Fusarium wilt caused by Fusarium oxysporum f. sp. cucumerinum (FOC) is one of the major destructive soil-borne diseases infecting cucumber. In this study, we screened 60 target strains isolated from vinegar waste compost, from which 10 antagonistic strains were identified to have the disease suppression capacity of bio-control agents. The 16S rDNA gene demonstrated that the biocontrol agents were Paenibacillus polymyxa (P. polymyxa), Bacillus amyloliquefaciens (B. amyloliquefaciens) and Bacillus licheniformis (B. licheniformis). Based on the results of antagonistic activity experiments and pot experiment, an interesting strain of P. polymyxa (named NSY50) was selected for further research. Morphological, physiological and biochemical characteristics indicated that this strain was positive for protease and cellulase and produced indole acetic acid (22.21 ± 1.27 μg mL−1) and 1-aminocyclopropane-1-carboxylate deaminase (ACCD). NSY50 can significantly up-regulate the expression level of defense related genes PR1 and PR5 in cucumber roots at the early stages upon challenge with FOC. However, the gene expression levels of a set of defense-related genes, such as the plant nucleotide-binding site (NBS)-leucine-rich repeat (LRR) gene family (e.g., Csa001236, Csa09775, Csa018159), 26 kDa phloem protein (Csa001568, Csa003306), glutathione-S-transferase (Csa017734) and phenylalanine ammonia-lyase (Csa002864) were suppressed by pretreatment with NSY50 compared with the single challenge with FOC after nine days of inoculation. Of particular interest was the reduced expression of these genes at disease progression stages, which may be required for F. oxysporum dependent necrotrophic disease development.

•The temperature difference inside greenhouse with different hollow block walls was revealed from analyses of heat flows.•Two layered walls were designed based on heat transfer characteristics of hollow block wall filled by soil or perlite.•A solar greenhouse thermal model was used to compare wall temperature profile and its effect on indoor air temperature.In the present study, the volumetric thermal capacity and thermal conductivity of hollow block wall were changed by filling soil and perlite in the cavities to improve wall thermal performance. The results show that filling soil or perlite in the cavities of hollow blocks is a feasible way of improving the thermal performance of north wall of Chinese solar greenhouses. Filling hollow blocks with soil increases the thermal capacity of wall, and more heat can be stored. Filling hollow blocks with perlite increases the thermal resistance of north wall so less heat is lost. Then, two layered composite walls (wall HB-2P4S and HB-4P2S) were designed and their performance was compared. Model simulation and experimental results suggest that the composite wall with 40 cm of hollow block filled with soil and 20 cm of hollow block filled with perlite had better thermal performance than the composite wall that was composed of with 40 cm hollow block filled with perlite and 20 cm filled with soil.

The present study aimed to investigate the effect of exogenous spermidine (Spd) on cucumber (Cucumis sativus L. cv. Jinyou No. 4) growth and carbon–nitrogen balance under 80 mM Ca(NO3)2 stress. The result showed that leaf-applied Spd (1 mM) treatment alleviated the growth inhibition caused by Ca(NO3)2 stress by regulating the carbon–nitrogen balance in cucumber seedlings. The application of exogenous Spd effectively regulated the transcription levels and activities of major carbon–nitrogen metabolism enzymes, resulting in a significant decrease of NO3− and NH4+ contents under Ca(NO3)2 stress. In addition, Spd treatment remarkably increased the accumulation of soluble carbohydrates (sucrose, fructose and glucose), thus protected enzyme activities related N metabolism and effectively promoted NO3− assimilation under Ca(NO3)2 stress. Exogenous Spd also enhanced total amino acids, which serve as the building blocks of protein, and promoted the biosynthesis of soluble protein. In the presence of Spd, total C content and the C/N ratio increased significantly, while total N content decreased in response to Ca(NO3)2 stress. Based on our results, we suggested that exogenous Spd could effectively accelerate nitrate transformation into amino acids and improve the accumulation of carbon assimilation production, thereby enhancing the ability of the plants to maintain their C–N balance, and eventually promote the cucumber Ca(NO3)2 stress tolerance.

Ca(NO3)2 stress is one of the most serious constraints to plants production and limits the plants growth and development. Application of polyamines is a convenient and effective approach for enhancing plant salinity tolerance. The present investigation aimed to discover the photosynthetic carbon–nitrogen (C–N) mechanism underlying Ca(NO3)2 stress tolerance by spermidine (Spd) of cucumber (Cucumis sativus L. cv. Jinyou No. 4). Seedling growth and photosynthetic capacity [including net photosynthetic rate (PN), stomatal conductance (Gs), intercellular CO2 concentration (Ci), and transpiration rate (Tr)] were significantly inhibited by Ca(NO3)2 stress (80 mM). However, a leaf-applied Spd (1 mM) treatment alleviated the reduction in growth and photosynthesis in cucumber caused by Ca(NO3)2 stress. Furthermore, the application of exogenous Spd significantly decreased the accumulation of NO3− and NH4+ caused by Ca(NO3)2 stress and remarkably increased the activities of N metabolism enzymes simultaneously. In addition, photosynthesis N-use efficiency (PNUE) and free amino acids were significantly enhanced by exogenous Spd in response to Ca(NO3)2 stress, thus promoting the biosynthesis of N containing compounds and soluble protein. Also, the amounts of several carbohydrates (including sucrose, fructose and glucose), total C content and the C/N radio increased significantly in the presence of Spd. Based on our results, we suggest that exogenous Spd could effectively accelerate nitrate transformation into amino acids and improve cucumber plant photosynthesis and C assimilation, thereby enhancing the ability of the plants to maintain their C/N balance, and eventually promote the growth of cucumber plants under Ca(NO3)2 stress.

The effects of foliar spraying with spermidine (Spd) on antioxidant system in tomato (Lycopersicon esculentum Mill.) seedlings were investigated under high temperature stress. The high temperature stress significantly inhibited plant growth and reduced chlorophyll (Chl) content. Application of exogenous 1 mM Spd alleviated the inhibition of growth induced by the high temperature stress. Malondialdehyde (MDA), hydrogen peroxide (H2O2) content and superoxide anion (O2) generation rate were significantly increased by the high temperature stress, but Spd significantly reduced the accumulation of reactive oxygen species (ROS) and MDA content under the stress. The high temperature stress significantly decreased glutathione (GSH) content and activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR), but increased contents of dehydroascorbic acid (DHA), ascorbic acid (AsA), and oxidized glutathione (GSSG) in tomato leaves. However, Spd significantly increased the activities of antioxidant enzymes, levels of antioxidants and endogenous polyamines in tomato leaves under the high temperature stress. In addition, to varying degrees, Spd regulated expression of MnSOD, POD, APX2, APX6, GR, MDHAR, DHAR1, and DHAR2 genes in tomato leaves exposed to the high temperature stress. These results suggest that Spd could change endogenous polyamine levels and alleviate the damage by oxidative stress enhancing the non-enzymatic and enzymatic antioxidant system and the related gene expression.

The effects of exogenous putrescine (8 mmol L−1) on the carbohydrate content, glycolytic metabolism, citric acid cycle intermediates, key enzyme activities and their corresponding gene expression in cucumber (cucumis sativus L. cv. Jingyou NO.4) subjected to salt stress (75 mmol L−1 NaCl) in solution culture was studied. Phosphofructokinase (PFK), pyruvate kinase (PK) and phosphoenolpyruvate pyruvate kinase (PEPC) activities were significantly decreased in leaves when exposed to salt stress. NaCl stress caused the PFK and PK gene expression and the content of pyruvate to significantly decrease, while PEPC gene expression increased. Salt stress also significantly reduced isocitrate dehydrogenase, malate dehydrogenase and succinate dehydrogenase and their levels of transcription, causing decreases in the citric acid, succinic acid and malic acid content in leaves. Exogenous putrescine reversed the salt stress and increased gene expression and, in turn, the key enzymes involved in the glycolysis pathway and Krebs cycle, which significantly increased the amounts of organic acids and pyruvate produced, promoting the release of more energy currency (ATP and ADP). These results suggest that Put effectively participate glycolysis pathway and Krebs cycle, reducing the excessive accumulation of carbohydrates in the leaves, and providing more material and energy for the defense salt-induced injury, thus enhances cucumber seedling to salt stress tolerance.

Our results based on proteomics data and physiological alterations proposed the putative mechanism of exogenous Spd enhanced salinity tolerance in cucumber seedlings.Current studies showed that exogenous spermidine (Spd) could alleviate harmful effects of salinity. It is important to increase our understanding of the beneficial physiological responses of exogenous Spd treatment, and to determine the molecular responses underlying these responses. Here, we combined a physiological analysis with iTRAQ-based comparative proteomics of cucumber (Cucumis sativus L.) leaves, treated with 0.1 mM exogenous Spd, 75 mM NaCl and/or exogenous Spd. A total of 221 differentially expressed proteins were found and involved in 30 metabolic pathways, such as photosynthesis, carbohydrate metabolism, amino acid metabolism, stress response, signal transduction and antioxidant. Based on functional classification of the differentially expressed proteins and the physiological responses, we found cucumber seedlings treated with Spd under salt stress had higher photosynthesis efficiency, upregulated tetrapyrrole synthesis, stronger ROS scavenging ability and more protein biosynthesis activity than NaCl treatment, suggesting that these pathways may promote salt tolerance under high salinity. This study provided insights into how exogenous Spd protects photosynthesis and enhances salt tolerance in cucumber seedlings.

The application of exogenous 24-epibrassinolide promotes Brassinosteroids intracellular signalling in cucumber, which leads to differentially expressed proteins that participate in different life process to relieve Ca(NO3)2damage. NO3− and Ca2+ are the main anion and cation of soil secondary salinization during greenhouse cultivation. Brassinosteroids (BRs), steroidal phytohormones, regulate various important physiological and developmental processes and are used against abiotic stress. A two-dimensional electrophoresis gel coupled with MALDI-TOF/TOF MS was performed to investigate the effects of exogenous 24-epibrassinolide (EBL) on proteomic changes in cucumber seedling roots under Ca(NO3)2 stress. A total of 80 differentially accumulated protein spots in response to stress and/or exogenous EBL were identified and grouped into different categories of biological processes according to Gene Ontology. Under Ca(NO3)2 stress, proteins related to nitrogen metabolism and lignin biosynthesis were induced, while those related to cytoskeleton organization and cell-wall neutral sugar metabolism were inhibited. However, the accumulation of abundant proteins involved in protein modification and degradation, defence mechanisms against antioxidation and detoxification and lignin biosynthesis by exogenous EBL might play important roles in salt tolerance. Real-time quantitative PCR was performed to investigate BR signalling. BR signalling was induced intracellularly under Ca(NO3)2 stress. Exogenous EBL can alleviate the root indices, effectively reduce the Ca2+ content and increase the K+ content in cucumber roots under Ca(NO3)2 stress. This study revealed the differentially expressed proteins and BR signalling-associated mRNAs induced by EBL in cucumber seedling roots under Ca(NO3)2 stress, providing a better understanding of EBL-induced salt resistance in cucumber seedlings. The mechanism for alleviation provides valuable insight into improving Ca(NO3)2 stress tolerance of other horticultural plants.

In this research, we investigated the effects of grafting on intermediate metabolites and key enzymes of glycolysis and the tricarboxylic acid (TCA) cycle in self-grafted and salt-tolerant pumpkin rootstock-grafted cucumber seedlings supplied with nutrient solution and subjected to 80 mM Ca(NO3)2 stress for 6 days. Ca(NO3)2 stress induced accumulation of 3-phosphoglycerate (3-PGA) and phosphoenolpyruvate (PEP) in the leaves of self-grafted cucumber seedlings and enhanced the activities of phosphoenolpyruvate carboxylase (PEPC) and enolase (ENO). Succinic acid and malic acid contents and isocitrate dehydrogenase, succinate dehydrogenase (SDH), and malate dehydrogenase (MDH) activities in self-grafted seedlings were significantly decreased by Ca(NO3)2 stress. In addition, activities of PEPC, ENO, SDH, and MDH and contents of glycolysis intermediate metabolites (citric, succinic, and malic acids) were significantly higher in leaves of rootstock-grafted seedlings compared with those in self-grafted seedlings under saline conditions. Furthermore, leaf adenosine triphosphate (ATP) content of rootstock-grafted seedlings was relatively higher than that in self-grafted plants under salt stress, with an opposite effect observed on adenosine diphosphate content. These results indicate that rootstock grafting alleviates Ca(NO3)2 stress-induced inhibition of the glycolytic pathway and the TCA cycle in cucumber seedling leaves, which may aid the respiratory metabolism of cucumber seedlings and help maintain a high ATP synthesis level, thereby increasing the biomass of cucumber seedlings and enhancing their salt tolerance.

To investigate the effects of NaCl stress on plant growth, nitrogen assimilation, proline and soluble protein contents, and gene expression of enzymes of nitrogen metabolism, a hydroponic experiment using cucumber (Cucumis sativus L.) seedlings was performed. The seedlings were grown in nutrient solution supplemented with 0 or 84 mM NaCl for up to 9 days. Plant biomass, especially root biomass, was significantly decreased under NaCl stress. Salinity significantly increased ammonium content, but decreased nitrate and soluble protein contents in leaves and roots. Salt stress caused a significant increase in proline content, which peaked on day 3 of NaCl treatment. Moreover, salt stress significantly decreased activities of nitrate reductase (NR), glutamine synthetase (GS), glutamate synthase (GOGAT), and glutamate dehydrogenase (GDH) in the roots and leaves on the 3rd, 6th, and 9th day of NaCl treatment. A semiquantitative RT-PCR approach showed that changes in NR, GS and GOGAT gene expression were consistent with the salt-induced changes in enzyme activities. These results suggest that salt stress-induced growth inhibition in cucumber seedlings may involve disruption of nitrogen absorption and decreased activities of enzymes associated with nitrogen assimilation.

The objective of this study was to identify the effects of exogenous putrescine (Put) on leaf traits, hormones levels, and carbohydrate metabolism in cucumber seedlings under salt stress. The stress of 75 mM NaCl decreased plant growth and photosynthetic carboxylation efficiency. NaCl also increased the leaf thickness but destroyed the leaf internal structure. The contents of indoleacetic acid, zeatin, and zeatin riboside were decreased and the abscisic acid level was increased by NaCl. In addition, NaCl stress caused the accumulation of sucrose and starch in leaves and led to the feedback inhibition of photosynthesis, which can be inferred from the lower maximum rate of RuBisCo-mediated carboxylation (Vcmax). The analysis of starch-metabolizing enzyme activities suggested that the largely improved amylopectin contents contributed to the starch accumulation. Put regulated the levels and ratios of endogenous hormones, lowered the leaf thickness, and protected the leaf structure from damage caused by NaCl. Furthermore, Put decreased the accumulation of sucrose and starch caused by NaCl in leaves through regulating the activities of enzymes involved in carbohydrate metabolism. In conclusion, Put regulates the hormones levels, leaf structure, and carbohydrate metabolism and therefore improves the photosynthesis and growth of NaCl-stressed cucumber plants.

The objective of this study was to identify the effects of exogenous putrescine on photosynthetic performance and heat dissipation capacity in cucumber seedlings under salt stress. The stress of 75 mM NaCl for 7 days caused a significant decrease in net photosynthetic rate (PN). The experiment employed a chlorophyll fluorescence imaging technique and demonstrated that the maximal quantum yield of photosystem II photochemistry (Fv/Fm) and the actual photochemical efficiency of photosystem II (ΦPSII) were reduced by salt stress. Moreover, salt stress markedly reduced the photochemical quenching coefficient (qP) and non-photochemical quenching coefficient (qN), and significantly increased non-regulated heat dissipation (ΦNO). However, stressed plants supplied with exogenous putrescine exhibited higher PN and ΦPSII, which indicated that putrescine can alleviate the detrimental effects on photosynthesis induced by salt stress. Putrescine sprayed on stressed plants significantly enhanced the regulated energy dissipation (ΦNPQ) and decreased ΦNO. Application of exogenous putrescine also changed the levels of xanthophyll cycle components and further enhanced the de-epoxidation state of xanthophyll cycle pigments under salt stress. Under control conditions, putrescine exerted little influence on the photosynthetic parameters in cucumber leaves. In conclusion, the application of exogenous putrescine may improve the heat dissipation capacity by promoting the xanthophyll cycle to reduce the damage caused by excess excitation energy, thus enhancing the salt tolerance of cucumber seedlings.

This study focuses on the impact of carbohydrate metabolism and endogenous polyamines levels in leaves of cucumber seedlings under salt stress by exogenous BRs.The effects of 24-epibrassinolide (EBL) on carbohydrate metabolism and endogenous content of polyamines were investigated in cucumber seedlings (Cucumis sativus L. cv. Jinyou No. 4) exposed to salinity stress [80 mM Ca(NO3)2]. Spraying of exogenous EBL partially enhanced the enzyme activities of sucrose phosphate synthase, sucrose synthase and acid invertase; thus, raising the level of sucrose, fructose and total soluble sugars. The amylase activity was also increased by EBL, companied by the rising of sucrose level. These results indicated that EBL improved the carbohydrate metabolism of cucumber under Ca(NO3)2 stress. Moreover, EBL raised the levels of soluble conjugated and insoluble bound polyamines while lowered the free polyamines content, particularly putrescine. Our experiment demonstrated that exogenous EBL elevated stability of cellular membrane and positively improve the carbohydrate metabolism in cucumber growing under Ca(NO3)2 stress.

Seedlings of the hypoxia-sensitive cucumber cultivar were hydroponically grown under hypoxia for 7 d in the presence or absence of 24-epibrassinolide (EBR, 2.1 nM). Hypoxia significantly inhibited growth, while EBR partially counteracted this inhibition. Leaf net photosynthetic rate (PN), stomatal conductance, transpiration rate, and water-use efficiency declined greatly, while the stomatal limitation value increased significantly. The maximum net photosynthetic rate was strongly reduced by hypoxia, indicating that stomatal limitation was not the only cause of the PN decrease. EBR markedly diminished the harmful effects of hypoxia on PN as well as on stomata openness. It also greatly stimulated CO2 fixation by the way of increasing the carboxylation capacity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), ribulose-1,5-bisphosphate regeneration, Rubisco activity, and the protection of Rubisco large subunit from degradation. Our data indicated that photosystem (PS) II was damaged by hypoxia, while EBR had the protective effect. EBR further increased nonphotochemical quenching that could reduce photodamage of the PSII reaction center. The proportion of absorbed light energy allocated for photochemical reaction (P) was reduced, while both nonphotochemical reaction dissipation of light energy and imbalanced partitioning of excitation energy between PSI and PSII increased. EBR increased P and alleviated this imbalance. The results suggest that both stomatal and nonstomatal factors limited the photosynthesis of cucumber seedlings under hypoxia. EBR alleviated the growth inhibition by improving CO2 asimilation and protecting leaves against PSII damage.

We investigated the effects of exogenous spermidine (Spd) on growth, photosynthesis and expression of the Calvin cycle-related genes in cucumber seedlings (Cucumis sativus L.) exposed to NaCl stress. Salt stress reduced net photosynthetic rates (PN), actual photochemical efficiency of PSII (ΦPSII) and inhibited plant growth. Application of exogenous Spd to salinized nutrient solution alleviated salinity-induced the inhibition of plant growth, together with an increase in PN and ΦPSII. Salinity markedly reduced the maximum carboxylase activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Vcmax), the maximal velocity of RuBP regeneration (Jmax), triose-phosphate utilization capacity (TPU) and carboxylation efficiency (CE). Spd alleviated the negative effects on CO2 assimilation induced by salt stress. Moreover, Spd significantly increased the activities and contents of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and fructose-1,6-biphosphate aldolase (ALD; aldolase) in the salt-stressed cucumber leaves. On the other hand, salinity up-regulated the transcriptional levels of ribulose-1,5-bisphosphate (RCA), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribrokinase (PRK) and down-regulated the transcriptional levels of ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (RbcL), ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS), ALD, triose-3-phosphate isomerase (TPI), fructose-1,6-bisphosphate phosphatase (FBPase) and 3-phosphoglyceric acid kinase (PGK). However, Spd application to salt-stressed plant roots counteracted salinity-induced mRNA expression changes in most of the above-mentioned genes. These results suggest that Spd could improve photosynthetic capacity through regulating gene expression and activity of key enzymes for CO2 fixation, thus confers tolerance to salinity on cucumber plants.

•We study that the plant growth-promoting bacteria (PGPB) alleviates the stress on plant.•The physiological metabolism was inhibited by hypoxia.•The regulation of proteins and metabolisms by PGPB under hypoxia was studied.•We compare the proteins and metabolisms between hypoxia with and without PGPB.Plant growth-promoting bacteria (PGPB) can both facilitate plant growth and improve plant resistance to a variety of environmental stresses. In order to investigate the mechanisms that PGPB use to protect plants under hypoxic conditions, the protein profiles of stressed and non-stressed cucumber roots, either treated or not treated with PGPB, were examined. Two dimensional difference in-gel electrophoresis (DIGE) was used to detect significantly up- or down-regulated proteins (p < 0.05, |ratio| > 1.5) in cucumber roots in response to hypoxia. There were 1980, 1893 and 1735 protein spots detected from cucumber roots in the absence of stress in the presence of the PGPB Pseudomonas putida UW4, following hypoxic stress, and following hypoxic stress in the presence of P. putida UW4, respectively. The numbers of significantly changed protein spots were 0, 106, and 147 in these three treatments respectively. Proteins were identified by LTQ-MS/MS and categorized into classes corresponding to transcription, protein synthesis, signal transduction, carbohydrate and nitrogen metabolism, defense stress, antioxidant, binding and others. The functions of the proteins whose expression changed significantly were analyzed in detail, contributing to a more thorough understanding of how PGPB mediate the stress response in plants.Biological significanceTo our knowledge, only a limited number of papers have addressed cucumber proteomics, this study is the first report to describe the effect of plant growth-promoting bacteria (P. putida UW4) on cucumber plants under hypoxic stress using a proteomic approach. Thus, this work provides new insights to understand the cross-reactivity between P. putida UW4 and cucumber plant. A model of cucumber roots in response to P. putida UW4 and hypoxia was proposed: P. putida UW4 and hypoxic stress caused changes of gene expression in cucumber roots, then transcription was stimulated, the proteins involved in carbohydrate metabolism, nitrogen metabolism, defense stress, antioxidant, binding and others were induced, these proteins might work cooperatively to release hypoxic stress and promote cucumber growth. These results describe a dynamic protein network to explain the promotion mechanism of P. putida UW4, and also provide a solid basis for further functional research of single nodes of this network.

The aim of this study was to evaluate the effects of different light quality of light emitting diode (LED) on the growth, concentration of chlorophyll and chlorophyll biosynthesis precursors of non-heading Chinese cabbage (Brassica campestris L.). Seedlings of the cultivar Te Ai Qing were cultured for 28 days under 6 treatments: red light (R), blue light (B), green light (G), yellow light (Y), red plus blue light (RB) and dysprosium lamp (CK). Lighting experiments were performed under controlled conditions (photon flux density 150 μmol m−2 s−1; 12 h photoperiod; 18–20 °C). The fresh and dry mass were the greatest under RB, which were significantly higher than other light treatments. The fresh mass under RB was almost twice higher compared to other light treatments. Plant height was highest under R treatment and was lowest under B. RB treatment also lowered the plant height significantly. The highest soluble sugar concentration was observed under B. The soluble protein concentration was the greatest under RB. The R treatment was adverse to pigment accumulation. The concentration of photosynthetic pigments and chlorophyll biosynthesis precursors were higher under RB. The RB treatment was beneficial to pigment accumulation.

The effects of 0.1 μM 24-epibrassinolide (EBL) on plant growth (plant height, leaf area, fresh weight, and dry weight), chlorophyll content, photosynthetic characteristics, antioxidant enzymes, and chloroplast ultrastructure were investigated using cucumber seedlings (Cucumis sativus L. cv. Jinyou No. 4) with 80 mM Ca(NO3)2 to induce stress. The presence of Ca(NO3)2 caused significant reductions in net photosynthetic rate (PN), stomatal conductance (Gs), intercellular CO2 concentration (Ci), and transpiration rate (Tr) of leaves. In addition, Ca(NO3)2 markedly reduced the chlorophyll content and inhibited photochemical activity, including the actual photochemical efficiency (ΦPSII). In contrast, EBL increased the chlorophyll content, especially chlorophyll b, and minimized the harmful effects on photosynthesis caused by the Ca(NO3)2. The application of EBL to the plants subjected to Ca(NO3)2-enhanced photochemical activity. EBL protected the photosynthetic membrane system from oxidative damage due to up-regulating the capacity of the antioxidant systems. Microscopic analyses revealed that Ca(NO3)2 affected the structure of the photosynthetic apparatus and membrane system and induced damage of granal thylakoid layers, while EBL recovered the typical shape of chloroplasts and promoted the formation of grana. Taken together, EBL compensated for damage/losses by Ca(NO3)2 due to the regulation of photosynthetic characteristics and the antioxidant system.

The effects of exogenous spermidine (Spd) application to hypoxic nutrient solution on the contents of endogenous polyamines (PAs) and respiratory metabolism in the roots of cucumber (Cucumis sativus L.) seedlings were investigated. Cucumber seedlings were grown hydroponically in control and hypoxic nutrient solutions with and without addition of Spd at a concentration of 0.05 mM. The activities of key enzymes involved in the tricarboxylic acid cycle (TCAC), such as succinate dehydrogenase (SDH) and isocitrate dehydrogenase (IDH), were significantly inhibited under root-zone hypoxia with dissolved oxygen (DO) at 1 mg/l. In contrast, the activities of enzymes involved in the process of fermentation, such as pyruvate decarboxylase (PDC), alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH), and alanine aminotransferase (AlaAT), were significantly increased. Thus, aerobic respiration was inhibited and fermentation was enhanced in the roots of cucumber seedlings as a result of decreasing ATP content to inhibit the dry weight of seedlings under hypoxic stress. Moreover, the contents of free, soluble conjugated, and insoluble bound putrescine (Put), Spd, and spermine (Spm) in the roots of cucumber seedlings were significantly increased under hypoxia stress. Interestingly, application of Spd to hypoxic roots markedly suppressed the accumulation of free Put and, in contrast, promoted an increase in free Spd and Spm, as well as soluble conjugated and insoluble bound Put, Spd, and Spm contents. From these data, we deduced that exogenous Spd promotes the conversion of free Put into free Spd and Spm, and soluble conjugated and insoluble bound PAs under hypoxia stress. Furthermore, the activities of LDH, PDC, and ADH were suppressed and, in contrast, the activities of SDH and IDH were enhanced by application of exogenous Spd to hypoxic roots. As a result, aerobic respiration was enhanced but fermentation metabolism was inhibited in the roots of cucumber seedlings, leading to an increase in ATP content to alleviate the inhibited dry weight of seedlings due to hypoxia stress. These results suggest that application of Spd to hypoxic nutrient solution promoted conversion of free Put into free Spd and Spm as well as soluble conjugated and insoluble bound PAs, further enhanced IDH and SDH activities, and inhibited ethanol fermentation and lactate fermentation, resulting in increased ATP content and eventually enhanced tolerance of cucumber plants to root-zone hypoxia.

The relationship between the activity of xanthophyll cycle and chlorophyll (Chl) metabolism was investigated using two cultivars, Helan No. 3 (seawater-tolerant cultivar) and Yuanye (seawater-sensitive cultivar), of spinach (Spinacia oleracea L.) plants cultured in Hoagland’s nutrient solution, with or without seawater (40%). The results showed that, in plants of two cultivars with seawater, the xanthophyll cycle seems to show a principal protection mechanism against photoinhibition under seawater stress. Furthermore, accumulation of reactive oxygen species (ROS) in chloroplasts of two cultivars was enhanced by seawater to lower the activity of porphobilinogen deaminase. Namely, the conversion of porphobilinogen into uroporphyrinogen III involved in Chl biosynthetic processes was inhibited by seawater. In Helan No. 3 spinach plants with seawater, higher activity of xanthophyll cycle in the leaves dissipated more excess light energy, which appeared to lower the levels of ROS in chloroplasts. As a consequence, the Chl biosynthesis in Helan No. 3 leaves with seawater showed only a weak inhibition and the activity of chlorophyllase (Chlase) was not affected by seawater stress. In contrast, a more pronounced accumulation of ROS in chloroplasts of Yuanye leaves, which possess lower xanthophyll cycle activity, severely inhibited Chl biosynthesis and remarkably enhanced the activity of Chlase, which aggravates the decomposition of Chl. These results suggest that higher activity of xanthophyll cycle in seawater-tolerant spinach plays a role in maintaining Chl metabolic processes, probably by decreasing the levels of ROS, when the plants are cultured in the nutrient solution with seawater (40%).

To understand metabolic modifications in plants under salt stress, the physiological and biochemical responses of cucumber (Cucumis sativus L. cv. Jinchun No. 2) seedlings to salt stress was investigated. The dry weight and fresh weight of cucumber seedling roots were significantly reduced by treatment with NaCl; Na+ and Cl− were increased, while K+ and K+/Na+ ratio were decreased. To identify components of salt stress signaling, we compared the high resolution two-dimensional gel electrophoresis (2-DE) protein profiles of control and NaCl-treated roots, and the intensity of 34 protein spots varied. Of these spots, the identities of 29 (21 up-regulated and 8 down-regulated protein spots induced after salt stress) were determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and liquid chromatography electro-spray ionization tandem mass spectrometry (LC–ESI-MS/MS). The majority of the proteins had functions related to metabolism, energy and transport, and are involved in regulating reactions and defending against stress. A semi-quantitative reverse transcriptional-polymerase chain reaction (PCR) approach based on peptide sequences was used to compare transcript and protein accumulation patterns for 10 candidate proteins. Of these proteins, 8 patterns of induced transcript accumulation were consistent with those of induced protein accumulation. It is therefore likely that the response of the plant’s proteome to NaCl stress is complex, and that the identified proteins may play an important role in regulating adaptation activities following exposure to NaCl stress in order to facilitate ion homeostasis.The biochemical responses of cucumber seedlings exposed to salt stress were investigated using a proteomics approach. The proteins identified may play an important role in regulating complex adaptation activities during salt stress to facilitate ion homeostasis.

The effects of 10 mM putrescine (Put) treated by spraying on leaves on growth, chlorophyll content, photosynthetic gas-exchange characteristics, and chlorophyll fluorescence were investigated by growing cucumber plants (Cucumis sativus L. cv. ChangChun mici) using hydroponics with or without 65 mM NaCl as a salt stress. Salt stress caused the reduction of growth such as leaf area, root volume, plant height, and fresh and dry weights. Furthermore, net photosynthesis rate (Pn), stomatal conductance (gs), intercellular CO2 concentration (Ci), and transpiration rate (Tr) were also reduced by NaCl, but water use efficiency (WUE; Pn/Tr) showed a tendency to be enhanced rather than reduced by NaCl. However, Put alleviated the reduction of Pn by NaCl, and showed a further reduction of Ci by NaCl. The reduction of gs and Tr by NaCl was not alleviated at all. The enhancement of WUE by NaCl was shown to have no alleviation at day 1 after starting the treatment, but after that, the enhancement was gradually reduced till the control level. Maximum quantum efficiency of PSII (Fv/Fm) showed no effects by any conditions based on the combination of NaCl and Put, and in addition, kept constant values in plants grown in each nutrient solution during this experimental period. The efficiency of excitation energy capture by open photosystem II (PSII) (Fv′/Fm′), actual efficiency of PSII (ΦPSII), and the coefficient on photochemical quenching (qP) of plants with NaCl were reduced with time, and the reduction was alleviated till the control level by treatment with Put. The Fv′/Fm′, ΦPSII, and qP of plants without NaCl and/or with Put showed no variation during the experiment. Non-photochemical quenching of the singlet excited state of chlorophyll a (NPQ) showed quite different manner from the others as mentioned above, namely, continued to enhance during the experiment.

The effects of 24-epibrassinolide (EBR) added to nutrient solution on growth of cucumber (Cucumis sativus L.) under root-zone hypoxia were investigated. Cucumber seedlings were hydroponically grown for 8 days in normoxic and hypoxic nutrient solutions with and without addition of EBR at 1 μg l−1. EBR exerted little influence on plant performance in the normoxic nutrient solution, while the chemical alleviated root-zone hypoxia-induced inhibition of root and shoot growth and net photosynthetic rate (Pn). EBR added to hypoxic nutrient solution caused an increase in the concentration of fructose, sucrose, and total soluble sugars in the roots but not in the leaves. Root-zone hypoxia enhanced the activities of lactate dehydrogenase (LDH), alcohol dehydrogenase (ADH), and pyruvate decarboxylase in the roots. Interestingly, EBR further enhanced ADH activity but lowered LDH activity in hypoxic roots. These results suggest that EBR added to hypoxic nutrient solution may stimulate the photosynthate allocation down to roots and the shift from lactate fermentation to alcohol fermentation in hypoxic roots, resulting in the increase in ATP production through glycolysis and the avoidance of cytosolic acidosis and eventually enhanced tolerance of cucumber plants to root-zone hypoxia.

With water culture, this paper studied the effects of exogenous spermidine (Spd) on the net photosynthetic rate (Pn), intercellular CO2 concentrations (Ci), stomatal conductance (Gs), transpiration rate (Tr), apparent quantum yield (Φc), and carboxylation efficiency (CE) of cucumber seedlings under hypoxia stress. The results showed that Pn decreased gradually under the hypoxia stress, and reached the minimum 10 days later, which was 63.33% of the control. Compared with that of the hypoxia-stressed plants, the Pn 10 days after the application of exogenous Spd increased by 1.25 times. A negative correlation (R2=0.473−0.7118) was found between Pn and Ci, and Gs and Tr changed in wider ranges, which decreased under the hypoxia-stress, but increased under the hypoxia-stress plus exogenous Spd application. There was a significant positive correlation between Gs and Tr (R2=0.7821−0.9458), but these two parameters had no significant correlation with Pn. The hypoxia stress induced a decrease of Φc and CE by 63.01% and 72.33%, respectively, while the hypoxia stress plus exogenous Spd application made Φc and CE increase by 23% and 14%, respectively. The photo-inhibition of cucumber seedlings under hypoxia stress was mainly caused by non-stomatal inhibition, while the exogenous Spd alleviating the hypoxia stress by repairing photosynthesis systems.

Seedlings from the salt-sensitive cucumber cultivar Jinchun No. 2 and the salt-tolerant cucumber cultivar Changchun Mici were exposed for 8 days to 50 mmol/L NaCl in the absence or in the presence of exogenous foliar spraying PAs [putrescine (Put), spermidine (Spd), and spermine (Spm) 1 mmol/L] to compare the effects of different kinds of polyamines (PAs) on plant tolerance to salinity. This paper studied the effects of exogenous PAs on K+, Na+ and Cl− in different organs of cucumber seedlings. The results showed that K+ content as well as the ratios of K/Na and Cl/Na decreased, while Na+ and Cl− concentrations increased in salt-treated cucumber seedlings. The differences in K+, Na+ and Cl− content and the K/Na and Cl/Na ratios were greater for the salt sensitive cultivar Jinchun No. 2 than for the salt-tolerant cultivar Changchun Mici. Cucumber seedlings treated with exogenous polyamines and combined with salinity exhibited a higher level of K+ accumulation and lower levels of Na+ and Cl− accumulation compared with the seedlings treated only with salt stress. Among the three kinds of polyamines, Spd and Spm were more effective in inhibiting the accumulation of Na+ and reduction of K+. However, Put was more effective in reducing Cl− accumulation. Furthermore, all of the three kinds of exogenous polyamines could increase the ratio of K/Na, improving the absorption and transport selectivities of K+ and Na+ from stems to leaves for both cultivars. In conclusion, exogenous polyamines could alleviate salt damage to some extent and enhance the accumulation of biomass. Among the three kinds of polyamines, spermidine was most effective. Exogenous polyamines could improve tolerance of cucumber seedlings under salt stress by regulating the absorption and distribution of ions in different organs.

•Low temperature and weak light stress cause the inhibition of photosynthesis and nitrogen metabolism in tomato seedlings.•Chlorophyll fluorescence imaging technique was used to analyze the photochemical efficiency of photosystem II.•EBR could alleviate the photoinhibition and improve the photochemical efficiency of stressed-tomato seedlings.•EBR could enhance the assimilation of nitrate and ammonium in the primary nitrogen metabolism pathway.•EBR could confer resistance to tomato plants against low temperature and weak light stress.In the present study, we aimed to investigate the effects of exogenous 24-epibrassinolide (EBR) on growth, photosynthetic characteristics, chlorophyll fluorescence imaging, and nitrogen metabolism of tomato leaves under low temperature and weak light conditions. The results showed that foliar application of EBR significantly alleviated the inhibition of plant growth, and increased the fresh and dry weights of tomato plants under a combined low temperature and weak light stress. Moreover, EBR also increased the net photosynthetic rate (Pn), light saturation point (LSP), maximal quantum yield of PSII photochemistry (Fv/Fm), actual photochemical efficiency of PSII (ФPSII), and photochemical quenching coefficient (qP), but decreased the intercellular CO2 concentration (Ci), light compensation point (LCP) and apparent quantum efficiency (AQE) under low temperature and weak light conditions. In addition, application of EBR to tomato leaves significantly enhanced the activities of nitrate reductase (NR), glutamate dehydrogenase (GDH), glutamine synthetase (GS), and glutamate synthase (GOGAT), but decreased the ammonium content and nitrite reductase (NiR) activity. We observed that EBR remarkably increased the contents of aspartic acid, threonine, serine, glycine, and phenylalanine, while decreasing the accumulation of cysteine, methionine, arginine, and proline under a combined low temperature and light stress. These results suggest that EBR could alleviate the combined stress-induced harmful effects on photosynthesis and nitrogen metabolism, thus leading to improved plant growth.

Nitric oxide (NO), an endogenous signaling molecule in plants and animals, mediates responses to abiotic and biotic stresses. This study was conducted in nutrient solution to investigate the effects of exogenous sodium nitroprusside (SNP), an NO donor, on plant growth and free polyamine content in cucumber leaves and roots under NaCl stress. The results showed that 100 μM SNP in solution significantly improved the growth of cucumber seedlings under NaCl stress for 8 days, as indicated by increased, plant height, stem thickness, fresh weight and increased dry matter accumulation. Further analysis demonstrated that the content of free polyamines and the activity of polyamine oxidase (PAO) in cucumber seedling leaves and roots initially increased dramatically under NaCl stress, although they decreased over a longer period of stress. Throughout the treatment period, the value of (spermine + spermidine)/putrescine [(Spd + Spm)/Put] also decreased under NaCl stress compared to the control. In contrast, the application of 100 μM SNP in the nutrient solution decreased the content of free Put, Spd, total free polyamines and PAO activity under NaCl stress. It also caused an increase in the content of Spm and the value of (Spd + Spm)/Put, adjusted the ratio of three kinds of free polyamines (Put, Spd, Spm) in cucumber seedlings. The high (Spd + Spm)/Put value and the accumulation of Spm were beneficial to improving the salt tolerance of plants. Therefore, NO alleviated the damage to cucumber seedlings caused by salt stress. NO enhanced the tolerance of cucumber seedlings to NaCl stress by regulating the content and proportions of the different types of free polyamines.Research highlights▶ Nitric oxide enhanced salt tolerance in cucumber seedlings. ▶ Enhanced tolerance related to an increase in the content of Spm and the value of (Spd + Spm)/Put. ▶ Nitric oxide adjusted the biosynthesis of polyamines and the ratio of three different polyamines (Put, Spd, Spm). ▶ Exogenous nitric oxide improved the accumulation of Spm content and the (Spd + Spm)/Put value.

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.

Cowpea (Vigna unguiculata L. cv. Lulutong 1) is a leguminous vegetable which is well adapted to infertile acid soil. The mechanisms of aluminum (Al) resistance in cowpea are poorly understood. This study aimed to analyze the possible Al resistance mechanisms of cowpea. After exposure to Al, both malate and citrate were secreted from cowpea roots, but the secretion of these organic acids was not specific to Al. Potassium (P) starvation also induced the secretion of malate and citrate. The secretion of malate was the dominant response to Al stress. Exposure to 50 μM La3+ (Lanthanum) did not induce organic acid secretion. Secretion of organic acid was detected after 3–6 h exposure to Al, and increased significantly after 6 h exposure, which suggested that this plant showed a pattern II-type organic acid secretion. This is supported by the finding that a protein-synthesis inhibitor, cycloheximide (CHM), significantly inhibited secretion. Two types of anion-channel inhibitors had different effects on Al-induced secretion of organic acids, 9-anthracene carboxylic acid (A-9-C), which completely inhibited secretion whereas niflumic acid (NIF) had no effect. In the presence of A-9-C, Al inhibited root elongation more significantly. Compared with the control, the root tips showed no significant change in organic acid content or in citrate synthase (CS), malate dehydrogenase (MDH) and succinate dehydrogenase (SDH) activities. Taken together, these results indicated that the secretion of malate played a dominant role in the response to Al stress by cowpea plants. The secretion pattern was type II, and protein synthesis might be involved in the secretion process.Highlights► In this study we analyzed the characteristic of organic acid secretion induced by Al in cowpea roots. ► The cowpea is well adapt to acid infertile soil. ► We found that malate secreted from cowpea roots is dominant in response to Al. ► There is a marked lag phase between the addition of Al and the onset of organic acid release. ► We thought that the synthesis of proteins involved in the transport of malate anions in root apices of cowpea may be induced by Al.

•Leaf morphology, structure and biochemical substance of plantlets in vitro were investigated to identify the effects of different LED-generated spectra on leaves under the same light intensity.•The effects of spectral quality on the endogenous hormone content and leaf anatomical features of illuminated in vitro leaves were studied.•Monochromic blue LEDs, monochromic red LEDs and three mixtures of blue plus red LEDs on the growth and morphogenesis of leaves on P. grandiflorum plantlets in vitro under the same light intensity were studied to select the best light source for this cultivation system.In many parts of Asia, balloon flower (Platycodon grandiflorum (Jacq.) A. DC., P. grandiflorum) is much valued because of its ornamental and medicinal attributes. Although previous studies have described techniques of plant regeneration and culture, the influence of different spectra has not been documented. We investigated the effect of light spectra on leaf morphology, anatomy and chemical composition using cultured nodal explants grown under light-emitting diodes (LEDs) at a photosynthetic photon flux density of 50 μmol m−2 s−1. Treatments were: monochromatic blue light (B), 75% blue + 25% red light (BR31), 50% blue + 50% red light (BR11), 25% blue + 75% red light (BR13) and monochromatic red light (R). Fluorescent white lamps (FL) were used as a control. Treatment B induced larger leaf area, leaf thickness and dry mass of whole plant and higher leaf number than treatment R or FL. Plantlets from treatment R had the highest specific leaf mass, soluble sugar and sucrose content. Chlorophyll and L-ascorbic acid content was higher in leaves grown under LED variants than in those grown under FL. The indoleacetic acid content in B- and BR31-treated leaves was higher than that in the R-treated leaves. In addition, B and BR31 induced large and nearly rectangular palisade parenchyma cells, while the palisade parenchyma cells were elliptical in shape and small in the FL, BR11 and BR13 treatments, or irregular in the R treatment. Leaves from the R and BR11 treatments had the highest stomata frequency, moreover, FL-, B- and BR31-treated leaves possessed normal elliptical stomata. Treatment B induced the highest ETR, ΦPSII, and qp, significantly higher than treatment R or FL. Our data revealed that light spectrum strongly affected the morphology and growth index of P. grandiflorum plantlets cultured in vitro.

We investigated the effects of short-term root-zone hypoxic stress and exogenous calcium application or deficiency in an anoxic nutrient solution on nitrogen metabolism in the roots of the muskmelon cultivar Xiyu No. 1. Seedlings grown in the nutrient solution under hypoxic stress for 6 d displayed significantly reduced plant growth and soluble protein concentrations. However, NO3− uptake rate and activities of nitrate reductase and glutamate synthase were significantly increased. We also found higher amounts of nitrate, ammonium, amino acids, heat-stable proteins, polyamines, H2O2, as well as higher polyamine oxidase activity in the roots. In comparison to the reactions seen under hypoxic stress, exogenous calcium application led to a marked increase in plant weights, photosynthesis parameters, NO3− uptake rate and contents of nitrate, ammonium, amino acids (e.g., glutamic acid, proline, glycine, cystine, γ-aminobutyric acid), soluble and heat-stable proteins, free spermine, and insoluble bound polyamines. Meanwhile, exogenous calcium application resulted in significantly increased activities for nitrate reductase, glutamine synthetase, and glutamate synthase but decreased activities for diamine and polyamine oxidase, as well as lower H2O2 content in roots during exposure to hypoxia. However, calcium deficiency in the nutrient solution decreased plant weight, photosynthesis parameters, NO3− reduction, amino acids (e.g., alanine, aspartic acid, glutamic acid, γ-aminobutyric acid), protein, all polyamines except for free putrescine, and the activities of glutamate synthase and glutamine synthetase. Additionally, there was an increase in the NO3− uptake rate, polyamine oxidase activity and H2O2 contents under hypoxia–Ca. Simultaneously, exogenous calcium had little effect on nitrate absorption and transformation, photosynthetic parameters, and plant growth under normoxic conditions. These results suggest that calcium confers short-term hypoxia tolerance in muskmelon, most likely by promoting nitrate uptake and accelerating its transformation into amino acids, heat-stable proteins or polyamines, as well as by decreasing polyamine degradation in muskmelon seedlings.

Previously, we found that the amelioration of photosynthetic capacity by bottle gourd (Lagenaria siceraria Standl.) rootstock in watermelon seedlings (Citrullus lanatus [Thunb.] Mansf.) with salt treatment might be closely related to the enzymes in Calvin cycle such as ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) (Yang et al., 2012). We confirmed this and showed more details in this study that improved photosynthesis of watermelon plants by bottle gourd rootstock was associated with the decreased stomata resistance and the increased photochemical activity and photosynthetic metabolism with or without 100 mM NaCl stress for 3 days. The analysis of gas exchange parameters showed that self-grafted plants suffered serious non-stomatal limitation to photosynthesis under salt stress while rootstock-grafted plants were mainly affected by stomata limitation in stress conditions. Further, results showed that NaCl stress markedly reduced the chlorophyll content, damaged the structure of photosynthetic apparatus, and inhibited photochemical activity and CO2 assimilation in self-grafted plants. In contrast, rootstock-grafting increased the chlorophyll content, especially chlorophyll b, and minimized the harmful effects on photosystem II (PSII) reaction center and the thylakoids structure induced by NaCl stress. Furthermore, rootstock-grafting enhanced the content and activity of Rubisco and thus elevated carbon fixation in the leaves of watermelon scions under salt stress. The gene expressions of enzymes related to ribulose-1,5-bisphosphate (RuBP) regeneration were also up-regulated by rootstock and this probably guaranteed the sufficient supply of RuBP for the operation of Calvin cycle in watermelon scions under salt stress. Thus, bottle gourd rootstock promoted photosynthesis by the activation of stomatal and non-stomatal abilities, especially the regulation of a variety of photosynthetic enzymes, including Rubisco in grafted watermelon plants under NaCl stress.

2,4-Epibrassinolide (EBL) is a plant hormone that plays a pivotal role in regulation of plants growth and development processes under abiotic stress. The investigation was carried out to study the effect of EBL on mineral nutrients uptake and distribution with ion element analysis and X-ray microanalysis in cucumber seedlings (Cucumis sativus L. cv. Jinyou No.4) under 80 mM Ca(NO3)2 stress. Our study found EBL significantly alleviated the inhibitory of P, K, Na, Mg, Fe, Mn, or Cl uptake in shoot or root by Ca(NO3)2 stress. Under Ca(NO3)2 stress, X-ray microanalysis showed that high levels of Ca by EBL treatment accumulated in the epidermal cells of root and gradually decreased from epidermal cells to stellar parenchyma. K+ levels were restored in different cross section of roots and high K+ level mostly accumulated in stellar parenchyma. The results of Ca2+ ultra-structural localization showed Ca2+ particles of antimonite precipitate by EBL were partly decreased in mesophyll and root cells, and Ca2+ precipitate distributed in intercellular spaces again. Increased Ca2+-ATPase activity and ATP content by EBL were also contributed to extrude excess Ca2+ from the cytoplasm. These results suggested that EBL could alleviate the ion damage from excess Ca2+ through regulating mineral nutrients uptake and distribution.

The plant growth, nitrogen absorption, and assimilation in watermelon (Citrullus lanatus [Thunb.] Mansf.) were investigated in self-grafted and grafted seedlings using the salt-tolerant bottle gourd rootstock Chaofeng Kangshengwang (Lagenaria siceraria Standl.) exposed to 100 mM NaCl for 3 d. The biomass and NO3− uptake rate were significantly increased by rootstock while these values were remarkably decreased by salt stress. However, compared with self-grafted plants, rootstock-grafted plants showed higher salt tolerance with higher biomass and NO3− uptake rate under salt stress. Salinity induced strong accumulation of nitrate, ammonium and protein contents and a significant decrease of nitrogen content and the activities of nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), and glutamate synthase (GOGAT) in leaves of self-grafted seedlings. In contrast, salt stress caused a remarkable decrease in nitrate content and the activities of GS and GOGAT, and a significant increase of ammonium, protein, and nitrogen contents and NR activity, in leaves of rootstock-grafted seedlings. Compared with that of self-grafted seedlings, the ammonium content in leaves of rootstock-grafted seedlings was much lower under salt stress. Glutamate dehydrogenase (GDH) activity was notably enhanced in leaves of rootstock-grafted seedlings, whereas it was significantly inhibited in leaves of self-grafted seedlings, under salinity stress. Three GDH isozymes were isolated by native gel electrophoresis and their expressions were greatly enhanced in leaves of rootstock-grafted seedlings than those of self-grafted seedlings under both normal and salt-stress conditions. These results indicated that the salt tolerance of rootstock-grafted seedlings might (be enhanced) owing to the higher nitrogen absorption and the higher activities of enzymes for nitrogen assimilation induced by the rootstock. Furthermore, the detoxification of ammonium by GDH when the GS/GOGAT pathway was inhibited under salt stress might play an important role in the release of salt stress in rootstock-grafted seedlings.