o Jian Xia

Brassinosteroids promote metabolism of pesticides in cucumber.

Brassinosteroids (BRs) are known to protect crops from the toxicity of herbicides, fungicides and insecticides. It is shown here that application of 24-epibrassinolide (EBR) accelerated metabolism of various pesticides and consequently reduced their residual levels in cucumber ( Cucumis sativus L). Chlorpyrifos, a widely used insecticide, caused significant reductions of net photosynthetic rate (Pn) and quantum yield of PSII (Phi(PSII)) in cucumber leaves. EBR pretreatment alleviated the declines of Pn and Phi(PSII) caused by chlorpyrifos application, and this effect of EBR was associated with reductions of chlorpyrifos residues. To understand how EBR promotes chlorpyrifos metabolism, the effects of EBR on activity and expression of enzymes involved in pesticide metabolism were analyzed. EBR had a positive effect on the activation of glutathione S-transferase (GST), peroxidase (POD), and glutathione reductase (GR) after treatment with chlorpyrifos, although the effect on GR was attenuated at later time points when plants were treated with 1 mM chlorpyrifos. In addition, EBR enhanced the expression of P450 and MRP, which encode P450 monooxygenase and ABC-type transporter, respectively. However, the expression of GST was consistently lower than that of plants treated with only chlorpyrifos. Importantly, the stimulatory effect of EBR on pesticide metabolism was also observed for cypermethrin, chlorothalonil, and carbendazim, which was attributed to the enhanced activity and genes involved in pesticide metabolism. The results suggest that BRs may be promising, environmentally friendly, natural substances suitable for wide application to reduce the risks of human and environment exposure to pesticides.

Hydrogen peroxide is involved in the cold acclimation-induced chilling tolerance of tomato plants

Cold acclimation increases plant tolerance to a more-severe chilling and in this process an accumulation of H(2)O(2) in plants is often observed. To examine the role of H(2)O(2) in cold acclimation in plants, the accumulation of H(2)O(2), antioxidant metabolism, the glutathione redox state, gas exchange and chlorophyll fluorescence were analyzed after cold acclimation at 12/10xa0°C and during the subsequent chilling at 7/4xa0°C in tomato (Solanum lycopersicum) plants. Cold acclimation modestly elevated the levels of H(2)O(2), the gene expression of respiratory burst oxidase homolog 1 (Rboh1) and NADPH oxidase activity, leading to the up-regulation of the expression and activity of antioxidant enzymes. In non-acclimated plants chilling caused a continuous rise in the H(2)O(2) content, an increase in the malondialdehyde (MDA) content and in the oxidized redox state of glutathione, followed by reductions in the CO(2) assimilation rate and the maximum quantum yield of photosystem II (F(v)/F(m)). However, in cold-acclimated plants chilling-induced photoinhibition, membrane peroxidation and reductions in the CO(2) assimilation rate were significantly alleviated. Furthermore, a treatment with an NADPH oxidase inhibitor or H(2)O(2) scavenger before the plants subjected to the cold acclimation abolished the cold acclimation-induced beneficial effects on photosynthesis and antioxidant metabolism, leading to a loss of the cold acclimation-induced tolerance against chilling. These results strongly suggest that the H(2)O(2) generated by NADPH oxidase in the apoplast of plant cells plays a crucial role in cold acclimation-induced chilling tolerance.

Light quality affects incidence of powdery mildew, expression of defence-related genes and associated metabolism in cucumber plants

To determine whether light quality affects the incidence of disease, we exposed cucumber (Cucumis sativus L. cv. Jinyan No. 4) plants at the 4-leaf stage to white and other monochromatic lights and tested the effects on plant response to Sphaerotheca fuliginea, defence-related gene expression and metabolic changes. Exposure to red light resulted in higher levels of H2O2 and salicylic acid (SA), and stronger expression of defence genes such as PR-1 than exposure to white or other monochromatic lights. In comparison, plants grown under purple and blue light had higher activities of phenylalanine ammonia-lyase (PAL) and polyphenoloxidase (PPO) and higher level of flavonoids than plants grown under other lights. Furthermore, plants grown under red light were more resistant whilst plants grown under other monochromatic lights were less resistant to Sphaerotheca fuliginea than plants grown under white light. These results suggest a role of red light in light-enhanced resistance, which correlates with enhanced SA-dependent signaling pathway.

Changes in electron transport, superoxide dismutase and ascorbate peroxidase isoenzymes in chloroplasts and mitochondria of cucumber leaves as influenced by chilling

In order to clarify the relationship between chill-induced disturbance in photosynthetic, respiratory electron transport and the metabolism of reactive oxygen species (ROS), leaf gas exchange, chlorophyll fluorescence quenching, respiration, and activities of superoxide dismutase (SOD) and ascorbate peroxidase (APX) were investigated in chloroplasts and mitochondria of cucumber (Cucumis sativus) leaves subjected to a chill (8 °C) for 4 d. Chilling decreased net photosynthetic rate (PN) and quantum efficiency of photosystem 2 (ΦPS2), but increased the ratio of ΦPS2 to the quantum efficiency of CO2 fixation (ΦCO2) and non-photochemical quenching (NPQ) in cucumber leaves. While chilling inhibited the activity of cytochrome respiration pathway, it induced an increase of alternative respiration pathway activity and the reduction level of Q-pool. Chilling also significantly increased O2• production rate, H2O2 content, and SOD and APX activities in chloroplasts and mitochondria. There was a more significant increase in SOD and APX activities in chloroplasts than in mitochondria with the increase of membrane-bound Fe-SOD and tAPX in chloroplasts being more significant than other isoenzymes. Taken together, chilling inhibited PN and cytochrome respiratory pathway but enhanced the photosynthetic electron flux to O2 and over-reduction of respiratory electron transport chain, resulting in ROS accumulation in cucumber leaves. Meanwhile, chilling resulted in an enhancement of the protective mechanisms such as thermal dissipation, alternative respiratory pathway, and ROS-scavenging mechanisms (SODs and APXs) in chloroplasts and mitochondria.

Melatonin enhances thermotolerance by promoting cellular protein protection in tomato plants

Melatonin is a pleiotropic signaling molecule that provides physiological protection against diverse environmental stresses in plants. Nonetheless, the mechanisms for melatonin‐mediated thermotolerance remain largely unknown. Here, we report that endogenous melatonin levels increased with a rise in ambient temperature and that peaked at 40°C. Foliar pretreatment with an optimal dose of melatonin (10 μmol/L) or the overexpression of N‐acetylserotonin methyltransferase (ASMT) gene effectively ameliorated heat‐induced photoinhibition and electrolyte leakage in tomato plants. Both exogenous melatonin treatment and endogenous melatonin manipulation by overexpression of ASMT decreased the levels of insoluble and ubiquitinated proteins, but enhanced the expression of heat‐shock proteins (HSPs) to refold denatured and unfolded proteins under heat stress. Meanwhile, melatonin also induced expression of several ATG genes and formation of autophagosomes to degrade aggregated proteins under the same stress. Proteomic profile analyses revealed that protein aggregates for a large number of biological processes accumulated in wild‐type plants. However, exogenous melatonin treatment or overexpression of ASMT reduced the accumulation of aggregated proteins. Aggregation responsive proteins such as HSP70 and Rubisco activase were preferentially accumulated and ubiquitinated in wild‐type plants under heat stress, while melatonin mitigated heat stress‐induced accumulation and ubiquitination of aggregated proteins. These results suggest that melatonin promotes cellular protein protection through induction of HSPs and autophagy to refold or degrade denatured proteins under heat stress in tomato plants.

HsfA1a upregulates melatonin biosynthesis to confer cadmium tolerance in tomato plants.

Melatonin regulates broad aspects of plant responses to various biotic and abiotic stresses, but the upstream regulation of melatonin biosynthesis by these stresses remains largely unknown. Herein, we demonstrate that transcription factor heat‐shock factor A1a (HsfA1a) conferred cadmium (Cd) tolerance to tomato plants, in part through its positive role in inducing melatonin biosynthesis under Cd stress. Analysis of leaf phenotype, chlorophyll content, and photosynthetic efficiency revealed that silencing of the HsfA1a gene decreased Cd tolerance, whereas its overexpression enhanced plant tolerance to Cd. HsfA1a‐silenced plants exhibited reduced melatonin levels, and HsfA1a overexpression stimulated melatonin accumulation and the expression of the melatonin biosynthetic gene caffeic acid O‐methyltransferase 1 (COMT1) under Cd stress. Both an in vitro electrophoretic mobility shift assay and in vivo chromatin immunoprecipitation coupled with qPCR analysis revealed that HsfA1a binds to the COMT1 gene promoter. Meanwhile, Cd stress induced the expression of heat‐shock proteins (HSPs), which was compromised in HsfA1a‐silenced plants and more robustly induced in HsfA1a‐overexpressing plants under Cd stress. COMT1 silencing reduced HsfA1a‐induced Cd tolerance and melatonin accumulation in HsfA1a‐overexpressing plants. Additionally, the HsfA1a‐induced expression of HSPs was partially compromised in COMT1‐silenced wild‐type or HsfA1a‐overexpressing plants under Cd stress. These results demonstrate that HsfA1a confers Cd tolerance by activating transcription of the COMT1 gene and inducing accumulation of melatonin that partially upregulates expression of HSPs.

Brassinosteroid regulates secondary metabolism in tomato towards enhanced tolerance to phenanthrene

We investigated the role of 24-epibrassinolide (EBR) in the amelioration of phenanthrene (PHE) stress in tomato (Solanum lycopersicum L.). Exposure to PHE (300 μM) significantly decreased shoot and root length (19 and 16 %, respectively), fresh mass (35 and 43 %, respectively), contents of chlorophyll a (26 %), chlorophyll b (27 %) and carotenoids (18 %) in tomato plants. In addition, PHE increased the malondialdehyde (MDA) content (57 %) and activity of secondary metabolism related enzymes glutathione-S-transferase (GST), glucose-6-phosphate dehydrogenase (G6PDH), shikimate dehydrogenase (SKDH), phenylalanine ammonia-lyase (PAL) and cinnamyl alcohol dehydrogenase (CAD). The expression levels of GST1, PPO, SKDH, PAL and CAD genes were also induced by PHE. Importantly, EBR (0.1 μM) alone and in combination with PHE increased the growth, biomass and activity of those enzymes significantly over control and PHE alone, respectively. Consistent with enzymes activities transcript levels of GST1, PPO, SKDH, PAL and CAD were further increased in PHE+EBR over PHE alone. However, MDA content was remarkably decreased in PHE+EBR than PHE alone. Meanwhile, content of phenols, flavonoids and antioxidant activity were increased by PHE and PHE+EBR further increased all those parameters. These observations suggest that EBR regulates secondary metabolism in tomato which might enhance tolerance to PHE.

Brassinosteroid-induced CO2 assimilation is associated with increased stability of redox-sensitive photosynthetic enzymes in the chloroplasts in cucumber plants

Brassinosteroids (BRs) play important roles in plant growth, development, photosynthesis and stress tolerance; however, the mechanism underlying BR-enhanced photosynthesis is currently unclear. Here, we provide evidence that an increase in the BR level increased the quantum yield of PSII, activities of Rubisco activase (RCA) and fructose-1,6-bisphosphatase (FBPase), and CO(2) assimilation. BRs upregulated the transcript levels of genes and activity of enzymes involved in the ascorbate-glutathione cycle in the chloroplasts, leading to an increased ratio of reduced (GSH) to oxidized (GSSG) glutathione in the chloroplasts. An increased GSH/GSSG ratio protected RCA from proteolytic digestion and increased the stability of redox-sensitive enzymes in the chloroplasts. These results strongly suggest that BRs are capable of regulating the glutathione redox state in the chloroplasts through the activation of the ascorbate-glutathione cycle. The resulting increase in the chloroplast thiol reduction state promotes CO(2) assimilation, at least in part, by enhancing the stability and activity of redox-sensitive photosynthetic enzymes through post-translational modifications.

Protective Role of Putrescine Against Salt Stress is Partially Related to the Improvement of Water Relation and Nutritional Imbalance in Cucumber

ABSTRACT Polyamines play a variety of physiological roles in plant growth and development. To investigate whether exogenous putrescine (Put) has roles in protecting plants against salt stress, Put (100 μ M) was added to nutrient solution three days before cucumber (Cucumis sativusL. cv. “Jinyan No.4) seedlings were exposed to 100 mM sodium chloride (NaCl) treatment. Putrescine treatment significantly ameliorated the detrimental effects of NaCl on root growth and this was associated with a decrease of Na uptake and an increase in potassium accumulation in roots. Manganese (Mn) content in roots was decreased by salinity stress but increased by Put pretreatment. Furthermore, osmotic stress associated with NaCl treatment decreased leaf water potential and water content, while these effects were alleviated by Put pretreatment. The decreases in net photosynthetic rate (Pn) and stomatal conductance (Gs) by NaCl were also diminished by Put treatment. The results indicate that Put may play an important role in protecting cucumber plants against salt stress.

Compensatory acclimated mechanisms of photoprotection in a Xa mutant of Lycopersicon esculentum Mill

To probe the role of xanthophylls in non-photochemical quenching (NPQ) and the compensatory acclimated photoprotection mechanisms, a tomato (Lycopersicon esculentum Mill. cv. Ailsa Craig) Xa mutant with deficit in lutein (L) and neoxanthin (N) contents was used. The Xa mutant showed lowered NPQ, an increased degree of de-epoxidation state [(A+Z)/(V+A+Z)], and decreases of photosystem 2 (PS2) antenna size. Although the Xa mutant had a CO2 assimilation rate similar to that of Ailsa Craig, it exhibited a much larger stomatal conductance (gs) than Ailsa Craig. Decreased electron flux in PS2 (JPS2) for the Xa mutant was associated with electron flux for photorespiratory carbon oxidation (Jo) and alternative electron flux in PS2 (Ja) while electron flux for photosynthetic carbon reduction (Jc) was not different from Ailsa Craig. Moreover, the Xa mutant also exhibited higher activities of antioxidant enzymes, higher contents of ascorbate and glutathione, and lower contents of reactive oxygen species. Hence some compensatory acclimated mechanisms of photoprotection operated properly in the lack of NPQ and xanthophylls.