Maize Mutant Research Articles

Brachytic2 mutation is able to counteract the main pleiotropic effects of brown midrib3 mutant in maize

Maize is the basis of nutrition of domesticated herbivores and one of the most promising energy crops. The presence of lignin in the cell wall, tightly associated to carbohydrates, prevents the physical access of enzymes such as cellulase, limiting the carbohydrate degradability and consequently the energy value. To increase the utilization of the biomass cellulose content, the challenge of breeding programs is to lower or modify the lignin components. In maize several mutations are able to modify the lignin content and in particular the mutation in brown midrib3 (bm3) gene appeared as one of the most promising in breeding programs. Unfortunately this mutation has several negative pleiotropic effects on various important agronomic traits such as stay green, lodging and susceptibility to several infections.The maize Brachyitic 2 (br2) gene encodes for a putative protein involved in polar movement of auxins. br2 mutant plants are characterized by shortening of lower stalk internodes, unusual stalk strength and tolerance to wind lodging, darker leaves persisting longer in the active green state in comparison to wild type plants, suggesting a possible utilization of br2 plants to counteract the negative effects of the bm3 mutation. In this work, we report the generation and a preliminary characterization of the double mutant bm3 br2, suggesting the potential use of this new genetic material to increase biomass cellulose utilization.

Phenotypical Characterization of a Mutant dea1 With Developmental Defect in Female Inflorescence and Mutantion Impact on Yield in Maize

The establishment of female inflorescence morphology is of great significance to the formation of final maize yield. defective ear1 (dea1) is a novel maize mutant with developmental defect of female inflorescence caused by natural variation. Morphological analysis revealed that the mutant dea1 was characterized as a “scar-like” crack on the adaxial side of the top of the ear, accounting for 28.6-100.0% of the ear length, with an average of 32.4%. The results of scanning electron microscope showed that there was collapse in the formation of paired spikelet primordium at the base of the axillary meristem. Most of investigated botanical and agronomical traits of dea1 were lower than those of wild type, except for ear length and hundred grain weight. The grain yield per ear of mutant dea1 was 35.93% lower than that of wild type, and the width of mutation crack contributed the most to the yield loss per ear. The identification of the mutant dea1 and the characteristically phenotypic analysis provide a theoretical basis for the study of the molecular regulation mechanism of ear development and the application of high-yield breeding in maize.The establishment of female inflorescence morphology is of great significance to the formation of final maize yield. defective ear1 (dea1) is a novel maize mutant with developmental defect of female inflorescence caused by natural variation. Morphological analysis revealed that the mutant dea1 was characterized as a “scar-like” crack on the adaxial side of the top of the ear, accounting for 28.6-100.0% of the ear length, with an average of 32.4%. The results of scanning electron microscope showed that there was collapse in the formation of paired spikelet primordium at the base of the axillary meristem. Most of investigated botanical and agronomical traits of dea1 were lower than those of wild type, except for ear length and hundred grain weight. The grain yield per ear of mutant dea1 was 35.93% lower than that of wild type, and the width of mutation crack contributed the most to the yield loss per ear. The identification of the mutant dea1 and the characteristically phenotypic analysis provide a theoretical basis for the study of the molecular regulation mechanism of ear development and the application of high-yield breeding in maize.

Morphological Characterization and Transcriptome Analysis of New Dwarf and Narrow-Leaf (dnl2) Mutant in Maize

Lodging is the primary factor limiting high yield under a high plant density. However, an optimal plant height and leaf shape can effectively decrease the lodging risk. Here we studied an ethyl methanesulfonate (EMS)-induced dwarf and a narrow-leaf mutant, dnl2. Gene mapping indicated that the mutant was controlled by a gene located on chromosome nine. Phenotypic and cytological observations revealed that dnl2 showed inhibited cell growth, altered vascular bundle patterning, and disrupted secondary cell wall structure when compared with the wild-type, which could be the direct cause of the dwarf and narrow-leaf phenotype. The phytohormone levels, especially auxin and gibberellin, were significantly decreased in dnl2 compared to the wild-type plants. Transcriptome profiling of the internodes of the dnl2 mutant and wild-type revealed a large number of differentially expressed genes enriched in the cell wall biosynthesis, remodeling, and hormone biosynthesis and signaling pathways. Therefore, we suggest that crosstalk between hormones (the altered vascular bundle and secondary cell wall structure) may contribute to the dwarf and narrow-leaf phenotype by influencing cell growth. These results provide a foundation for DNL2 gene cloning and further elucidation of the molecular mechanism of the regulation of plant height and leaf shape in maize.

ENB1 encodes a cellulose synthase 5 that directs synthesis of cell wall ingrowths in maize basal endosperm transfer cells.

Development of the endosperm is strikingly different in monocots and dicots: it often manifests as a persistent tissue in the former and transient tissue in the latter. Little is known about the controlling mechanisms responsible for these different outcomes. Here we characterized a maize (Zea mays) mutant, endosperm breakdown1 (enb1), in which the typically persistent endosperm (PE) was drastically degraded during kernel development. ENB1 encodes a cellulose synthase 5 that is predominantly expressed in the basal endosperm transfer layer (BETL) of endosperm cells. Loss of ENB1 function caused a drastic reduction in formation of flange cell wall ingrowths (ingrowths) in BETL cells. Defective ingrowths impair nutrient uptake, leading to premature utilization of endosperm starch to nourish the embryo. Similarly, developing wild-type kernels cultured in vitro with a low level of sucrose manifested early endosperm breakdown. ENB1 expression is induced by sucrose via the BETL-specific Myb-Related Protein1 transcription factor. Overexpression of ENB1 enhanced development of flange ingrowths, facilitating sucrose transport into BETL cells and increasing kernel weight. The results demonstrated that ENB1 enhances sucrose supply to the endosperm and contributes to a PE in the kernel.

Maize PHYTOMELATONIN RECEPTOR1 functions in plant tolerance to osmotic and drought stress.

Phytomelatonin is a universal signal molecule that regulates plant growth and stress responses; however, only one receptor that can directly bind with and perceive melatonin signaling has been identified so far, namely AtPMTR1/CAND2 in Arabidopsis. Whether other plants contain a similar receptor and, if so, how it functions is still unknown. In this study, we identified a new phytomelatonin receptor in the monocot maize (Zea mays), and investigated its role in plant responses to osmotic and drought stress. Using homology searching, we identified a plasma membrane-localized protein, Zm00001eb214610/ZmPMTR1, with strong binding activity to melatonin as a potential phytomelatonin receptor in maize. Overexpressing ZmPMTR1 in Arabidopsis Col-0 promoted osmotic stress tolerance, and rescued osmotic stress sensitivity of the Arabidopsis cand2-1 mutant. Furthermore, ZmPMTR1 also largely rescued defects in melatonin-induced stomatal closure in the cand2-1 mutant, thereby reducing water loss rate and increasing tolerance to drought stress. In addition, we identified a maize mutant of ZmPMTR1, EMS4-06e2fl, with a point-mutation causing premature termination of protein translation, and found that this mutant had lower leaf temperatures, increased rate of water loss, and enhanced drought stress sensitivity. Thus, we present ZmPMTR1 as the first phytomelatonin receptor to be identified and examined in a monocot plant, and our results indicate that it plays an important function in the response of maize to drought stress.

Phenotypic effects from the expression of a deregulated AtGAD1 transgene and GABA pathway suppression mutants in maize.

Glutamate decarboxylase (GAD; EC 4.1.1.15) catalyzes the irreversible decarboxylation of glutamate to produce γ-aminobutyric acid (GABA); a ubiquitous non-protein amino acid involved in the regulation of several aspects of plant metabolism and physiology. To study the function of GAD and GABA in maize, we have; 1) introduced native and deregulated forms of AtGAD1 into maize with the intent of increasing the synthesis of GABA and 2) introduced constructs into maize designed to suppress the activity of several GABA shunt, GABA transport and GABA pathway genes. Maize plants expressing the deregulated AtGAD1 exhibit a severe chlorosis and retarded growth phenotype and have high levels of GABA, and Ca++/CaM-independent GAD activity. Plants expressing the suppression constructs for GABA biosynthetic and transport pathway genes had no observable phenotype whereas a knockout of GABA catabolic pathway genes led to growth and developmental defects under standard growth conditions. The implications of this study to our understanding of the action and function of GABA and GAD in crops are discussed.

Ground tissue circuitry regulates organ complexity in maize and Setaria.

Most plant roots have multiple cortex layers that make up the bulk of the organ and play key roles in physiology, such as flood tolerance and symbiosis. However, little is known about the formation of cortical layers outside of the highly reduced anatomy of Arabidopsis. Here, we used single-cell RNA sequencing to rapidly generate a cell-resolution map of the maize root, revealing an alternative configuration of the tissue formative transcription factor SHORT-ROOT (SHR) adjacent to an expanded cortex. We show that maize SHR protein is hypermobile, moving at least eight cell layers into the cortex. Higher-order SHR mutants in both maize and Setaria have reduced numbers of cortical layers, showing that the SHR pathway controls expansion of cortical tissue to elaborate anatomical complexity.

ZmTE1 promotes plant height by regulating intercalary meristem formation and internode cell elongation in maize.

SummaryMaize height is determined by the number of nodes and the length of internodes. Node number is driven by intercalary meristem formation and internode length by intercalary cell elongation, respectively. However, mechanisms regulating establishment of nodes and internode growth are unclear. We screened EMS‐induced maize mutants and identified a dwarf mutant zm66, linked to a single base change in TERMINAL EAR 1 (ZmTE1). Detailed phenotypic analysis revealed that zm66 (zmte1‐2) has shorter internodes and increased node numbers, caused by decreased cell elongation and disordered intercalary meristem formation, respectively. Transcriptome analysis showed that auxin signalling genes are also dysregulated in zmte1‐2, as are cell elongation and cell cycle‐related genes. This argues that ZmTE1 regulates auxin signalling, cell division, and cell elongation. We found that the ZmWEE1 kinase phosphorylates ZmTE1, thus confining it to the nucleus and probably reducing cell division. In contrast, the ZmPP2Ac‐2 phosphatase promotes dephosphorylation and cytoplasmic localization of ZmTE1, as well as cell division. Taken together, ZmTE1, a key regulator of plant height, is responsible for maintaining organized formation of internode meristems and rapid cell elongation. ZmWEE1 and ZmPP2Ac‐2 might balance ZmTE1 activity, controlling cell division and elongation to maintain normal maize growth.

Tomato and cotton G protein beta subunit mutants display constitutive autoimmune responses.

Heterotrimeric G protein Gβ‐deficient mutants in rice and maize display constitutive immune responses, whereas Arabidopsis Gβ mutants show impaired defense, suggesting the existence of functional differences between monocots and dicots. Using CRISPR/Cas9, we produced one hemizygous tomato line with a mutated SlGB1 Gβ gene. Homozygous slgb1 knockout mutants exhibit all the hallmarks of autoimmune mutants, including development of necrotic lesions, constitutive expression of defense‐related genes, and high endogenous levels of salicylic acid (SA) and reactive oxygen species, resulting in early seedling lethality. Virus‐induced silencing of Gβ in cotton reproduced the symptoms observed in tomato mutants, confirming that the autoimmune phenotype is not limited to monocot species but is also shared by dicots. Even though multiple genes involved in SA and ethylene signaling are highly induced by Gβ silencing in tomato and cotton, co‐silencing of SA or ethylene signaling components in cotton failed to suppress the lethal phenotype, whereas co‐silencing of the oxidative burst oxidase RbohD can repress lethality. Despite the autoimmune response observed in slgb1 mutants, we show that SlGB1 is a positive regulator of the pathogen‐associated molecular pattern (PAMP)‐triggered immunity (PTI) response in tomato. We speculate that the phenotypic differences observed between Arabidopsis and tomato/cotton/rice/maize Gβ knockouts do not necessarily reflect divergences in G protein‐mediated defense mechanisms.

The SUMO ligase MMS21 profoundly influences maize development through its impact on genome activity and stability.

The post-translational addition of SUMO plays essential roles in numerous eukaryotic processes including cell division, transcription, chromatin organization, DNA repair, and stress defense through its selective conjugation to numerous targets. One prominent plant SUMO ligase is METHYL METHANESULFONATE-SENSITIVE (MMS)-21/HIGH-PLOIDY (HPY)-2/NON-SMC-ELEMENT (NSE)-2, which has been connected genetically to development and endoreduplication. Here, we describe the potential functions of MMS21 through a collection of UniformMu and CRISPR/Cas9 mutants in maize (Zea mays) that display either seed lethality or substantially compromised pollen germination and seed/vegetative development. RNA-seq analyses of leaves, embryos, and endosperm from mms21 plants revealed a substantial dysregulation of the maize transcriptome, including the ectopic expression of seed storage protein mRNAs in leaves and altered accumulation of mRNAs associated with DNA repair and chromatin dynamics. Interaction studies demonstrated that MMS21 associates in the nucleus with the NSE4 and STRUCTURAL MAINTENANCE OF CHROMOSOMES (SMC)-5 components of the chromatin organizer SMC5/6 complex, with in vitro assays confirming that MMS21 will SUMOylate SMC5. Comet assays measuring genome integrity, sensitivity to DNA-damaging agents, and protein versus mRNA abundance comparisons implicated MMS21 in chromatin stability and transcriptional controls on proteome balance. Taken together, we propose that MMS21-directed SUMOylation of the SMC5/6 complex and other targets enables proper gene expression by influencing chromatin structure.

Suppressing chlorophyll degradation by silencing OsNYC3 improves rice resistance to Rhizoctonia solani, the causal agent of sheath blight.

SummaryNecrotrophic fungus Rhizoctonia solani Kühn (R. solani) causes serious diseases in many crops worldwide, including rice and maize sheath blight (ShB). Crop resistance to the fungus is a quantitative trait and resistance mechanism remains largely unknown, severely hindering the progress on developing resistant varieties. In this study, we found that resistant variety YSBR1 has apparently stronger ability to suppress the expansion of R. solani than susceptible Lemont in both field and growth chamber conditions. Comparison of transcriptomic profiles shows that the photosynthetic system including chlorophyll biosynthesis is highly suppressed by R. solani in Lemont but weakly in YSBR1. YSBR1 shows higher chlorophyll content than that of Lemont, and inducing chlorophyll degradation by dark treatment significantly reduces its resistance. Furthermore, three rice mutants and one maize mutant that carry impaired chlorophyll biosynthesis all display enhanced susceptibility to R. solani. Overexpression of OsNYC3, a chlorophyll degradation gene apparently induced expression by R. solani infection, significantly enhanced ShB susceptibility in a high‐yield ShB‐susceptible variety ‘9522’. However, silencing its transcription apparently improves ShB resistance without compromising agronomic traits or yield in field tests. Interestingly, altering chlorophyll content does not affect rice resistance to blight and blast diseases, caused by biotrophic and hemi‐biotrophic pathogens, respectively. Our study reveals that chlorophyll plays an important role in ShB resistance and suppressing chlorophyll degradation induced by R. solani infection apparently improves rice ShB resistance. This discovery provides a novel target for developing resistant crop to necrotrophic fungus R. solani.

PPR647 Protein Is Required for Chloroplast RNA Editing, Splicing and Chloroplast Development in Maize.

Chloroplasts play an essential role in plant growth and development. Any factors affecting chloroplast development will lead to abnormal plant growth. Here, we characterized a new maize mutant, albino seedling mutant 81647 (as-81647), which exhibits an entirely albino phenotype in leaves and eventually died before the three-leaf stage. Transmission electron microscopy (TEM) demonstrated that the chloroplast thylakoid membrane was impaired and the granum lamellae significantly decreased in as-81647. Map-based cloning and transgenic analysis confirmed that PPR647 encodes a new chloroplast protein consisting of 11 pentratricopeptide repeat domains. Quantitative real-time PCR (qRT-PCR) assays and transcriptome analysis (RNA-seq) showed that the PPR647 mutation significantly disrupted the expression of PEP-dependent plastid genes. In addition, RNA splicing and RNA editing of multiple chloroplast genes showed severe defects in as-81647. These results indicated that PPR647 is crucial for RNA editing, RNA splicing of chloroplast genes, and plays an essential role in chloroplast development.

Phosphorus application improves grain yield in low phytic acid maize synthetic populations

Maize mutants with low phytic acid have a compromised overall agronomic performance that results in low yields. This study was conducted to investigate the effect of P (18, 26 and 34 mg/kg) on the agronomic performance of low and high phytic acid (LPA and HPA) maize synthetic populations of tropical origin, compared to two commercial hybrids (SC701 and LS8520). Subsequently, a germination test was performed on the seeds produced from the different levels of P fertilizer application rates. The germination test was conducted in the laboratory, using a germination paper towel, while the agronomic study was conducted in a controlled environment. The measured parameters included days to 50% flowering, plant height, and grain yield, as well as the final germination and germination velocity index. The results found that the grain yield increased by 1.30, 0.51, 2.41 and 1.87 t/ha in LPA, HPA, SC701 and LS8520, from the application of 18–26 mg/kg of P, respectively. However, there were non-significant differences (p > 0.05) in the grain yields of all varieties at a P application of 26 and 34 mg/kg. The final germination increased by 4% and 2% in LPA and LS8520, respectively, with the increase in the P application rate being from 18 to 26 mg/kg. However, no significant differences (p > 0.05) were found in the final germination percentage of all varieties at 26 mg/kg of P. This study indicated that the optimum application of P at planting enhances the overall performance of the LPA maize synthetic population to a level that is comparable to commercially-grown varieties.

English

Maize (Zea mays), is a plant of economic and nutritional interest, which is often confronted with unfavorable environmental conditions, including water stress. This phenomenon forces the plants to considerably reduce their production by disturbing their metabolism. Seeds of maize of the variety EV8728 were subjected to different doses of gamma radiation (100, 200 and 300 grays) in order to induce mutations in plants that could lead to resistance to water stress. Thus, plants from gamma irradiated seeds were used to evaluate the impact of water stress on leaf physiological parameters (leaf area, density and pore area of stomata and assimilatory pigments). Water stress had a depressive effect on all leaf parameters in maize plants. The degree of sensitivity or tolerance of the plants depends on the dose of irradiation and the intensity of stress applied. The increase in the level of water stress reduces the leaf surface and that of the stomata pores. This in turn increases the density of stomata. In addition, chlorophyll a  was more sensitive to the effect of water stress than chlorophyll b. Key words: Maize mutants, water stress, gamma irradiation, leaves, stomata ostioles.

Genetic and molecular characterisation of subtropically adapted low-phytate genotypes for utilisation in mineral biofortification of maize (

Phytic acid (PA) is a major anti-nutritional factor in maize grains and significantly reduces the bioavailability of minerals such as iron and zinc in humans. Reduction of PA in maize (Zea mays L.) thus assumes great significance. Despite the availability of low phytic-acid (lpa) mutants in maize, their utilisation in breeding has been limited because of non-availability of well adapted lpa-based inbreds. A set of 24 subtropically adapted lpa1-1-based maize inbreds derived through molecular breeding was evaluated along with seven wild-type inbreds at multilocation. The lpa1-1 inbreds possessed 35.8% lower PA (1.68 mg/g) than the wild-type inbreds (2.61 mg/g). Mean proportion of PA was lower in the lpa1-1-based inbreds (62.8%) than in the wild-type inbreds (91.6%). The lpa1-1 inbreds showed similar performance for plant- and ear-height besides flowering behaviour as did wild types. Average grain yield among lpa1-1 inbreds (2735.9 kg/ha) was at par with the original versions (2907.3 kg/ha). Molecular profiling of these lpa1-1 inbreds using 60 genome-wide single-sequence repeats generated 172 alleles, with a mean of 2.87 alleles per locus. Mean polymorphism information content and mean gene diversity were 0.41 and 0.48 respectively. Genetic dissimilarity ranged from 0.23 to 0.81, with an average of 0.64. Cluster analyses grouped 24 lpa1-1 genotypes into three major clusters, and principal-coordinate analysis depicted the diverse nature of genotypes. The study also identified a set of potential hybrid combinations with low PA for their direct utilisation in biofortification program. This is the first study on comprehensive characterisation of lpa1-1-based inbreds adapted to subtropical conditions.

The Synthesis of Pentyl Leaf Volatiles and Their Role in Resistance to Anthracnose Leaf Blight.

Volatiles are important airborne chemical messengers that facilitate plant adaptation to a variety of environmental challenges. Lipoxygenases (LOXs) produce a bouquet of non-volatile and volatile oxylipins, including C6 green leaf volatiles (GLVs), which are involved in a litany of plant physiological processes. GLVs are emitted by a diverse array of plant species, and are the best-known group of LOX-derived volatiles. Five-carbon pentyl leaf volatiles (PLVs) represent another widely emitted group of LOX-derived volatiles that share structural similarity to GLVs, however, relatively little is known about their biosynthesis or biological activity. In this study, we utilized PLV-deficient mutants of maize and Arabidopsis and exogenous PLV applications to elucidate the biosynthetic order of individual PLVs. We further measured PLVs and GLVs after tissue disruption of leaves by two popular methods of volatile elicitation, wounding and freeze-thawing. Freeze-thawing distorted the volatile metabolism of both GLVs and PLVs relative to wounding, though this distortion differed between the two groups of volatiles. These results suggest that despite the structural similarity of these two volatile groups, they are differentially metabolized. Collectively, these results have allowed us to produce the most robust PLV pathway to date. To better elucidate the biological activity of PLVs, we show that PLVs induce maize resistance to the anthracnose pathogen, Colletotrichum graminicola, the effect opposite to that conferred by GLVs. Further analysis of PLV-treated and infected maize leaves revealed that PLV-mediated resistance is associated with early increases of oxylipin α- and γ-ketols, and later increases of oxylipin ketotrienes, hydroxytrienes, and trihydroxydienes. Ultimately, this study has produced the most up-to-date pathway for PLV synthesis, and reveals that PLVs can facilitate pathogen resistance through induction of select oxylipins.

Genetic control of kernel compositional variation in a maize diversity panel.

Maize (Zea mays L.) is a multi-purpose row crop grown worldwide, which, over time, has often been bred for increased yield at the detriment of lower composition grain quality. Some knowledge of the genetic factors that affect quality traits has been discovered through the study of classical maize mutants; however, much of the underlying genetic control of these traits and the interaction between these traits remains unknown. To better understand variation that exists for grain compositional traits in maize, we evaluated 501 diverse temperate maize inbred lines in five unique environments and predicted 16 compositional traits (e.g., carbohydrates, protein, and starch) based on the output of near-infrared (NIR) spectroscopy. Phenotypic analysis found substantial variation for compositional traits and the majority of variation was explained by genetic and environmental factors. Correlations and trade-offs among traits in different maize types (e.g., dent, sweetcorn, and popcorn) were explored, and significant differences and meaningful correlations were detected. In total, 22.9-71.0% of the phenotypic variation across these traits could be explained using 2,386,666 single nucleotide polymorphism (SNP) markers generated from whole-genome resequencing data. A genome-wide association study (GWAS) was conducted using these same markers and found 72 statistically significant SNPs for 11 compositional traits. This study provides valuable insights in the phenotypic variation and genetic control underlying compositional traits that can be used in breeding programs for improving maize grain quality.

E183K Mutation in Chalcone Synthase C2 Causes Protein Aggregation and Maize Colorless.

Flavonoids give plants their rich colors and play roles in a number of physiological processes. In this study, we identified a novel colorless maize mutant showing reduced pigmentation throughout the whole life cycle by EMS mutagenesis. E183K mutation in maize chalcone synthase C2 (ZmC2) was mapped using MutMap strategy as the causal for colorless, which was further validated by transformation in Arabidopsis. We evaluated transcriptomic and metabolic changes in maize first sheaths caused by the mutation. The downstream biosynthesis was blocked while very few genes changed their expression pattern. ZmC2-E183 site is highly conserved in chalcone synthase among Plantae kingdom and within species’ different varieties. Through prokaryotic expression, transient expression in maize leaf protoplasts and stable expression in Arabidopsis, we observed that E183K and other mutations on E183 could cause almost complete protein aggregation of chalcone synthase. Our findings will benefit the characterization of flavonoid biosynthesis and contribute to the body of knowledge on protein aggregation in plants.

Effect of Hormonal Regime and Explant Type on Cell Clusters Expression of Maize Mutants (Zea Mays L) Derived from Gamma Irradiated Seeds of Ev8728 variety

Maize (Zea mays L.) seeds irradiated with gamma or not from the fourth self-fertilisation cycle were selected for tissue culture. For this purpose, MS medium supplemented with 30 g.L-1 sucrose, 100 mg.L-1 casein hydrolysate, 100 mg.L-1 myo-inositol and 6 g.L-1 agar was used. For this purpose, three auxins type (2.4-dichlorophenoxyacetic acid (2.4-D), 2-methoxy-3.6-dichlorobenzoic acid (Dicamba) and Indole-3-Acetic Acid (AIA)) and explants (root, epicotyl and leaf) were tested. The results showed that 2.4-D more precisely at 3.5 mg.L-1 was the best auxin for callus induction in the different maize mutants studied. The induction rate, dry matter weight and water content of callus varied according to the type, age, explant position and the maize mutants studied (control, 200 and 300 grays). Thus, the 7-day root and more precisely its basal part was better for callogenesis. In addition, mutants of the 200 grays dose were more expressive in the ability to induce callus in EV8728maize variety.

Aldoximes are precursors of auxins in Arabidopsis and maize.

Two natural auxins, phenylacetic acid (PAA) and indole-3-acetic acid (IAA), play crucial roles in plant growth and development. One route of IAA biosynthesis uses the glucosinolate intermediate indole-3-acetaldoxime (IAOx) as a precursor, which is thought to occur only in glucosinolate-producing plants in Brassicales. A recent study showed that overproducing phenylacetaldoxime (PAOx) in Arabidopsis increases PAA production. However, it remains unknown whether this increased PAA resulted from hydrolysis of PAOx-derived benzyl glucosinolate or, like IAOx-derived IAA, is directly converted from PAOx. If glucosinolate hydrolysis is not required, aldoxime-derived auxin biosynthesis may occur beyond Brassicales. To better understand aldoxime-derived auxin biosynthesis, we conducted an isotope-labelled aldoxime feeding assay using an Arabidopsis glucosinolate-deficient mutant sur1 and maize, and transcriptomics analysis. Our study demonstrated that the conversion of PAOx to PAA does not require glucosinolates in Arabidopsis. Furthermore, maize produces PAA and IAA from PAOx and IAOx, respectively, indicating that aldoxime-derived auxin biosynthesis also occurs in maize. Considering that aldoxime production occurs widely in the plant kingdom, aldoxime-derived auxin biosynthesis is likely to be more widespread than originally believed. A genome-wide transcriptomics study using PAOx-overproduction plants identified complex metabolic networks among IAA, PAA, phenylpropanoid and tryptophan metabolism.