CsSHMT3 gene enhances the growth and development in cucumber seedlings under salt stress.

Salt stress is one of the major factors limiting plant growth and productivity. Many studies have shown that serine hydroxymethyltransferase (SHMT) gene play an important role in growth, development and stress response in plants. However, to date, there have been few studies on whether SHMT3 can enhance salt tolerance in plants. Therefore, the effects of overexpression or silencing of CsSHMT3 gene on cucumber seedling growth under salt stress were investigated in this study. The results showed that overexpression of CsSHMT3 gene in cucumber seedlings resulted in a significant increase in chlorophyll content, photosynthetic rate and proline (Pro) content, and antioxidant enzyme activity under salt stress condition; whereas the content of malondialdehyde (MDA), superoxide anion (H2O2), hydrogen peroxide (O2·-) and relative conductivity were significantly decreased when CsSHMT3 gene was overexpressed. However, the content of chlorophyll and Pro, photosynthetic rate, and antioxidant enzyme activity of the silenced CsSHMT3 gene lines under salt stress were significantly reduced, while MDA, H2O2, O2·- content and relative conductivity showed higher level in the silenced CsSHMT3 gene lines. It was further found that the expression of stress-related genes SOD, CAT, SOS1, SOS2, NHX, and HKT was significantly up-regulated by overexpressing CsSHMT3 gene in cucumber seedlings; while stress-related gene expression showed significant decrease in silenced CsSHMT3 gene seedlings under salt stress. This suggests that overexpression of CsSHMT3 gene increased the salt tolerance of cucumber seedlings, while silencing of CsSHMT3 gene decreased the salt tolerance. In conclusion, CsSHMT3 gene might positively regulate salt stress tolerance in cucumber and be involved in regulating antioxidant activity, osmotic adjustment, and photosynthesis under salt stress. KEY MESSAGE: CsSHMT3 gene may positively regulate the expression of osmotic system, photosynthesis, antioxidant system and stress-related genes in cucumber.

Optimizing Cucumis sativus seedling vigor: the role of pistachio wood vinegar and date palm compost in nutrient mobilization.

BACKGROUND: The goal of this research is to enhance the quality of cucumber seedlings grown in greenhouses by experimenting with various soilless culture mediums (CMs) and the application of pistachio wood vinegar (WV). The experimental setup was designed as a factorial experiment within a randomized complete block design (RCBD), in greenhouse conditions featuring three replications to assess the effects of different culture media (CMs) and concentrations of pistachio wood vinegar (WV) on cucumber seedling growth. Cucumber seeds were planted in three CMs: coco peat-peat moss, coco peat-vermicompost, and date palm compost-vermicompost mixed in a 75:25 volume-to-volume ratio. These were then treated with pistachio WV at concentrations of 0, 0.5, and 1%, applied four times during irrigation following the emergence of the third leaf. RESULTS: The study revealed that treating seedlings with 0.5% WV in the date palm compost-vermicompost CM significantly enhanced various growth parameters. Specifically, it resulted in a 90% increase in shoot fresh mass, a 59% increase in shoot dry mass, an 11% increase in root fresh mass, a 36% increase in root dry mass, a 65% increase in shoot length, a 62% increase in leaf area, a 25% increase in stem diameter, a 41% increase in relative water content (RWC), and a 6% improvement in membrane stability index (MSI), all in comparison to untreated seedlings grown in coco peat-peat moss CM. Furthermore, chlorophyll a, b, total chlorophyll, and carotenoid levels were 2.3, 2.7, 2.6, and 2.7 times higher, respectively, in seedlings treated with 0.5% WV and grown in the date palm compost-vermicompost CM, compared to those treated with the same concentration of WV but grown in coco peat-peat moss CM. Additionally, the Fv/Fm ratio saw a 52% increase. When plant nutrition was enhanced with the date palm compost-vermicompost CM and 1% WV, auxin content rose by 130% compared to seedlings grown in coco peat-peat moss CM and treated with 0.5% WV. CONCLUSIONS: The study demonstrates that using 0.5% WV in conjunction with date palm compost-vermicompost CM significantly betters the quality of cucumber seedlings, outperforming other treatment combinations.

CsHSFA1d Promotes Drought Stress Tolerance by Increasing the Content of Raffinose Family Oligosaccharides and Scavenging Accumulated Reactive Oxygen Species in Cucumber.

Drought is the most severe form of stress experienced by plants worldwide. Cucumber is a vegetable crop that requires a large amount of water throughout the growth period. In our previous study, we identified that overexpression of CsHSFA1d could improve cold tolerance and the content of endogenous jasmonic acid in cucumber seedlings. To explore the functional diversities of CsHSFA1d, we treat the transgenic plants under drought conditions. In this study, we found that the heat shock transcription factor HSFA1d (CsHSFA1d) could improve drought stress tolerance in cucumber. CsHSFA1d overexpression increased the expression levels of galactinol synthase (CsGolS3) and raffinose synthase (CsRS) genes, encoding the key enzymes for raffinose family oligosaccharide (RFO) biosynthesis. Furthermore, the lines overexpressing CsHSFA1d showed higher enzymatic activity of GolS and raffinose synthase to increase the content of RFO. Moreover, the CsHSFA1d-overexpression lines showed lower reactive oxygen species (ROS) accumulation and higher ROS-scavenging enzyme activity after drought treatment. The expressions of antioxidant genes CsPOD2, CsAPX1 and CsSOD1 were also upregulated in CsHSFA1d-overexpression lines. The expression levels of stress-responsive genes such as CsRD29A, CsLEA3 and CsP5CS1 were increased in CsHSFA1d-overexpression lines after drought treatment. We conclude that CsHSFA1d directly targets and regulates the expression of CsGolS3 and CsRS to promote the enzymatic activity and accumulation of RFO to increase the tolerance to drought stress. CsHSFA1d also improves ROS-scavenging enzyme activity and gene expression indirectly to reduce drought-induced ROS overaccumulation. This study therefore offers a new gene target to improve drought stress tolerance in cucumber and revealed the underlying mechanism by which CsHSFA1d functions in the drought stress by increasing the content of RFOs and scavenging the excessive accumulation of ROS.

Cell wall invertase 3 plays critical roles in providing sugars during pollination and fertilization in cucumber.

In plants, pollen-pistil interactions during pollination and fertilization mediate pollen hydration and germination, pollen tube growth, and seed set and development. Cell wall invertases (CWINs) help provide the carbohydrates for pollen development; however, their roles in pollination and fertilization have not been well established. In cucumber (Cucumis sativus), CsCWIN3 showed the highest expression in flowers, and we further examined CsCWIN3 for functions during pollination to seed set. Both CsCWIN3 transcript and CsCWIN3 protein exhibited similar expression patterns in the sepals, petals, stamen filaments, anther tapetum, and pollen of male flowers, as well as in the stigma, style, transmitting tract, and ovule funiculus of female flowers. Notably, repression of CsCWIN3 in cucumber did not affect the formation of parthenocarpic fruit but resulted in an arrested growth of stigma integuments in female flowers and a partially delayed dehiscence of anthers with decreased pollen viability in male flowers. Consequently, the pollen tube grew poorly in the gynoecia after pollination. In addition, CsCWIN3-RNA interference plants also showed affected seed development. Considering that sugar transporters could function in cucumber fecundity, we highlight the role of CsCWIN3 and a potential close collaboration between CWIN and sugar transporters in these processes. Overall, we used molecular and physiological analyses to determine the CsCWIN3-mediated metabolism during pollen formation, pollen tube growth, and plant fecundity. CsCWIN3 has essential roles from pollination and fertilization to seed set but not parthenocarpic fruit development in cucumber.

Attenuation of Zucchini mosaic virus disease in cucumber plants by mycorrhizal symbiosis.

KEY MESSAGE: Arbuscular mycorrhizal fungi generated systemic acquired resistance in cucumber to Zucchini yellow mosaic virus, indicating their prospective application in the soil as a sustainable, environmentally friendly approach to inhibit the spread of pathogens. The wide spread of plant pathogens affects the whole world, causing several plant diseases and threatening national food security as it disrupts the quantity and quality of economically important crops. Recently, environmentally acceptable mitigating practices have been required for sustainable agriculture, restricting the use of chemical fertilizers in agricultural areas. Herein, the biological control of Zucchini yellow mosaic virus (ZYMV) in cucumber (Cucumis sativus L.) plants using arbuscular mycorrhizal (AM) fungi was investigated. Compared to control plants, ZYMV-infected plants displayed high disease incidence (DI) and severity (DS) with various symptoms, including severe yellow mosaic, mottling and green blisters of leaves. However, AM fungal inoculation exhibited 50% inhibition for these symptoms and limited DS to 26% as compared to non-colonized ones. The detection of ZYMV by the Enzyme-Linked Immunosorbent Assay technique exhibited a significant reduction in AM-inoculated plants (5.23-fold) compared with non-colonized ones. Besides, mycorrhizal root colonization (F%) was slightly reduced by ZYMV infection. ZYMV infection decreased all growth parameters and pigment fractions and increased the malondialdehyde (MDA) content, however, these parameters were significantly enhanced and the MDA content was decreased by AM fungal colonization. Also, the protein, proline and antioxidant enzymes (POX and CAT) were increased with ZYMV infection with more enhancements due to AM root colonization. Remarkably, defence pathogenesis-related (PR) genes such as PR-a, PR-b, and PR-10 were quickly expressed in response to AM treatment. Our findings demonstrated the beneficial function of AM fungi in triggering the plant defence against ZYMV as they caused systemic acquired resistance in cucumber plants and supported their potential use in the soil as an environment-friendly method of hindering the spread of pathogenic microorganisms sustainably.

A quick and effective method for thermostability differentiation in cucumber (Cucumis sativus L.).

High temperature affects the growth and production of cucumber. Selecting thermotolerant cucumber cultivars is conducive to coping with high temperatures and improving production. Thus, a quick and effective method for screening thermotolerant cucumber cultivars is needed. In this study, four cucumber cultivars were used to identify heat resistance indexes. The morphological, physiological and biochemical indexes were measured. When exposed to high temperatures, thermotolerant cucumber had a more stable photosystem, membrane, and oxidation-reduction systems. The impact of high temperatures on plants is multifaceted, and the accurate discrimination of heat resistance cannot be achieved solely based on a single or multiple indicators. Therefore, principal component analysis (PCA) was employed to comprehensively evaluate the heat resistance of cucumber plants. The results showed that the heat resistance obtained by PCA was significantly correlated with the heat injury index. In addition, the stepwise regression equation identified two heat-related indices, hydrogen peroxide content (H2 O2 ) and photosynthetic operating efficiency (Fq'/Fm'), and they can quickly distinguish the heat resistance of the other 8 cucumber cultivars. These results will help to accelerate the selection of thermotolerant resources and assist in cucumber breeding.

The differential expressions of aquaporins underline the diverse strategies of cucumber and tomato against salinity and zinc stress.

Salinity and excess zinc are two main problems that have limited agriculture in recent years. Aquaporins are crucial in regulating the passage of water and solutes through cells and may be essential for mitigating abiotic stresses. In the present study, the adaptive response to moderate salinity (60 mM NaCl) and excess Zn (1 mM ZnSO4 ) were compared alone and in combination in Cucumis sativus L. and Solanum lycopersicum L. Water relations, gas exchange and the differential expression of all aquaporins were analysed. The results showed that cucumber plants under salinity maintained the internal movement of water through osmotic adjustment and the overexpression of specific PIPs aquaporins, following a "conservation strategy". As tomato has a high tolerance to salinity, the physiological parameters and the expression of most aquaporins remained unchanged. ZnSO4 was shown to be stressful for both plant species. While cucumber upregulated 7 aquaporin isoforms, the expression of aquaporins increased in a generalized manner in tomato. Despite the differences, water relations and transpiration were adjusted in both plants, allowing the RWC in the shoot to be maintained. The aquaporin regulation in cucumber plants facing NaCl+ZnSO4 stress was similar in the two treatments containing NaCl, evidencing the predominance of salt in stress. However, in tomato, the induced expression of specific isoforms to deal with the combined stress differed from independent stresses. The results clarify the key role of aquaporin regulation in facing abiotic stresses and their possible use as markers of tolerance to salinity and heavy metals in plants.

Deep learning-based association analysis of root image data and cucumber yield.

The root system is important for the absorption of water and nutrients by plants. Cultivating and selecting a root system architecture (RSA) with good adaptability and ultrahigh productivity have become the primary goals of agricultural improvement. Exploring the correlation between the RSA and crop yield is important for cultivating crop varieties with high-stress resistance and productivity. In this study, 277 cucumber varieties were collected for root system image analysis and yield using germination plates and greenhouse cultivation. Deep learning tools were used to train ResNet50 and U-Net models for image classification and segmentation of seedlings and to perform quality inspection and productivity prediction of cucumber seedling root system images. The results showed that U-Net can automatically extract cucumber root systems with high quality (F1_score ≥ 0.95), and the trained ResNet50 can predict cucumber yield grade through seedling root system image, with the highest F1_score reaching 0.86 using 10-day-old seedlings. The root angle had the strongest correlation with yield, and the shallow- and steep-angle frequencies had significant positive and negative correlations with yield, respectively. RSA and nutrient absorption jointly affected the production capacity of cucumber plants. The germination plate planting method and automated root system segmentation model used in this study are convenient for high-throughput phenotypic (HTP) research on root systems. Moreover, using seedling root system images to predict yield grade provides a new method for rapidly breeding high-yield RSA in crops such as cucumbers.

Preliminary study on the diagnosis of NK stress based on the puncture mechanical characteristics of cucumber stem.

To investigate the relationship between stem puncture mechanical characteristics and NK stress diagnosis, the microstructure, surface morphology, cellulose and lignin content, puncture mechanical characteristics, and epidermal cell morphology of cucumber stems were measured herein. The results indicated that the middle stem, which had a diameter of approximately 7000 µm, was more suitable for puncturing due to its lower amount of epidermal hair, and its gradual regularity in shape. Further, the cucumber stems were protected from puncture damage due to their ability to rapidly heal within 25 h.. The epidermal penetration of the cucumber stems increased with the increase in cellulose and lignin, though cellulose played a more decisive role. The epidermal break distance increased with an increase in N application and decreased with an increase in K+ application, but the change in intercellular space caused by K+ supply was the most critical factor affecting the epidermal break distance. In addition, a decrease in K+ concentration led to a decrease in epidermal brittleness, whereas the factors affecting epidermal toughness were more complex. Finally, we found that although the detection of epidermal brittleness and toughness on nutrient stress was poor under certain treatment, the puncture mechanical characteristics of the stem still had a significant indicative effect on N application rate. Therefore, elucidating of the relationship between the puncture mechanical characteristics of the stems and crop nutritional stress is not only beneficial for promoting stem stress physiology research but also for designing on-site nutritional testing equipment in the future.

Combined toxicity influence of polypropylene microplastics and di-2-ethylhexyl phthalate on physiological-biochemical characteristics of cucumber (Cucumis sativus L.).

Microplastics and di-2-ethylhexyl phthalate (DEHP) are prevalent and emerging pollutants in agro-ecosystem, raising concerns due to their widespread co-presence. Nevertheless, their combined toxicity on terrestrial plants remains largely unexplored. This study investigated the impact of polypropylene microplastics (MPs), DEHP, and their mixture on the physiological and biochemical characteristics of cucumber seedlings. The changes of membrane stability index (MSI), antioxidase activities, photosynthetic pigments and chlorophyll fluorescence in cucumber seedlings were assessed. The results demonstrated that MPs alone significantly inhibited MSI, photosynthetic pigments (Chl a, Chl b, and Chl a + b), Fm and qp of cucumber seedlings, and significantly promoted the carotene content and antioxidant enzyme activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) in cucumber seedlings. While DEHP alone significantly inhibited MSI and photosynthetic pigments of cucumber seedlings, and significantly promoted antioxidant enzyme activities in cucumber seedlings. Moreover, the combined toxicity of MPs and DEHP was found to be less pronounced than that of the single action of MPs and DEHP. The interaction between DEHP and MPs may contribute to the reduced toxicity. Abbott's modeling revealed that the combined toxicity systems were all antagonistic (RI < 1). Two-factor analysis and principal component analysis further confirmed that the treatment of MPs alone contributed the most to the toxicological effects of the physiological properties of cucumbers. In summary, this study highlighted the importance of understanding the combined effects of MPs and DEHP on plant physiology, providing insights for the development of effective treatments for emerging pollutants in agricultural ecosystems.

Nanomaterial Size and Surface Modification Mediate Disease Resistance Activation in Cucumber (Cucumis sativus).

Crop disease represents a serious and increasing threat to global food security. Lanthanum oxide nanomaterials (La2O3 NMs) with different sizes (10 and 20 nm) and surface modifications (citrate, polyvinylpyrrolidone [PVP], and poly(ethylene glycol)) were investigated for their control of the fungal pathogen Fusarium oxysporum (Schl.) f. sp cucumerinum Owen on six-week-old cucumber (Cucumis sativus) in soil. Seed treatment and foliar application of the La2O3 NMs at 20-200 mg/kg (mg/L) significantly suppressed cucumber wilt (decreased by 12.50-52.11%), although the disease control efficacy was concentration-, size-, and surface modification-dependent. The best pathogen control was achieved by foliar application of 200 mg/L PVP-coated La2O3 NMs (10 nm); disease severity was decreased by 67.6%, and fresh shoot biomass was increased by 49.9% as compared with pathogen-infected control. Importantly, disease control efficacy was 1.97- and 3.61-fold greater than that of La2O3 bulk particles and a commercial fungicide (Hymexazol), respectively. Additionally, La2O3 NMs application enhanced cucumber yield by 350-461%, increased fruit total amino acids by 295-344%, and improved fruit vitamin content by 65-169% as compared with infected controls. Transcriptomic and metabolomic analyses revealed that La2O3 NMs: (1) interacted with calmodulin, subsequently activating salicylic acid-dependent systemic acquired resistance; (2) increased the activity and expression of antioxidant and related genes, thereby alleviating pathogen-induced oxidative stress; and (3) directly inhibited in vivo pathogen growth. The findings highlight the significant potential of La2O3 NMs for suppressing plant disease in sustainable agriculture.

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Fulvic acid (FA) participates in the regulation of drought stress tolerance in plants, but the underlying mechanisms remain unclear. We carried out an experiment with cucumber cultivar 'Jinyou 35' as the test material and the polyethylene glycol (PEG-6000) being used to simulate drought stress. The concentration effect of FA on drought alleviation of cucumber as well as the effect of FA on photosynthetic enzymes activities, chloroplast ultrastructure, fluorescence parameters, water use efficiency, yield and quality of cucumber plants were studied through spraying FA with different concentrations (0, 100, 300, 500, 700 and 900 mg·L-1). The results showed that FA with different concentrations significantly promoted the relative water content and leaf area and decreased drought injure index as well as the electrolyte leakage and malondialdehyde (MDA) content, compared with the control (0 mg·L-1) under drought stress. The mitigative effect of FA increased first and then decreased with the increases of FA concentration, with 700 mg·L-1 FA showing the best effect. FA significantly enhanced the chlorophyll content, Rubiasco and Rubisco activase (RCA) activities and gene expression, net photosynthesis (Pn), maximal photochemical efficiency of PSⅡin darkness, actual photochemical efficiency, absorbed light energy per unit area, captured light energy per unit area, quantum yield of electron transport and PSⅠ activity, decreased the increase of K point and maintained chloroplast ultrastructure. The experiment in solar-greenhouse showed that FA obviously increased water use efficiency, promoted dry matter accumulation as well as the contents of Vc, soluble sugar, soluble protein and free amino acid, and decreased tannin content. Our results suggested that FA could improve the yield and quality of cucumber in solar greenhouse under drought stress.

Bacterial type III effector-induced plant C8 volatiles elicit antibacterial immunity in heterospecific neighbouring plants via airborne signalling.

Upon sensing attack by pathogens and insect herbivores, plants release complex mixtures of volatile compounds. Here, we show that the infection of lima bean (Phaseolus lunatus L.) plants with the non-host bacterial pathogen Pseudomonas syringae pv. tomato led to the production of microbe-induced plant volatiles (MIPVs). Surprisingly, the bacterial type III secretion system, which injects effector proteins directly into the plant cytosol to subvert host functions, was found to prime both intra- and inter-specific defense responses in neighbouring wild tobacco (Nicotiana benthamiana) plants. Screening of each of 16 effectors using the Pseudomonas fluorescens effector-to-host analyser revealed that an effector, HopP1, was responsible for immune activation in receiver tobacco plants. Further study demonstrated that 1-octen-3-ol, 3-octanone and 3-octanol are novel MIPVs emitted by the lima bean plant in a HopP1-dependent manner. Exposure to synthetic 1-octen-3-ol activated immunity in tobacco plants against a virulent pathogen Pseudomonas syringae pv. tabaci. Our results show for the first time that a bacterial type III effector can trigger the emission of C8 plant volatiles that mediate defense priming via plant-plant interactions. These results provide novel insights into the role of airborne chemicals in bacterial pathogen-induced inter-specific plant-plant interactions.

Triacontanol modulates salt stress tolerance in cucumber by altering the physiological and biochemical status of plant cells.

Cucumber is an important vegetable but highly sensitive to salt stress. The present study was designed to investigate the comparative performance of cucumber genotypes under salt stress (50 mmol L-1) and stress alleviation through an optimized level of triacontanol @ 0.8 mg L-1. Four cucumber genotypes were subjected to foliar application of triacontanol under stress. Different physiological, biochemical, water relations and ionic traits were observed to determine the role of triacontanol in salt stress alleviation. Triacontanol ameliorated the lethal impact of salt stress in all genotypes, but Green long and Marketmore were more responsive than Summer green and 20252 in almost all the attributes that define the genetic potential of genotypes. Triacontanol performs as a good scavenger of ROS by accelerating the activity of antioxidant enzymes (SOD, POD, CAT) and compatible solutes (proline, glycinebetaine, phenolic contents), which lead to improved gas exchange attributes and water relations and in that way enhance the calcium and potassium contents or decline the sodium and chloride contents in cucumber leaves. Furthermore, triacontanol feeding also shows the answer to yield traits of cucumber. It was concluded from the results that the salinity tolerance efficacy of triacontanol is valid in enhancing the productivity of cucumber plants under salt stress. Triacontanol was more pronounced in green long and marketer green than in summer green and 20252. Hence, the findings of this study pave the way towards the usage of triacontanol @ 0.8 mg L-1, and green long and marketer genotypes may be recommended for saline soil.

Functional Characterization of a Cucumber (Cucumis sativus L.) Vacuolar Invertase, CsVI1, Involved in Hexose Accumulation and Response to Low Temperature Stress.

Cucumber (Cucumis sativus L.), an important vegetable plant species, is susceptible to low temperature stress especially during the seedling stage. Vacuolar invertase (VI) plays important roles in plant responses to abiotic stress. However, the molecular and biochemical mechanisms of VI function in cucumber, have not yet been completely understood and VI responses to low temperature stress and it functions in cold tolerance in cucumber seedlings are also in need of exploration. The present study found that hexose accumulation in the roots of cucumber seedlings under low temperature stress is closely related to the observed enhancement of invertase activity. Our genome-wide search for the vacuolar invertase (VI) genes in cucumber identified the candidate VI-encoding gene CsVI1. Expression profiling of CsVI1 showed that it was mainly expressed in the young roots of cucumber seedlings. In addition, transcriptional analysis indicated that CsVI1 expression could respond to low temperature stress. Recombinant CsVI1 proteins purified from Pichia pastoris and Nicotiana benthamiana leaves could hydrolyze sucrose into hexoses. Further, overexpression of CsVI1 in cucumber plants could increase their hexose contents and improve their low temperature tolerance. Lastly, a putative cucumber invertase inhibitor was found could form a complex with CsVI1. In summary, these results confirmed that CsVI1 functions as an acid invertase involved in hexose accumulation and responds to low temperature stress in cucumber seedlings.

TERMINAL FLOWER 1 and TERMINAL FLOWER 1d respond to temperature and photoperiod signals to inhibit determinate growth in cucumber.

Plants monitor environmental cues to balance their vegetative and productive growth by optimizing their inflorescence architecture. TERMINAL FLOWER 1 (TFL1) and its orthologs regulate the inflorescence structure in cucumber, yet the mechanisms underlying their responses to environmental factors and the formation of terminal flowers remain elusive. Here, we performed map-based cloning to identify the gene that controls a season-dependent determinate growth phenotype and found that it was caused by the complete deletion of CsTFL1 in the genome of cucumber line WI1983Hde. In the CsTFL1 deletion plants (CsTFL1del ), determinate growth could be partially rescued by high-temperature and long-day conditions. The expressions of CsTFL1 and its ortholog CsTFL1d could be upregulated by long-day and high-temperature signals. Knockdown of CsTFL1d resulted in determinate growth and the formation of terminal flowers in WT. These results indicate that the induction of CsTFL1d expression by long-day and high-temperature might partially rescue determinate growth of CsTFL1del . Furthermore, biochemical analyses showed that CsTFL1d interacts directly with CsNOT2a, which indicated that CsTFL1d and CsTFL1 function via similar regulatory mechanism. Our data suggest that CsTFL1 and CsTFL1d co-contribute to inhibit determinate growth by responding to temperature and photoperiod signals. It provides mechanistic insights into how environmental cues sculpt the inflorescence architecture of cucumber.

The amino acid permease (AAP) genes CsAAP2A and SlAAP5A/B are required for oomycete susceptibility in cucumber and tomato.

Cucurbit downy mildew (DM), caused by the obligate biotroph Pseudoperonospora cubensis, is a destructive disease in cucumber. A valuable source of DM resistance is the Indian cucumber accession PI 197088, which harbours several quantitative trait loci (QTLs) contributing to quantitatively inherited DM resistance. With a combination of fine-mapping and transcriptomics, we identified Amino Acid Permease 2A (CsAAP2A) as a candidate gene for QTL DM4.1.3. Whole-genome and Sanger sequencing revealed the insertion of a Cucumis Mu-like element (CUMULE) transposon in the allele of the resistant near-isogenic line DM4.1.3. To confirm whether loss of CsAAP2A contributes to partial DM resistance, we performed targeting induced local lesions in genomes on a DM-susceptible cucumber genotype to identify an additional csaap2a mutant, which indeed was partially DM resistant. In view of the loss of the putative function as amino acid transporter, we measured amino acids in leaves. We found that DM-inoculated leaves of line DM4.1.3 (with the csaap2a mutation) contained significantly fewer amino acids than wild-type cucumber. The decreased flow of amino acids towards infected leaves in csaap2a plants compared to the wild type might explain the resistant phenotype of the mutant, as this would limit the available nutrients for the pathogen and thereby its fitness. To examine whether AAP genes play a conserved role as susceptibility factors in plant-oomycete interactions, we made targeted mutations in two AAP genes from tomato and studied the effect on susceptibility to Phytophthora infestans. We conclude that not only CsAAP2A but also SlAAP5A/SlAAP5B are susceptibility genes for oomycete pathogens.

Melatonin alleviates imidacloprid phytotoxicity to cucumber (Cucumis sativus L.) through modulating redox homeostasis in plants and promoting its metabolism by enhancing glutathione dependent detoxification.

Melatonin (Mel), a powerful antioxidant that has the ability to regulate physiological and biochemical processes in plants under abiotic stresses. However, its roles in pesticide detoxification is poorly understood. Herein, selecting leaf spraying insecticide imidacloprid (IMD) as the model, we demonstrated the detoxification mechanism underlying root pretreatment of Mel on IMD in cucumber. IMD treatment affected the primary light conversion efficiency of photosystem II (Fv/Fm), reduced the quantum yield, and increased hydrogen peroxide and superoxide anions contents as well as the levels of membrane lipid peroxidation, indicating that excessive IMD treatment induces oxidative stress. Nonetheless, by increasing the appropriate levels of exogenous Mel, the photosynthesis of cucumber under IMD treatment reached the control levels, effectively removing reactive oxygen species. Furthermore, the content and ratio of ascorbate (AsA) and glutathione (GSH) were decreased under IMD treatment; Mel treatment enhanced the AsA/DHA and GSH/GSSG ratios, as well as the activities of MDHAR, DHAR and GR, suggesting that Mel could alleviate oxidative stress of cucumber treated with IMD by regulating the ascorbic acid-glutathione cycle. Importantly, IMD degradation rate and glutathione S-transferase (GST) activity increased after Mel treatment. The levels of transcripts encoding antioxidant enzymes GPX and GST (GST1,2 and 3) were also increased, indicating that Mel accelerated IMD degradation. These results suggest that Mel plays an important role in the detoxification of IMD by promoting GST activity and transcription and the AsA-GSH cycle, thus providing an approach for plants to reduce IMD residue through the plant's own detoxification mechanism.

Dose-dependent methyl jasmonate effects on photosynthetic traits and volatile emissions: biphasic kinetics and stomatal regulation.

Exogenous application of methyl jasmonate (MeJA) has been extensively used to study jasmonate-dependent signaling events triggered by biotic stresses. MeJA application leads to complex jasmonate-dependent physiological responses, including changes in stomatal openness and induction of emissions of a multitude of volatile compounds. Whether the alterations in stomatal conductance and emissions of MeJA-induced volatiles are quantitatively associated with MeJA dose, and whether the induced volatile emissions are regulated by modifications in stomatal conductance had been poorly known until recently. Our latest studies highlighted a biphasic kinetics of jasmonate-dependent volatile emissions induced by MeJA treatment in the model species cucumber (Cucumis sativus), indicating induction of an immediate stress response and subsequent gene-expression level response. Both the immediate and delayed responses were MeJA dose-dependent. The studies further demonstrated that stomata modulated the kinetics of emissions of water-soluble volatiles in a MeJA dose-dependent manner. These studies contribute to understanding of plant short- and long-term responses to different biotic stress severities as simulated by treatments with a range of MeJA doses corresponding to mild to acute stress.

Adaptation of cucumber seedlings to low temperature stress by reducing nitrate to ammonium during it's transportation.

BACKGROUND: Low temperature severely depresses the uptake, translocation from the root to the shoot, and metabolism of nitrate and ammonium in thermophilic plants such as cucumber (Cucumis sativus). Plant growth is inhibited accordingly. However, the availability of information on the effects of low temperature on nitrogen transport remains limited. RESULTS: Using non-invasive micro-test technology, the net nitrate (NO3-) and ammonium (NH4+) fluxes in the root hair zone and vascular bundles of the primary root, stem, petiole, midrib, lateral vein, and shoot tip of cucumber seedlings under normal temperature (NT; 26 °C) and low temperature (LT; 8 °C) treatment were analyzed. Under LT treatment, the net NO3- flux rate in the root hair zone and vascular bundles of cucumber seedlings decreased, whereas the net NH4+ flux rate in vascular bundles of the midrib, lateral vein, and shoot tip increased. Accordingly, the relative expression of CsNRT1.4a in the petiole and midrib was down-regulated, whereas the expression of CsAMT1.2a-1.2c in the midrib was up-regulated. The results of 15N isotope tracing showed that NO3--N and NH4+-N uptake of the seedlings under LT treatment decreased significantly compared with that under NT treatment, and the concentration and proportion of both NO3--N and NH4+-N distributed in the shoot decreased. Under LT treatment, the actual nitrate reductase activity (NRAact) in the root did not change significantly, whereas NRAact in the stem and petiole increased by 113.2 and 96.2%, respectively. CONCLUSIONS: The higher net NH4+ flux rate in leaves and young tissues may reflect the higher NRAact in the stem and petiole, which may result in a higher proportion of NO3- being reduced to NH4+ during the upward transportation of NO3-. The results contribute to an improved understanding of the mechanism of changes in nitrate transportation in plants in response to low-temperature stress.