Multivariate analysis compares and evaluates drought and flooding tolerances of maize germplasm.

Drought and flooding are the two most important environmental factors limiting maize (Zea mays L.) production globally. This study aimed to investigate the physiological mechanisms and accurate evaluation indicators and methods of maize germplasm involved in drought and flooding stresses. The twice replicated pot experiments with 60 varieties, combined with the field validation experiment with 3 varieties, were conducted under well-watered, drought, and flooding conditions. Most varieties exhibited stronger tolerance to drought than flooding due to higher antioxidant enzyme activities, osmotic adjustment substances, and lower reactive oxygen species. In contrast, flooding stress resulted in higher levels of reactive oxygen species (particularly O2-), ascorbate peroxidase, catalase, peroxidase, and soluble sugars but lower levels of superoxide dismutase, proline, and soluble protein compared with well-watered conditions. Superoxide dismutase, peroxidase, catalase, ascorbate peroxidase, proline, soluble sugars, and protein contents, in addition to plant height, leaf area/plant, and stem diameter, were accurate and representative indicators for evaluating maize tolerance to drought and flooding stresses and could determine a relatively high mean forecast accuracy of 100.0% for the comprehensive evaluation value. A total of 4 principal components were extracted, in which different principal components played a vital role in resisting different water stresses. Finally, the accuracy of the 3 varieties screened by multivariate analysis was verified in the field. This study provides insights into the different physiological mechanisms and accurate evaluation methods of maize germplasm involved in drought and flooding stresses, which could be valuable for further research and breeding.

CsPAO2 Improves Salt Tolerance of Cucumber through the Interaction with CsPSA3 by Affecting Photosynthesis and Polyamine Conversion.

Polyamine oxidases (PAOs) are key enzymes in polyamine metabolism and are related to the tolerance of plants to abiotic stresses. In this study, overexpression of cucumber (Cucumis sativus L.) PAO2 (CsPAO2) in Arabidopsis resulted in increased activity of the antioxidant enzyme and accelerated conversion from Put to Spd and Spm, while malondialdehyde content (MDA) and electrolyte leakage (EL) was decreased when compared with wild type, leading to enhanced plant growth under salt stress. Photosystem Ⅰ assembly 3 in cucumber (CsPSA3) was revealed as an interacting protein of CsPAO2 by screening yeast two-hybrid library combined with in vitro and in vivo methods. Then, CsPAO2 and CsPSA3 were silenced in cucumber via virus-mediated gene silencing (VIGS) with pV190 as the empty vector. Under salt stress, net photosynthetic rate (Pn) and transpiration rate (Tr) of CsPAO2-silencing plants were lower than pV190-silencing plants, and EL in root was higher than pV190-silencing plants, indicating that CsPAO2-silencing plants suffered more serious salt stress damage. However, photosynthetic parameters of CsPSA3-silencing plants were all higher than those of CsPAO2 and pV190-silencing plants, thereby enhancing the photosynthesis process. Moreover, CsPSA3 silencing reduced the EL in both leaves and roots when compared with CsPAO2-silencing plants, but the EL only in leaves was significantly lower than the other two gene-silencing plants, and conversion from Put to Spd and Spm in leaf was also promoted, suggesting that CsPSA3 interacts with CsPAO2 in leaves to participate in the regulation of salt tolerance through photosynthesis and polyamine conversion.

CsCDPK6, a CsSAMS1-Interacting Protein, Affects Polyamine/Ethylene Biosynthesis in Cucumber and Enhances Salt Tolerance by Overexpression in Tobacco.

S-adenosylmethionine synthetase (SAMS) plays a crucial role in regulating stress responses. In a recent study, we found that overexpression of the cucumber gene CsSAMS1 in tobacco can affect the production of polyamines and ethylene, as well as enhancing the salt stress tolerance of tobacco, but the exact underlying mechanisms are elusive. The calcium-dependent protein kinase (CDPK) family is ubiquitous in plants and performs different biological functions in plant development and response to abiotic stress. We used a yeast two-hybrid system to detect whether the protein CDPK6 could interact with SAMS1 and verified their interaction by bimolecular fluorescence complementation (BiFC) and co-immunoprecipitation (Co-IP) assays. To further explore the function of cucumber CDPK6, we isolated and characterized CsCDPK6 in cucumber. CsCDPK6 is a membrane protein that is highly expressed under various abiotic stresses, including salt stress. It was also observed that ectopic overexpression of CsCDPK6 in tobacco enhanced salt tolerance. Under salt stress, CsCDPK6-overexpressing lines enhanced the survival rate and reduced stomatal apertures in comparison to wild-type (WT) lines, as well as lowering malondialdehyde (MDA) and hydrogen peroxide (H2O2) contents and causing less relative electrolyte leakage. Moreover, repression of CsCDPK6 expression by virus-induced gene silencing (VIGS) in cucumber seedling cotyledons under salt stress increased ethylene production and promoted the transformation from putrescine (Put) to spermidine (Spd) and spermine (Spm). These findings shed light on the interaction of CsSAMS1 and CsCDPK6, which functions positively to regulate salt stress in plants.

GABA: A Key Player in Drought Stress Resistance in Plants.

γ-aminobutyric acid (GABA) is a non-protein amino acid involved in various physiological processes; it aids in the protection of plants against abiotic stresses, such as drought, heavy metals, and salinity. GABA tends to have a protective effect against drought stress in plants by increasing osmolytes and leaf turgor and reducing oxidative damage via antioxidant regulation. Guard cell GABA production is essential, as it may provide the benefits of reducing stomatal opening and transpiration and controlling the release of tonoplast-localized anion transporter, thus resulting in increased water-use efficiency and drought tolerance. We summarized a number of scientific reports on the role and mechanism of GABA-induced drought tolerance in plants. We also discussed existing insights regarding GABA's metabolic and signaling functions used to increase plant tolerance to drought stress.

Ozone Induced Stomatal Regulations, MAPK and Phytohormone Signaling in Plants.

Ozone (O3) is a gaseous environmental pollutant that can enter leaves through stomatal pores and cause damage to foliage. It can induce oxidative stress through the generation of reactive oxygen species (ROS) like hydrogen peroxide (H2O2) that can actively participate in stomatal closing or opening in plants. A number of phytohormones, including abscisic acid (ABA), ethylene (ET), salicylic acid (SA), and jasmonic acid (JA) are involved in stomatal regulation in plants. The effects of ozone on these phytohormones' ability to regulate the guard cells of stomata have been little studied, however, and the goal of this paper is to explore and understand the effects of ozone on stomatal regulation through guard cell signaling by phytohormones. In this review, we updated the existing knowledge by considering several physiological mechanisms related to stomatal regulation after response to ozone. The collected information should deepen our understanding of the molecular pathways associated with response to ozone stress, in particular, how it influences stomatal regulation, mitogen-activated protein kinase (MAPK) activity, and phytohormone signaling. After summarizing the findings and noting the gaps in the literature, we present some ideas for future research on ozone stress in plants.

Melatonin alleviates nickel phytotoxicity by improving photosynthesis, secondary metabolism and oxidative stress tolerance in tomato seedlings.

Arable land contamination with nickel (Ni) has become a major threat to worldwide crop production. Recently, melatonin has appeared as a promising stress-relief substance that can alleviate heavy metal-induced phytotoxicity in plants. However, the plausible underlying mechanism of melatonin function under Ni stress has not been fully substantiated in plants. Herein, we conducted an experiment that unveiled critical mechanisms in favor of melatonin-mediated Ni-stress tolerance in tomato. Ni stress markedly inhibited growth and biomass by impairing the photosynthesis, photosystem function, mineral homeostasis, root activity, and osmotic balance. In contrast, melatonin application notably reinforced the plant growth traits, increased photosynthesis efficiency in terms of chlorophyll content, upregulation of chlorophyll synthesis genes, i.e. POR, CAO, CHL G, gas exchange parameters, and PSII maximum efficiency (Fv/Fm), decreased Ni accumulation and increased mineral nutrient homeostasis. Moreover, melatonin efficiently restricted the hydrogen peroxide (H2O2) and superoxide radical production and increased RBOH expression and restored cellular integrity (less malondialdehyde and electrolyte leakage) through triggering the antioxidant enzyme activities and modulating AsA-GSH pools. Notably, oxidative stress was effectively mitigated by upregulation of several defense genes (SOD, CAT, APX, GR, GST, MDHAR, DHAR) and melatonin biosynthesis-related genes (TDC, T5S, SNAT, ASMT). Besides, melatonin treatment enhanced secondary metabolites (phenols, flavonoids, and anthocyanin) contents along with their encoding genes (PAL, CHS) expression, and these metabolites potentially restricted excess H2O2 accumulation. In conclusion, our findings deciphered the potential functions of melatonin in alleviating Ni-induced phytotoxicity in tomato through boosting the biomass production, photosynthesis, nutrient uptake, redox balance, and secondary metabolism.

Fine-tuning of soil water and nutrient fertilizer levels for the ecological restoration of coal-mined spoils using Elaeagnus angustifolia.

Coal mining activities remain of great environmental concern because of several negative impacts on soil ecosystems. Appropriate revegetation interventions of coal-spoiled lands can provide environmental management solutions to restore soil degraded ecosystems. The present study addressed the potential of the pioneer woody species, Elaeagnus angustifolia, in the restoration of coal-mined spoils under a range of different water (W) levels and nitrogen (N) and phosphorus (P) applications. Our results show how moderate applications of N (N60 = 60 mg N kg-1 soil) and P (P90 = 90 mg P kg-1 soil) fertilizers led either to maximum or minimum growth performance of E. angustifolia depending on whether W was applied at very high (W80 = 80% field capacity) or very low (W40 = 40% field capacity) levels suggesting that W was the main limiting factor for plant growth. Very low-W regime (W40N60P90) also caused significant reduction of photosynthetic parameters, including net photosynthetic rate, transpiration rate and water use efficiency. The combination of high W-N doses with low P doses (W70N96P36) positively influenced gas-exchange parameters, chlorophyll and carotenoid contents. Seedlings treated with low-W and -N doses (W50N24P144) showed highest increases in malondialdehyde content and lowest levels of relative water content (RWC). Decreases in malondialdehyde content and increases in RWC were observed following a gradual increment of W and N doses, indicating that high W and N doses contributed to drought tolerance of E. angustifolia by protecting cell membranes and increasing water status. Low-W and -N applications considerably increased the activities of antioxidant enzymes (superoxide dismutase, catalase, and peroxidase) and the contents of proline and soluble sugars, suggesting that E. angustifolia developed defensive strategies to avoid damage induced by water scarcity. Results from heatmap and principal component analyses confirmed that W and N were the main clustering factors, and both N and P performed well at high-W dose. The optimum growth performance of E. angustifolia was found under a combination of W level at 66.0% of field capacity, N dose of 74.0 mg kg-1 soil, and P dose of 36.0 mg kg-1 soil. Our findings demonstrate how optimum growth performance of E. angustifolia can be achieved by fine-tuning doses of W, N, and P resources, and how this in turn could greatly support the ecological restoration of coal-mined degraded environments.

Melatonin alleviates heat-induced damage of tomato seedlings by balancing redox homeostasis and modulating polyamine and nitric oxide biosynthesis.

BACKGROUND: Melatonin is a pleiotropic signaling molecule that plays multifarious roles in plants stress tolerance. The polyamine (PAs) metabolic pathway has been suggested to eliminate the effects of environmental stresses. However, the underlying mechanism of how melatonin and PAs function together under heat stress largely remains unknown. In this study, we investigated the potential role of melatonin in regulating PAs and nitric oxide (NO) biosynthesis, and counterbalancing oxidative damage induced by heat stress in tomato seedlings. RESULTS: Heat stress enhanced the overproduction of reactive oxygen species (ROS) and damaged inherent defense system, thus reduced plant growth. However, pretreatment with 100 µM melatonin (7 days) followed by exposure to heat stress (24 h) effectively reduced the oxidative stress by controlling the overaccumulation of superoxide (O2•-) and hydrogen peroxide (H2O2), lowering the lipid peroxidation content (as inferred based on malondialdehyde content) and less membrane injury index (MII). This was associated with increased the enzymatic and non-enzymatic antioxidants activities by regulating their related gene expression and modulating the ascorbate-glutathione cycle. The presence of melatonin induced respiratory burst oxidase (RBOH), heat shock transcription factors A2 (HsfA2), heat shock protein 90 (HSP90), and delta 1-pyrroline-5-carboxylate synthetase (P5CS) gene expression, which helped detoxify excess ROS via the hydrogen peroxide-mediated signaling pathway. In addition, heat stress boosted the endogenous levels of putrescine, spermidine and spermine, and increased the PAs contents, indicating higher metabolic gene expression. Moreover, melatonin-pretreated seedlings had further increased PAs levels and upregulated transcript abundance, which coincided with suppression of catabolic-related genes expression. Under heat stress, exogenous melatonin increased endogenous NO content along with nitrate reductase- and NO synthase-related activities, and expression of their related genes were also elevated. CONCLUSIONS: Melatonin pretreatment positively increased the heat tolerance of tomato seedlings by improving their antioxidant defense mechanism, inducing ascorbate-glutathione cycle, and reprogramming the PAs metabolic and NO biosynthesis pathways. These attributes facilitated the scavenging of excess ROS and increased stability of the cellular membrane, which mitigated heat-induced oxidative stress.

Comparative Metabolome and Transcriptome Analyses Reveal the Regulatory Mechanism of Purple Leafstalk Production in Taro (Colocasia esculenta L. Schott).

Taro is a plant in the Araceae family, and its leafstalk possesses significant botanical and culinary value owing to its noteworthy medicinal and nutritional attributes. Leafstalk colour is an essential attribute that significantly influences its desirability and appeal to both breeders and consumers. However, limited information is available about the underlying mechanism responsible for the taro plant's colouration. Thus, the purpose of the current study was to elucidate the information on purple leafstalks in taro through comprehensive metabolome and transcriptome analysis. In total, 187 flavonoids, including 10 anthocyanins, were identified. Among the various compounds analysed, it was observed that the concentrations of five anthocyanins (keracyanin chloride (cyanidin 3-O-rutinoside chloride), cyanidin 3-O-glucoside, tulipanin (delphinidin 3-rutinoside chloride), idaein chloride (cyanidin 3-O-galactoside), and cyanidin chloride) were found to be higher in purple taro leafstalk compared to green taro leafstalk. Furthermore, a total of 3330 differentially expressed genes (DEGs) were identified by transcriptome analysis. Subsequently, the correlation network analysis was performed to investigate the relationship between the expression levels of these differentially expressed genes and the content of anthocyanin. There were 18 DEGs encoding nine enzymes detected as the fundamental structural genes contributing to anthocyanin biosynthesis, along with seven transcription factors (3 MYB and 4 bHLH) that may be promising candidate modulators of the anthocyanin biosynthesis process in purple taro leafstalk. The findings of the current investigation not only provide a comprehensive transcriptional code, but also give information on anthocyanin metabolites as well as beneficial insights into the colour mechanism of purple taro leafstalk.

Comparative analysis of the medicinal and nutritional components of different varieties of Pueraria thomsonii and Pueraria lobata.

Pueraria thomsonii and Pueraria lobata are important medicinal plants with unique chemical compositions that are widely used in traditional Chinese medicine. To compare the nutritional and medicinal profiles of these two species, we analyzed the flavonoid, dietary fiber, total starch, and crude protein contents of one P. lobata and three P. thomsonii varieties using ultra-performance liquid chromatography-tandem mass spectrometry, enzyme weight, acid hydrolysis, and Kjeldahl methods. Furthermore, we used principal component analysis and hierarchical clustering heatmap analysis to separate the data obtained from the P. thomsonii and P. lobata samples. We detected 279 flavonoid compounds in the two Pueraria species, including 90 isoflavones and 78 flavonoids. A large proportion of isoflavones and flavonoids were more abundant in P. lobata than in P. thomsonii. The total starch content was significantly higher in P. thomsonii than in P. lobata. By contrast, the soluble dietary fiber, insoluble dietary fiber, and crude protein contents were substantially lower in P. thomsonii than in P. lobata. Taken together, our results demonstrate that P. lobata is better suited for use as a medicine, whereas P. thomsonii is better suited as an edible food, and provide a theoretical foundation for developing P. thomsonii and P. lobata germplasm resources.

Physiological mechanism of melatonin attenuating to osmotic stress tolerance in soybean seedlings.

Drought is one of the most significant abiotic stress threatening to crop production worldwide. Soybean is a major legume crop with immense economic significance, but its production is highly dependent on optimum rainfall or abundant irrigation. As the global climate changes, it is more important to find solutions to make plants more resilient to drought. The prime aimed of the study is to investigate the effect of melatonin on drought tolerance in soybean and its potential mechanisms. Soybean seedlings were treated with 20% polyethylene glycol 6000 (PEG 6000) and subjected to osmotic stress (14 days) with or without 100 µM melatonin treatment. Our results revealed that melatonin supplementation significantly mitigated PEG-induced growth retardation and increased water absorption ability. Foliar application of melatonin also increased gas exchange and the chlorophyll fluorescence attributes by the mitigation of the osmotic-induced reduction of the reaction activity of photosystems I and II, net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), electron transport activity, and photosynthetic efficiency. In addition, PEG-induced elevated production of reactive oxygen species (ROS) and malondialdehyde (MDA) content were significantly reversed by melatonin treatment. Equally important, melatonin boosted the antioxidant activities of soybean plants. Moreover, osmotic stress substantially increased abscisic acid (ABA) accumulation in roots and leaves, while melatonin-received plant leaves accumulated less ABA but roots content higher ABA. Similarly, melatonin significantly suppressed ABA biosynthesis and signaling gene expression in soybean exposed to drought stress. Furthermore, osmotic stress significantly suppressed plasmalemma (GmPIPs) and tonoplast aquaporin (GmTIPs) genes expression, and their transcript abundance was up-regulated by melatonin co-addition. Taken together, our results indicated that melatonin potentially improves drought tolerance of soybean through the regulation of ABA and aquaporin gene expression, increasing photosynthetic efficiency as well as enhancing water uptake efficiency.

Physiological and transcriptomic study reveal SeNPs-mediated AsIII stress detoxification mechanisms involved modulation of antioxidants, metal transporters, and transcription factors in Glycine max L. (Merr.) roots.

Physiological changes and genome-wide alteration in gene expression were performed in soybean (Glycine max [L.] Merr.) roots exposed to AsⅢ (25 µmol/L) alone and supplemented with selenium nanoparticles (SeNPs) at the concentration of 10 and 25 µmol/L at the V2 growth stage. Excessive arsenic in the root zone poses a potential threat to soybean yield, particularly to roots, due to the limited translocation of AsIII from root to shoot in the case of soybean. We hypothesized that SeNPs can relieve AsⅢ toxicity to soybean root by reducing the AsⅢ uptake and regulating the internal tolerance mechanism of the plants. Results accomplished that SeNPs had positive impact on soybean dry weight and roots parameters under AsⅢ stress. Then, we further evaluated physiological indexes, whole genome transcriptomic analysis and quantitative real-time PCR to elucidate the underlying mechanism of AsⅢ tolerance under SeNPs supplementation. Under the condition of AsⅢ-stress, SeNPs exposure significantly reduced the electrolyte leakage, O2-•, H2O2 and MDA accumulation while increasing the antioxidants level. The RNA-seq dataset revealed total of 5819 up and 7231 down expressed DEGs across all libraries. The number of exclusively regulated genes were higher under As + SeNP10 (4909) treatment than in the AsⅢ-alone (4830) and As + SeNP25 (3311) treatments. The KEGG and GO analyses revealed that stress responsive DEGs such as glutathione S-transferase, glutathione peroxidase, ascorbate, glutaredoxin, thioredoxin, and phytochelatins synthase are responsible for AsⅢ tolerance under the SeNPs supplementation. Similarly, sulfate transporter, and ABC transporters (ATP-binding cassettes) expression were induced, and aquaporin channels related DEGs expression were reduced under SeNPs application in AsⅢ exposure condition. Furthermore, the expression of molecular chaperones (HSP) and transcription factors (MYB, bZIP, bHLH, and HSFs) were increased in SeNPs treatment groups. These results provide vital information of AsⅢ tolerance mechanism in response to SeNPs in soybean. We suggest that functional characterization of these genes will help us learn more about the SeNPs responsive arsenic tolerance mechanism in soybean.

ABA activated SnRK2 kinases: an emerging role in plant growth and physiology.

Members of the SNF1-related protein kinase 2 (SnRK2) family are plant-specific serine or threonine kinases that play a pivotal role in the response of plants to abiotic stresses. Members of this plant-specific kinase family have included a critical regulator (SnRK2) of abscisic acid (ABA) response in plants. Plant organ development is governed substantially by the interaction of the SnRK2 and the phytohormone abscisic acid (ABA). Recent research has revealed a synergistic link between SnRK2 and ABA signaling in a plant's response to stress such as drought and shoot growth. SnRK2 kinases play a dual role in the control of SnRK1 and the development of a plant. The dual role of SnRK2 kinases promotes plant growth under optimal conditions and in the absence of ABA while inhibiting the growth of plants in response to ABA. In this review, we have uncovered the roles of ABA-activated SnRK2 kinases in plants, as well as their physiological mechanisms.

Optimizing Nitrogen and Seed Rate Combination for Improving Grain Yield and Nitrogen Uptake Efficiency in Winter Wheat.

Nitrogen (N) supply and seed rate (SR) are two essential factors that affect the accumulation and partitioning of N and dry matter (DM) and, therefore, grain yield (GY) and N use efficiency (NUE). The objective of this experiment was to optimize N application and SR to regulate wheat growth and increase both GY and NUE. The results revealed that net photosynthetic rate (Pn), stomatal conductance (Gs), chlorophyll content, and activities of metabolic enzymes (NR and GS) significantly increased with increasing of N levels while decreasing SR. Plant tillers, GY, DM before anthesis, and N translocation, N agronomic efficiency (NAE), N recovery efficiency (NRE), and N uptake efficiency (NUPE) were highest in a combined treatment of N235 and SR180. However, N levels beyond 235 kg ha-1 significantly decreased NAE, NRE, and NUPE. By increasing SR from 135 to 180 kg ha-1 an increase of 12.9 % and 9.1% GY and NUPE, respectively, was observed. Based on this result, we estimate that 1 kg N ha-1 might be replaced by an increase of approximately 0.6 kg ha-1 SR. Our study suggested that using a combination of N and SR (N235 + SR180) could attain maximum GY and improve NUE parameters.

Putrescine regulates stomatal opening of cucumber leaves under salt stress via the H2O2-mediated signaling pathway.

The stomatal aperture is imperative for photosynthesis in higher plants. The function of polyamines (PAs) in stomatal regulation under a stressful environment has not been fully determined. In this study, we demonstrated the mechanism by which putrescine (Put) regulates stomatal changes in cucumber leaves under salt stress. The results showed that foliar application of Put alleviated the decrease of stomatal aperture and photosynthesis caused by salt stress and promoted plant growth. Exogenous Put caused a significant increase in endogenous PAs and hydrogen peroxide (H2O2) levels by 105.43% and 27.97%, respectively, while decreased abscisic acid (ABA) content by 67.68% under salt stress. However, application of inhibitors of aminoguanidine hydrochloride (AG), 1, 8-diaminooctane (1, 8-DO), diphenyleneiodonium chloride (DPI) and salicylhydroxamic acid (SHAM) upregulated the 9-cis-cyclocarotenoid dioxygenase (NCED) gene and downregulated the reduced glutathione synthetase (GSHS) gene. These inhibitors also decreased the stomatal aperture, levels of H2O2 and reduced glutathione (GSH), but increased the ABA content under salt stress and Put treatment conditions. The order of influence is AG > 1, 8-DO > DPI > SHAM. However, Put-induced downregulation of ABA content and upregulation of GSH content under salt stress were effectively blocked by N, N'-dimethylthiourea (DMTU, H2O2 scavenger) and 1-chloro-2,4-dinitrobenzene (CDNB, GSH scavenger) treatments. Taken together, these results suggest that Put induced the formation of H2O2 signaling mediates the degradation of PAs by diamine oxidase (DAO), increasing GSH content and inhibiting the accumulation of ABA in leaves, thereby promoting stomatal opening in salt-stressed cucumber leaves.

Nitrogen fertilization coupled with foliar application of iron and molybdenum improves shade tolerance of soybean under maize-soybean intercropping.

Maize-soybean intercropping is practiced worldwide because of some of the anticipated advantages such as high crop yield and better utilization of resources (i.e., water, light, nutrients and land). However, the shade of the maize crop has a detrimental effect on the growth and yield of soybean under the maize-soybean intercropping system. Hence, this experiment was conducted to improve the shade tolerance of such soybean crops with optimal nitrogen (N) fertilization combined with foliar application of iron (Fe) and molybdenum (Mo). The treatments comprised five (5) maize-soybean intercropping practices: without fertilizer application (F0), with N fertilizer application (F1), with N fertilizer combined with foliar application of Fe (F2), with N fertilizer coupled with foliar application of Mo (F3) and with N fertilizer combined with foliar application of Fe and Mo (F4). The findings of this study showed that maize-soybean intercropping under F4 treatment had significantly (p< 0.05) increased growth indices such as leaf area (cm2), plant height (cm), stem diameter (mm), stem strength (g pot-1), and internode length (cm) and yield indices (i.e., No of pods plant-1, grain yield (g plant-1), 100-grain weight (g), and biomass dry matter (g plant-1)) of the soybean crop. Moreover, intercropping under F4 treatment enhanced the chlorophyll SPAD values by 26% and photosynthetic activities such as Pn by 30%, gs by 28%, and Tr by 28% of the soybean crops, but reduced its CO2 by 11%. Furthermore, maize-soybean intercropping under F4 treatment showed improved efficiency of leaf chlorophyll florescence parameters of soybean crops such as Fv/Fm (26%), qp (17%), ϕPSII (20%), and ETR (17%), but reduced NPQ (12%). In addition, the rubisco activity and soluble protein content of the soybean crop increased by 18% in maize-soybean intercropping under F4 treatment. Thus, this suggested that intercropping under optimal N fertilization combined with foliar application of Fe and Mo can improve the shade tolerance of soybean crops by regulating their chlorophyll content, photosynthetic activities, and the associated enzymes, thereby enhancing their yield and yield traits.

Exogenous spermidine modulates polyamine metabolism and improves stress responsive mechanisms to protect tomato seedlings against salt stress.

Salt stress negatively affects plant growth, development, and crop productivity causing serious economic loss to agricultural production. Here, we investigated the exogenous application of spermidine (Spd) on tomato seedlings grown under salt stress. Salt stress reduced plant growth, biomass accumulation and chlorophyll contents, thus negatively affecting photosynthesis. Alternatively, Spd application effectively reduced the salinity-induced adverse effects in tomato seedlings by activating the H2O2 mediated signaling involving the enhanced expression of RBOH1 and salt stress-responsive genes SlMYB102, SlHKT1, SlWRKY1 and SlDREB2, and improving detoxification through higher antioxidative activity and osmolyte (proline) accumulation under salt stress. It was further confirmed by significantly lower amount of H2O2, malondialdehyde and electrolyte leakage, and better ion homeostasis (Na+/K+ ratio) and photosynthetic performance of Spd-treated seedlings under salt stress. Furthermore, Spd application modulated endogenous polyamines and enhanced the biosynthesis of endogenous Spd and spermine from putrescine. Altogether, these results confirm the important role of Spd against salt stress and suggest that the increased endogenous Spd content in plants could regulate a number of stress-responsive mechanisms to protect tomato seedlings against salt stress. These results provide a good direction for further elucidation of the detailed interplay between polyamine metabolism and H2O2-mediated signaling, which would help to improve abiotic stress tolerance in plants.

Exogenous Putrescine Increases Heat Tolerance in Tomato Seedlings by Regulating Chlorophyll Metabolism and Enhancing Antioxidant Defense Efficiency.

Crops around the world are facing a diversity of environmental problems, of which high temperatures are proving to be the most serious threat to crops. Polyamine putrescine (Put) acts as a master growth regulator that contributes to optimal plant growth and development and increased stress tolerance. Here, the current study aimed to elucidate how Put functions in regulating chlorophyll (Chl) metabolism, oxidative stress, and antioxidant defense, as well as to characterize the expression of genes related to heat stress in tomato seedlings under such stress. The results revealed that Put treatment significantly attenuates heat-induced damage by promoting biomass production, increasing photosynthetic efficiency, and inhibiting excessive production of oxidative stress markers. Heat stress markedly decreased the Chl content in the tomato leaf and accelerated the leaf yellowing process. However, Put-treated tomato seedlings showed a higher Chl content, which could be associated with the functions of Put in elevating PBGD activity (Chl biosynthesis enzyme) and suppressing the activity of the Chl catabolic enzyme (Chlase and MDCase). Under high-temperature stress, the expression levels of the gene encoding factors involved in Chl biosynthesis and Chl catabolism were significantly down- and upregulated, respectively, and this trend was reversed in Put-treated heat-stressed seedlings. In addition, exogenous application of Put boosted the activity of antioxidant enzymes, along with the levels of expression of their encoding genes, only in plants that were heat stressed. Furthermore, the expression levels of heat-shock-related genes (HSP90, HSP70, and HsfA1) were elevated in Put-treated, high-temperature-stressed tomato seedlings. Taken together, our results indicate that Put treatment significantly increases the heat tolerance of tomato seedlings, by elevating Chl concentrations and suppressing Chl catabolic enzyme activity, modulating endogenous free PA content, increasing antioxidant defense efficiency, and upregulating the expression of heat-shock-related genes.

Melatonin Improves Drought Stress Tolerance of Tomato by Modulating Plant Growth, Root Architecture, Photosynthesis, and Antioxidant Defense System.

Tomato is an important vegetable that is highly sensitive to drought (DR) stress which impairs the development of tomato seedlings. Recently, melatonin (ME) has emerged as a nontoxic, regulatory biomolecule that regulates plant growth and enhances the DR tolerance mechanism in plants. The present study was conducted to examine the defensive role of ME in photosynthesis, root architecture, and the antioxidant enzymes' activities of tomato seedlings subjected to DR stress. Our results indicated that DR stress strongly suppressed growth and biomass production, inhibited photosynthesis, negatively affected root morphology, and reduced photosynthetic pigments in tomato seedlings. Per contra, soluble sugars, proline, and ROS (reactive oxygen species) were suggested to be improved in seedlings under DR stress. Conversely, ME (100 µM) pretreatment improved the detrimental-effect of DR by restoring chlorophyll content, root architecture, gas exchange parameters and plant growth attributes compared with DR-group only. Moreover, ME supplementation also mitigated the antioxidant enzymes [APX (ascorbate peroxidase), CAT (catalase), DHAR (dehydroascorbate reductase), GST (glutathione S-transferase), GR (glutathione reductase), MDHAR (monodehydroascorbate reductase), POD (peroxidase), and SOD (superoxide dismutase)], non-enzymatic antioxidant [AsA (ascorbate), DHA (dehydroascorbic acid), GSH (glutathione), and GSSG, (oxidized glutathione)] activities, reduced oxidative damage [EL (electrolyte leakage), H2O2 (hydrogen peroxide), MDA (malondialdehyde), and O2•- (superoxide ion)] and osmoregulation (soluble sugars and proline) of tomato seedlings, by regulating gene expression for SOD, CAT, APX, GR, POD, GST, DHAR, and MDHAR. These findings determine that ME pretreatment could efficiently improve the seedlings growth, root characteristics, leaf photosynthesis and antioxidant machinery under DR stress and thereby increasing the seedlings' adaptability to DR stress.

Overexpression of 7-hydroxymethyl Chlorophyll a Reductase from Cucumber in Tobacco Accelerates Dark-Induced Chlorophyll Degradation.

7-hydroxymethyl chlorophyll (Chl) a reductase (HCAR) plays critical roles in the Chl cycle and degradation during leaf senescence, however, its function in horticultural crops remains unknown. Here, we identified an HCAR gene (CsHCAR) from cucumber (Cucumis sativus L.) and investigated its roles in response to dark-induced Chl degradation. CsHCAR encoded 459 amino acids, which were orthologous to Arabidopsis HCAR, had the conserved domains, and localized in the chloroplast. Gene expression analysis showed that CsHCAR expression was the highest in senescent leaves and was responsive to different stresses and phytohormone treatments. Overexpression of CsHCAR in tobacco accelerated dark-induced Chl degradation through enhancing the expression of Chl catabolic genes. After 10 d of darkness treatment, the biomass of CsHCAR overexpression plants was reduced. Furthermore, the value of net photosynthetic rate, maximum quantum yield of photosystem II, and effective quantum yield of photosystem II in CsHCAR overexpression plants was significantly reduced in comparison to that in wild-type (WT) plants. The photosynthetic protein content, including Lhcb1, Lhcb2, Lhcb4, RbcS, and RbcL in CsHCAR overexpression plants exhibited a lower level as compared to that observed in WT plants. In addition, the expression of genes encoding these proteins in CsHCAR overexpression plants was significantly lower than that in WT plants. Moreover, CsHCAR overexpression plants inhibited the dark-induced accumulation of reactive oxygen species (ROS). These results indicate that CsHCAR affects the stability of photosynthetic proteins in chloroplasts, positively regulates Chl degradation, and plays an important role in maintaining ROS homeostasis in leaves.