Induction of chilling tolerance in cucumber (Cucumis sativus) seedlings by endogenous and applied ethanol

Five-day-old etiolated cucumber ( Cucumis sativus L.) seedlings cv. Marketmore held at 2°C for 72 h developed chilling injury, resulting in desiccation and collapse of the hypocotyl tissue and eventual plant death. Hypoxia-induced accumulation of ethanol and acetaldehyde led to tolerance of subsequent chilling, as evidenced by continued hypocotyl growth and freedom from injury. Attenuated accumulation of volatiles by applied bisulfite reduced the development of hypoxia-induced chilling tolerance in seedlings. In seedlings held in normoxia cold tolerance was induced by applied ethanol vapors, whereas acetaldehyde had a marginal effect, suggesting that hypoxia-induced cold tolerance may arise from the accumulation and activity of ethanol. Cold tolerance was also induced by exposure of seedlings to volatile anesthetics including n -propanol, n -butanol, chloroform and halothane, suggesting that ethanol activity may result from fluidization of membrane lipids. This view is consistent with results which showed that ethanol activity was not associated with lipid metabolism. However, development of cold tolerance in ethanol-enriched tissues was time dependent, indicating that ethanol activity probably also entails biosynthetic event(s).     

Induction by Electric Currents of Ethylene Biosynthesis in Cucumber (Cucumis sativus L.) Fruit

The effects of an electric current on ethylene biosynthesis were investigated in cucumber (Cucumis sativus L.) fruit that were producing almost no ethylene. Direct currents at 0.5 to 3.0 milliamperes induced much ethylene synthesis, with a rapid continuous increase in the rate, which reached a peak within 5 to 6 hours and then decreased. The rate of production was greater with a stronger current. Ethylene production was not observed after the use of a sine-wave alternating current (60 hertz) at 3 milliamperes, the magnitude at which a direct current had the greatest effect. The activity of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase and ethylene forming enzyme (EFE) increased before the rise in ethylene production. ACC synthase and EFE were activated sixfold and fourfold, respectively, by 2 hours. The concentration of ACC increased linearly up to 6 hours and then decreased. Ethylene induction by an electric current was suppressed almost completely by the infiltration of the cucumbers with 5 millimolar aminooxyacetic acid, an inhibitor of ACC synthase, and was also suppressed 70% by 5 millimolar salicylic acid, an inhibitor of EFE. The results indicate that the ethylene induced by the direct current was synthesized via the ACC-ethylene pathway as a result of electrical stress, a new kind of stress to be identified.     

Role of leaf apoplast in silicon-mediated manganese tolerance of Cucumis sativus L.

Silicon (Si) supplied as sodium silicate (1·8 mm ) clearly decreased symptoms of manganese (Mn) toxicity in Cucumis sativus L. (cv. Chinesische Schlange) grown in nutrient solution with low to elevated Mn concentrations (0·5–1000 µm ). Despite approximately the same total Mn content in the leaves, plants not treated with Si had higher Mn concentrations in the intercellular washing fluid (IWF) compared with plants treated with Si, especially in the BaCl₂‐ and DTPA‐exchangeable fraction of the leaf apoplast. The Mn concentration of the IWF correlated positively with the severity of Mn‐toxicity symptoms and negatively with the Si supply. Furthermore, in Si‐treated plants less Mn was located in the symplast (< 10%) and more Mn was bound to the cell wall (> 90%) compared with non‐Si‐treated plants (about 50% in each compartment). Manganese present in Si‐treated plants is therefore less available and for this reason less toxic than in plants not treated with Si. It is concluded that Si‐mediated tolerance of Mn in C. sativus is a consequence of stronger binding of Mn to cell walls and a lowering of Mn concentration within the symplast. These results support the role of Si as an important beneficial element in plant nutrition.     

Genetic Transformation of Cucumis sativus by Agrobacterium rhizogenes

Hairy roots were obtained in vitro 10 days after inoculation of cucumber ( Cucumis sativus L. ) cotyledon explants with the strains of Agrobacterium rhiwgenes R1000 and R1601. The frequency of the cotyledon explants transformed by R1000 and R1601 was up to 87.5% and 88.9%, respectively. All hairy roots induced by the strains of R1000 and R1601 grew rapidly on solid hormone-free MS medium. The roots incited by A. rhizogenes R1000 could be divided into three phenotypes. The roots of phenotype Ⅰ were similar to the normal ones, but had more numerous lateral roots. Roots of phenotype m were much stouter and shorter, they elongated very slowly and were more highly branched than roots of phenotype Ⅰ . Roots of phenotype Ⅱ were of intermediate in appearance. However, the roots incited by A. rhizogenes R1601 appeared similar to phenotype Ⅰ roots incited by A. rhizogenes R1000. Transformation was confirmed by opine detection.     

Induction of abiotic stress tolerance in plants by endophytic microbes

Endophytes are micro‐organisms including bacteria and fungi that survive within healthy plant tissues and promote plant growth under stress. This review focuses on the potential of endophytic microbes that induce abiotic stress tolerance in plants. How endophytes promote plant growth under stressful conditions, like drought and heat, high salinity and poor nutrient availability will be discussed. The molecular mechanisms for increasing stress tolerance in plants by endophytes include induction of plant stress genes as well as biomolecules like reactive oxygen species scavengers. This review may help in the development of biotechnological applications of endophytic microbes in plant growth promotion and crop improvement under abiotic stress conditions.

Significance and Impact of the Study

Increasing human populations demand more crop yield for food security while crop production is adversely affected by abiotic stresses like drought, salinity and high temperature. Development of stress tolerance in plants is a strategy to cope with the negative effects of adverse environmental conditions. Endophytes are well recognized for plant growth promotion and production of natural compounds. The property of endophytes to induce stress tolerance in plants can be applied to increase crop yields. With this review, we intend to promote application of endophytes in biotechnology and genetic engineering for the development of stress‐tolerant plants.     

Water relations and photosynthesis in Cucumis sativus L. leaves under salt stress

Hydroponically grown cucumber plants were exposed to 14-d period of salinity (0, 50, 100 mM NaCl). NaCl caused reduction in the relative water content in the leaves. The Na⁺ content increased and the K⁺ content decreased. The net photosynthetic rate, stomatal conductance and transpiration rate were markedly decreased by all of the salt treatments. Salinity decreased also the maximum quantum efficiency of photosystem 2 (PS 2) determined as the variable to maximum fluorescence ratio, the photochemical quantum yield of PS 2 and the photochemical fluorescence quenching, while the non-photochemical quenching increased. Above results indicate that NaCl affects photosynthesis through both stomata closure and non-stomatal factors.     

Oxidative stress in cucumber (Cucumis sativus L) seedlings treated with acifluorfen

Treatment of diphenyl ether herbicide acifluorfen-Na (AF-Na) to intact cucumber (Cucumis sativus L cv Poinsette) seedlings induced overaccumulation of protoporphyrin IX in light (75 mumole m-2 s-1). The extra-plastidic protoporphyrin IX accumulated during the light exposure disappeared within two hours of transfer of acifluorofen-treated seedlings to darkness. The dark disappearance was due to re-entry of migrated protoporphyrin IX into the plastid and its subsequent conversion to protochlorophyllide. In light, protoporphyrin IX acted as a photosensitizer and caused generation of active oxygen species. The latter caused damage to the cellular membranes by peroxidation of membrane lipids that resulted in production of malondialdehyde. Damage to the plastidic membranes resulted in damage to photosystem I and photosystem II reactions. Dark-incubation of herbicide-sprayed plants before their exposure to light enhanced photodynamic damage due to diffusion of the herbicide to the site of action. Compared to control, in treated samples the cation-induced increases in variable fluorescence/maximum fluorescence ratio and increase in photosystem II activity was lower due to reduced grana stacking in herbicide-treated and light-exposed plants.     

离体培养黄瓜子叶诱导雌花的研究

采用添加Spd和IAA的MS培养基培养离体黄瓜子叶,研究了Spd和IAA对雌花诱导的协同作用,及昼夜温差、培养基中N素和pH值对雌花诱导的影响。结果表明,分别添加Spd、IAA时的雌花诱导率和单株雌花数偏低或为0,12 mg·L^-1 Spd与0.01mg·L^-1 IAA 配合时的诱导效果明显高于单独处理的,而对照组未见雌花,说明Spd和IAA对雌花诱导的协同作用显著。在0、2、6、10℃昼夜温差,60、70、80、90 mmol·L^-1的N素含量和pH 5.4、5.8、6.2、6.6的培养条件下,70 mmol·L^-1 N、6℃温差和pH 6.2时的雌花诱导效果较好,表明适当提高昼夜温差、培养基中N素和pH值有利于黄瓜子叶的雌花诱导。     

Exogenous spermidine affects polyamine metabolism in salinity-stressed Cucumis sativus roots and enhances short-term salinity tolerance

We investigated the effects of short-term salinity stress and spermidine application to salinized nutrient solution on polyamine metabolism and various stress defense reactions in the roots of two cucumber (Cucumis sativus L.) cultivars, Changchun mici and Jinchun No. 2. Seedlings grown in nutrient solution salinized with 50 mM NaCl for 8 d displayed reduced relative water content, net photosynthetic rates and plant growth, together with increased lipid peroxidation and electrolyte leakage in the roots. These changes were more marked in cv. Jinchun No. 2 than in cv. Changchun mici, confirming that the latter cultivar is more salinity-tolerant than the former. Salinity stress caused an increase in superoxide and hydrogen peroxide production, particularly in cv. Jinchun No. 2 roots, while the salinity-induced increase in antioxidant enzyme activities and proline contents in the roots was much larger in cv. Changchun mici than in cv. Jinchun No. 2. In comparison to cv. Jinchun No. 2, cv. Changchun mici showed a marked increase in arginine decarboxylase, ornithine decarboxylase, S-adenosylmethionine decarboxylase and diamine oxidase activities, as well as free spermidine and spermine, soluble conjugated and insoluble bound putrescine, spermidine and spermine contents in the roots during exposure to salinity. On the other hand, spermidine application to salinized nutrient solution resulted in alleviation of the salinity-induced membrane damage in the roots and plant growth and photosynthesis inhibition, together with an increase in polyamine and proline contents and antioxidant enzyme activities in the roots of cv. Jinchun No. 2 but not of cv. Changchun mici. These results suggest that spermidine confers short-term salinity tolerance on cucumber probably through inducing antioxidant enzymes and osmoticants.     

Arbuscular mycorrhizal fungi regulate the oxidative system,hormones and ionic equilibrium to trigger salt stress tolerance in Cucumis sativus L.

Arbuscular mycorrhizal fungi (AMF) association increases plant stress tolerance. This study aimed to determine the mitigation effect of AMF on the growth and metabolic changes of cucumbers under adverse impact of salt stress. Salinity reduced the water content and synthesis of pigments. However, AMF inoculation ameliorated negative effects by enhancing the biomass, synthesis of pigments, activity of antioxidant enzymes, including superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase, and the content of ascorbic acid, which might be the result of lower level lipid peroxidation and electrolyte leakage. An accumulation of phenols and proline in AMF-inoculated plants also mediated the elimination of superoxide radicals. In addition, jasmonic acid, salicylic acid and several important mineral elements (K, Ca, Mg, Zn, Fe, Mn and Cu) were enhanced with significant reductions in the uptake of deleterious ions like Na+. These results suggested that AMF can protect cucumber growth from salt stress.     

Photosynthesis Regulation by Glucohexaose Through Redox Changes in Cucumis sativus

To investigate the effects of glucohexaose (P6) on cucumber, leaf CO₂ assimilation, chlorophyll fluorescence parameters, chlorophyll content, and carbohydrate metabolism were examined in cucumber plants. The net photosynthetic rate (P _n ) of cucumber leaves was enhanced after being treated with 10 μg mL^?1 P6. The increase was correlated with increases in transpiration rate (E) and stomatal conductance (G _s), whereas the intercellular CO₂ concentration (C _i) was not different from the control plants. Chlorophyll content, absorption of light energy per unit area (ABS/CS), capture of light energy per unit area (TR_o/CS), quantum yield of electron transport per unit area (ET_o/CS), maximum photochemical efficiency of PSII (φP _o), quantum yield of photosynthetic institution electron transfer (φE _o), probability of other electron acceptors that captured exciton-transferred electrons to the electronic chain which exceeds QA (ψ _o), number of reaction centers per unit leaf area (RC/CS_o), and the performance index on absorption basis (PI_ABS) were improved, but heat dissipation per unit area (DI_o/CS) and maximum quantum yield of non-chemical quenching (φD _o) were reduced. In addition, increases in sucrose, soluble sugars, and starch contents were observed in P6-treated plants. However, H₂O₂ scavenger (DMTU) or NADPH oxidase inhibitor (DPI) pretreatment significantly abolished the effect of P6 on photosynthesis. The results demonstrated that ROS played a critical role in P6-induced photosynthesis. The increase in chlorophyll content together with efficient light absorption, transmission, and conversion in P6-treated plants is important for increasing photosynthesis.     

Effects of oxidative stress on chlorophyll biosynthesis in cucumber (Cucumis sativus) cotyledons

We conducted a series of experiments to assess the effects of oxidative stress on chlorophyll biosynthesis in the vascular plant Cucumis sativus (cucumber). Specifically, cucumber cotyledons were treated with 100 μ M methyl viologen (MV) and subsequently exposed to dark (0 μE m⁻² s⁻¹), low light (40–45 μE m⁻² s⁻¹), or high light (1500–1600 μE m⁻² s⁻¹). Following treatment, extracts of these samples were subjected to high-performance liquid chromatography (HPLC) to quantitate the accumulation of chlorophyll biosynthetic pathway intermediates. The results of these analyses revealed significant accumulation of Mg-protoporphyrin IX monomethyl ester (Mg-proto IX ME) in green (14-h illuminated) as well as in etiolated cotyledons with MV treatment. These data suggest that MV-induced oxidative stress may have inhibited Mg-proto IX ME cyclase activity. Upon exposure to high light, in the presence or absence of MV, both green and etiolated cotyledons predominantly accumulated protoporphyrin IX (Proto IX). These elevated levels of Proto IX might be attributable to attenuated activity of any or all of the following enzymes: Mg-chelatase, Fe-chelatase and protoporphyrinogen IX oxidase. We also observed that MV-induced oxidative stress impacts on chlorophyll biosynthesis to a greater extent than on photosystem II. These results demonstrate that oxidative stress impedes key steps in chlorophyll biosynthesis by either directly or indirectly inhibiting the activity of these enzymes.     

温度对黄瓜幼苗光合生理弱光耐受性的影响

以不耐弱光的津研2号和较耐弱光的戴多星黄瓜(Cucumis sativus L.)为试材,在人工气候室内研究适温25 ℃/18 ℃（昼/夜）、亚适温15 ℃/9 ℃和低温9 ℃/7 ℃对弱光(75～85 μmol·m^-2·s^-1)耐受性的影响.结果表明：弱光下黄瓜叶片的SPAD、净光合速率（P_n）、气孔导度（G_s）、蒸腾速率（T_r）、水分利用效率（WUE）、实际光化学效率（Φ_PSⅡ）、光化学猝灭（q_P）等指标下降,下降程度随温度的降低而加剧,而超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性上升.逆境解除后的恢复过程中,光合和荧光参数逐渐恢复,荧光参数恢复速度快于气体交换参数.弱光下温度越低对黄瓜幼苗叶片光合机构造成的伤害越重,低温降低了叶片对弱光的耐受性.在低温、弱光处理过程中,津研2号P_n、Φ_PSⅡ、q_P等下降程度较戴多星明显,而在随后的恢复过程中其回升速度较戴多星迟缓,表明弱光下戴多星对低温的耐受性强于津研2号.     

The uptake of ferulic and p-hydroxybenzoic acids by Cucumis sativus

The uptake of ferulic acid (FA) and p-hydroxybenzoic acid (p-HBA) from solutions (0.1–1.0 mM, pH 4.0–7.0), was determined for intact and ex     

黄瓜韧皮部的类血影蛋白

以黄瓜 (CucumissativusL .)叶柄为实验材料 ,应用胶体金免疫电镜技术证明类血影蛋白存在于韧皮部的筛管_伴胞复合体中 ,广泛分布于筛分子中的韧皮蛋白纤丝以及筛分子网络结构上 ,并且分布在伴胞的细胞质和线粒体膜以及筛分子与伴胞之间的分支状胞间连丝上 ,表明该蛋白可能由伴胞合成并经由二者之间的胞间连丝运输到筛分子中。用免疫印迹技术证明 ,黄瓜韧皮部汁液蛋白中存在类血影蛋白 ,其分子量约为 2 6 0kD ,与动物细胞中血影蛋白的分子量接近     

Acclimation of chlorophyll biosynthetic reactions to temperature stress in cucumber (Cucumis sativus L.)

The adaptive responses of the greening process of plants to temperature stress were studied in cucumber (Cucumis sativus L. cv. Poinsette) seedlings grown at ambient (25 °C), low (7 °C) and high (42 °C) temperatures. Plastids isolated from these seedlings were incubated at different temperatures and the net syntheses of various tetrapyrroles were monitored. In plastids isolated from control seedlings grown at 25 °C, the optimum temperature for synthesis of Mg-protoporphyrin IX monoester or protochlorophyllide was 35 °C. Temperature maxima for Mg-protoporphyrin IX monoester and protochlorophyllide syntheses were shifted to 30 °C in chill-stressed seedlings. The net synthesis of total tetrapyrroles was severely reduced in heat-stressed seedlings and the optimum temperature for Mg-protoporphyrin IX monoester or protochlorophyllide synthesis shifted slightly towards higher temperatures, i.e. a broader peak was observed. To further study the temperature acclimation of seedlings with respect to the greening process, tetrapyrrole biosynthesis was monitored at 25 °C after pre-heating the plastids (28–70 °C) isolated from control, chill- and heat-stressed seedlings. In comparison to 28 °C-pre-heated plastids the percent inhibition of protochlorophyllide synthesis in 40 °C-pre-heated plastids was higher than for the control (25 °C-grown) in chill-stressed seedlings and lower than for the control in heat-stressed seedlings. Maximum synthesis of total tetrapyrroles and protoporphyrin IX was observed when chloroplasts were heated at 50 °C, which was probably due to heat-induced activation of the enzymes involved in protoporphyrin IX synthesis. Prominent shoulders towards lower or higher temperatures were seen in chill-stressed or heat-stressed seedlings, respectively. The shift in optimum temperature for tetrapyrrole biosynthesis in chill- and heat-stressed seedlings was probably due to acclimation of membranes possibly undergoing desaturation or saturation of membrane lipids. Proteins synthesized in response to temperature-stress may also play an important role in conferring stress-tolerance in plants. Received: 8 October 1998 / Accepted: 19 November 1998     

黄瓜韧皮部的类血影蛋白

以黄瓜(Cucumis sativus L.)叶柄为实验材料,应用胶体金免疫电镜技术证明类血影蛋白存在于韧皮部的筛管-伴胞复合体中,广泛分布于筛分子中的韧皮蛋白纤丝以及筛分子网络结构上,并且分布在伴胞的细胞质和线粒体膜以及筛分子与伴胞之间的分支状胞间连丝上,表明该蛋白可能由伴胞合成并经由二者之间的胞间连丝运输到筛分子中.用免疫印迹技术证明,黄瓜韧皮部汁液蛋白中存在类血影蛋白,其分子量约为260 kD,与动物细胞中血影蛋白的分子量接近.