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Otoni, W. C.

Publications and source records attributed to Otoni, W. C..

4 recordsLinked to original sources

Optimization of somatic embryogenesis in Euterpe edulis Martius using auxin analogs and atomic force microscopy

Euterpe edulis Martius is an endangered species of the Atlantic Forest, whose fruits have high antioxidant potential, and propagated exclusively by seeds. The present study assessed the ability of different auxin inducers and picloram analogs to trigger somatic embryogenesis in E. edulis. Immature seeds were harvested, and their zygotic embryos were excised and grown in MS culture medium supplemented with 2,4-D dichlorophenoxyacetic acid (2,4-D) or picloram at 150, 300, 450, 600 {micro}M. The activity of picloram analogs triclopyr and clopyralid was evaluated in semisolid MS medium. At maturation and germination, picloram-derived calli and somatic embryos isolated from triclopyr-grown cultures were first transferred to pre-maturation medium and, after 30 days, to basal MS or MS medium supplemented with either 5 {micro}M abscisic acid or 0.53 {micro}M 1-naphthaleneacetic acid plus 12.3 {micro}M 2-isopentenyladenine. Finally, somatic embryos with root protrusions were transferred to MS medium devoid of sucrose for 30 days and then acclimatized ex vitro. Scanning, transmission, and atomic force microscopy revealed that picloram was superior to 2,4-D but less effective than triclopyr (100 {micro}M) in inducing embryogenesis. Maturation and germination of somatic embryos in E. edulis can be maximized by 5 {micro}M abscisic acid, and selecting calli via atomic force microscopy. HighlightThis work opens novel roads for embryogenic induction, using a new and more efficient inducer than the usual ones, and an innovative evaluation technique based on AFM.

plant biology↗

Plant age-dependent dynamics of annatto pigment (bixin) biosynthesis in Bixa orellana L.

Age affects the production of secondary metabolites, but how developmental cues regulate secondary metabolism remains poorly understood. Annatto (Bixa orellana L.) is a source of bixin, an apocarotenoid used in the worlds food industry worldwide. Understanding how age-dependent mechanisms control bixin biosynthesis is of great interest for plant biology and for pharmaceutical, cosmetic, and textile industries. Here, we used genetic and molecular tools to unravel the role of the annatto age regulated miRNA156 (miR156) targeted SQUAMOSA PROMOTER BINDING PROTEIN LIKE (BoSPL) genes in secondary metabolism. Low expression of several BoSPL genes in miR156 overexpressing annatto plants (OE::156) impacted leaf ontogeny, reducing bixin production and increasing abscisic acid (ABA) levels. Modulation of BoCCD4;4 and BoCCD1 expression, key genes in lycopene cleavage, was associated with diverting the carbon flux from bixin to ABA, whereas upregulation of lycopene {beta} cyclase genes implies the xanthophyll biosynthetic pathway acted as a carbon sink in OE::156 plants. Proteomic analyses revealed low accumulation of most secondary metabolite-related enzymes in OE::156 plants, suggesting that miR156 targeted BoSPLs are required to activate several annatto secondary metabolic pathways. Our findings suggest that carbon flux in B. orellana OE::156 leaves was redirected from bixin to ABA production, indicating an age-dependent leaf dynamics of bixin biosynthesis. Importantly, our study opened a new venue to future annatto breeding programs aiming to improve bixin output.

plant biology↗

Dynamics of annatto pigment synthesis and accumulation revealed by integrated chemical, anatomical, and RNA-Seq analyses

Bixin is a commercially valuable apocarotenoid pigment found in the seed aril of Bixa orellana. The dynamics and regulation of its biosynthesis and accumulation during seed development remain largely unknown. Here, we combined chemical, anatomical, and transcriptomic data to provide stage-specific resolution of the cellular and molecular events occurring during B. orellana seed development. Seeds at five developmental stages (S1-S5) were used for analysis of bixin content and seed anatomy, and three of them (S1, S3 and S4) selected for Illumina HiSeq sequencing. Bixin accumulated sharply during seed development, particularly during the S2 stage, peaking at the S4 stage, and then decreasing slightly in the S5 stage. Anatomical analysis revealed that bixin accumulated in the large central vacuole of specialized cells, which were scattered throughout the developing mesotesta at the S2 stage, but enlarged progressively at later stages, until they occupied most of the parenchyma in the aril. A total of 13 million reads were generated and assembled into 73,381 protein-encoding contigs, from which 312 were identified as containing 1-deoxy-D-xylulose-5-phosphate/2-C-methyl-D-erythritol-4-phosphate (DOXP/MEP), carotenoid, and bixin pathways genes. Differential expression analysis of these genes revealed that 50 of them were differentially expressed between all the seed developmental stages, including seven carotenoid cleavage dioxygenases, eight aldehyde dehydrogenases and 22 methyltransferases. Taken together, these results provide a comprehensive description of the cellular and molecular events related to the dynamics of bixin synthesis and accumulation during seed development in B. orellana.

plant biology↗

Auxin-driven ecophysiological diversification of leaves in domesticated tomato

Heterobaric leaves have bundle sheath extensions (BSEs) that compartmentalise the parenchyma, whereas homobaric leaves do not. The presence of BSEs affects leaf hydraulics and photosynthetic rate. The tomato (Solanum lycopersicum) obscuravenosa (obv) mutant lacks BSEs. Here we identify the obv gene and the causative mutation, a non-synonymous amino acid change that disrupts a C2H2 zinc finger motif in a putative transcription factor. This mutation exists as a rare polymorphism in the natural range of wild tomatoes, but has increased in frequency in domesticated tomatoes, suggesting that the latter diversified into heterobaric and homobaric leaf types. The obv mutant displays reduced vein density, leaf hydraulic conductance and photosynthetic assimilation rate. We show that these and other effects on plant development, including changes in leaf insertion angle, leaf margin serration, minor vein density and fruit shape, are controlled by OBV via changes in auxin signalling. Loss of function of the transcriptional regulator AUXIN RESPONSE FACTOR (ARF4) also results in defective BSE development, revealing an additional component of a novel genetic module controlling aspects of leaf development important for ecological adaptation and subject to breeding selection. One sentence summarydistribution of tomato heterobaric and homobaric leaves is controlled by a single-nucleotide polymorphism in an auxin-related transcription factor

plant biology↗