Maize Roots Research Articles

Fermentation broth of a novel endophytic fungus enhanced maize salt tolerance by regulating sugar metabolism and phytohormone biosynthesis or signaling

Soil salinization is a major environmental factor that severely affects global agriculture. Root endophytes can enter root cells, and offer various ecological benefits, such as promoting plant growth, improving soil conditions, and enhancing plant resistance. Su100 is a novel strain of endophytic fungus that was characterized from blueberry roots. In this study, we focused on evaluating the effects of Su100 secretion on maize growth. The results demonstrated that maize treated with Su100 fermentation broth (SFB) exhibited significantly stronger salt tolerance than the control. It is worth mentioning that the treated root system not only had an advantage in terms of biomass but also a change in root structure with a significant increase in lateral roots (LRs) compared to the control. Transcriptome analysis combined with hormone content measurements indicated that SFB upregulated the auxin signaling pathway, and also caused alterations in brassinosteroids (BR) and jasmonic acid (JA) biosynthesis and signaling pathways. Transcriptome analyses also indicated that SFB caused significant changes in the sugar metabolism of maize roots. The major changes included: enhancing the conversion and utilization of sucrose in roots; increasing carbon flow to uridine diphosphate glucose (UDPG), which acted as a precursor for producing more cell wall polysaccharides, mainly pectin and lignin; accelerating the tricarboxylic acid cycle, which were further supported by sugar content determinations. Taken together, our results indicated that the enhanced salt tolerance of maize treated with SFB was due to the modulation of sugar metabolism and phytohormone biosynthesis or signaling pathways. This study provided new insights into the mechanisms of action of endophytic fungi and highlighted the potential application of fungal preparations in agriculture.

Mechanisms of ROS-mediated interactions between Bacillus aryabhattai LAD and maize roots to promote plant growth

BackgroundPlant growth-promoting rhizobacteria (PGPR), as a group of environmentally friendly bacteria growing in the rhizosphere of plants, play an important role in plant growth and development and resistance to environmental stresses. However, their limited understanding has led to the fact that their large-scale use in agriculture is still scarce, and the mechanisms by which beneficial bacteria are selected by plants and how they interact with them are still unclear.MethodIn this study, we investigated the interaction between the auxin-producing strain Bacillus aryabhattai LAD and maize roots, and performed transcriptomic and metabolomic analyses of Bacillus aryabhattai LAD after treatment with maize root secretions(RS).ResultsOur results show that there is a feedback effect between the plant immune system and bacterial auxin. Bacteria activate the immune response of plant roots to produce reactive oxygen species(ROS), which in turn stimulates bacteria to synthesize IAA, and the synthesized IAA further promotes plant growth. Under the condition of co-culture with LAD, the main root length, seedling length, root surface area and root volume of maize increased by 197%, 107%, 89% and 75%, respectively. In addition, the results of transcriptome metabolome analysis showed that LAD was significantly enriched in amino acid metabolism, carbohydrate metabolism and lipid metabolism pathways after RS treatment, including 93 differentially expressed genes and 45 differentially accumulated metabolites.ConclusionOur findings not only provide a relevant model for exploring the effects of plant-soil microbial interactions on plant defense functions and thereby promoting plant growth, but also lay a solid foundation for the future large-scale use of PGPR in agriculture for sustainable agricultural development.

Rhizosphere flavonoids alleviates the inhibition of soybean nodulation caused by shading under maize-soybean strip intercropping

The flavonoids produced by legume roots are signal molecules that induce nod genes for symbiotic rhizobium. Nevertheless, the promoting effects of flavonoids in root exudates in intercropping system on soybean nodulation are still unknown. A two years of field experiments was carried with maize soybean strip intercropping, i.e., the interspecific row spacing of 30 cm (MS30), 45 cm (MS45), 60 cm (MS60), and sole soybean/maize:SS/MM, and root interaction, i.e., root no barrier (NB) and root polythene-plastic barrier (PB), to evaluate relationships between flavonoids in root exudates and nodulation. We found that root-root interaction between soybean and maize enhances the nodules number and fresh weight in intercropped soybean. This enhancement increase gradually with expansion of interspecific distance. Proportion of nodules with diameter greater than 0.4cm was higher in intercropped soybean with NB than with PB. The expressions of nodules-related genes (GmENOD40, GmNIN2b and GmEXPB2) were up-regulated. Furthermore, compared with monocropping, isoflavones secretion of soybean roots reduced, flavonoids and flavonols secretion of maize and soybean roots increased under intercropping. The secretion of differential metabolites of flavonoids in the rhizosphere of maize and soybean declined with root barrier. The expressions of GmCHS8 and GmIFS1 in soybean roots were up-regulated and GmICHG was down-regulated under root interaction. The most of the flavonoids and flavonol compounds were positively correlated with nodule diameter. The nodules number, the nodules fresh weight and the proportion of nodules with a diameter greater than 0.2 cm increased in different genotypes of soybean treated with maize root exudate, which promoted the improvement of nitrogen fixation capacity. Therefore, maize-soybean strip intercropping combined with reasonable spacing to enhance the positive effect of underground root interaction, and improve the nodulation and nitrogen fixation capacity of intercropping soybean.

Root Anatomy: Preparing, Imaging, and Analyzing Maize Root Cross-Sections.

Root anatomy plays a critical structural and functional role in the maize root system, and regulates edaphic stress tolerance. The function and genetic basis of several maize root anatomical traits for stress tolerance have been demonstrated. Leveraging root anatomical traits in maize thus holds great potential for developing cultivars with greater nutrient and water efficiency. Key for such approaches is the ability to characterize the root anatomy of plants of interest. Here, we outline a systematic method for preparing, imaging, and analyzing maize root cross-sections. The protocol describes root sectioning (by hand or using a vibratome), preparation of microscope slides and toluidine blue staining, imaging under a light microscope, and both manual and semiautomated methods for anatomical feature extraction from images. The protocol enables the visualization and quantification of various anatomical tissues and traits, and its simplicity, adaptability, and accessibility make it an ideal choice for both small- and large-scale phenotyping studies in maize and other plant species. This standardized protocol provides researchers with a comprehensive methodology to accurately dissect root structures, enabling in-depth analyses that are essential for understanding plant growth, development, and adaptive value for stress tolerance.

Root Anatomical Imaging and Phenotyping in Maize.

Root anatomy plays a crucial role in regulating essential processes such as the absorption and movement of water and nutrients in plants. Root anatomy also impacts the energy costs of building and sustaining root tissues, tissue mechanics, and interactions with other organisms. Although several studies in maize have confirmed the functional utility of numerous root anatomical traits, such as that of cortical cell size and number for stress adaptation, there have been significant obstacles in measuring and analyzing root anatomical characteristics. This has resulted in gaps in our understanding of the genetic control and range of phenotypic variations among different cultivars, and how this diversity relates to overall fitness. Here, we review root anatomical phenotypes in maize and their function in stress adaptation, and briefly discuss phenotyping methods available for root anatomy. We further introduce a simple and accessible phenotyping approach that enables a comprehensive investigation of maize root anatomy. Detailed characterization of root traits and the implementation of robust methods for root anatomical phenotyping could have wide-ranging benefits across various areas of plant science, from fundamental research to enhancing crop breeding efforts.

Improving the productivity of intercropping through above and below ground separation: A case study on photosynthetic characteristics and root distribution

Soybean and maize have important industrial applications, as their yield and production distribution plays an important role in supporting the global agricultural industries. In maize and soybean intercropping system, yield optimization relies on a comprehensive understanding of the complex interactions between above-ground and below-ground processes. Therefore, to investigate the impacts of various planting and separation methods on the spatial distribution of roots, soil moisture, nutrient uptake, photosynthetic physiology of leaves at different positions as well as yield advantages in soybean and maize intercropping, a two-year field experiment was conducted. The results indicted that maize root length density (RLD) increased by 24.3 %-25.5 % in non-separated maize rows and by 25.5 %-30.2 % in 10 cm from the maize rows as compared to the below-ground separated. While soybean RLD was reduced by 13.2–16.5 % in soybean rows and by 34.5 %-36.5 % in 10 cm from soybean rows. The change of the above parameters resulted in a noteworthy decrease in soil water content within the rows of maize, which was effectively mitigated by the below-ground separation. The photosynthetic rate (Pn) in middle and lower leaves of maize increased by 51.1 % and 39.2 % in above and below-ground separation, while less in upper leaves. Above-ground separation can alleviate the decrease of Pn in the upper and middle leaves of soybean, which was only decreased by 2.7 % and 3.0 % as compared with monocropping. The main constraint on maize production is the Pn of the middle and lower leaves, the upper leaves being secondary factors. Intercropping enhanced chlorophyll fluorescence parameters, while below-ground separation decreased it in the middle and lower leaves of maize. However, above-ground separation increased fluorescence characteristics in soybeans across all leaves. Intercropped maize benefited more from the below-ground than above-ground advantages, while the opposite was true for soybeans. The above-ground separation improves the overall LER and contributes the most to the yield advantages.

TopoRoot+: computing whorl and soil line traits of field-excavated maize roots from CT imaging

BackgroundThe use of 3D imaging techniques, such as X-ray CT, in root phenotyping has become more widespread in recent years. However, due to the complexity of the root structure, analyzing the resulting 3D volumes to obtain detailed architectural root traits remains a challenging computational problem. When it comes to image-based phenotyping of excavated maize root crowns, two types of root features that are notably missing from existing methods are the whorls and soil line. Whorls refer to the distinct areas located at the base of each stem node from which roots sprout in a circular pattern (Liu S, Barrow CS, Hanlon M, Lynch JP, Bucksch A. Dirt/3D: 3D root phenotyping for field-grown maize (zea mays). Plant Physiol. 2021;187(2):739–57. https://doi.org/10.1093/plphys/kiab311.). The soil line is where the root stem meets the ground. Knowledge of these features would give biologists deeper insights into the root system architecture (RSA) and the below- and above-ground root properties.ResultsWe developed TopoRoot+, a computational pipeline that produces architectural traits from 3D X-ray CT volumes of excavated maize root crowns. Building upon the TopoRoot software (Zeng D, Li M, Jiang N, Ju Y, Schreiber H, Chambers E, et al. Toporoot: A method for computing hierarchy and fine-grained traits of maize roots from 3D imaging. Plant Methods. 2021;17(1). https://doi.org/10.1186/s13007-021-00829-z.) for computing fine-grained root traits, TopoRoot + adds the capability to detect whorls, identify nodal roots at each whorl, and compute the soil line location. The new algorithms in TopoRoot + offer an additional set of fine-grained traits beyond those provided by TopoRoot. The addition includes internode distances, root traits at every hierarchy level associated with a whorl, and root traits specific to above or below the ground. TopoRoot + is validated on a diverse collection of field-grown maize root crowns consisting of nine genotypes and spanning across three years. TopoRoot + runs in minutes for a typical volume size of \\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\:40{0}^{3}$$\\end{document} on a desktop workstation. Our software and test dataset are freely distributed on Github.ConclusionsTopoRoot + advances the state-of-the-art in image-based phenotyping of excavated maize root crowns by offering more detailed architectural traits related to whorls and soil lines. The efficiency of TopoRoot + makes it well-suited for high-throughput image-based root phenotyping.

Cold mediates maize root hair developmental plasticity via epidermis-specific transcriptomic responses.

Cold stress during early development limits maize (Zea mays L.) production in temperate zones. Low temperatures restrict root growth and reprogram gene expression. Here, we provide a systematic transcriptomic landscape of maize primary roots, their tissues, and cell types in response to cold stress. The epidermis exhibited a unique transcriptomic cold response, and genes involved in root hair formation were dynamically regulated in this cell type by cold. Consequently, activation of genes involved in root hair tip growth contributed to root hair recovery under moderate cold conditions. The maize root hair defective mutants roothair defective 5 (rth5) and roothair defective 6 (rth6) displayed enhanced cold tolerance with respect to primary root elongation. Furthermore, dehydration response element-binding protein 2.1 (dreb2.1) was the only member of the dreb subfamily of AP2/EREB transcription factor genes upregulated in primary root tissues and cell types but exclusively downregulated in root hairs upon cold stress. Plants overexpressing dreb2.1 significantly suppressed root hair elongation after moderate cold stress. Finally, the expression of rth3 was regulated by dreb2.1 under cold conditions, while rth6 transcription was regulated by dreb2.1 irrespective of the temperature regime. We demonstrated that dreb2.1 negatively regulates root hair plasticity at low temperatures by coordinating the expression of root hair defective genes in maize.

Spent coffee waste-derived biochar improves physical properties, water retention, and maize (Zea mays L.) growth in sandy soil

Adding organic soil amendments can improve the physical and hydrological properties of soil, subsequently enhancing fertility for better crop production. In this study, spent Arabica and Columbian coffee wastes and their respective biochars were evaluated as soil amendments to improve the physical and hydrological properties of loamy sand soil and enhance maize (Zea mays L.) crop growth. Spent Arabica coffee (AC) and Columbian coffee (CC) wastes were collected and transformed into biochar through pyrolysis process at 550 °C with a residence time of 3 h and pyrolysis rate of 5 °C per minute. The AC and CC derived biochar were termed as ABC and CBC, respectively. The produced soil amendments were applied to soil at 0% (control), 1%, 3%, and 5% in a column setup. The moisture characteristics, including water infiltration, evaporation, and water retention, were investigated. Thereafter, the prepared amendments were applied to loamy sand soils at 0% (control), 1%, 3%, and 5% (w/w) application rates. Maize growth was then observed for a period of 30 days under greenhouse conditions. Results of the column trials showed that ABC and CBC applied at 5% reduced the cumulative water evaporation by 57%–66% and cumulative infiltration by 124%–181% compared to control. Likewise, 5% application of ABC and CBC resulted in 101 to 130% higher water retention in loamy sand soil. Results of the greenhouse experiment showed that 5% application of ABC and CBC amendments resulted in root biomass of 2.12 and 2.38 g, respectively, as compared to 0.51 g in control treatment. Similar treatments resulted in shoot biomass of 9.70 and 9.93 g respectively, as compared to 7.37 g in control. Likewise, 5% application of CBC and ABC increase plant height from 15.71 to 30.94 cm in ABC and 33.23 cm in CBC. Overall, 5% application of coffee waste-derived biochars significantly reduced water evaporation and infiltration, while increasing soil water retention and maize plant height, root biomass, and shoot biomass. Therefore, spent coffee waste-derived biochar could effectively be employed to improve physical and hydrological properties of loamy sand soils for better crop productivity.

MiRNA Sequencing Analysis in Maize Roots Treated with Neutral and Alkaline Salts.

Soil salinization/alkalization is a complex environmental factor that includes not only neutral salt NaCl but also other components like Na2CO3. miRNAs, as small molecules that regulate gene expression post-transcriptionally, are involved in plant responses to abiotic stress. In this study, maize seedling roots were treated for 5 h with 100 mM NaCl, 50 mM Na2CO3, and H2O, respectively. Sequencing analysis of differentially expressed miRNAs under these conditions revealed that the Na2CO3 treatment group had the most differentially expressed miRNAs. Cluster analysis indicated their main involvement in the regulation of ion transport, binding, metabolism, and phenylpropanoid and flavonoid biosynthesis pathways. The unique differentially expressed miRNAs in the NaCl treatment group were related to the sulfur metabolism pathway. This indicates a significant difference in the response patterns of maize to different treatment groups. This study provides theoretical evidence and genetic resources for further analysis of the molecular mechanisms behind maize's salt-alkali tolerance.

Short-term microbial community dynamics induced by 13C-labeled maize root, its derived biochar and NPK in long-term amended soil

Crop residues and their derived biochar are frequently used for their potential to improve grain yield, soil fertility and carbon (C) sequestration. However, the effects of root are often overlooked, and the effects of chemical fertilizer (NPK) combined with root or its biochar on microbial community structure need further study. This study used 13C-labeled maize root, its biochar and soil with different fertilization for 8 years as materials and substrates. A 112-day incubation experiment was conducted to explore the effects of microbial community on the C processing. During incubation, the root-C (54.9%) mineralized significantly more than biochar-C (12.8%), while NPK addition significantly increased the root-C mineralization. Adding biochar alone did not significantly change the microbial community. Compared to the biochar treatment (BC), the root treatment (R) notably increased the contents of total phospholipid fatty acids (PLFAs), 13C-PLFA and the proportion of fungi and Gram-negative bacteria, but reduced the proportion of actinomycetes. The root mineralization was significantly correlated with the relative content of 13C-Gram-positive bacteria and 13C-fungi, while biochar mineralization was significantly correlated with the relative content of 13C-Gram-positive bacteria and 13C-actinomycetes. Notably, NPK addition significantly increased the contribution of biochar-C to PLFA-C pool, while decreasing the contribution of root-C. In summary, due to microbial adaptation to the lack of bioavailable C in biochar-amended soil, biochar can act as a buffer against the significant disturbance caused by NPK to microbial communities and native soil organic carbon (SOC), which contributes to the steady enhancement in soil C storage.Graphical

Analysis of Bacterial Community Characteristics in Maize Root Zones Under Maize-soybean Compound Planting Mode

Maize-soybean compound intercropping has the potential to increase yield and is being tested for spreading in Huang-Huai-hai Plain. However, the main regulatory regions of this cropping pattern on soil microbial communities have not been clarified. In the present study, the tested samples were collected from three maize root zones of bulk soil, rhizosphere soil, and roots under mono- and intercropping planting modes, respectively. The non-rhizosphere soil chemical properties and enzyme activities were determined, and bacterial communities were characterized using high-throughput sequencing of the 16S rRNA gene V3-V4 region. Compared with monocropping, the maize bulk soil electric conductivity （EC）, soil organic matter （SOM）, available potassium （AK）, available phosphorus （AP）, total nitrogen （TN）, and enzyme activities of intercropping were significantly increased. The α diversities and β diversity of the bacterial community in rhizosphere soil were significantly different between the two planting modes. There were 11 bacteria genera with significantly higher abundance in the rhizosphere soil of compound planting than that of monoculture, and TN, AP, and catalase were the three most important factors contributing to their distribution. The abundances of 8 genera among the 11 genera mentioned above, unclassified Vicinamibacterales, unclassified Geminicoccaceae, MND1, unclassified Gemmatimonadaceae, Acidibacter, unclassified Vicinamibacteraceae, Sphingomonas, and unclassified Comamonadaceae were significantly positively correlated with TN. As for the bacteria distribution in maize root, AK contributed the most and had a significantly negative correlation with unclassified Rhizobiaceae and unclassified Microscillaceae and a positive correlation with Haliangium. Maize-soybean compound intercropping affected mainly the bacterial community of maize rhizosphere and had an evident effect on soil fertilizer cultivation and microbial diversity regulation, which provides a theoretical basis and practical guidance for rational intercropping to maintain agroecosystem biodiversity.

The phytosiderophore analogue proline-2′-deoxymugineic acid is more efficient than conventional chelators for improving iron nutrition in maize

ABSTRACT Iron (Fe) is an essential nutrient but has poor bioavailability because of its low solubility under conditions of high pH, such as calcareous soils. Previously, we synthesized an analogue of the natural Fe(III) chelator 2′-deoxymugineic acid secreted by graminaceous plants for efficient Fe uptake, designated as proline-2′-deoxymugineic acid (PDMA). Soil application of Fe(III)-PDMA ameliorated symptoms of Fe deficiency in rice. In the present study, we explored the potential of PDMA as Fe nutrition supplement in maize, a major graminaceous crop, and rice for comparison. In calcareous soil pots, Fe deficiency chlorosis of maize was efficiently recovered by a single application of Fe(III)-PDMA. In contrast, maize plants treated with conventional Fe(III)-chelator complexes, such as Fe(III)-ethylenediaminetetraacetic acid (EDTA) or Fe(III)-N,N′-di(2-hydroxybenzyl)ethylenediamine-N,N′-diacetic acid monohydrochloride (HBED), showed little or no recovery. Similarly, rice Fe chlorosis in calcareous soil pots was efficiently recovered by a single application of Fe(III)-PDMA, but not by other conventional Fe(III)-chelator complexes. Application of Fe(III)-PDMA was also effective in ameliorating Fe deficiency chlorosis in hydroponically grown maize seedlings; other Fe(III)-chelator complexes showed minimal or no efficacy. Addition of low concentrations of metal-free PDMA recovered Fe chlorosis of maize or rice in the presence of Fe(III)-EDTA, suggesting possible ligand exchange from EDTA to PDMA for subsequent Fe(III)-PDMA uptake by ZmYS1 and OsYSL15 transporters. Similar effects were observed with Fe(III)-HBED, which has a higher stability constant, but to a lesser extent than Fe(III)-EDTA, suggesting that ligand exchange from HBED to PDMA might be less prone to occur than for chelators with moderate stability constants, such as EDTA. Ferric-chelate reductase assays of maize roots showed substantial reduction of Fe(III)-PDMA, but this reduction activity was not increased under Fe-deficient condition. These results suggested that PDMA is an efficient reagent for improvement of Fe nutrition in graminaceous crops, including maize, because of suitability of Fe(III)-PDMA as a substrate for chelation-based Fe uptake systems.

The lactonase BxdA mediates metabolic specialisation of maize root bacteria to benzoxazinoids

Root exudates contain specialised metabolites that shape the plant’s root microbiome. How host-specific microbes cope with these bioactive compounds, and how this ability affects root microbiomes, remains largely unknown. We investigated how maize root bacteria metabolise benzoxazinoids, the main specialised metabolites of maize. Diverse and abundant bacteria metabolised the major compound in the maize rhizosphere MBOA (6-methoxybenzoxazolin-2(3H)-one) and formed AMPO (2-amino-7-methoxy-phenoxazin-3-one). AMPO forming bacteria were enriched in the rhizosphere of benzoxazinoid-producing maize and could use MBOA as carbon source. We identified a gene cluster associated with AMPO formation in microbacteria. The first gene in this cluster, bxdA encodes a lactonase that converts MBOA to AMPO in vitro. A deletion mutant of the homologous bxdA genes in the genus Sphingobium, did not form AMPO nor was it able to use MBOA as a carbon source. BxdA was identified in different genera of maize root bacteria. Here we show that plant-specialised metabolites select for metabolisation-competent root bacteria. BxdA represents a benzoxazinoid metabolisation gene whose carriers successfully colonize the maize rhizosphere and thereby shape the plant’s chemical environmental footprint.

The impact of water hyacinth biochar on maize growth and soil properties: The influence of pyrolysis temperature

AbstractIntroductionOptions for managing water hyacinths (WHs) include converting the biomass into biochar for soil amendment. However, less has been known about the impact of WH‐based biochar developed in varying pyrolysis temperatures on plant growth and soil qualities.Materials and MethodsA pot experiment was undertaken in a factorial combination of WH biochars (WHBs) developed at three temperatures (350°C, 550°C and 750°C) and two application rates (5 and 20 t ha−1), plus a control without biochar. Maize was grown as a test crop for 2 months under natural conditions.ResultsOur study showed that applying WHB developed between 350°C and 750°C at 20 t ha−1 increased maize shoot and root dry biomass by 47.7% to 17.6% and 78.4% to 54.1%, respectively. Nevertheless, raising the biochar pyrolysis temperature decreased maize growth, whereas increasing the application rate displayed a positive effect. The application of WHB generated at 350°C and 550°C at 20 t ha−1 resulted in significant improvements in soil total nitrogen (17.9% to 25%), cation exchange capacity (27.3% to 20.2%), and ammonium‐nitrogen (60.7% to 59.6%), respectively, over the control. Additionally, applying WHB produced from 350°C to 750°C at 20 t ha−1 enhanced soil carbon by 38.5%–56.3%, compared to the control. Conversely, applying biochar produced at 750°C resulted in higher soil pH (6.3 ± 0.103), electrical conductivity (0.23 ± 0.01 dS m−1) and available phosphorus (21.8 ± 2.53 mg kg−1).ConclusionWHBs developed at temperatures of 350°C and 550°C with an application rate of 20 t ha−1 were found to be optimal for growing maize and improving soil characteristics. Our study concludes that pyrolysis temperature significantly governs the effectiveness of biochar produced from a specific biomass source.

Arbuscular Mycorrhizal Fungal Interacted with Biochar and Enhanced Phosphate-Solubilizing Microorganism Abundance and Phosphorus Uptake in Maize

In agricultural production, it is crucial to increase the availability of phosphorus (P) in cultivated soil to solve the P limitation. Arbuscular mycorrhizal fungi (AMF) have been proven to promote crop nutrient absorption effectively, while biochar can lead to improvements in soil properties. However, the possible synergistic effect of AMF and biochar on P uptake by crops as well as its underlying mechanisms are unclear. In this study, we conducted a pot experiment to explore the effects of biochar and AMF (Glomus etunicatum) on the community of rhizospheric phosphate-solubilizing microorganisms (PSMs) of maize (Zea mays L. Xianyu-335) using metagenomic methods. The experiment used 0 mg P2O5 g·kg−1 soil (P0) and 30 mg P2O5 g·kg−1 soil (P30) application rates. Each P application rate included 0 (NC), 20 g·kg−1 biochar (BC) addition, inoculation AMF, and without AMF treatments (NM) for a total of eight treatments. During the experiment, both the P uptake and the biomass of maize were measured. The study found that the combination of AMF and biochar significantly increased the mycorrhizal colonization rate of maize roots, regardless of P application level. It was observed that the P uptake by maize was significantly increased when exposed to a combination of AMF and biochar. The increase in P uptake in P0 treatments was 67% higher than the sum of the effects of biochar and AMF inoculation alone. The increase was only 35% higher in P30 treatments, demonstrating a substantially higher interactive effect under P0 than under P30 conditions. The AM-BC treatments significantly increased the abundance of Streptomyces, Bacillus, and Pseudomonas, genera that are known to contain PSMs. In addition, the abundance of genes related to P-cycling (gcd, phoD, and ugpQ) in PSMs increased significantly by 1.5–1.8 times in AM-BC treatments compared with NM-BC and AM-NC treatments under P0 conditions. This increase was significantly and positively correlated with the P uptake. Overall, the results suggest that biochar can help AMF colonize the roots, increasing the functional roles of PSMs in the rhizosphere, which in turn promotes P uptake and biomass in maize. This study provides a new way to improve P-use efficiency and reduce the need for P-fertilizer application in agricultural production.

A phased small interfering RNA-derived pathway mediates lead stress tolerance in maize.

Phased small interfering RNAs (phasiRNAs) are a distinct class of endogenous small interfering RNAs, which regulate plant growth, development and environmental stress response. To determine the effect of phasiRNAs on maize (Zea mays L.) tolerance to lead (Pb) stress, the roots of 305 maize lines under Pb treatment were subjected to generation of individual databases of small RNAs. We identified 55 high-confidence phasiRNAs derived from 13 PHAS genes (genes producing phasiRNAs) in this maize panel, of which 41 derived from nine PHAS loci were negatively correlated with Pb content in the roots. The potential targets of the 41 phasiRNAs were enriched in ion transport and import. Only the expression of PHAS_1 (ZmTAS3j, Trans-Acting Short Interference RNA3) was regulated by its cis-expression quantitative trait locus and thus affected the Pb content in the roots. Using the Nicotiana benthamiana (N. benthamiana) transient expression system, 5'-rapid amplification of cDNA ends, and Arabidopsis heterologously expressed, we verified that ZmTAS3j was cleaved by zma-miR390 and thus generated tasiRNA targeting ARF genes (tasiARFs), and that the 5' and 3' zma-miR390 target sites of ZmTAS3j were both necessary for efficient biosynthesis and functional integrity of tasiARFs. We validated the involvement of the zma-miR390-ZmTAS3j-tasiARF-ZmARF3-ZmHMA3 pathway in Pb accumulation in maize seedlings using genetic, molecular, and cytological methods. Moreover, the increased Pb tolerance in ZmTAS3j-overexpressed lines was likely attributed to the zma-miR390-ZmTAS3j-tasiARF-ZmARF3-SAURs pathway, which elevated indole acetic acid levels and thus reactive oxygen species scavenging capacity in maize roots. Our study reveals the importance of the TAS3-derived tasiRNA pathway in plant adaptation to Pb stress.

Phenylpropanoid-rich maize root extract serves as a natural antidepressant

BackgroundDepression is a serious and complex mental disease that has attracted worldwide attention because of its high incidence rate, high disability rate and high mortality. Excitotoxicity is one of the most important mechanisms involved in the pathophysiological process of depression. In our previous studies, n-butanol extract from maize roots was found to have good neuroprotective effects due to its antioxidative activity. However, the antidepressive effective constituents, efficacy in vivo and mechanism of action of maize root extracts have not been determined. PurposeThis study aimed to determine the main active neuroprotective compound in maize root extract and investigate its antidepressant effects and possible underlying mechanism in vitro and in vivo. MethodsSixteen extracts were isolated and purified from maize roots. The active components of the most active extracts of maize roots (hereafter referred to as EM 2) were identified using UF-HPLC-QTOF/MS. In vitro cell models of NMDA-induced excitotoxicity in SH-SY5Y cells were used to analyze the anti-excitatory activity of the extracts. The MTT assay and Annexin V-FITC/PI Apoptosis Detection were used to evaluate cell viability. Several network pharmacological strategies have been employed to investigate the potential mechanism of action of EM 2. The effects of EM 2 on depressive-like behaviors were evaluated in CUMS mice. Changes in the levels of related proteins were detected via western blotting. ResultsAmong the 16 extracts extracted by n-butanol, EM 2 was determined to be the most active extract against NMDA-induced excitotoxicity by n-butanol extraction. Meanwhile, seventeen compounds were further identified as the main active components of EM 2. Mechanistically, EM 2 inhibited NMDA-induced excitatory injury in SH-SY5Y cells and alleviated the depressive-like behaviors of CUMS mice by suppressing NR2B and subsequently mediating the downstream CREB/TRKB/BDNF, PI3K/Akt and MAPK pathways, as well as the Nrf2/HO-1 antioxidant signaling pathway. ConclusionThe study indicated that EM 2 could potentially be developed as a potential therapeutic candidate to cure depression in NMDA-induced excitatory damage.

PIP family-based association studies uncover ZmPIP1;6 involved in Pb accumulation and water absorption in maize roots

Excessive lead (Pb) in the soil affects crop growth and development, thus threatening human beings via food chains. Plasma membrane intrinsic proteins (PIPs) facilitate the transport of substrates across cell membranes. Herein, we characterized maize PIPs and identified eight Pb accumulation-associated PIP genes using association studies. Among these, ZmPIP1;6 was simultaneously correlated with root Pb concentrations under various Pb treatment stages. Significant correlations were observed between the ZmPIP1;6 expression abundance and Pb accumulation in maize roots. Ectopic expression in yeast showed that ZmPIP1;6 conferred Pb accumulation in the cells and affected Pb tolerance in yeast. Overexpression in maize demonstrated that ZmPIP1;6 altered the Pb concentration performance and root moisture content under Pb stress. Meanwhile, protein interaction analyses suggested that ZmPIP1; 6 and three PIP2 members formed isoforms and facilitate water uptake in maize roots. However, ZmPIP1; 6 improved Pb absorption in maize roots probably by interacting with CASP-like protein 2C3 and/or another metal transporter. Moreover, the significant variants in the ZmPIP1;6 promoter caused the variations in ZmPIP1;6 expression level and Pb accumulation among various maize germplasms. Our study will contribute to understanding of PIP family-mediated Pb accumulation in crops and bioremediation of Pb-polluted soils.

Effects of AMF on Maize Yield and Soil Microbial Community in Sandy and Saline Soils.

This study aimed to investigate the effects of applying arbuscular mycorrhizal fungi (AMF) on maize root growth and yield formation under different soil conditions. This study was conducted under sandy soil (S) and saline-alkali soil (Y), with treatments of AMF application (AM) and no AMF application (CK). The root characteristics, yield, and quality of maize were measured. High-throughput sequencing technology was employed to assess the impact of AMF on the soil microbial community structure, and the correlation between soil microbes and soil physicochemical properties was elucidated. The results show that under both sandy and saline-alkali soil conditions, AMF application significantly enhanced maize root growth, yield, grain quality, and soil available nitrogen (AN), available phosphorus (AP), and available potassium (AK) contents compared to the CK treatment. Soil microbial Alpha diversity analysis indicated that AMF application effectively increased soil microbial diversity and richness. Principal coordinate analysis (PCoA) and microbial community structure analysis revealed significant differences in bacterial communities between AM treatment in sandy soil (SAM) and CK in sandy soil (SCK), and significant differences in both bacterial and fungal communities between AM treatment in saline-alkali soil (YAM) and CK in saline-alkali soil (YCK). Furthermore, significant correlations between microbial communities and soil physicochemical properties were found, such as AN, AP, AK, soil salinity (SS), and organic matter (OM) content. AMF application had a greater impact on bacterial communities than on fungal communities. This study demonstrated that the use of AMF as a bio-fungal fertilizer was effective in improving spring maize yields, especially in terms of yield increase and quality stability in sandy and saline soils, thereby contributing to safe and sustainable cropping practices.