Tolerant Maize Research Articles

Integrated physiological, biochemical and transcriptomic analyses reveal the mechanism of salt tolerance induced by a halotolerant Serratia sp. NTN6 in maize

Salt stress is a serious environmental challenge that hinders crop growth and yield. Plant growth-promoting rhizobacteria (PGPR) can successfully and environmentally friendly boost plant development in saline soils. In this study, a halotolerant Serratia sp. NTN6 was isolated from the rhizosphere soil of halophyte Puccinellia tenuiflora and its plant growth-promoting properties were determined. NTN6 inoculation significantly promoted seed germination and seedling growth in maize under salt stress. Shoot height, stem thickness, root fresh weight, shoot fresh weight, root dry weight, and shoot dry weight of salt-stressed seedlings increased by 36.3%, 32.5%, 146.4%, 193.6%, 25.0%, and 109.6%, respectively, after inoculation with NTN6. NTN6-mediated maize salt tolerance was revealed via transcriptomic, physiological, and biochemical analyses. Under salt stress, NTN6 regulated the transcript levels of genes related to chlorophyll synthesis, photosynthesis apparatus, and carbon assimilation pathways, and thus increasing chlorophyll content, Fv/Fm, and net photosynthetic rate; NTN6 reduced the content of O2•–, H2O2, and MDA by regulating the transcripts and activities of antioxidant enzymes; NTN6 affected the transcriptional response of genes involved in ABA synthesis and signaling to salt stress and reduced ABA level. Finally, NTN6 boosted photosynthetic capacity in salt-stressed maize by modulating photochemical activity, C4 pathway and Calvin cycle as well as mitigating reactive oxygen species damage to the photosystem reaction center. This study first investigated the mechanism of Serratia sp.-induced salt tolerance in maize and these findings demonstrate the valuable contribution of halotolerant PGPR strain from halophytes in improving salinity tolerance in non-halophytic crops.

Metabolism of the 4-Hydroxyphenylpyruvate Dioxygenase Inhibitor, Mesotrione, in Multiple-Herbicide-Resistant Palmer amaranth (Amaranthus palmeri).

Metabolic resistance to the maize-selective, HPPD-inhibiting herbicide, mesotrione, occurs via Phase I ring hydroxylation in resistant waterhemp and Palmer amaranth; however, mesotrione detoxification pathways post-Phase I are unknown. This research aims to (1) evaluate Palmer amaranth populations for mesotrione resistance via survivorship, foliar injury, and aboveground biomass, (2) determine mesotrione metabolism rates in Palmer amaranth populations during a time course, and (3) identify mesotrione metabolites including and beyond Phase I oxidation. The Palmer amaranth populations, SYNR1 and SYNR2, exhibited higher survival rates (100%), aboveground biomass (c.a. 50%), and lower injury (25-30%) following mesotrione treatment than other populations studied. These two populations also metabolized mesotrione 2-fold faster than sensitive populations, PPI1 and PPI2, and rapidly formed 4-OH-mesotrione. Additionally, SYNR1 and SYNR2 formed 5-OH-mesotrione, which is not produced in high abundance in waterhemp or naturally tolerant maize. Metabolite features derived from 4/5-OH-mesotrione and potential Phase II mesotrione-conjugates were detected and characterized by liquid chromatography-mass spectrometry (LCMS).

Heat-Resistant Inbred Lines Coordinate the Heat Response Gene Expression Remarkably in Maize (Zea mays L.).

High temperatures are increasingly becoming a prominent environmental factor accelerating the adverse influence on the growth and development of maize (Zea mays L.). Therefore, it is critical to identify the key genes and pathways related to heat stress (HS) tolerance in maize. Great challenges have been faced in dissecting genetic mechanisms and uncovering master genes for HS tolerance. Here, Z58D showed more thermotolerance than AF171 at the seedling stage with a lower wilted leaf rate and H2O2 accumulation under HS conditions. Transcriptomic analysis identified 3006 differentially expressed genes (DEGs) in AF171 and 4273 DEGs in Z58D under HS treatments, respectively. Subsequently, GO enrichment analysis showed that commonly upregulated genes in AF171 and Z58D were significantly enriched in the following biological processes, including protein folding, response to heat, response to temperature stimulus and response to hydrogen peroxide. Moreover, the comparison between the two inbred lines under HS showed that response to heat and response to temperature stimulus were significantly over-represented for the 1234 upregulated genes in Z58D. Furthermore, more commonly upregulated genes exhibited higher expression levels in Z58D than AF171. In addition, maize inbred CIMBL55 was verified to be more tolerant than B73, and more commonly upregulated genes also showed higher expression levels in CIMBL55 than B73 under HS. These consistent results indicate that heat-resistant inbred lines may coordinate the remarkable expression of genes in order to recover from HS. Additionally, 35 DEGs were conserved among five inbred lines via comparative transcriptomic analysis. Most of them were more pronounced in Z58D than AF171 at the expression levels. These candidate genes may confer thermotolerance in maize.

The Synergistic Impact of Arbuscular Mycorrhizal Fungi and Compost Tea to Enhance Bacterial Community and Improve Crop Productivity under Saline-Sodic Condition.

Soil salinity has a negative impact on the biochemical properties of soil and on plant growth, particularly in arid and semi-arid regions. Using arbuscular mycorrhizal fungi (Glomus versiform) and foliar spray from compost tea as alleviating treatments, this study aimed to investigate the effects of alleviating salt stress on the growth and development of maize and wheat grown on a saline-sodic soil during the period of 2022/2023. Six treatments were used in the completely randomized factorial design experiment. The treatments included Arbuscular mycorrhizal fungus (AMF0, AMF1) and varied concentrations of compost tea (CT0, CT50, and CT100). AMF colonization, the bacterial community and endosphere in the rhizosphere, respiration rate, growth parameters, and the productivity were all evaluated. The application of AMF and CT, either separately or in combination, effectively mitigated the detrimental effects caused by soil salinity. The combination of AMF and CT proved to be highly efficient in improving the infection rate of AMF, the bacterial community in the rhizosphere and endosphere, growth parameters, and grain yield of maize and wheat. Therefore, it can be proposed that the inoculation of mycorrhizal fungi with compost tea in saline soils is an important strategy for enhancing salt tolerance in maize and wheat plants through improving microbial activity, the infection rate of AMF, and overall maize and wheat productivity.

ZmADF5, a Maize Actin-Depolymerizing Factor Conferring Enhanced Drought Tolerance in Maize.

Drought stress is seriously affecting the growth and production of crops, especially when agricultural irrigation still remains quantitatively restricted in some arid and semi-arid areas. The identification of drought-tolerant genes is important for improving the adaptability of maize under stress. Here, we found that a new member of the actin-depolymerizing factor (ADF) family; the ZmADF5 gene was tightly linked with a consensus drought-tolerant quantitative trait locus, and the significantly associated signals were detected through genome wide association analysis. ZmADF5 expression could be induced by osmotic stress and the application of exogenous abscisic acid. Its overexpression in Arabidopsis and maize helped plants to keep a higher survival rate after water-deficit stress, which reduced the stomatal aperture and the water-loss rate, as well as improved clearance of reactive oxygen species. Moreover, seventeen differentially expressed genes were identified as regulated by both drought stress and ZmADF5, four of which were involved in the ABA-dependent drought stress response. ZmADF5-overexpressing plants were also identified as sensitive to ABA during the seed germination and seedling stages. These results suggested that ZmADF5 played an important role in the response to drought stress.

The ZmNF-YC1-ZmAPRG pathway modulates low phosphorus tolerance in maize.

Phosphorus (P) is an essential nutrient for plant growth and yield. Low phosphate use efficiency makes it important to clarify the molecular mechanism of low P stress. In our previous studies, a P efficiency gene ZmAPRG was identified. Here, we further screened the upstream regulator ZmNF-YC1 of ZmAPRG by yeast one hybrid (Y1H) assay, and found it was a low inorganic phosphorus (Pi)-inducible gene. The results of dual luciferase assays, expression analysis, and ChIP-qPCR assays showed that ZmNF-YC1 is a positive regulator of ZmAPRG. Overexpression of ZmNF-YC1 improved low P tolerance, whereas knockout of ZmNF-YC1 decreased low P tolerance in maize. Bimolecular fluorescence complementation (BiFC), yeast two hybrid (Y2H) assay, and yeast three hybrid (Y3H) assay further showed that ZmNF-YC1 can interact with ZmNF-YB14, and recruit ZmNF-YA4/10 to form NF-Y complexes. Transcriptional activation assay confirmed that the NF-Y complexes can activate the promoters of ZmAPRG. Meanwhile, transcriptome and metabolome analyses indicated that overexpression of ZmAPRG improves low P tolerance by regulating lipid composition and photosynthetic capacity, and chlorophyll fluorescence parameters provided evidence in support of this hypothesis. Furthermore, overexpression of ZmAPRG increased grain yield in inbred and hybrid maize under low P conditions. Taken together, our research revealed a low P tolerance mechanism of the ZmNF-YC1-ZmAPRG pathway.

Strigolactone roles in maize tolerance to low nitrogen involve shifts in acquisition and partitioning of protein, sulfur, and iron

Abstract Background and Aims Nitrogen (N) is an essential macronutrient that can limit plant development and crop yield through widespread physiological and molecular impacts. In maize, N-starvation enhances biosynthesis and exudation of strigolactones (SLs) in a process reversible by nitrate addition and consequent repression of genes for SL biosynthesis. Methods In the present study, a maize mutant deficient in SL biosynthesis (zmccd8) allowed an in-depth analysis of SL contributions under low N. Both hydroponic and field conditions were used to better characterize the response of the mutant to N availability. Results The severity of responses to N-limitation by the SL-deficient zmccd8 mutant extended from growth parameters to content of iron, sulfur, protein, and photosynthetic pigments, as well as pronounced impacts on expression of key genes, which could be crucial molecular target for the SL-mediated acclimatation to N shortage. Conclusions Our results demonstrate that SLs are critical for physiological acclimation to N deficiency by maize and identify central players in this action. Further contributions by iron and sulfur are implicated in the complex pathway underlying SL modulation of responses to N-deprivation, thus widening our knowledge on SL functioning and providing new hints on their potential use in agriculture.

Insights into the mechanism of abscisic acid-induced 2, 4-D tolerance in maize (Zea mays L.) seedlings: Study on the regulation of osmolytes, fluorescence, photosynthetic carbon reactions, and antioxidant metabolism

In this experiment, maize plants were subjected to different concentrations (0, 50, 100, and 250 µM) of the herbicide 2, 4-dichlorophenoxyacetic acid (2, 4-D), with some groups primed with abscisic acid (ABA) and others not. Initial observations showed that after five days of incubation, seedlings exposed to the highest concentration of 2, 4-D exhibited stunted growth and a reduction in chlorophyll accumulation. Key parameters such as relative water content, relative growth rate, and net assimilation rate decreased (by 25%, 39%, and 35% respectively) under this condition. However, these parameters partially recovered with ABA priming, showing average improvements (8%, 14%, and 16%). Recovery in tissue hydration was also noted, coinciding with increased concentrations of proline (Pro) and glycine betaine (GB), and a decrease in the Na+/K+ ratio. When exposed to the highest concentration of the herbicide, fluorescence activity was reduced, but this too was reversed with ABA treatment. A notable inhibition of photosynthesis occurred, particularly at the decarboxylation reactions involving NADP-malic enzyme and phosphoenolpyruvate carboxylase, affecting total sugar concentrations. Under the influence of 2, 4-D, there was a significant increase in reactive oxygen species, such as superoxide (35%) and hydrogen peroxide (78%), but these levels significantly diminished with ABA priming. Abscisic acid priming also mitigated oxidative stress, evidenced by a reduction in malondialdehyde (by 10%) and carbonyl content (by 11%), along with lower electrolyte leakage. The regulation of oxidative pathways through ABA priming was apparent as the plants showed increased activity of enzymes such as peroxidases, catalases, and those involved in glutathione metabolism, including glutathione reductase and glutathione S-transferase. The study also discussed how ABA metabolism regulates sensitivity to 2, 4-D and highlighted certain parameters for selecting redox homeostasis against herbicidal toxicity.

Chitosan-fulvic acid nanoparticles enhance drought tolerance in maize via antioxidant defense and transcriptional reprogramming.

Nanoparticles are promising alternatives to synthetic fertilizers in the context of climate change and sustainable agriculture. Maize plants were grown under gradient concentrations (50 μM, 100 μM, 200 μM, 500 μM, and 1 mM) of chitosan (Ch), fulvic acid (FA) or chitosan-fulvic acid nanoparticles (Ch-FANPs). Based on the overall phenotypic assessment, 100 μM was selected for downstream experiments. Maize plants grown under this optimized concentration were thereafter subjected to drought stress by water withholding for 14 days. Compared to the individual performances, the combined treatment of Ch-FANPs supported the best plant growth over chitosan, fulvic acid, or sole watered plants and alleviated the adverse effects of drought by enhancing root and shoot growth, and biomass by an average 20%. In addition, Ch-FANPs-treated plants exhibited a significant reduction in hydrogen peroxide (H2O2) content (~10%), with a concomitant increase in ascorbate peroxidase (APX) activity (>100%) while showing a reduced lipid peroxidation level observed by the decrease in malondialdehyde (MDA) content (~100%) and low electrolyte leakage level. Furthermore, chlorophyll content increased significantly (>100%) in maize plants treated with Ch-FANPs compared to Ch or FA and control in response to drought. The expression of drought-induced transcription factors, ZmDREB1A, ZmbZIP1, and ZmNAC28, and the ABA-dependent ZmCIPK3 was upregulated by Ch-FANPs. Owing to the above, Ch-FANPs are proposed as a growth-promoting agent and elicitor of drought tolerance in maize via activation of antioxidant machinery and transcriptional reprogramming of drought-related genes.

Identification of maize genotypes tolerance to acid soil stress using multiple criteria

This study aims to select maize hybrids that can tolerate acidic soil stress using several tolerance indexes. The research took place at two sites. Muneng Experimental Farm has average soil (pH=6.8), and Moncongloe Experimental Farm has acid soil (pH=5.8) from January to April 2023. A Randomized Complete Block Design (RCBD) was employed to organize ten maize hybrids, namely TH 1, TH 2, TH 3, TH 4, TH 5, TH 6, TH 7, TH 8, NK 6172, and P 32, each replicated three times. The variable observation was yield, standardized to 15% moisture content. To determine the tolerance of maize, six tolerance indexes were applied: The Stress Tolerant Index (STI), the Stress Susceptibility Index, the Mean Productivity (MP), the Harmonic Mean, the Stress Relative index (SI), and the Stress Susceptibility Percentage Index (SSPI). The result showed that hybrids that tolerate acidic soil are TH 1 and TH 2, as indicated by their average rating and standard deviation. In contrast, TH 5 and TH 7 hybrids are sensitive to acidic soil conditions. The STI and MP are suitable tolerant indexes that can select acid-tolerant hybrids with high yields under optimum and acidic conditions.

Integrated Transcriptomics and Metabolomics Analysis of Two Maize Hybrids (ZD309 and XY335) under Heat Stress at the Flowering Stage.

High temperature around flowering has a serious impact on the growth and development of maize. However, few maize genes related to flowering under heat stress have been confirmed, and the regulatory mechanism is unclear. To reveal the molecular mechanism of heat tolerance in maize, two maize hybrids, ZD309 and XY335, with different heat resistance, were selected to perform transcriptome and metabolomics analysis at the flowering stage under heat stress. In ZD309, 314 up-regulated and 463 down-regulated differentially expressed genes (DEGs) were detected, while 168 up-regulated and 119 down-regulated DEGs were identified in XY335. By comparing the differential gene expression patterns of ZD309 and XY335, we found the "frontloaded" genes which were less up-regulated in heat-tolerant maize during high temperature stress. They included heat tolerance genes, which may react faster at the protein level to provide resilience to instantaneous heat stress. A total of 1062 metabolites were identified via metabolomics analysis. Lipids, saccharides, and flavonoids were found to be differentially expressed under heat stress, indicating these metabolites' response to high temperature. Our study will contribute to the identification of heat tolerance genes in maize, therefore contributing to the breeding of heat-tolerant maize varieties.

Nanopore Direct RNA Sequencing Reveals the Short-Term Salt Stress Response in Maize Roots

Transcriptome analysis, relying on the cutting-edge sequencing of cDNA libraries, has become increasingly prevalent within functional genome studies. However, the dependence on cDNA in most RNA sequencing technologies restricts their ability to detect RNA base modifications. To address this limitation, the latest Oxford Nanopore Direct RNA Sequencing (ONT DRS) technology was employed to investigate the transcriptome of maize seedling roots under salt stress. This approach aimed to unveil both the RNA transcriptional profiles and alterations in base modifications. The analysis of the differential expression revealed a total of 1398 genes and 2223 transcripts that exhibited significant variation within the maize root system following brief exposure to salt stress. Enrichment analyses, such as the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway assessments, highlighted the predominant involvement of these differentially expressed genes (DEGs) in regulating ion homeostasis, nitrogen metabolism, amino acid metabolism, and the phytohormone signaling pathways. The protein–protein interaction (PPI) analysis showed the participation of various proteins related to glycolytic metabolism, nitrogen metabolism, amino acid metabolism, abscisic acid signaling, and the jasmonate signaling pathways. It was through this intricate molecular network that these proteins collaborated to safeguard root cells against salt-induced damage. Moreover, under salt stress conditions, the occurrence of variable shear events (AS) in RNA modifications diminished, the average length of poly(A) tails underwent a slight decrease, and the number of genes at the majority of the variable polyadenylation (APA) sites decreased. Additionally, the levels of N5-methylcytosine (m5C) and N6-methyladenosine (m6A) showed a reduction. These results provide insights into the mechanisms of early salt tolerance in maize.

Kısıtlı Sulama Uygulamalarının Mısırda Bazı Biyo – Aktif Bileşenler, Kalite Özellikleri ve Tane Verimine Etkileri

As a result of environmental difficulties and water scarcity, maize production has been challenged in arid and semi – arid regions. A management strategy for stabilizing corn production under water shortage conditions involves the use of drought – tolerant hybrids and suitable irrigation regimes. The study was conducted in the Prof. Dr. Abdulkadir AKÇIN trial area of “Selcuk University, Agriculture Faculty, Crop Science Department, Konya, TÜRKİYE” during 2019 and 2020 growing seasons. The aim of the current work is determining the effects of deficit irrigation and grain colour factors on water shortage tolerance of maize in terms of bio – active compounds, quality traits and grain yield. The factors of the trial were irrigations [50% (I1), 75% (I2) and 100% (I3) of evaporation from Class A evaporation PAN] and genotypes [DKC 5783, red corn and Sakarya]. It was noted that DKC 5783 had the highest values at two (thousand grain weight, grain yield), Sakarya had at two (total phenolic compounds and grain crude oil) and red corn had at four (total anthocyanin content, total antioxidant activity, grain crude protein and starch) properties among eight under I1 during 2019 as well as red corn had at two (total anthocyanin content and total phenolic compounds), Sakarya had at two (grain crude protein and grain crude oil) and DKC 5783 had highest values at four (thousand grain weight, total antioxidant activity, starch and grain yield) features among eight under I1 during 2020. Thousand grain weight values of I3 were 6.71% and 0.57% more than I2 while starch was 9.19% and 3.96% more than I2 during 2019 – 2020. Red corn had better contents of bio – active compounds than other two varieties during both years of the trial. The mean yield of I3 was 6.87% more than I2 in 2019 and 9.05% more than I2 in 2020 which revealed that regulated deficit irrigation might help growers to cope with decline in water availability also 25% water restriction caused tolerable decreases in grain yield and some yield compounds of the current work.

Calcineurin B-like protein ZmCBL8-1 promotes salt stress resistance in Arabidopsis.

ZmCBL8-1 enhances salt stress tolerance in maize by improving the antioxidant system to neutralize ROS homeostasis and inducing Na+/H+ antiporter gene expressions of leaves. Calcineurin B-like proteins (CBLs) as plant-specific calcium sensors have been explored for their roles in the regulation of abiotic stress tolerance. Further, the functional variations in ZmCBL8, encoding a component of the salt overly sensitive pathway, conferred the salt stress tolerance in maize. ZmCBL8-1 is a transcript of ZmCBL8 found in maize, but its function in the salt stress response is still unclear. The present study aimed to characterize the protein ZmCBL8-1 that was determined to be composed of 194 amino acids (aa) with three conserved EF hands responsible for binding Ca2+. However, a 20-aa fragment was found to be missing from its C-terminus relative to another transcript of ZmCBL8. Results indicated that it harbored a dual-lipid modification motif MGCXXS at its N-terminus and was located on the cell membrane. The accumulation of ZmCBL8-1 transcripts was high in the roots but relatively lower in the leaves of maize under normal condition. In contrast, its expression was significantly decreased in the roots, while increased in the leaves under NaCl treatment. The overexpression of ZmCBL8-1 resulted in higher salt stress resistance of transgenic Arabidopsis in a Ca2+-dependent manner relative to that of the wild type (WT). In ZmCBL8-1-overexpressing plants exposed to NaCl, the contents of malondialdehyde and hydrogen peroxide were decreased in comparison with those in the WT, and the expression of key genes involved in the antioxidant defense system and Na+/H+ antiporter were upregulated. These results suggested that ZmCBL8-1 played a positive role in the response of leaves to salt stress by inducing the expression of Na+/H+ antiporter genes and enhancing the antioxidant system to neutralize the accumulation of reactive oxygen species. These observations further indicate that ZmCBL8-1 confers salt stress tolerance, suggesting that transcriptional regulation of the ZmCBL8 gene is important for salt tolerance.

ZmMYB56 regulates stomatal closure and drought tolerance in maize seedlings through the transcriptional regulation of ZmTOM7

The growth and yield of essential crops, including maize, are significantly endangered by drought. Closing stomata, limiting water dissipation, and improving water use efficiency are important components of plant drought responses. In our study, the MYB-like transcription factor ZmMYB56, expressed in maize guard cells, played important roles in regulating stomatal closure and drought tolerance. Mutations in ZmMYB56 triggered elevated stomatal conductance, rapid water loss in isolated leaves, and severe drought sensitivity in plants. ZmMYB56 possesses transcriptional activation activity, and is expressed specifically in stomatal guard cells. As an R2R3 transcription factor, ZmMYB56 can bind the cis-acting element on the ZmTOM7 promoter sequence, activating its expression. Correspondingly, the ZmTOM7 transcript level is downregulated in Zmmyb56 seedlings. Transgenic Arabidopsis plants overexpressing ZmTOM7 exhibit limited stomatal conductance and elevated drought tolerance, while the ZmTOM7 mutation is linked to higher stomatal conductance and substantial drought sensitivity in maize seedlings. According to these findings, we conclude that ZmTOM7 operates as a key target gene of ZmMYB56 and is involved in ZmMYB56-regulated stomatal closure and maize drought tolerance. Our findings regarding the functional mechanisms of maize ZmMYB56 transcription factors in stomatal closure and drought stress enable a potential genetic resource for improving the drought resistance of maize.

Overexpression of ZmSUS1 increased drought resistance of maize (Zea mays L.) by regulating sucrose metabolism and soluble sugar content.

ZmSUS1 improved drought tolerance of maize by regulating sucrose metabolism and increasing soluble sugar content, and endowing transgenic maize with higher relative water content and photosynthesis levels. Sucrose synthase (SUS), a key enzyme of sugar metabolism, plays an important role in the regulation of carbon partitioning in plant, and affects important agronomic traits and abiotic responses to adversity. However, the function of ZmSUS1 in plant drought tolerance is still unknown. In this study, the expression patterns of ZmSUS1 in different tissues and under drought stress were analyzed in maize (Zea mays L.). It was found that ZmSUS1 was highly expressed during kernel development but also in leaves and roots of maize, and ZmSUS1 was induced by drought stress. Homozygous transgenic maize lines overexpressing ZmSUS1 increased the content and activity of SUS under drought stress and exhibited higher relative water content, proline and abscisic acid content in leaves. Specifically, the net photosynthetic rate and the soluble sugar contents including sucrose, glucose, fructose and SUS decomposition products including UDP-glucose (UDP-G) and ADP-glucose (ADP-G) in transgenic plants were significantly improved after drought stress. RNA-seq analysis showed that overexpressing of ZmSUS1 mainly affected the expression level of carbon metabolism-related genes. Especially the expression level of sucrose metabolism-related genes including sucrose phosphatase gene (SPP), sucrose phosphate synthase gene (SPS) and invertase gene (INV) were significantly up-regulated in transgenic maize. Overall, these results suggested that ZmSUS1 improved drought tolerance by regulating sucrose metabolism and increasing the soluble sugar content, and endowing transgenic maize with higher relative water content and photosynthesis levels, which can serve as a new gene candidate for cultivating drought-resistant maize varieties.

Identification of the Maize PP2C Gene Family and Functional Studies on the Role of ZmPP2C15 in Drought Tolerance

The protein phosphatase PP2C plays an important role in plant responses to stress. Therefore, the identification of maize PP2C genes that respond to drought stress is particularly important for the improvement and creation of new drought-resistant assortments of maize. In this study, we identified 102 ZmPP2C genes in maize at the genome-wide level. We analyzed the physicochemical properties of 102 ZmPP2Cs and constructed a phylogenetic tree with Arabidopsis. By analyzing the gene structure, conserved protein motifs, and synteny, the ZmPP2Cs were found to be strongly conserved during evolution. Sixteen core genes involved in drought stress and rewatering were screened using gene co-expression network mapping and expression profiling. The qRT-PCR results showed 16 genes were induced by abscisic acid (ABA), drought, and NaCl treatments. Notably, ZmPP2C15 exhibited a substantial expression difference. Through genetic transformation, we overexpressed ZmPP2C15 and generated the CRISPR/Cas9 knockout maize mutant zmpp2c15. Overexpressing ZmPP2C15 in Arabidopsis under drought stress enhanced growth and survival compared with WT plants. The leaves exhibited heightened superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and catalase (CAT) activities, elevated proline (Pro) content, and reduced malondialdehyde (MDA) content. Conversely, zmpp2c15 mutant plants displayed severe leaf dryness, curling, and wilting under drought stress. Their leaf activities of SOD, POD, APX, and CAT were lower than those in B104, while MDA was higher. This suggests that ZmPP2C15 positively regulates drought tolerance in maize by affecting the antioxidant enzyme activity and osmoregulatory substance content. Subcellular localization revealed that ZmPP2C15 was localized in the nucleus and cytoplasm. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) experiments demonstrated ZmPP2C15’s interaction with ZmWIN1, ZmADT2, ZmsodC, Zmcab, and ZmLHC2. These findings establish a foundation for understanding maize PP2C gene functions, offering genetic resources and insights for molecular design breeding for drought tolerance.

The maize WRKY transcription factor ZmWRKY64 confers cadmium tolerance in Arabidopsis and maize (Zea mays L.).

ZmWRKY64 positively regulates Arabidopsis and maize Cd stress through modulating Cd uptake, translocation, and ROS scavenging genes expression. Cadmium (Cd) is a highly toxic heavy metal with severe impacts on crops growth and development. The WRKY transcription factor is a significant regulator influencing plant stress response. Nevertheless, the function of the WRKY protein in maize Cd stress response remains unclear. Here, we identified a maize WRKY gene, ZmWRKY64, the expression of which was enhanced in maize roots and leaves under Cd stress. ZmWRKY64 was localized in the nucleus and displayed transcriptional activity in yeast. Heterologous expression of ZmWRKY64 in Arabidopsis diminished Cd accumulation in plants by negatively regulating the expression of AtIRT1, AtZIP1, AtHMA2, AtNRAMP3, and AtNRAMP4, which are involved in Cd uptake and transport, resulting in Cd stress tolerance. Knockdown of ZmWRKY64 in maize led to excessive Cd accumulation in leaf cells and in the cytosol of the root cells, resulting in a Cd hypersensitive phenotype. Further analysis confirmed that ZmWRKY64 positively regulated ZmABCC4, ZmHMA3, ZmNRAMP5, ZmPIN2, ZmABCG51, ZmABCB13/32, and ZmABCB10, which may influence Cd translocation and auxin transport, thus mitigating Cd toxicity in maize. Moreover, ZmWRKY64 could directly enhance the transcription of ZmSRG7, a reported key gene regulating reactive oxygen species homeostasis under abiotic stress. Our results indicate that ZmWRKY64 is important in maize Cd stress response. This work provides new insights into the WRKY transcription factor regulatory mechanism under a Cd-polluted environment and may lead to the genetic improvement of Cd tolerance in maize.

Early-season maize responses to salt stress: Morpho-physiological, leaf reflectance, and mineral composition

Salt stress is a major environmental factor that affects maize production. The impact of salt stress during the early vegetative stage inhibits growth and development. In this study, two maize hybrids, A6659 and P1316, were subjected to five salinity treatments with electrical conductivity 0, 3, 6, 9, and 12 dS m−1 for 28 days. A total of 26 traits were evaluated, including morpho-physiological, biomass, leaf reflectance, and mineral composition in two independent studies. Under salt stress, stomatal conductance and transpiration were reduced, resulting in a canopy temperature rise of 4 °C. A higher salinity level (12 dS m−1) reduced plant height (47%) and total leaf area (44%), consequently reducing total shoot dry weight (37%) and total root dry weight (24%). Leaf minerals such as potassium, magnesium, and phosphorous declined with increasing salt concentrations. The elevated salt concentration in the rhizosphere caused an increase in leaf reflectance at the near-infrared (NIR) region, which is attributed to macronutrient deficiency under stress conditions. The results demonstrated that maize could withstand salt stress up to 3 dS m−1, beyond which plant performance declines depending on the genetics. This study provides insights into key traits that can be used for screening or breeding maize for early-season salt stress tolerance.

Physiological Characteristic Changes and Transcriptome Analysis of Maize (Zea mays L.) Roots under Drought Stress.

Water deficit is a key limiting factor for limiting yield in maize (Zea mays L.). It is crucial to elucidate the molecular regulatory networks of stress tolerance for genetic enhancement of drought tolerance. The mechanism of drought tolerance of maize was explored by comparing physiological and transcriptomic data under normal conditions and drought treatment at polyethylene glycol- (PEG-) induced drought stress (5%, 10%, 15%, and 20%) in the root during the seedling stage. The content of saccharide, SOD, CAT, and MDA showed an upward trend, proteins showed a downward trend, and the levels of POD first showed an upward trend and then decreased. Compared with the control group, a total of 597, 2748, 6588, and 5410 differentially expressed genes were found at 5%, 10%, 15%, and 20% PEG, respectively, and 354 common DEGs were identified in these comparisons. Some differentially expressed genes were remarkably enriched in the MAPK signaling pathway and plant hormone signal transduction. The 50 transcription factors (TFs) divided into 15 categories were screened from the 354 common DEGs during drought stress. Auxin response factor 10 (ARF10), auxin-responsive protein IAA9 (IAA9), auxin response factor 14 (ARF14), auxin-responsive protein IAA1 (IAA1), auxin-responsive protein IAA27 (IAA27), and 1 ethylene response sensor 2 (ERS2) were upregulated. The two TFs, including bHLH 35 and bHLH 96, involved in the MAPK signal pathway and plant hormones pathway, are significantly upregulated in 5%, 10%, 15%, and 20% PEG stress groups. The present study provides greater insight into the fundamental transcriptome reprogramming of grain crops under drought.