Abstract:

Nicotinate degradation has hitherto been elucidated only in bacteria. In the ascomycete Aspergillus nidulans, six loci, hxnS/AN9178 encoding the molybdenum cofactor-containing nicotinate hydroxylase, AN11197 encoding a Cys2/His2 zinc finger regulator HxnR, together with AN11196/hxnZ, AN11188/hxnY, AN11189/hxnP and AN9177/hxnT, are clustered and stringently co-induced by a nicotinate derivative and subject to nitrogen metabolite repression mediated by the GATA factor AreA. These genes are strictly co-regulated by HxnR. Within the hxnR gene, constitutive mutations map in two discrete regions. Aspergillus nidulans is capable of using nicotinate and its oxidation products 6-hydroxynicotinic acid and 2,5-dihydroxypyridine as sole nitrogen sources in an HxnR-dependent way. HxnS is highly similar to HxA, the canonical xanthine dehydrogenase (XDH), and has originated by gene duplication, preceding the origin of the Pezizomycotina. This cluster is conserved with some variations throughout the Aspergillaceae. Our results imply that a fungal pathway has arisen independently from bacterial ones. Significantly, the neo-functionalization of XDH into nicotinate hydroxylase has occurred independently from analogous events in bacteria. This work describes for the first time a gene cluster involved in nicotinate catabolism in a eukaryote and has relevance for the formation and evolution of co-regulated primary metabolic gene clusters and the microbial degradation of N-heterocyclic compounds.

Author affiliation: Ámon, Judit. University of Szeged. Faculty of Science and Informatics; Hungría

Author affiliation: Fernández y Martín, Rafael. Université Paris Sud; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Author affiliation: Bokor, Eszter. University of Szeged. Faculty of Science and Informatics; Hungría

Author affiliation: Cultrone, Antonietta. Université Paris Sud; Francia

Author affiliation: Kelly, Joan M.. University of Essex; Reino Unido

Author affiliation: Flipphi, Michel. Université Paris Sud; Francia

Author affiliation: Scazzocchio, Claudio. Université Paris Sud; Francia. University of Essex; Reino Unido. Imperial College London; Reino Unido

Author affiliation: Hamari, Zsuzsanna. Université Paris Sud; Francia. University of Szeged. Faculty of Science and Informatics; Hungría

Abstract:

Germinating oilseeds convert stored lipids into sugars and thereafter in metabolic energy that is used in seedling growth and establishment. During germination, the induced lipolysis linked to the glyoxylate pathway and gluconeogenesis produces sucrose, which is then transported to the embryo and driven through catabolic routes. Herein, we report that the sunflower transcription factor HaWRKY10 regulates carbon partitioning by reducing carbohydrate catabolism and increasing lipolysis and gluconeogenesis. HaWRKY10 was regulated by abscisic acid and gibberellins in the embryo leaves 48 hours after seed imbibition and highly expressed during sunflower seed germination and seedling growth, concomitantly with lipid mobilization. Sunflower leaf discs overexpressing HaWRKY10 showed repressed the expression of genes related to sucrose cleavage and glycolysis compared to controls. Moreover, HaWRKY10 constitutive expression in Arabidopsis seeds produced higher decrease in lipid reserves, whereas starch and sucrose were more preserved compared to wild type. Gene transcripts abundance and enzyme activities involved in stored lipid mobilization and gluconeogenesis increased more in transgenic than in wild type seeds 36 hours after imbibition whereas the negative regulator of lipid mobilization, ABI4, was repressed. Altogether, the results point out a functional parallelism between tissues and plant species, and reveal HaWRKY10 as a positive regulator of storage reserve mobilization in sunflower.

Author affiliation: Raineri, Jesica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina

Author affiliation: Hartman, Matias Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina

Author affiliation: Chan, Raquel Lia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina

Author affiliation: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina

Author affiliation: Ribichich, Karina Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina

Abstract:

Interval timing within the seconds-to-minutes range involves the interaction of the prefrontal cortex and basal ganglia via dopaminergic-glutamatergic pathways. Because the secreted protein BDNF is able to modulate dopamine release as well as glutamatergic activity, we hypothesized that BDNF may be important for these timing mechanisms. Recently the transcription factor CaRF was identified as an important modulator of BDNF expression in the cerebral cortex. In the present study, a strain of Carf knockout mice were evaluated for their ability to acquire the "Start" and "Stop" response thresholds under sequential and simultaneous training conditions, using multiple (15-s and 45-s) or single (30-s) target durations using the peak-interval procedure. Both Carf+/- and Carf-/- mice were impaired in their ability to acquire timed response thresholds relative to Carf+/+ mice. These results suggest that the inhibitory processes required to set response thresholds and exert temporal control of behavior may be dependent on CaRF regulation of genes including Bdnf in cortico-striatal circuits.

Author affiliation: Agostino, Patricia. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Cronobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Author affiliation: Cheng, C. K.. A*STAR/Duke-NUS Neuroscience Research Partnership; Singapur

Author affiliation: Williams, C. L.. Duke University. Department of Psychology and Neuroscience; Estados Unidos

Author affiliation: West, A. E.. Duke University. Department of Neurobiology; Estados Unidos

Author affiliation: Meck, W. H.. Duke University. Department of Psychology and Neuroscience; Estados Unidos

Abstract:

UV-B radiation inhibits plant growth, and this inhibition is, to a certain extent, regulated by miR396-mediated repression of Growth Regulating Transcription factors (GRFs). Moreover, E2Fe transcription factor also modulates Arabidopsis leaf growth. Here, we provide evidence that, at UV-B intensities that induce DNA damage, E2Fc participates in the inhibition of cell proliferation. We demonstrate that E2Fc-deficient plants show a lower inhibition of leaf size under UV-B conditions that damage DNA, decreased cell death after exposure and altered SOG1 and ATR expression. Interestingly, the previously reported participation of E2Fe in UV-B responses, which is a transcriptional target of E2Fc, is independent and different from that described for E2Fc. Conversely, we here demonstrate that E2Fc has an epistatic role over the miR396 pathway under UV-B conditions. Finally, we show that inhibition of cell proliferation by UV-B is independent of the regulation of class II TCP transcription factors. Together, our results demonstrate that E2Fc is required for miR396 activity on cell proliferation under UV-B, and that its role is independent of E2Fe, probably modulating DNA damage responses through the regulation of SOG1 and ATR transcript levels.

Author affiliation: Gomez, Maria Sol. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina

Author affiliation: Falcone Ferreyra, María Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina

Author affiliation: Sheridan, Maria Luján. Universidad Nacional de Rosario; Argentina

Author affiliation: Casati, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina

Abstract:

Arabidopsis thaliana HomeoBox 1 (AtHB1) is a homeodomain-leucine zipper transcription factor described as a transcriptional activator with unknown function. Its role in A. thaliana development was investigated. AtHB1 expression was analyzed in transgenic plants bearing its promoter region fused to reporter genes. Knock-down mutant and overexpressor plant phenotypes were analyzed in different photoperiod regimes. AtHB1 was mainly expressed in hypocotyls and roots and up-regulated in seedlings grown under a short-day photoperiod. AtHB1 knock-down mutants and overexpressors showed shorter and longer hypocotyls, respectively, than wild type (WT). AtHB1 transcript levels were lower in PHYTOCHROME-INTERACTING FACTOR 1 (PIF1) mutants than in controls, suggesting that AtHB1 is regulated by PIF1 in hypocotyls. β-glucuronidase (GUS) activity in Nicotiana benthamiana leaves cotransformed with PromAtHB1::GUS and 35S::PIF1 indicated that PIF1 induces AtHB1 expression. Hypocotyl lenght was measured in seedlings of athb1, pif1, or double athb1/pif1 mutants and PIF1 or AtHB1 overexpressors in WT, athb1 or pif1 backgrounds, both in short- or long-day. These analyses allowed us to determine that AtHB1 is a factor acting downstream of PIF1. Finally, a transcriptome analysis of athb1 mutant hypocotyls revealed that AtHB1 regulates genes involved in cell wall composition and elongation. The results suggest that AtHB1 acts downstream of PIF1 to promote hypocotyl elongation, especially in response to short-day photoperiods.

Author affiliation: Capella, Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina

Author affiliation: Ribone, Pamela Anahí. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina

Author affiliation: Arce, Agustín Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina

Author affiliation: Chan, Raquel Lia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina

Abstract:

IMPORTANCE: Disseminated retinoblastoma is usually fatal. Identification of small amounts (minimal dissemination [MD]) of tumor cells in extraocular sites might be a tool for designing appropriate treatments. OBJECTIVE: To test cone-rod homeobox (CRX) transcription factor as a lineage-specific molecular marker for metastatic retinoblastoma and for evaluation of MD. DESIGN, SETTING, AND PARTICIPANTS: In a prospective cohort design study, we evaluated CRX messenger RNA (mRNA) by retrotranscription followed by real-time polymerase chain reaction as a diagnostic test in samples obtained from bone marrow, peripheral blood, and cerebrospinal fluid (CSF) at diagnosis, after induction chemotherapy, and during follow-up. The study was conducted from June 30, 2008, to June 30, 2014. Seventeen retinoblastoma primary tumors, 2 retinoblastoma cell lines, and 47 samples of bone marrow from other cancers (controls) were studied. Seventeen patients with metastatic retinoblastoma (9 at diagnosis, 8 at relapse; age range: 18-41 months) were included. MAIN OUTCOMES AND MEASURES: Detection of CRX mRNA as a marker for metastatic retinoblastoma and MD in bone marrow and CSF and its correlation with clinical findings. RESULTS: Cone-rod homeobox mRNA was expressed in all tumors (relative expression levels range, 8.1 × 10-5 to 5.6) and cell lines. In control samples, there was no amplification of CRX; only the housekeeping gene (GAPDH) demonstrated amplification. Bone marrow metastatic cells showed expression of CRX mRNA in all 9 children presenting with metastasis at the diagnosis (relative expression levels, 6.0 × 10-5 to 0.67). After induction chemotherapy, no evidence of MD of tumor cells was seen in any of the 8 responding children since only GAPDH showed amplification. In the CSF of children who had a metastatic relapse, CRX mRNA detection was positive in 2 patients in whom no conclusive results were reached by immunocytology for disialoganglioside GD2. Minimal dissemination in the CSF was associated with a clinical relapse in 2 cases. No concomitant MD was evident in the bone marrow in any case. CONCLUSIONS AND RELEVANCE: These data suggest that CRX mRNA is a novel marker for retinoblastoma at extraocular sites. In this study among patients with bone marrow metastasis, there was a quick, complete, and sustained molecular response after induction chemotherapy. In all patients with secondary metastasis, CSF relapse occurred independently from the bone marrow, suggesting a sanctuary site.

Author affiliation: Torbidoni, Ana Vanesa. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Author affiliation: Laurent, Viviana Eunice. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentina

Author affiliation: Sampor, Claudia. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentina

Author affiliation: Ottaviani, Daniela. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentina

Author affiliation: Vazquez, Valeria. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentina

Author affiliation: Gabri, Mariano Rolando. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina

Author affiliation: Vazquez Rossi, Jorge Eduardo. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentina

Author affiliation: De Dávila, María T.. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentina

Author affiliation: Alonso, Cristina. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentina

Author affiliation: Alonso, Daniel Fernando. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentina

Author affiliation: Chantada, Guillermo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentina

Abstract:

Los factores de transcripción (FTs) HD-Zip I de Arabidopsis thaliana, AtHB12 y AtHB7, presentan una alta similitud de secuencia con NaHD20 de Nicotiana attenuata. La expresión de NaHD20 se induce por tratamientos relacionados al estrés y se expresa fuertemente en corolas. NaHD20 controla el aumento de los niveles de ABA de las hojas cuando las plantas se encuentran en condiciones de deshidratación y regula la producción y apertura de flores tanto en condiciones normales como de estrés hídrico severo. NaHD20 induce el desarrollo y la apertura de las flores, controlando los niveles de la hormona ABA. Durante la apertura de la corola, este FT induce la emisión del volátil acetona de bencilo, responsable de atraer polinizadores. AtHB12 se expresa en estadios tempranos, controlando el desarrollo foliar y radicular durante el estadio vegetativo. AtHB7 aumenta su expresión en el estadio reproductivo e induce el desarrollo foliar, la producción de clorofila y la fotosíntesis. AtHB12 y AtHB7 se regulan entre sí, afectado la expresión de su parálogo. En estrés hídrico, estos FTs se inducen fuertemente aunque actúan de manera diferente: AtHB12 regula el consumo mientras que AtHB7 controla la pérdida de agua a través del cierre estomático. AtHB12 tiene un efecto positivo en la producción de semillas estrés hídrico leve. Los tres FTs participan en mecanismos del desarrollo vegetal relacionados al estrés hídrico y a la hormona ABA. AtHB12 y AtHB7, considerados duplicaciones génicas, afectan la expresión de su parálogo, coordinando finamente la regulación de los procesos en los que están involucrados.

The Arabidopsis thaliana AtHB12 and AtHB7 are paralogues TFs from HD-Zip I family, known to be involved in water stress responses. NaHD20 from Nicotiana attenuata presents high sequence similarity with the first two. NaHD20 expression was induced by phytohormones like jasmonic acid, salicylic acid and abscisic acid (ABA), and by water stress. This TF was highly expressed in corollas. NaHD20 controls ABA levels in leaves during water stress and also regulates flowers production and opening in both control and stress conditions. NaHD20 also regulates flowers development and opening, by controlling ABA levels in corollas. During the opening of the corolla, NaHD20 regulates the release of the volatile compound, bencyl acetone, responsible for pollinators attraction. AtHB12 controls root and leaf development in seedlings under standard growth conditions and water uptake and seed production in water-stressed plants. On the other hand, AtHB7 controls leaf development, stomatal conductance, chlorophyll levels and photosynthesis only in mature plants. Importantly, AtHB7 and AtHB12 are part of a feedback loop that regulates each other expression either positively or negatively, depending on the developmental stage of the plant and environmental conditions, probably involving other factors. The results suggested that AtHB7 and AtHB12 diverged along the evolution to fine tune processes associated with development and responses to water stress. These three TFs participate in mechanisms involved in water stress and ABA responses. Regarding AtHB12 and AtHB7, considered as gene duplications, a feed-back loop allows them to regulate each other expression finely coordinating the functions in which they are involved.

Consejo Nacional de Investigaciones Científicas y Técnicas

Universidad Nacional del Litoral

Abstract:

Arabidopsis COX5b-1 encodes an isoform of the zinc binding subunit 5b of mitochondrial cytochrome c oxidase. A promoter region required for expression and induction by sucrose of this gene was analyzed using plants stably transformed with mutagenized promoter fragments fused to the gus reporter gene. Promoter dependent expression is absolutely dependent on a G-box present at -228 from the translation start site. This element interacts in vitro and in vivo with transcription factors from the bZip family, preferentially with the abscisic acid-responsive element binding factor AREB2/ABF4. A region located upstream of the G-box (-333/-259) contains elements with the core sequence ATCATT and distalB-like sequences (CCACTTG) that are required for expression in vegetative tissues. These sequences bind different sets of proteins present in plant nuclear extracts and participate in induction by sucrose (ATCATT) and abscisic acid (distalB) of the COX5b-1 promoter. We propose that the COX5b-1 promoter has acquired novel regulatory mechanisms during evolution after gene duplication. These novel mechanisms have allowed the diversification of expression patterns, but also the conservation of some responses that, as induction by sucrose, are shared by COX5b-1 and other genes encoding components of the mitochondrial respiratory chain. Conservation of these responses may be a pre-requisite for the successful incorporation of new regulatory elements in this class of genes.

Author affiliation: Comelli, Raul Nicolas. Universidad Nacional del Litoral; Argentina

Author affiliation: Viola, Ivana Loren. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina

Author affiliation: Gonzalez, Daniel Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina

Abstract:

BACKGROUND: The initiation of flowering is an important developmental transition as it marks the beginning of the reproductive phase in plants. The MADS-box transcription factors (TFs) FLOWERING LOCUS C (FLC) and SHORT VEGETATIVE PHASE (SVP) form a complex to repress the expression of genes that initiate flowering in Arabidopsis. Both TFs play a central role in the regulatory network by conferring seasonal patterns of flowering. However, their interdependence and biological relevance when acting as a complex have not been extensively studied. RESULTS: We characterized the effects of both TFs individually and as a complex on flowering initiation using transcriptome profiling and DNA-binding occupancy. We find four major clusters regulating transcriptional responses, and that DNA binding scenarios are highly affected by the presence of the cognate partner. Remarkably, we identify genes whose regulation depends exclusively on simultaneous action of both proteins, thus distinguishing between the specificity of the SVP:FLC complex and that of each TF acting individually. The downstream targets of the SVP:FLC complex include a higher proportion of genes regulating floral induction, whereas those bound by either TF independently are biased towards floral development. Many genes involved in gibberellin-related processes are bound by the SVP:FLC complex, suggesting that direct regulation of gibberellin metabolism by FLC and SVP contributes to their effects on flowering. CONCLUSIONS: The regulatory codes controlled by SVP and FLC were deciphered at the genome-wide level revealing substantial flexibility based on dependent and independent DNA binding that may contribute to variation and robustness in the regulation of flowering.

Author affiliation: Mateos, Julieta Lisa. Fundación Instituto Leloir; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Max Planck Institute for Plant Breeding Research; Alemania

Author affiliation: Madrigal, Pedro. Polish Academy of Sciences; Polonia

Author affiliation: Tsuda, Kenichi. Max Planck Institute for Plant Breeding Research; Alemania

Author affiliation: Rawat, Vimal. Max Planck Institute for Plant Breeding Research; Alemania

Author affiliation: Richter, René. Max Planck Institute for Plant Breeding Research; Alemania

Author affiliation: Romera Branchat, Maida. Max Planck Institute for Plant Breeding Research; Alemania

Author affiliation: Fornara, Fabio. Max Planck Institute for Plant Breeding Research; Alemania

Author affiliation: Schneeberger, Korbinian. Max Planck Institute for Plant Breeding Research; Alemania

Author affiliation: Krajewski, Pawel. Polish Academy of Sciences; Polonia

Author affiliation: Coupland, George. Max Planck Institute for Plant Breeding Research; Alemania

Abstract:

The role of four cysteines present within the homeodomain of the homeodomain-leucine zipper (HD-Zip) class III protein Athb-9 has been studied. DNA binding by the Athb-9 HD-Zip domain was only observed after incubation in the presence of reducing agents or the thioredoxin system, suggesting that the protein is sensitive to redox conditions. A similar behavior was observed for proteins that show the same binding specificity of Athb-9 present in nuclear extracts. The use of single and double mutants indicated that two out of three of the cysteines at positions 23, 38 and 42 are required for redox sensitivity, while Cys58 is not involved. A role of Cys23 and Cys58 in determining the DNA binding efficiency and specificity, respectively, of the reduced Athb-9 HD-Zip domain was also evident from these studies. It can be postulated that redox conditions may modulate the function of Athb-9 in plant development. Sequence conservation suggests that the results can be extended to all HD-Zip III transcription factors.

Author affiliation: Comelli, Raul Nicolas. Universidad Nacional del Litoral; Argentina

Author affiliation: Gonzalez, Daniel Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina