Browsing by Author "Moyano, Tomás"

Now showing 1 - 3 of 3
  • Thumbnail Image
    Item
    Differences in berry primary and secondary metabolisms identified by transcriptomic and metabolic profiling of two table grape color somatic variants
    (2019) Santibáñez, Claudia; Meyer Regueiro, Carlos José; Martínez, Litsy; Moyano, Tomás; Lunn, John; Feil, Regina; Dai, Zhanwu; Carrasco, David; Arroyo García, Rosa; Hilbert,Ghislaine; Renaud, Christel; Delrot, Serge; Manke Nachtigall, Fabiane; Gutiérrez Ilabaca, Rodrigo Antonio; Matus, José Tomás; Gomès, Eric; Arce Johnson, Jorge Patricio
    Anthocyanins are flavonoids responsible for the color of berries in skin-pigmented grapevine (Vitis vinifera L.). Due to the widely adopted vegetative propagation of this species, somatic mutations occurring in meristematic cell layers can be fixed and passed into the rest of the plant when cloned. In this study we focused on the transcriptomic and metabolic differences between two color somatic variants. Using microscopic, metabolic and mRNA profiling analyses we compared the table grape cultivar (cv.) ‘Red Globe’ (RG, with purplish berry skin) and cv. ‘Chimenti Globe’ (CG, with a contrasting reddish berry skin color). As expected, significant differences were found in the composition of flavonoids and other phenylpropanoids, but also in their upstream precursors’ shikimate and phenylalanine. Among primary metabolites, sugar phosphates related with sucrose biosynthesis were less accumulated in cv. ‘CG’. The red-skinned cv. ‘CG’ only contained di-hydroxylated anthocyanins (i.e. peonidin and cyanidin) while the tri-hydroxylated derivatives malvidin, delphinidin and petunidin were absent, in correlation to the reddish cv. ‘CG’ skin coloration. Transcriptomic analysis showed alteration in flavonoid metabolism and terpenoid pathways and in primary metabolism such as sugar content. Eleven flavonoid 3’5’-hydroxylase gene copies were down-regulated in cv. ‘CG’. This family of cytochrome P450 oxidoreductases are key in the biosynthesis of tri-hydroxylated anthocyanins. Many transcription factors appeared down-regulated in cv. ‘CG’ in correlation to the metabolic and transcriptomic changes observed. The use of molecular markers and its confirmation with our RNA-seq data showed the exclusive presence of the null MYBA2 white allele (i.e. homozygous in both L1 and L2 layers) in the two somatic variants. Therefore, the differences in MYBA1 expression seem sufficient for the skin pigmentation differences and the changes in MYBA target gene expression in cv. ‘Chimenti Globe’.
  • No Thumbnail Available
    Item
    Genome-wide analysis in response to nitrogen and carbon identifies regulators for root AtNRT2 transporters
    (2021) Ruffel, Sandrine; Chaput, Valentín; Przybyla Toscano, Jonathan; Fayos, Ian; Ibarra, Catalina; Moyano, Tomás; Fizames, Cécile; Tillard, Pascal; O’Brien, José Antonio; Gutiérrez, Rodrigo A.; Gojon, Alain; Lejay, Laurence
    In Arabidopsis (Arabidopsis thaliana), the High-Affinity Transport System (HATS) for root nitrate (⁠NO3−⁠) uptake depends mainly on four NRT2 NO3− transporters, namely NRT2.1, NRT2.2, NRT2.4, and NRT2.5. The HATS is the target of many regulations to coordinate nitrogen (N) acquisition with the N status of the plant and with carbon (C) assimilation through photosynthesis. At the molecular level, C and N signaling pathways control gene expression of the NRT2 transporters. Although several regulators of these transporters have been identified in response to either N or C signals, the response of NRT2 gene expression to the interaction of these signals has never been specifically investigated, and the underlying molecular mechanisms remain largely unknown. To address this question we used an original systems biology approach to model a regulatory gene network targeting NRT2.1, NRT2.2, NRT2.4, and NRT2.5 in response to N/C signals. Our systems analysis of the data identified three transcription factors, TGA3, MYC1, and bHLH093. Functional analysis of mutants combined with yeast one-hybrid experiments confirmed that all three transcription factors are regulators of NRT2.4 or NRT2.5 in response to N or C signals. These results reveal a role for TGA3, MYC1, and bHLH093 in controlling the expression of root NRT2 transporter genes.
  • No Thumbnail Available
    Item
    Loop assembly: a simple and open system for recursive fabrication of DNA circuit
    (2019) Pollak, Bernardo; Cerda, Ariel; Delmans, Mihails; Álamos, Simón; Moyano, Tomás; West, Anthony; Gutiérrez, Rodrigo A.; Patron, Nicola J.; Federici, Fernán; Haseloff, Jim
    High-efficiency methods for DNA assembly have enabled the routine assembly of syntheticDNAs of increased size and complexity. However, these techniques require customization,elaborate vector sets or serial manipulations for the different stages of assembly. We have developed Loop assembly based on a recursive approach to DNA fabrication. Thesystem makes use of two Type IIS restriction endonucleases and corresponding vector sets forefficient and parallel assembly of large DNA circuits. Standardized level 0 parts can be assem-bled into circuits containing 1, 4, 16 or more genes by looping between the two vector sets.The vectors also contain modular sites for hybrid assembly using sequence overlap methods. Loop assembly enables efficient and versatile DNA fabrication for plant transformation. Weshow the construction of plasmids up to 16 genes and 38 kb with high efficiency (> 80%).We have characterized Loop assembly on over 200 different DNA constructs and validatedthe fidelity of the method by high-throughput Illumina plasmid sequencing. Our method provides a simple generalized solution for DNA construction with standardizedparts. The cloning system is provided under an OpenMTA license for unrestricted sharing andopen access.