnuo no. 1, but decreased in IR-50. The results indicated that higher activities of SOD and GR and higher contents of AsA and GSH in Xiangnuo no. 1 under paraquat induced oxidative stress were associated with its tolerance to paraquat, while paraquat induced damage to IR-50 was related to decreased activities of SOD, APX and GR and contents of AsA and GSH. F _v/F _m, Φ _PSII, and qP remained high in Xiangnuo no. 1, while they decreased greatly in IR-50 under paraquat induced oxidative stress.     

Preliminary study on the effects of heavy metals on the growth and some antioxidant enzymes in Arthrospira platensis‐M2 strain

In this study, we have analyzed superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR) activities, biomass accumulation and chlorophyll‐a content in the Arthrospira platensis ‐M2 strain grown at different concentrations of zinc (Zn), tin (Sn) and mercury (Hg). We found that there is a close relationship between chlorophyll‐a content and biomass accumulation in A. platensis ‐M2 strain as a result of Zn, Sn and Hg exposures. Sn was found to be the most toxic heavy metal among others because of the continious inhibition of both biomass and chlorophyll‐a accumulation at 500 and 1000 μg mL^?1 concentrations after the third day of the study, while they represented continuous increases at each Zn and Hg concentration over 7 days. Lower concentrations of Zn and Sn stimulate SOD and GR activities remarkably, probably due to oxidative stress caused by heavy metal toxicity. APX activity was significantly lowered by higher concentrations of the three metals used in this study. Our results suggest that higher heavy metal concentrations inhibited SOD, APX and GR activities but biomass and chlorophyll‐a accumulation endured in a time‐dependent manner, possibly due to some different defence mechanisms, which remain to be investigated.     

Comparison of plantain plantlets propagated in temporary immersion bioreactors and gelled medium during in vitro growth and acclimatization

The current work compared the physiological characteristics of plantain (Musa AAB) plantlets micropropagated in temporary immersion bioreactors (TIB) and on a gelled medium (GM). The plantlets were evaluated during in vitro growth (in the shoot elongation phase) and at the end of ex vitro acclimatization. TIB improved rooting and gave rise to longer shoots and higher dry mass. Respiration rate was the highest at the beginning of shoot elongation in both the TIB and GM plantlets. Photosynthetic rate in TIB was significantly higher than in GM from the midpoint of acclimatization, whereas a pyruvate kinase (PK) activity was lower. Starch accumulation was ca. two fold higher in corms than in leaves and always higher in the TIB than GM plantlets. The higher expression of genes coding for carbon metabolism enzymes PK and phosphoenolpyruvate carboxylase (PEPC) in TIB than in PM indicates a more important role of an autotrophic metabolism in the TIB plantlets when compared to the GM ones. The accumulated reserves were used during the first days of acclimatization leading to the higher survival rates and to the better plant quality of the TIB plantlets.     

Differential Responses of Two Wheat Varieties Differing in Salt Tolerance to the Combined Stress of Mn and Salinity

The effect of Mn and NaCl on growth, mineral nutrients and antioxidative enzymes in two tetroploid wheat genotypes differing in salt tolerance was investigated in this study. 100 mM NaCl and Mn stress significantly inhibited plant growth, photosynthesis and Ca uptake, while stimulated ROS accumulation, MDA and proline content in wheat plants, Mn stress also increased SOD, APX, GR and DHAR activities. Durum wheat (AS780) was less affected by 100 mM NaCl and Mn stress than emmer wheat (AS847) due to more proline production, higher antioxidative enzymes activities and less-affected mineral nutrients. Application of 10 mM NaCl to Mn-stressed durum wheat alleviated Mn-induced damage by reducing Mn accumulation and translocation, while promoting proline accumulation and SOD, APX and GR activities. Irrespective of NaCl level, the combined stress of Mn and NaCl caused more severe oxidative stress, result in further reduction of photosynthetic rate and plant growth in emmer wheat as compared to Mn stress alone. The additively negative effects of NaCl and Mn stress on growth of emmer wheat results from reduced SOD and APX activities as well as Ca, Cu and Fe accumulation in both shoots and roots. These results suggest that salt-tolerant durum wheat is superior to emmer in adapting to Mn stress and the combined stress of salinity and Mn.

     

Cadmium effects on mineral nutrition and stress in <Emphasis Type="Italic">Potamogeton crispus</Emphasis>

The mechanisms of plant tolerance to cadmium stress were studied by short-term exposure of Potamogeton crispus L. to various concentrations of Cd ranging from 0 to 0.09 mM. The accumulation of Cd and its influence on nutrient elements, chlorophyll pigments, ultrastructure, proline and MDA contents, and free radical production, as well as the activities of superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and glutathione reductase (GR) were investigated. The higher Cd concentration in the medium resulted in a significant enhancement of Cd accumulation. Photosynthetic pigment content decreased and ultrastructural damage to the leaf cells was aggravated with the increase in the Cd concentrations. Disruption of chloroplasts and mitochondria was observed even at the lowest concentration of Cd. Meantime, the rate of O₂^*− generation and the contents of H₂O₂ and MDA significantly increased under Cd stress, suggesting that Cd caused oxidative stress. In addition, the antioxidant system was clearly activated following Cd exposure. SOD and POD activities increased initially and then decreased, while APX and GR activities markedly increased. Simultaneously, mineral nutrition was disturbed. While K, P, Ca, and Cu contents decreased, Na, Fe, and Mn contents increased. Induction of antioxidant enzyme activities in leaves exposed to elevated Cd concentrations may be involved in Cd tolerance of P. crispus.     

Silicon increases boron tolerance and reduces oxidative damage of wheat grown in soil with excess boron

The effect of silicon on the growth, boron concentrations, malondialdehyde (MDA) content, lipoxygenase (LOX) activity, proline (PRO) and H₂O₂ accumulation, and the activities of major antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX)] and non-enzymatic antioxidants (AA) of wheat grown in soil originally with toxic B concentrations were investigated. Applied of 5.0 and 10.0 mM Si to the B toxic soil significantly increased Si concentration of the wheat and counteracted the deleterious effects of B on shoot growth. The contents of PRO, H₂O₂, MDA, and LOX activity of wheat grown in B toxic soil were significantly reduced by Si treatments. Compared with control plants, the activities of SOD, CAT, APX and content of AA were decreased by applied Si. Based on the present work, it can be concluded that Si alleviates B toxicity of wheat by preventing oxidative membrane damage and also translocation of B from root to shoot and/or soil to plant.     

嫁接对低温弱光下辣椒幼苗膜脂过氧化及抗氧化酶活性的影响

以‘卫士’为砧木,以‘赤峰特选’为接穂进行嫁接,在光照培养箱内对辣椒自根苗(对照)和嫁接苗进行低温 (8 ℃/5 ℃) 弱光(100 μmol·m^-2·s^-1)处理,处理7 d后在正常条件(25 ℃/18 ℃,550～600 μmol·m^-2·s^-1)下恢复3 d,研究低温弱光下辣椒嫁接苗和自根苗电解质渗漏率(EL)、丙二醛(MDA)含量、抗氧化酶活性及根系活力的变化.结果表明：低温弱光胁迫初期,辣椒幼苗叶片与根系的EL、MDA含量和超氧化物歧化酶(SOD)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性均显著升高,而根系活力大幅降低;1～3 d后EL和MDA含量趋于平稳,SOD、POD、APX、GR活性逐渐降低,根系活力呈上升趋势.恢复3 d后,嫁接苗EL、MDA含量、抗氧化酶活性及根系活力多达到或超过胁迫前水平(根系的MDA含量较胁迫前略高);而自根苗的EL和MDA含量仍显著高于胁迫前.与自根苗相比,嫁接苗在各处理阶段的EL和MDA含量显著降低,而SOD、POD、APX、GR活性及根系活力明显升高,说明嫁接可有效降低辣椒植株的膜脂过氧化,减轻低温弱光对其细胞膜的伤害.     

Mass multiplication of protocorm-like bodies using bioreactor system and subsequent plant regeneration in Phalaenopsis

Protocorm-like bodies (PLBs) formed on leaf segmentsin vitro were used as explants for bioreactor cultures. Continuous immersion cultures (air lift column and air lift-balloon bioreactor), and temporary immersion cultures (with or without charcoal filter attached) were used for the culture of PLB sections. A temporary immersion culture with charcoal filter attached was most suitable for PLB culture. About 18,000 PLBs were harvested from 20 g of inoculum (∼1000 PLB sections) in 2 l Hyponex medium after 8 weeks of incubation. Aeration in a bioreactor at 0.5 or 2.0 volume of air per volume of medium min⁻¹ (vvm) yielded similar levels of biomass production. PLBs grown in bioreactors were cultured on solid Murashige and Skoog, Vacin and Went, Knudson C, Lindemann and Hyponex media. Hyponex medium was found to be suitable for conversion of PLBs into plantlets and 83% of PLBs transformed into plantlets on this medium. The feasibility of using PLBs for large-scale micropropagation was evaluated for scaled-up liquid cultures in bioreactors, rate of proliferation, and regeneration. This revised version was published online in June 2006 with corrections to the Cover Date.     

Salicylic acid and calcium-induced protection of wheat against salinity

Soil salinity is one of the important environmental factors that produce serious agricultural problems. The objective of the present study was to determine the interactive effect of salicylic acid (SA) and calcium (Ca) on plant growth, photosynthetic pigments, proline (Pro) concentration, carbonic anhydrase (CA) activity and activities of antioxidant enzymes of Triticum aestivum L. (cv. Samma) under salt stress. Application of 90 mM of NaCl reduced plant growth (plant height, fresh weight (FW) and dry weight (DW), chlorophyll (Chl) a, Chl b, CA activity) and enhanced malondialdehyde (MDA) and Pro concentration. However, the application of SA or Ca alone as well as in combination markedly improved plant growth, photosynthetic pigments, Pro concentration, CA activity and activities of antioxidant enzymes peroxidase (POD), catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR) and ascorbate peroxidase (APX) under salt stress. It was, therefore, concluded that application of SA and Ca alone as well as in combination ameliorated the adverse effect of salinity, while combined application proved more effective to reduce the oxidative stress generated by NaCl through reduced MDA accumulation, Chl a/b ratio and Chls degradation and enhanced activities of antioxidant enzymes.     

Exogenous proline alleviates the effects of H2O2-induced oxidative stress in wild almond species

The effect of proline on the antioxidant system in the leaves of eight species of wild almond (Prunus spp.) exposed to H₂O₂-mediated oxidative stress was studied. The levels of endogenous proline (Pro) and hydrogen peroxide, and the activities of total superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), and guaiacol peroxidase (POD) were measured. The degradation of chlorophyll but not carotenoids occurred in leaves in the solution of 5 mM H₂O₂. An increase in membrane lipid peroxidation was observed in H₂O₂ treatment, as assessed by MDA level and percentage of membrane electrolyte leakage (EL). Significant increases in total SOD and CAT activities, as well as decreases in APX and POD activities, were detected in H₂O₂-treated leaves. The three SOD isoforms showed different behavior, as Mn-SOD activity was enhanced by H₂O₂, whereas Fe-SOD and Cu/Zn-SOD activities were inhibited. In addition, Pro accumulation up to 0.1 ??mol/g fr wt, accompanied by significant decreases in ascorbate and glutathione levels, was observed in H₂O₂-treated leaves. After two different treatments with 10 mM Pro + 5 mM H₂O₂, total SOD and CAT activities were similar to the levels in control plants, while POD and APX activities were higher if compared to the leaves exposed only to H₂O₂. Pro + H₂O₂ treatments also caused a strong reduction in the cellular H₂O₂ and MDA contents and EL. The results showed that Pro could have a key role in protecting against oxidative stress injury of wild almond species by decreasing membrane oxidative damage.