Maize miRNAs and their putative target genes involved in chilling stress response in 5-day old seedlings.

BACKGROUND: In the context of early sowing of maize as a promising adaptation strategy that could significantly reduce the negative effects of climate change, an in-depth understanding of mechanisms underlying plant response to low-temperature stress is demanded. Although microRNAs (miRNAs) have been recognized as key regulators of plant stress response, research on their role in chilling tolerance of maize during early seedling stages is scarce. Therefore, it is of great significance to explore chilling-responsive miRNAs, reveal their expression patterns and associated target genes, as well as to examine the possible functions of the conserved and novel miRNAs. In this study, the role of miRNAs was examined in 5d-old maize seedlings of one tolerant and one sensitive inbred line exposed to chilling (10/8 °C) stress for 6 h and 24 h, by applying high throughput sequencing. RESULTS: A total of 145 annotated known miRNAs belonging to 30 families and 876 potentially novel miRNAs were identified. Differential expression (DE) analysis between control and stress conditions identified 98 common miRNAs for both genotypes at one time point and eight miRNAs at both time points. Target prediction and enrichment analysis showed that the DE zma-miR396, zma-miR156, zma-miR319, and zma-miR159 miRNAs modulate growth and development. Furthermore, it was found that several other DE miRNAs were involved in abiotic stress response: antioxidative mechanisms (zma-miR398), signal transduction (zma-miR156, zma-miR167, zma-miR169) and regulation of water content (zma-miR164, zma-miR394, zma-miR396). The results underline the zma-miRNAs involvement in the modulation of their target genes expression as an important aspect of the plant's survival strategy and acclimation to chilling stress conditions. CONCLUSIONS: To our understanding, this is the first study on miRNAs in 5-d old seedlings' response to chilling stress, providing data on the role of known and novel miRNAs post-transcriptional regulation of expressed genes and contributing a possible platform for further network and functional analysis.

Utilizing transcriptomics and metabolomics to unravel key genes and metabolites of maize seedlings in response to drought stress.

BACKGROUND: Drought stress can substantially restrict maize growth and productivity, and global warming and an increasing frequency of extreme weather events are likely to result in more yield losses in the future. Therefore, unraveling the molecular mechanism underlying the response to drought stress is essential for breeding drought-resilient crops. RESULTS: In this study, we subjected the 3-leaf-period plants of two maize inbred lines, a drought-tolerant line (si287) and a drought-sensitive line (X178), to drought stress for seven days while growing in a chamber. Subsequently, we measured physiological traits and analyzed transcriptomic and metabolic profiles of two inbred lines. Our KEGG analysis of genes and metabolites revealed significant differences in pathways related to glycolysis/gluconeogenesis, flavonoid biosynthesis, starch and sucrose metabolism, and biosynthesis of amino acids. Additionally, our joint analysis identified proline, tryptophan and phenylalanine are crucial amino acids for maize response to drought stress. Furthermore, we concentrated on tryptophan (Trp), which was found to enhance tolerance via IAA-ABA signaling, as well as SA and nicotinamide adenine dinucleotide (NAD) consequent reactive oxygen species (ROS) scavenging. We identified three hub genes in tryptophan biosynthesis, indole-3-acetaldehyde oxidase (ZmAO1, 542,228), catalase 1 (ZmCAT1, 542,369), and flavin-containing monooxygenase 6 (ZmYUC6, 103,629,142), High expression of these genes plays a significant role in regulating drought tolerance. Two metabolites related to tryptophan biosynthesis, quinolinic acid, and kynurenine improved maize tolerance to drought stress by scavenging reactive oxygen species. CONCLUSIONS: This study illuminates the mechanisms underlying the response of maize seedlings to drought stress. Especially, it identifies novel candidate genes and metabolites, enriching our understanding of the role of tryptophan in drought stress. The identification of distinct resistance mechanisms in maize inbred lines will facilitate the exploration of maize germplasm and the breeding of drought-resilient hybrids.

Intraspecific alternative phenotypes contribute to variation in species' strategies for growth.

Ecologists have historically sought to identify the mechanisms underlying the maintenance of local species diversity. High-dimensional trait-based relationships, such as alternative phenotypes, have been hypothesized as important for maintaining species diversity such that phenotypically dissimilar individuals compete less for resources but have similar performance in a given environment. The presence of alternative phenotypes has primarily been investigated at the community level, despite the importance of intraspecific variation to diversity maintenance. The aims of this research are to (1) determine the presence or absence of intraspecific alternative phenotypes in three species of tropical tree seedlings, (2) investigate if these different species use the same alternative phenotypes for growth success, and (3) evaluate how findings align with species co-occurrence patterns. We model species-specific relative growth rate with individual-level measurements of leaf mass per area (LMA) and root mass fraction (RMF), environmental data, and their interactions. We find that two of the three species have intraspecific alternative phenotypes, with individuals within species having different functional forms leading to similar growth. Interestingly, individuals within these species use the same trait combinations, high LMA × low RMF and low LMA × high RMF, in high soil nutrient environments to acquire resources for higher growth. This similarity among species in intraspecific alternative phenotypes and variables that contribute most to growth may lead to their negative spatial co-occurrence. Overall, we find that multiple traits or interactions between traits and the environment drive species-specific strategies for growth, but that individuals within species leverage this multi-dimensionality in different ways for growth success.

Microplastic influences the ménage à trois among the plant, a fungal pathogen, and a plant growth-promoting fungal species.

Microplastics (MP) can influence a plethora of fungal species within the rhizosphere. Nevertheless, there are few studies on the direct impacts of MPs on soil fungi and their intricate interplay with plants. Here, we investigated the impact of polyethylene microspheres (PEMS) on the ecological interactions between Fusarium solani, a plant pathogenic fungus, and Trichoderma viride, a fungal plant growth promotor, within the rhizosphere of Solanum lycopersicum (tomato). Spores of F. solani and T. viride were pre-incubated with PEMS at two concentrations, 100 and 1000 mg L-1. Mycelium growth, sporulation, spore germination, and elongation were evaluated. Tomato seeds were exposed to fungal spore suspensions treated with PEMS, and plant development was subsequently assessed after 4 days. The results showed that PEMS significantly enhanced the sporulation (106.0 % and 70.1 %) but compromised the spore germination (up to 27.3 % and 32.2 %) and radial growth (up to -5.2% and -21.7 %) of F. solani and T. viride, respectively. Furthermore, the 100 and 1000 mg L-1 concentrations of PEMS significantly (p<0.05) enhanced the mycelium density of T. viride (9.74 % and 22.30 %, respectively), and impaired the germ-tube elongation of F. solani after 4 h (16.16 % and 11.85 %, respectively) and 8 h (4 % and 17.10 %, respectively). In addition, PEMS amplified the pathogenicity of F. solani and boosted the bio-enhancement effect of T. viride on tomato root growth. Further, PEMS enhanced the bio-fungicidal effect of T. viride toward F. solani (p<0.05). In summary, PEMS had varying effects on F. solani and T. viride, impacting their interactions and influencing their relationship with tomato plants. It intensified the beneficial effects of T. viride and increased the aggressiveness of F. solani. This study highlights concerns regarding the effects of MPs on fungal interactions in the rhizosphere, which are essential for crop soil colonization and resource utilization.

Role of organic and inorganic amendments on physiological attributes of germinating pea seedlings under arsenic stress.

There are scarce data regarding the effects of soil amendments on biophysicochemical responses of plants at the early stages of growth/germination. This study critically compares the effects of ethylene-diamine-tetra-acetic-acid (EDTA) and calcium (Ca) on biophysicochemical responses of germinating pea seedlings under varied arsenic levels (As, 25, 125, 250 µM). Arsenic alone enhanced hydrogen peroxide (H2O2) level in pea roots (176%) and shoot (89%), which significantly reduced seed germination percentage, pigment contents, and growth parameters. Presence of EDTA and Ca in growth culture minimized the toxic effects of As on pea seedlings, EDTA being more pertinent than Ca. Both the amendments decreased H2O2 levels in pea tissues (16% in shoot and 13% in roots by EDTA, and 7% by Ca in shoot), and maintained seed germination, pigment contents, and growth parameters of peas close to those of the control treatment. The effects of all As-treatments were more pronounced in the pea roots than in the shoot. The presence of organic and inorganic amendments can play a useful role in alleviating As toxicity at the early stages of pea growth. The scarcity of data demands comparing plant biophysicochemical responses at different stages of plant growth (germinating vs mature) in future studies.

Till date, abundant research has focused on plant biophysicochemical responses to different types of pollutants. However, the majority of these studies dealt with pollutant exposure to mature plants (generally after a vegetative growth period of 1­2 weeks). Despite significant research, there are still limited data regarding the biophysicochemical responses of plants at their early stages of germination and growth. In fact, stresses at germination or at an early stage of growth can be highly fatal and may significantly affect the ultimate plant growth and potential to remediate the contaminated sites. Therefore, the current study deals with the exposure of germinating pea seedlings to arsenic (As) stress under varied amendments. This experimental plan helped to understand the initial biophysicochemical changes induced in pea plants under As stress.

Metabolic and Functional Interactions of H2S and Sucrose in Maize Thermotolerance through Redox Homeodynamics.

Hydrogen sulfide (H2S) is a novel gasotransmitter. Sucrose (SUC) is a source of cellular energy and a signaling molecule. Maize is the third most common food crop worldwide. However, the interaction of H2S and SUC in maize thermotolerance is not widely known. In this study, using maize seedlings as materials, the metabolic and functional interactions of H2S and SUC in maize thermotolerance were investigated. The data show that under heat stress, the survival rate and tissue viability were increased by exogenous SUC, while the malondialdehyde content and electrolyte leakage were reduced by SUC, indicating SUC could increase maize thermotolerance. Also, SUC-promoted thermotolerance was enhanced by H2S, while separately weakened by an inhibitor (propargylglycine) and a scavenger (hypotaurine) of H2S and a SUC-transport inhibitor (N-ethylmaleimide), suggesting the interaction of H2S and SUC in the development of maize thermotolerance. To establish the underlying mechanism of H2S-SUC interaction-promoted thermotolerance, redox parameters in mesocotyls of maize seedlings were measured before and after heat stress. The data indicate that the activity and gene expression of H2S-metabolizing enzymes were up-regulated by SUC, whereas H2S had no significant effect on the activity and gene expression of SUC-metabolizing enzymes. In addition, the activity and gene expression of catalase, glutathione reductase, ascorbate peroxidase, peroxidase, dehydroascorbate reductase, monodehydroascorbate reductase, and superoxide dismutase were reinforced by H2S, SUC, and their combination under non-heat and heat conditions to varying degrees. Similarly, the content of ascorbic acid, flavone, carotenoid, and polyphenol was increased by H2S, SUC, and their combination, whereas the production of superoxide radicals and the hydrogen peroxide level were impaired by these treatments to different extents. These results imply that the metabolic and functional interactions of H2S and sucrose signaling exist in the formation of maize thermotolerance through redox homeodynamics. This finding lays the theoretical basis for developing climate-resistant maize crops and improving food security.

Physiological and Proteome Analysis of the Effects of Chitosan Oligosaccharides on Salt Tolerance of Rice Seedlings.

Rice (Oryza sativa L.) is an important social-economic crop, and rice seedlings are easily affected by salt stress. Chitosan oligosaccharide (COS) plays a positive role in promoting plant growth and development. To gain a better understanding of the salt tolerance mechanism of rice under the action of COS, Nipponbare rice seedlings were selected as the experimental materials, and the physiological and biochemical indexes of rice seedlings in three stages (normal growth, salt stress and recovery) were measured. Unlabelled quantitative proteomics technology was used to study differential protein and signaling pathways of rice seedlings under salt stress, and the mechanism of COS to improve rice tolerance to salt stress was elucidated. Results showed that after treatment with COS, the chlorophyll content of rice seedlings was 1.26 times higher than that of the blank group (CK). The root activity during the recovery stage was 1.46 times that of the CK group. The soluble sugar in root, stem and leaf increased by 53.42%, 77.10% and 9.37%, respectively. The total amino acid content increased by 77% during the stem recovery stage. Furthermore, the malondialdehyde content in root, stem and leaf increased by 21.28%, 26.67% and 32.69%, respectively. The activity of oxide dismutase (SOD), peroxidase (POD) and oxygenase (CAT) were increased. There were more differentially expressed proteins in the three parts of the experimental group than in the CK group. Gene Ontology (GO) annotation of these differentially expressed proteins revealed that the experimental group was enriched for more entries. Then, through the Kyoto Encyclopedia of Genes and Genomes (KEGG), the top ten pathways enriched with differentially expressed proteins in the two groups (COS and CK groups) were utilized, and a detailed interpretation of the glycolysis and photosynthesis pathways was provided. Five key proteins, including phosphofructokinase, fructose bisphosphate aldolases, glycer-aldehyde-3-phosphate dehydrogenase, enolase and pyruvate kinase, were identified in the glycolysis pathway. In the photosynthesis pathway, oxygen evolution enhancement proteins, iron redox proteins and ferredoxin-NADPH reductase were the key proteins. The addition of COS led to an increase in the abundance of proteins, a response of rice seedlings to salt stress. COS helped rice seedlings resist salt stress. Furthermore, using COS as biopesticides and biofertilizers can effectively increase the utilization of saline-affected farmland, thereby contributing to the alleviating of the global food crisis.

Antifungal Properties of Bio-AgNPs against D. pinodes and F. avenaceum Infection of Pea (Pisum sativum L.) Seedlings.

Ascochyta blight and Fusarium root rot are the most serious fungal diseases of pea, caused by D. pinodes and F. avenaceum, respectively. Due to the lack of fully resistant cultivars, we proposed the use of biologically synthesized silver nanoparticles (bio-AgNPs) as a novel protecting agent. In this study, we evaluated the antifungal properties and effectiveness of bio-AgNPs, in in vitro (poisoned food technique; resazurin assay) and in vivo (seedlings infection) experiments, against D. pinodes and F. avenaceum. Moreover, the effects of diseases on changes in the seedlings' metabolic profiles were analyzed. The MIC for spores of both fungi was 125 mg/L, and bio-AgNPs at 200 mg/L most effectively inhibited the mycelium growth of D. pinodes and F. avenaceum (by 45 and 26%, respectively, measured on the 14th day of incubation). The treatment of seedlings with bio-AgNPs or fungicides before inoculation prevented the development of infection. Bio-AgNPs at concentrations of 200 mg/L for D. pinodes and 100 mg/L for F. avenaceum effectively inhibited infections' spread. The comparison of changes in polar metabolites' profiles revealed disturbances in carbon and nitrogen metabolism in pea seedlings by both pathogenic fungi. The involvement of bio-AgNPs in the mobilization of plant metabolism in response to fungal infection is also discussed.

Comparative analysis of two kinds of garlic seedings: qualities and transcriptional landscape.

BACKGROUND: Facility cultivation is widely applied to meet the increasing demand for high yield and quality, with light intensity and light quality being major limiting factors. However, how changes in the light environment affect development and quality are unclear in garlic. When garlic seedlings are grown, they can also be exposed to blanching culture conditions of darkness or low-light intensity to ameliorate their appearance and modify their bioactive compounds and flavor. RESULTS: In this study, we determined the quality and transcriptomes of 14-day-old garlic and blanched garlic seedlings (green seedlings and blanched seedlings) to explore the mechanisms by which seedlings integrate light signals. The findings revealed that blanched garlic seedlings were taller and heavier in fresh weight compared to green garlic seedlings. In addition, the contents of allicin, cellulose, and soluble sugars were higher in the green seedlings. We also identified 3,872 differentially expressed genes between green and blanched garlic seedlings. The Kyoto Encyclopedia of Genes and Genomes analysis suggested enrichment for plant-pathogen interactions, phytohormone signaling, mitogen-activated protein kinase signaling, and other metabolic processes. In functional annotations, pathways related to the growth and formation of the main compounds included phytohormone signaling, cell wall metabolism, allicin biosynthesis, secondary metabolism and MAPK signaling. Accordingly, we identified multiple types of transcription factor genes involved in plant-pathogen interactions, plant phytohormone signaling, and biosynthesis of secondary metabolites among the differentially expressed genes between green and blanched garlic seedlings. CONCLUSIONS: Blanching culture is one facility cultivation mode that promotes chlorophyll degradation, thus changing the outward appearance of crops, and improves their flavor. The large number of DEGs identified confirmed the difference of the regulatory machinery under two culture system. This study increases our understanding of the regulatory network integrating light and darkness signals in garlic seedlings and provides a useful resource for the genetic manipulation and cultivation of blanched garlic seedlings.

Regulatory effects of silicon nanoparticles on the growth and photosynthesis of cotton seedlings under salt and low-temperature dual stress.

BACKGROUND: Silicon nanoparticles (SiO2-NPs) play a crucial role in plants mitigating abiotic stress. However, the regulatory mechanism of SiO2-NPs in response to multiple stress remains unclear. The objectives of this study were to reveal the regulatory mechanism of SiO2-NPs on the growth and photosynthesis in cotton seedlings under salt and low-temperature dual stress. It will provide a theoretical basis for perfecting the mechanism of crop resistance and developing the technology of cotton seedling preservation and stable yield in arid and high salt areas. RESULTS: The results showed that the salt and low-temperature dual stress markedly decreased the plant height, leaf area, and aboveground biomass of cotton seedlings by 9.58%, 15.76%, and 39.80%, respectively. While SiO2-NPs alleviated the damage of the dual stress to cotton seedling growth. In addition to reduced intercellular CO2 concentration, SiO2-NPs significantly improved the photosynthetic rate, stomatal conductance, and transpiration rate of cotton seedling leaves. Additionally, stomatal length, stomatal width, and stomatal density increased with the increase in SiO2-NPs concentration. Notably, SiO2-NPs not only enhanced chlorophyll a, chlorophyll b, and total chlorophyll content, but also slowed the decrease of maximum photochemical efficiency, actual photochemical efficiency, photochemical quenching of variable chlorophyll, and the increase in non-photochemical quenching. Moreover, SiO2-NPs enhanced the activities of ribulose-1,5-bisphosphate carboxylase/oxygenase and phosphoenolpyruvate carboxylase, improved leaf water potential, and decreased abscisic acid and malondialdehyde content. All the parameters obtained the optimal effects at a SiO2-NPs concentration of 100 mg L- 1, and significantly increased the plant height, leaf area, and aboveground biomass by 7.68%, 5.37%, and 43.00%, respectively. Furthermore, significant correlation relationships were observed between photosynthetic rate and stomatal conductance, stomatal length, stomatal width, stomatal density, chlorophyll content, maximum photochemical efficiency, actual photochemical efficiency, photochemical quenching of variable chlorophyll, and Rubisco activity. CONCLUSION: The results suggested that the SiO2-NPs improved the growth and photosynthesis of cotton seedlings might mainly result from regulating the stomatal state, improving the light energy utilization efficiency and electron transport activity of PSII reaction center, and inducing the increase of Rubisco activity to enhance carbon assimilation under the salt and low-temperature dual stress.

Phosphorus limitation of early growth differs between nitrogen-fixing and nonfixing dry tropical forest tree species.

Tropical forests are often characterized by low soil phosphorus (P) availability, suggesting that P limits plant performance. However, how seedlings from different functional types respond to soil P availability is poorly known but important for understanding and modeling forest dynamics under changing environmental conditions. We grew four nitrogen (N)-fixing Fabaceae and seven diverse non-N-fixing tropical dry forest tree species in a shade house under three P fertilization treatments and evaluated carbon (C) allocation responses, P demand, P-use, investment in P acquisition traits, and correlations among P acquisition traits. Nitrogen fixers grew larger with increasing P addition in contrast to non-N fixers, which showed fewer responses in C allocation and P use. Foliar P increased with P addition for both functional types, while P acquisition strategies did not vary among treatments but differed between functional types, with N fixers showing higher root phosphatase activity (RPA) than nonfixers. Growth responses suggest that N fixers are limited by P, but nonfixers may be limited by other resources. However, regardless of limitation, P acquisition traits such as mycorrhizal colonization and RPA were nonplastic across a steep P gradient. Differential limitation among plant functional types has implications for forest succession and earth system models.

Maize WRKY28 interacts with the DELLA protein D8 to affect skotomorphogenesis and participates in the regulation of shade avoidance and plant architecture.

Competition for light from neighboring vegetation can trigger the shade-avoidance response (SAR) in plants, which is detrimental to their yield. The molecular mechanisms regulating SAR are well established in Arabidopsis, and some regulators of skotomorphogenesis have been found to be involved in the regulation of the SAR and plant architecture. However, the role of WRKY transcription factors in this process has rarely been reported, especially in maize (Zea mays). Here, we report that maize Zmwrky28 mutants exhibit shorter mesocotyls in etiolated seedlings. Molecular and biochemical analyses demonstrate that ZmWRKY28 directly binds to the promoter regions of the Small Auxin Up RNA (SAUR) gene ZmSAUR54 and the Phytochrome-Interacting Factor (PIF) gene ZmPIF4.1 to activate their expression. In addition, the maize DELLA protein Dwarf Plant8 (D8) interacts with ZmWRKY28 in the nucleus to inhibit its transcriptional activation activity. We also show that ZmWRKY28 participates in the regulation of the SAR, plant height, and leaf rolling and erectness in maize. Taken together, our results reveal that ZmWRKY28 is involved in GA-mediated skotomorphogenic development and can be used as a potential target to regulate SAR for breeding of high-density-tolerant cultivars.

Developmental dynamics of crassulacean acid metabolism (CAM) in Opuntia ficus-indica.

BACKGROUND AND AIMS: The relative contributions of C3 photosynthesis and crassulacean acid metabolism (CAM) during the earliest stages of development were investigated to assess how much each might contribute to cactus pear (Opuntia ficus-indica) productivity. METHODS: The developmental progression of C3 photosynthesis and CAM was assessed in seedlings and daughter cladodes of mature plants by titratable acidity, δ13C isotopic values and diel gas exchange measurements. KEY RESULTS: Nocturnal acidification was observed in seedling cladodes and cotyledons at the earliest stages of development and became highly significant by 75 days of development. Seedling cotyledons showed mean δ13C values of -21.4 and -17.1 ‰ at 30 and 100 days of age, respectively. Seedling cladodes showed mean δ13C values of -19.4 and -14.5 ‰ at 30 and 100 days of age, respectively. These values are typical of CAM plants. Net CO2 assimilation was negative, then occurred in both the day and the night, with nighttime fixation becoming predominant once the primary cladode reached 5 cm in size. Emergent daughter cladodes growing on mature plants showed nocturnal titratable acidity at the earliest stages of development, which became significant when daughter cladodes were >2.5-5 cm in height. Emergent daughter cladodes showed mean δ13C values of -14.5 to -15.6 ‰, typical of CAM plants. CO2 assimilation studies revealed that net CO2 uptake was negative in daughter cladodes <12 cm in length, but then exhibited net positive CO2 assimilation in both the day and the night, with net nocturnal CO2 assimilation predominating once the daughter cladode grew larger. CONCLUSIONS: Developing O. ficus-indica primary and daughter cladodes begin as respiring sink tissues that transition directly to performing CAM once net positive CO2 fixation is observed. Overall, these results demonstrate that CAM is the primary form of photosynthetic carbon assimilation for O. ficus-indica even at the earliest stages of seedling or daughter cladode development.

Potential of yeasts as biocontrol agents against Fusarium graminearum in vitro and on corn.

AIMS: The antifungal effect of the yeast species Kluyveromyces marxianus, Meyerozyma caribbica, and Wickerhamomyces anomalus was evaluated against two Fusarium graminearum strains (FRS 26 and FSP 27) in vitro and on corn seeds. METHODS AND RESULTS: The antifungal effect of the yeasts against F. graminearum was evaluated using scanning electron microscopy and extracellular chitinase and glucanase production to further elucidate the biocontrol mode of action. In addition, the germination percentage and vigor test were investigated after applying yeast on corn seeds. All the yeast strains inhibited fungal growth in vitro (57.4%-100.0%) and on corn seeds (18.9%-87.2%). In co-culture with antagonistic yeasts, F. graminearum showed collapsed hyphae and turgidity loss, which could be related to the ability of yeasts to produce chitinases and glucanases. The three yeasts did not affect the seed corn germination, and W. anomalus and M. caribbica increased corn seed growth parameters (germination percentage, shoot and root length, and shoot dry weight). CONCLUSION: Meyerozyma caribbica and W. anomalus showed satisfactory F. graminearum growth inhibition rates and did not affect seed growth parameters. Further studies are required to evaluate the application of these yeasts to the crop in the field.

Modulation of carotenoid biosynthesis in maize (Zea mays L.) seedlings by exogenous abscisic acid and salicylic acid under low temperature.

KEY MESSAGE: Abscisic acid could regulate structural genes in the carotenoid biosynthesis pathway and alleviate the decrease of carotenoids in maize seedlings under low-temperature stress. Low temperature often hampers the development of maize seedlings and hinders the accumulation of carotenoids, which are functional against chilling stress for plants and providing health benefits for human. To explore effective approaches in reducing chilling stress and enhancing the potential nutritional values of maize seedlings, exogenous plant hormones abscisic acid (ABA) and salicylic acid (SA) that may affect carotenoid biosynthesis were applied on low-temperature-stressed maize seedlings. Results showed that low temperature significantly reduced the carotenoid levels in maize seedlings, only preserving 62.8% in comparison to the control. The applied ABA probably interacted with the ABA-responsive cis-acting elements (ABREs) in the promoter regions of PSY3, ZDS and CHYB and activated their expressions. Consequently, the total carotenoid concentration was apparently increased to 1121 ± 47 ng·g-1 fresh weight (FW), indicating the stress alleviation by ABA. The application of SA did not yield positive results in alleviating chilling stress in maize seedlings. However, neoxanthin content could be notably boosted to 52.12 ± 0.45 ng·g-1 FW by SA, offering a biofortification strategy for specific nutritional enhancement. Structural gene PSY1 demonstrated positive correlations with ß-carotene and zeaxanthin (r = 0.93 and 0.89), while CRTISO was correlated with total carotenoids (r = 0.92), indicating their critical roles in carotenoid accumulation. The present study exhibited the effectiveness of ABA to mitigate chilling stress and improve the potential nutritional values in low-temperature-stressed maize seedlings, thereby promoting the production of plant-based food sources.

Elicitor Activity of Low-Molecular-Weight Alginates Obtained by Oxidative Degradation of Alginates Extracted from Sargassum muticum and Cystoseira myriophylloides.

Alginates extracted from two Moroccan brown seaweeds and their derivatives were investigated for their ability to induce phenolic metabolism in the roots and leaves of tomato seedlings. Sodium alginates (ALSM and ALCM) were extracted from the brown seaweeds Sargassum muticum and Cystoseira myriophylloides, respectively. Low-molecular-weight alginates (OASM and OACM) were obtained after radical hydrolysis of the native alginates. Elicitation was carried out by foliar spraying 20 mL of aqueous solutions (1 g/L) on 45-day-old tomato seedlings. Elicitor capacities were evaluated by monitoring phenylalanine ammonia-lyase (PAL) activity, polyphenols, and lignin production in the roots and leaves after 0, 12, 24, 48, and 72 h of treatment. The molecular weights (Mw) of the different fractions were 202 kDa for ALSM, 76 kDa for ALCM, 19 kDa for OACM, and 3 kDa for OASM. FTIR analysis revealed that the structures of OACM and OASM did not change after oxidative degradation of the native alginates. These molecules showed their differential capacity to induce natural defenses in tomato seedlings by increasing PAL activity and through the accumulation of polyphenol and lignin content in the leaves and roots. The oxidative alginates (OASM and OACM) exhibited an effective induction of the key enzyme of phenolic metabolism (PAL) compared to the alginate polymers (ALSM and ALCM). These results suggest that low-molecular-weight alginates may be good candidates for stimulating the natural defenses of plants.

Responses of maize (Zea mays L.) seedlings growth and physiological traits triggered by polyvinyl chloride microplastics is dominated by soil available nitrogen.

As a burgeoning pollutant, microplastics (MPs) has elicited global concern. However, ecological effects and mechanisms of MPs on plant-soil system are still poorly understood. In the present study, the impacts of polyvinyl chloride microplastics (PVC-MPs) on maize (Zea mays L.) seedlings growth and physiological traits and soil properties were discussed through a 30-day pot experiment. Results showed that PVC-MPs had greater toxicity effect on seedlings shoot biomass than root biomass. To defense the impact of PVC-MPs, the superoxide dismutase and catalase activities in seedlings leaf were stimulated. Moreover, the adhesion of MPs on soil particles increased, and soil microorganism, enzymes, and nutrients were altered significantly with increasing content of PVC-MPs. Notably, soil nitrate nitrogen decreased significantly with increasing content of PVC-MPs, whereas soil ammonium nitrogen was promoted under lower contents (0.1% and 1%) of PVC-MPs. Redundancy analysis indicated that soil nitrate nitrogen and ammonium nitrogen can explain 87.4% and 7.7% of variation in maize seedlings growth and physiological traits, respectively. These results display that maize seedlings shoot is more susceptible to the impact of PVC-MPs and soil available nitrogen is the primary limiting factor on maize seedlings growth and physiological traits triggered by PVC-MPs. Impacts of PVC-MPs on maize seedlings growth and physiological traits by nitrogen depletion lead to the possible yield and economic loess and potential risks due to the over use of nitrogen fertilizers.

First Report of Neopestalotiopsis clavispora causing Leaf Spot on Coconut Seedling in Malaysia.

Coconut (Cocos nucifera) is a high economic value cash crop in Malaysia. In December 2021, irregular spots with dotted rust-like appearance were observed mainly on the tip of the leaves of MATAG variety coconut seedlings at the nursery in Perak state. More than 90% of the coconut seedlings surveyed were infected with leaf spot symptoms. These symptoms could bring huge economic losses due to the downgrade value of the seedlings. 15 symptomatic leaves were obtained from the nursery, 10 mm2 of cut leaves were disinfected with 10% sodium hypochlorite for 10 minutes and rinsed with sterile distilled water before plated on potato dextrose agar (PDA). A total of 4 single-spore isolates were obtained and were observed morphologically. The isolates had white cotton-like appearance with undulate edge. Black acervuli were seen after 7 days of incubation at 26 °C. The conidia were fusiform and contained five cells with four septate and three versicolor cells in between the apical and basal cell. The conidia were 17.2 µm long and 5.9 µm wide (n=30). Conidia consisted of two to three apical appendages and one basal appendage. These morphological characters were consistent with the original description of Neopestalotiopsis clavispora (Santos et al., 2019; Abbas et al., 2022). Species identification was done by amplifying internal transcribed spacer (ITS) region using primers ITS 4 and ITS 5 (White et al., 1990) and beta-tubulin (TUB2) using primers Bt2a and Bt2b (Glass & Donaldson et al., 1995) of the representative isolate LKR1, then sequenced. The 488 bp ITS and 409 bp TUB2 sequences were deposited in GenBank under the accession numbers ON844193 and OP004810, respectively. Isolate LKR1 shares 99.8% identity with the ITS sequence (MH860736.1) of the reference pathogenic N. clavispora strain CBS:447.73 and 100% identity with the TUB2 sequence (KM199443.1) of the reference pathogenic N. clavispora strain CBS 447.73. The phylogenetic analysis confirmed that the isolate LKR1 belonged to N. clavispora when a supported clade is formed with 98% and 94% bootstrap support for ITS and TUB2 respectively with other related N. clavispora. Pathogenicity test was conducted by using five replicates of 8 month old seedlings, they were incubated under greenhouse condition and were watered daily. The leaves of the seedlings were injured with sterile needles and were sprayed with conidial suspension (1 x 10^6 conidia/ml). The control plants were also injured but sprayed with sterile distilled water. After a month, signature symptoms of spots on the leaves appear but none on the control seedling. N. clavispora was successfully re-isolated only from the inoculated symptomatic leaves and identified morphologically. No fungus was re-isolated from the control seedlings. The result was consistent even after repeating the test one more time. N. clavispora has been reported causing leaf spot on Macadamia integrifolia (Santos et al., 2019), Phoenix dactylifera L. (Basavand et al., 2020) and Musa acuminata (Qi et al., 2022). N. clavispora has also been reported causing rust-like appearance of leaves on strawberry (Fragaria × ananassa Duch.) (Obregón et al., 2018). To our knowledge, this is the first report of N. clavispora causing leaf spot disease on coconut seedlings in Malaysia. Through the identification of N. clavispora as the causal agent of leaf spot on coconut, this can help coconut growers to tackle the disease problem earlier thus, preventing the disease from spreading until the adult phase.

Study on Contact Stress Distribution Characteristics and Damage of Plug Seedlings Based on Flexible Pressure Sensor.

At present, there is a problem that the growth quality is reduced due to damage to the plug seedling pot during the transplanting process. In this study, the pressure distribution measurement system was used to measure the contact area of plug seedlings when they collided with the ground. The effects of seedling age and forward speed on the characteristics of contact stress distribution and potting damage were investigated through a single-factor experiment. The results were comprehensively considered based on the single-factor test, and the Box-Behnken test was used to optimize the design. The matrix loss rate was used as the evaluation index to determine the optimal parameter combination for transplanting: the tray specification was 72, the seedling age was 30 d, and the forward speed was 1.25 km·h-1. This study can provide a reference and technical support for further research on pot damage in plug seedling transplanting. The optimized parameters can provide practical guidance for reducing pot damage and improving growth quality during transplanting plug seedlings.

Multiomics Reveals the Regulatory Mechanisms of Arabidopsis Tissues under Heat Stress.

Understanding the mechanisms of responses to high temperatures in Arabidopsis will provide insights into how plants may mitigate heat stress under global climate change. And exploring the interconnections of different modification levels in heat stress response could help us to understand the molecular mechanism of heat stress response in Arabidopsis more comprehensively and precisely. In this paper, we combined multiomics analyses to explore the common heat stress-responsive genes and specific heat-responsive metabolic pathways in Arabidopsis leaf, seedling, and seed tissues. We found that genes such as AT1G54050 play a role in promoting proper protein folding in response to HS (Heat stress). In addition, it was revealed that the binding profile of A1B is altered under elevated temperature conditions. Finally, we also show that two microRNAs, ath-mir156h and ath-mir166b-5p, may be core regulatory molecules in HS. Also elucidated that under HS, plants can regulate specific regulatory mechanisms, such as oxygen levels, by altering the degree of CHH methylation.