Silicon confers cucumber resistance to salinity stress through regulation of proline and cytokinins.

Salt stress is a continuous threat to global crop production. Here, we studied the alleviation role of exogenous silicon (Si) in NaCl-stressed cucumber, with special emphasis on plant growth, proline (Pro) and hormone metabolisms. The results showed that Si supplementation ameliorated the adverse effects of NaCl on plants growth, biomass, and oxidative stress. Salt stress greatly increased the content of Pro throughout the experiment, while Si regulated Pro content in two distinct ways. Si promoted the salt-induced Pro levels after 3 and 6 days of treatment, but decreased it after 9 and 12 days of treatment. Moreover, P5CS and ProDH activities and P5CS gene play important roles in Si and salt-regulated Pro levels in different stress phase. Under stress condition, Si addition tend to revert the content of ABA, IAA, cytokinin and SA to the control levels in most cases. Further correlation analysis revealed a negative correlation between the root cytokinin and Pro content after 3 days of treatment, suggesting the interaction between cytokinin and Pro metabolism. Exogenous application of Pro and ProDH competitive inhibitor D-Lactate confirmed the possible interplay between Pro and cytokinin metabolism. Further study identified several CKX (Csa4G647490 and Csa1G589070) and IPT (Csa7G392940 and Csa3G150100) genes that may be responsible for the regulation of cytokinin accumulation by Si and/or Pro after short-term of treatment. The results suggested that Pro is a key factor in Si-induced salt tolerance, and Si-increased Pro content may participate in the regulation of cytokinin metabolism under short-term of salt stress.

Response of a Predatory ant to Volatiles Emitted by Aphid- and Caterpillar-Infested Cucumber and Potato Plants.

In response to herbivory by insects, various plants produce volatiles that attract enemies of the herbivores. Although ants are important components of natural and agro-ecosystems, the importance of herbivore-induced plant volatiles (HIPVs) as cues for ants for finding food sources have received little attention. We investigated responses of the ant Formica pratensis to volatiles emitted by uninfested and insect-infested cucumber (Cucumis sativus) and potato (Solanum tuberosum) plants. Cucumber plants were infested by the phloem-feeding aphid Aphis gossypii, the leaf chewer Mamestra brassicae or simultaneously by both insects. Potato plants were infested by either Aphis gossypii, by the leaf chewer Chrysodeixis chalcites or both. In olfactometer experiments, ants preferred volatile blends emitted by cucumber plants infested with M. brassicae caterpillars alone or combined with A. gossypii to volatiles of undamaged plants or plants damaged by A. gossypii only. No preference was recorded in choice tests between volatiles released by aphid-infested plants over undamaged plants. Volatiles emitted by potato plants infested by either C. chalcites or A. gossypii were preferred by ants over volatiles released by undamaged plants. Ants did not discriminate between potato plants infested with aphids and caterpillars over plants infested with aphids only. Plant headspace composition showed qualitative and/or quantitative differences between herbivore treatments. Multivariate analysis revealed clear separation between uninfested and infested plants and among herbivore treatments. The importance of HIPVs in indirect plant defence by ants is discussed in the context of the ecology of ant-plant interactions and possible roles of ants in pest management.

Integration of Hormonal and Nutritional Cues Orchestrates Progressive Corolla Opening.

Flower opening is essential for pollination and thus successful sexual reproduction; however, the underlying mechanisms of its timing control remain largely elusive. We identify a unique cucumber (Cucumis sativus) line '6457' that produces normal ovaries when nutrients are under-supplied, and super ovaries (87%) with delayed corolla opening when nutrients are oversupplied. Corolla opening in both normal and super ovaries is divided into four distinct phases, namely the green bud, green-yellow bud, yellow bud, and flowering stages, along with progressive color transition, cytological tuning, and differential expression of 14,282 genes. In the super ovary, cell division and cell expansion persisted for a significantly longer period of time; the expressions of genes related to photosynthesis, protein degradation, and signaling kinases were dramatically up-regulated, whereas the activities of most transcription factors and stress-related genes were significantly down-regulated; concentrations of cytokinins (CKs) and gibberellins were higher in accordance with reduced cytokinin conjugation and degradation and increased expression of gibberellin biosynthesis genes. Exogenous CK application was sufficient for the genesis of super ovaries, suggesting a decisive role of CKs in controlling the timing of corolla opening. Furthermore, 194 out of 11,127 differentially expressed genes identified in pairwise comparisons, including critical developmental, signaling, and cytological regulators, contained all three types of cis-elements for CK, nitrate, and phosphorus responses in their promoter regions, indicating that the integration of hormone modulation and nutritional regulation orchestrated the precise control of corolla opening in cucumber. Our findings provide a valuable framework for dissecting the regulatory pathways for flower opening in plants.

Staurosporine from the endophytic Streptomyces sp. strain CNS-42 acts as a potential biocontrol agent and growth elicitor in cucumber.

Chinese medicinal plants and their surrounding rhizospheric soil serve as promising sources of actinobacteria. A total of 180 actinobacteria strains were isolated from the rhizosphere soil, leaves, stems, and roots of nine selected plants and have been identified as potential biocontrol agents against Fusarium oxysporum f. sp. cucumerinum. An endophytic strain CNS-42 isolated from Alisma orientale showed the largest zone of inhibition demonstrating a potent effect against F. oxysporum f. sp. cucumerinum and a broad antimicrobial activity against bacteria, yeasts, and other pathogenic fungi. The in vivo biocontrol assays showed that the disease severity index was significantly reduced (P < 0.05), and plant shoot fresh weight and height increased greatly (P < 0.05) in plantlets treated with strain CNS-42 compared to the negative control. This isolate was identified as Streptomyces sp. based on cultural, physiological, morphological characteristics, and 16S rRNA gene analysis. Further bioassay-guided isolation and purification revealed that staurosporine was responsible for its antifungal and plant growth promoting activities and the latter property of staurosporine is reported for the first time. The in vivo assay was further performed and indicated that staurosporine showed good growth promoting effect on the plant shoot biomass of cucumber. This is the first critical evidence identifying CNS-42 as a biocontrol agent for the soil borne pathogen, F. oxysporum f. sp. cucumerinum.

Common arbuscular mycorrhizal networks amplify competition for phosphorus between seedlings and established plants.

Common mycorrhizal networks (CMNs) influence competition between plants, but reports regarding their precise effect are conflicting. We studied CMN effects on phosphorus (P) uptake and growth of seedlings as influenced by various disruptions of network components. Tomato (Solanum lycopersicon) seedlings grew into established networks of Rhizophagus irregularis and cucumber (Cucumis sativus) in two experiments. One experiment studied seedling uptake of (32)P in the network in response to cutting of cucumber shoots; the other analysed seedling uptake of P and nitrogen (N) in the presence of intact or severed arbuscular mycorrhizal fungus networks and at two soil P concentrations. Pre-established and intact networks suppressed growth of tomato seedlings. Cutting of cucumber shoots mitigated P deficiency symptoms of seedlings, which obtained access to P in the extraradical mycelium and thereby showed improved growth. Solitary seedlings growing in a network patch that had been severed from the CMN also grew much better than seedlings of the corresponding CMN. Interspecific and size-asymmetric competition between plants may be amplified rather than relaxed by CMNs that transfer P to large plants providing most carbon and render small plants P deficient. It is likely that grazing or senescence of the large plants will alleviate the network-induced suppression of seedling growth.

Effects of above- and belowground herbivory on growth, pollination, and reproduction in cucumber.

Plants experience unique challenges due to simultaneous life in two spheres, above- and belowground. Interactions with other organisms on one side of the soil surface may have impacts that extend across this boundary. Although our understanding of plant-herbivore interactions is derived largely from studies of leaf herbivory, belowground root herbivores may affect plant fitness directly or by altering interactions with other organisms, such as pollinators. In this study, we investigated the effects of leaf herbivory, root herbivory, and pollination on plant growth, subsequent leaf herbivory, flower production, pollinator attraction, and reproduction in cucumber (Cucumis sativus). We manipulated leaf and root herbivory with striped cucumber beetle (Acalymma vittatum) adults and larvae, respectively, and manipulated pollination with supplemental pollen. Both enhanced leaf and root herbivory reduced plant growth, and leaf herbivory reduced subsequent leaf damage. Plants with enhanced root herbivory produced 35% fewer female flowers, while leaf herbivory had no effect on flower production. While leaf herbivory reduced the time that honey bees spent probing flowers by 29%, probing times on root-damaged plants were over twice as long as those on control plants. Root herbivory increased pollen limitation for seed production in spite of increased honey bee preference for plants with root damage. Leaf damage and hand-pollination treatments had no effect on fruit production, but plants with enhanced root damage produced 38% fewer fruits that were 25% lighter than those on control plants. Despite the positive effect of belowground damage on honey bee visitation, root herbivory had a stronger negative effect on plant reproduction than leaf herbivory. These results demonstrate that the often-overlooked effects of belowground herbivores may have profound effects on plant performance.

[Alleviation effects of LaCl3 on photosynthetic characteristics of cucumber seedlings under nitrate stress].

By the method of hydroponic culture, this paper studied the alleviation effects of LaCl3 on the photosynthetic characteristics of cucumber seedlings under nitrate (140 mmol NO3(-) x L(-1)) stress, with the related mechanisms discussed. Under nitrate stress, the seedlings leaf chlorophyll and carotenoids contents decreased significantly, and the leaf Mg2+-ATPase, Ca2+-ATPase activities also decreased. On the 7th day of nitrate stress, the decrease of seedlings photosynthetic rate was mainly due to stomatal limitation; but on the 12th day of nitrate stress, the decrease was mainly due to no-stomatal limitation. Supplement with LaCl3 could make the cucumber seedlings keep relatively higher leaf Mg2+-ATPase and Ca2+-ATPase activities and chlorophyll and carotenoids contents, and applying 20 micromol x L(-1) of LaCl3 could increase the carotenoids content significantly. LaCl3 could also improve the leaf gas exchange, and alleviate the decrease of leaf Fv/Fm, PhiPSII, AQY, CE, and qp under nitrate stress, which helped the leaves making good use of light energy and maintaining higher CO2 assimilation capacity. An additional 20 micromol x L(-1) of LaCl3 could alleviate the nitrate stress on the photosynthesis of cucumber seedlings efficiently, but an additional 200 micromol x L(-1) of LaCl3 only had the alleviation effect at the initial period of nitrate stress. Our results could benefit to the improvement of greenhouse soil.

The DefH9-iaaM-containing construct efficiently induces parthenocarpy in cucumber.

Parthenocarpy (seedless fruits) is a desirable trait that has been achieved in many plant cultivars. We generated parthenocarpic cucumber fruits by introducing the chimeric DefH9-iaaM construct into the cucumber genome using an Agrobacterium tumefaciens-mediated protocol. The construct consists of the DefH9 promoter from Antirrhinum majus and the iaaM coding sequence from Pseudomonas syringae. Transgenic plants were obtained from nine independent transformation events: half of these were tetraploid and did not produce seeds following self-pollination, while the remaining half were capable of displaying parthenocarpy in the subsequent reproductive generation. Of the fruits produced by the transgenic lines, 70-90% were parthenocarpic. The segregation of the marker gene in the transgenic T(1) progeny indicated single gene inheritance. The seed set in the transgenic lines and their F(1) hybrids was lower than in the non-transgenic control plants. Some of the methodological details and the practical significance of the results are discussed.

Simultaneous quantitative LC-ESI-MS/MS analyses of salicylic acid and jasmonic acid in crude extracts of Cucumis sativus under biotic stress.

Salicylic acid (SA) and jasmonic acid (JA) are plant hormones involved in basal resistance against plant pathogens and also in induced resistance. The aim of this study is to develop a fast and sensitive method to determine simultaneously the levels of both these hormones. The present paper proposes a method that includes hormone extraction with MeOH-H(2)O-HOAc (90:9:1, v/v), evaporation of the extracts, and injection into the liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) system in multiple reaction monitoring (MRM). Endogenous SA and JA levels in noninfested control cucumber cotyledons were 30.96 and 0.73ngg(-1) fresh weight, respectively. In roots, the levels were 8.31 and 15.82ngg(-1) FW, respectively. In plants treated with the biological control agent Trichoderma asperellum strain T-34, the levels of SA and JA did not differ from control plants. Rhizoctonia solani-diseased cucumber plants showed higher levels of SA and JA compared to noninfested controls (up to 2 and 13-fold higher, respectively). Detection limits for SA and JA were 0.45 and 0.47ngg(-1) fresh weight, respectively. The results of our research include the development of a method that is both fast and highly sensitive in the simultaneous quantitation of SA and JA from crude cucumber plant extracts, avoiding any purification and derivatization steps.

The accumulation of phytoalexin in cucumber plant after stress.

During the course of pathogens penetrating the plant cell, besides of chemical secretion, the pathogens may cause mechanical signal by the physical pressure on the plant cell. In the current study, we use the pressure as the stress signal to study the induction in plant resistance and the effect of accumulation of phytoalexin. We found that stress can induce the resistance in cucumber seeding significantly. Peptides contained RGD motif can specific block the adhesion between plant cell wall and plasma membrane. When breaking the plant cell wall and plasma membrane by using RGD peptides, the stress induction effect is almost absolutely eliminated. The results of assay with TLC and HPLC showed that stress stimulation could increase the accumulation of cucumber seeding phytoalexin. So, we can conclude that the accumulation of phytoalexin is one possible reason of improve the stress induced resistance. When block the adhesion between plant cell wall and plasma membrane by RGD, there are only part of accumulation of phytoalexin. The results suggest that stress induced resistance and accumulation of phytoalexin of plant is required for the adhesion of plant cell wall-plasma membrane.

Herbivory differentially affects male and female reproductive traits of Cucumis sativus.

Herbivory is an important selection pressure in the life history of plants. Most studies use seed or fruit production as an indication of plant fitness, but the impact of herbivory on male reproductive success is usually ignored. It is possible that plants compensate for resources lost to herbivory by shifting the allocation from seed production to pollen production and export, or vice versa. This study examined the impact of herbivory by Helix aspersa on both male and female reproductive traits of a monoecious plant, Cucumis sativus. The effects of herbivory on the relative allocation to male and female flowers were assessed through measurements of the number and size of flowers of both sexes, and the amount of pollinator visitation. We performed two glasshouse experiments; the first looked at the impact of three levels of pre-flowering herbivory, and the second looked at four levels of herbivory after the plants had started to flower. We found that herbivory during the flowering phase led to a significant increase in the number of plants without male flowers. As a consequence there was significantly less pollen export from this population, as estimated by movement of a pollen analog. The size of female flowers was reduced by severe herbivory, but there was no affect on pollen receipt by the female flowers of damaged plants. The decrease in allocation to male function after severe herbivory may be adaptive when male reproductive success is very unpredictable.

The use of an electric field in increasing the resistance of plants to the action of unfavorable space flight factors.

The key role in increasing the resistance of plants to unfavorable space flight factors is assigned to biomembranes of root cells. It is these biomembranes in which numerous biochemical and biophysical processes determining the adaptive capacity of plant organisms occur. In the initial period of exposure to unfavorable space flight factors the adaptation reactions of the plant organism undoubtedly increase its resistance. But the intensification of removal of H+ ions through the plasmalemma with an increase of the external influence sharply raises the quantity of cations leaving the cell, which leads to the accumulation of a considerable quantity of intracellular negative charges. These charges together with negative charges built in the membrane force protons to concentrate on the external surface of the membrane. Since protons have a very strong electric field, they form such a charge of which the electric field is about from several to hundreds of V/cm. The concentration of positive charges of protons entails the formation of a double electric field which extremely impedes the diffusion of other ions. Thus, a proton barrier is formed. Its length can be very considerable due to which the whole process of transmembrane energy and mass-transfer is disturbed. The proton barrier is easily destroyed by a weak electric field created in the root zone. In experiments on electrostimulation of different plants under space flight conditions at the orbital station MIR the absorption of nutrient elements by the root system increased to the optimal level, the ratio of physiologically active substances in the rhizosphere was normalized, the content of chlorophyll, carotin, and ascorbic acid in leaves corresponded to the ground-based control. Understanding of the mechanism of formation of a proton barrier on the plasmalemma of root cells as a result of the response of plants to the negative action of external factors (microgravity) is of great importance. It allows the possibility of life support of the vegetable kingdom in extreme conditions to be estimated in a new way.

Systemic inhibition of arbuscular mycorrhiza development by root exudates of cucumber plants colonized by Glomus mosseae.

The arbuscular mycorrhizal (AM) non-host plants mustard, sugar beet, lupin and the AM host plant cucumber were used as test plants. Cucumber plants were grown either in the absence of the AM fungus (AMF) Glomus mosseae or in a split-root system, with one side mycorrhizal and one side non-mycorrhizal. Root exudates of the AM non-host plants, the non-mycorrhizal cucumber plants and the mycorrhizal and the non-mycorrhizal side of the split-root system of mycorrhizal cucumber plants were collected and applied to cucumber plants inoculated with the AMF. Root exudates of non-mycorrhizal cucumber plants showed a significant stimulatory effect on root colonization, whereas root exudates from the mycorrhizal and the non-mycorrhizal sides of a split-root system of a mycorrhizal cucumber plant did not show this stimulatory effect and were even slightly inhibitory. Root exudates of the two AM non-host plants mustard and sugar beet significantly reduced root colonization in cucumber plants, whereas no such effect was observed when root exudates of the AM non-host plant lupin were applied.

Effect of Mentha x piperita essential oil and monoterpenes on cucumber root membrane potential.

Peppermint (Mentha piperita L.) essential oil and its main components were assessed for their ability to interfere with plant plasma membrane potentials. Tests were conducted on root segments isolated from etiolated seedlings of cucumber (Cucumis sativus L.). Increasing the concentration of peppermint essential oil from 5 to 50 ppm caused a decrease in membrane potential (Vm) hyperpolarization of 10-3 mV, whereas concentrations from 100 up to 900 ppm caused an increasing depolarization of Vm (from 5 to 110 mV). When tested at 300 ppm, (+)-menthyl acetate, (-)-limonene and 1,8-cineole did not exert any significant effect on V(m), whereas (+)-menthofuran (73 mV), (+)-pulegone (85 mV), (+)-neomenthol (96 mV), (-)-menthol (105 mV) and (-)-menthone (111 mV) showed increased ability to depolarize V(m). A plot of log of octanol-water partition coefficient (K(ow)) against their depolarizing effect showed a significant negative correlation, suggesting that among all monoterpenoids increased membrane depolarization depends on lower K(ow). However, among monoterpene ketones, alcohols and furans, increased membrane depolarization is associated with a decline in water solubility. The possible effect of monoterpenoids on membrane ion fluxes is also discussed, since changes in the bioelectric potential of cells imply changes in the flux of ions across the plasma membrane

Dehydration-induced redistribution of amphiphilic molecules between cytoplasm and lipids is associated with desiccation tolerance in seeds.

This study establishes a relationship between desiccation tolerance and the transfer of amphiphilic molecules from the cytoplasm into lipids during drying, using electron paramagnetic resonance spectroscopy of amphiphilic spin probes introduced into imbibed radicles of pea (Pisum sativum) and cucumber (Cucumis sativa) seeds. Survival following drying and a membrane integrity assay indicated that desiccation tolerance was present during early imbibition and lost in germinated radicles. In germinated cucumber radicles, desiccation tolerance could be re-induced by an incubation in polyethylene glycol (PEG) before drying. In desiccation-intolerant radicles, partitioning of spin probes into lipids during dehydration occurred at higher water contents compared with tolerant and PEG-induced tolerant radicles. The difference in partitioning behavior between desiccation-tolerant and -intolerant tissues could not be explained by the loss of water. Consequently, using a two-phase model system composed of sunflower or cucumber oil and water, physical properties of the aqueous solvent that may affect the partitioning of amphiphilic spin probes were investigated. A significant relationship was found between the partitioning of spin probes and the viscosity of the aqueous solvent. Moreover, in desiccation-sensitive radicles, the rise in cellular microviscosity during drying commenced at higher water contents compared with tolerant or PEG-induced tolerant radicles, suggesting that the microviscosity of the cytoplasm may control the partitioning behavior in dehydrating seeds.

Behavior of methamidophos residues in peppers, cucumbers, and cherry tomatoes grown in a greenhouse: evaluation by decline curves.

Residue levels of methamidophos were determined in peppers, cucumbers, and cherry tomatoes grown in commercial greenhouses, up to 6 weeks after being sprayed with Monitor (methamidophos 60%). Mathematically defined decline curves were established by determining optimal relationships between methamidophos residues and time, using different models. Model functions that best fit experimental data were 1st-order function for cucumber, 1.5th-order function for pepper, and 1st-order root function for tomato. However, in all cases, the 1st-order function was legitimized statistically. Half-life times determined from the optimal functions were 8.68 days (cucumber), 13.28 days (pepper), and 2.77 days (tomato), whereas half-life times determined from the 1st-order reaction function were 8.68 days (cucumber), 17.04 days (pepper), and 7.47 days (tomato). In this work, some experiments to determine residue levels of methamidophos in these vegetables after multiple applications were also carried out. The unexpected high residue levels found in all cases after five successive applications seem to indicate that methamidophos presents certain long-term accumulative effects in the three studied vegetables.

Phytochrome A enhances the promotion of hypocotyl growth caused by reductions in levels of phytochrome B in its far-red-light-absorbing form in light-grown Arabidopsis thaliana.

We sought to determine if phytochrome B (phyB)-mediated responses to the red light (R)/far-red light (FR) ratio are affected by phytochrome A (phyA) activity in light-grown seedlings of Arabidopsis thaliana. Pulses of FR delayed into the dark period were less effective than end-of-day (EOD) FR in promoting hypocotyl growth over a given period in darkness. White light minus blue light interposed instead of darkness between the end of the white-light photoperiod and the FR pulse was sufficient to maintain responsivity to the decrease in phyB in FR-light-absorbing form in wild-type (WT) seedlings, but not in the phyA mutant. Compared with EOD R, hourly R+FR pulses provided throughout the night caused a stronger promotion of stem growth than a single EOD R+FR pulse in WT Arabidopsis, cucumber, mustard, sunflower, tobacco, and tomato, but not in phyA Arabidopsis or in the aurea mutant of tomato. WT seedlings of Arabidopsis responded to a range of high EOD R/FR ratios, whereas the phyA mutant required stronger reductions in the EOD R/FR ratio. In sunlight, phyA seedlings of Arabidopsis showed no response to the "early warning" signals of neighboring vegetation, and hypocotyl-growth promotion occurred at higher plant densities than in the WT. Thus, under a series of light conditions, the sensitivity or responsivity to reductions in the R/FR ratio were larger in WT than in phyA seedlings. A product of phyA is therefore proposed to enhance the hypocotyl-growth response to decreases in phyB in FR-light-absorbing form in light grown seedlings.

Expansin mode of action on cell walls. Analysis of wall hydrolysis, stress relaxation, and binding.

The biochemical mechanisms underlying cell wall expansion in plants have long been a matter of conjecture. Previous work in our laboratory identified two proteins (named "expansins") that catalyze the acid-induced extension of isolated cucumber cell walls. Here we examine the mechanism of expansin action with three approaches. First, we report that expansins did not alter the molecular mass distribution or the viscosity of solutions of matrix polysaccharides. We conclude that expansins do not hydrolyze the major pectins or hemicelluloses of the cucumber wall. Second, we investigated the effects of expansins on stress relaxation of isolated walls. These studies show that expansins account for the pH-sensitive and heat-labile components of wall stress relaxation. In addition, these experiments show that expansins do not cause a progressive weakening of the walls, as might be expected from the action of a hydrolase. Third, we studied the binding of expansins to the cell wall and its components. The binding characteristics are consistent with this being the site of expansin action. We found that expansins bind weakly to crystalline cellulose but that this binding is greatly increased upon coating the cellulose with various hemicelluloses. Xyloglucan, either solubilized or as a coating on cellulose microfibrils, was not very effective as a binding substrate. Expansins were present in growing cell walls in low quantities (approximately 1 part in 5000 on a dry weight basis), suggesting that they function catalytically. We conclude that expansins bind at the interface between cellulose microfibrils and matrix polysaccharides in the wall and induce extension by reversibly disrupting noncovalent bonds within this polymeric network. Our results suggest that a minor structural component of the matrix, other than pectin and xyloglucan, plays an important role in expansin binding to the wall and, presumably, in expansin action.