Rapid transcriptome responses of maize (Zea mays) to UV-B in irradiated and shielded tissues

BACKGROUND: Depletion of stratospheric ozone has raised terrestrial levels of ultraviolet-B radiation (UV-B), an environmental change linked to an increased risk of skin cancer and with potentially deleterious consequences for plants. To better understand the processes of UV-B acclimation that result in altered plant morphology and physiology, we investigated gene expression in different organs of maize at several UV-B fluence rates and exposure times. RESULTS: Microarray hybridization was used to assess UV-B responses in directly exposed maize organs and organs shielded by a plastic that absorbs UV-B. After 8 hours of high UV-B, the abundance of 347 transcripts was altered: 285 were increased significantly in at least one organ and 80 were downregulated. More transcript changes occurred in directly exposed than in shielded organs, and the levels of more transcripts were changed in adult compared to seedling tissues. The time course of transcript abundance changes indicated that the response kinetics to UV-B is very rapid, as some transcript levels were altered within 1 hour of exposure. CONCLUSIONS: Most of the UV-B regulated genes are organ-specific. Because shielded tissues, including roots, immature ears, and leaves, displayed altered transcriptome profiles after exposure of the plant to UV-B, some signal(s) must be transmitted from irradiated to shielded tissues. These results indicate that there are integrated responses to UV-B radiation above normal levels. As the same total UV-B irradiation dose applied at three intensities elicited different transcript profiles, the transcriptome changes exhibit threshold effects rather than a reciprocal dose-effect response. Transcriptome profiling highlights possible signaling pathways and molecules for future research

Editorial: Plant Genome-Epigenome Integrity Under Environmental Stress

Plants, due to their sessile life style, need to cope with different environmental stresses to allow proper growth and development. DNA, in association with histones, is packaged into chromatin. Several changes in chromatin structure, such as post-translational modifications of histones and DNA methylation strongly affect chromatin accessibility. In past years, it has been demonstrated that biotic and abiotic stresses trigger changes in chromatin structure, allowing modification of transcriptional programs and modulation of genome structure. Chromatin compaction and its accessibility to various factors, such as transcription factors or DNA repair enzymes, regulate transcriptional activity and also DNA repair efficiency. Chromatin remodelers, histone readers/modifiers and DNA methylases/demethylases are among the predominant factors that contribute to genome and epigenome dynamics.Fil: 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; ArgentinaFil: Roldán Arjona, Teresa. Universidad de Córdoba; EspañaFil: Molinier, Jean. Institut de Biologie Moléculaire Des Plantes; Francia. Centre National de la Recherche Scientifique; Franci

DDM1 and ROS1 have a role in UV-B induced- and oxidative DNA damage in A. thaliana

Absorption of UV-B by DNA induces the formation of covalent bonds between adjacent pyrimidines. In maize and arabidopsis, plants deficient in chromatin remodeling show increased DNA damage compared to WT plants after a UV-B treatment. However, the role of enzymes that participate in DNA methylation in DNA repair after UV-B damage was not previously investigated. In this work, we analyzed how chromatin remodeling activities that have an effect on DNA methylation affects the repair of UV-B damaged DNA using plants deficient in the expression of DDM1 and ROS1. First, we analyzed their regulation by UV-B radiation in arabidopsis plants. Then, we demonstrated that ddm1 mutants accumulated more DNA damage after UV-B exposure compared to Col0 plants. Surprisingly, ros1 mutants show less CPDs and 6-4PPs than WT plants after the treatment under light conditions, while the repair under dark conditions is impaired. Transcripts for two photolyases are highly induced by UV-B in ros1 mutants, suggesting that the lower accumulation of photoproducts by UV-B is due to increased photorepair in these mutants. Finally, we demonstrate that oxidative DNA damage does not occur after UV-B exposure in arabidopsis plants; however, ros1 plants accumulate high levels of oxoproducts, while ddm1 mutants have less oxoproducts than Col0 plants, suggesting that both ROS1 and DDM1 have a role in the repair of oxidative DNA damage. Together, our data provide evidence that both DDM1 and ROS1, directly or indirectly, participate in UV-B induced- and oxidative DNA damage repair.Fil: Qüesta, Julia I.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); Argentina. Universidad Nacional de Rosario; ArgentinaFil: Fina, Julieta Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); Argentina. Universidad Nacional de Rosario; ArgentinaFil: Casati, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); Argentina. Universidad Nacional de Rosario; Argentin

E2Fb and E2Fa transcription factors independently regulate the DNA damage response after ultraviolet B exposure in Arabidopsis

Ultraviolet (UV)B radiation affects plant growth inhibiting cell proliferation. This inhibition is in part controlled by the activity of transcription factors from the E2F family. In particular, the participation of E2Fc and E2Fe in UV-B responses in Arabidopsis plants was previously reported. However, the E2Fa and E2Fb contribution to these processes has still not been investigated. Thus, in this work, we provide evidence that, in Arabidopsis, both E2Fa and E2Fb control leaf size under UV-B conditions without participating in the repair of cyclobutane pyrimidine dimers in the DNA. Nevertheless, in UV-B-exposed seedlings, E2Fa, but not E2Fb, regulates primary root elongation, cell proliferation, and programmed cell death in the meristematic zone. Using e2fa mutants that overexpress E2Fb, we showed that the role of E2Fa in the roots could not be replaced by E2Fb. Finally, our results show that E2Fa and E2Fb differentially regulate the expression of genes that activate the DNA damage response and cell cycle progression, both under conditions without UV-B and after exposure. Overall, we showed that both E2Fa and E2Fb have different and non-redundant roles in developmental and DNA damage responses in Arabidopsis plants exposed to UV-B.Fil: Gómez, María Sol. Universidad Nacional de Rosario; Argentina. CENTRO DE BIOLOGIA MOLECULAR SEVERO OCHOA (CBMSO) ; UNIVERSIDAD AUTONOMA DE MADRID;Fil: Sheridan, María Luján. 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; ArgentinaFil: 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; Argentin

Ultraviolet-B Radiation Represses Primary Root Elongation by Inhibiting Cell Proliferation in the Meristematic Zone of Arabidopsis Seedlings

In Arabidopsis thaliana plants, exposure to UV-B induces an inhibition of primary root elongation. Different mutants have been isolated that are deficient in this response; however, little is known about the cellular and molecular mechanisms that regulate inhibition of root elongation in seedlings exposed to UV-B. In this work, we investigated the effect UV-B irradiation of different organs on primary root elongation. Our results demonstrate that irradiation of the leaves and shoots only induce a partial inhibition of primary root elongation, while when only roots are exposed to this radiation, primary root inhibition is similar as that measured when the complete seedling is irradiated. The consequences of exposure at different root developmental stages and times after the end of the treatment was also studied. We here show that inhibition of primary root elongation is a consequence of a decrease in cell proliferation in the meristematic zone of the primary roots, while the elongation zone size is not affected by the treatment. The decrease in cell number after UV-B exposure is partially compensated by an increase in cell length in the root meristem; however, this compensation is not enough to maintain the meristem size. We also here demonstrate that, similarly as what occurs in developing leaves, GROWTH REGULATING FACTOR 3 (GRF3) transcription factor regulates cell proliferation in UV-B irradiated roots; however, and in contrast to what occurs in the leaves, this response does not depend on the presence of MITOGEN ACTIVATED PROTEIN KINASE 3 (MPK3). Inhibition of primary root elongation by UV-B under our experimental conditions is also independent of the UV-B photoreceptor UV RESISTANT LOCUS 8 (UVR8) or ATAXIA TELANGIECTASIA MUTATED (ATM); but a deficiency in ATM AND RAD3-RELATED (ATR) expression increases UV-B sensitivity in the roots. Finally, our data demonstrate that UV-B affects primary root growth in various Arabidopsis accessions, showing different sensitivities to this radiation.Fil: Sheridan, María Luján. 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; ArgentinaFil: Simonelli, Lucio. 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; ArgentinaFil: Giustozzi, Marisol. 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; ArgentinaFil: 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; Argentin

Flavonols protect Arabidopsis plants against UV-B deleterious effects

Flavonols are synthesized by flavonol synthase (FLS) enzymes (Martens et al., 2010). These compounds absorb UV-B light in the 280?320 nm region, and their concentration increases in plants exposed to environmental abiotic and biotic stresses, including UV-B; consequently, flavonols are thought to act as UV-B filters (Agati et al., 2011). It has been also suggested that these metabolites function as reactive oxygen species (ROS) scavengers, as they contain an OH- group in the 3-position of the flavonoid skeleton, which allows them to chelate metals, inhibiting the formation of free radicals and ROS accumulation, once formed (Agati et al., 2009). For these reasons, it has been suggested that flavonols play uncharacterized roles in UV responses (Verdan et al., 2011). Nevertheless, despite the fact that the role of flavonols in UV-B protection has been inferred; the protection conferred by flavonols on the target sites of UV-B-damage has not been directly proven in planta. Recently, we demonstrated that maize FLS1 (ZmFLS1) complements the flavonol deficiency of the Arabidopsis fls1 mutant and decreases its high level of anthocyanins, characteristic of this mutant plant (Falcone Ferreyra et al., 2010). In order to demonstrate that flavonols protect plants against UV-B damage, we generated Arabidopsis transgenic plants overexpressing the maize FLS1 cDNA (35S:ZmFLS1), and evaluated different responses of these transgenic plants against UV-B damage.Fil: Emiliani, Julia.Fil: Grotewold, Erich.Fil: Falcone Ferreyra, María Lorena.Fil: Casati, Paula

Rapid Maize Leaf and Immature Ear Responses to UV-B Radiation

Because of their sessile lifestyle, plants have evolved adaptations to environmental factors, including UV-B present in solar radiation. To gain a better understanding of the initial events in UV-B acclimation, we have analyzed a 10 min to 1 h time course of transcriptome responses in irradiated and shielded leaves, and immature maize ears to unravel the systemic physiological and developmental responses in exposed and shielded organs. After 10 min of UV-B exposure, 262 transcripts are changed by at least two-fold in irradiated leaves, and this number doubles after 1 h. Indicative of the rapid modulation of transcription, 130 transcripts are only changed after 10 min. This is true not only in irradiated leaves, but also in shielded tissues. After 10 min of exposure, the overlap in transcriptome changes in irradiated and shielded organs is significant; however, after 30 min of UV-B, there are only two transcripts showing similar UV-B regulation between the three organs; 35 are similarly regulated in both IL and SL. Therefore, at longer irradiation times, there is more specificity of responses, and these are organ-specific. We suggest that early signaling in different tissues may be elicited by common signaling pathways, while at longer exposure times responses become more specific. To identify metabolites as possible signaling molecules, we looked for compounds that increased within 5–90 min in both irradiated and shielded leaves, to explain the kinetics of profound transcript changes within 1 h. We found that myoinositol is one such candidate metabolite; and we also demonstrate that if 0.1 mM myoinositol is applied to leaves of greenhouse maize, some metabolites that are changed by UV-B are also changed similarly by the chemical treatment. Therefore, this metabolite can partially mimic UV irradiation

ASF1 Proteins are Involved in UV-induced DNA Damage Repair and are Cell Cycle Regulated by E2F Transcription Factors in Arabidopsis thaliana

ASF1 is a key histone H3/H4 chaperone that participates in a variety of DNA and chromatin-related processes, including DNA repair, where chromatin assembly and disassembly is of primaryrelevance. Information concerning the role of ASF1 proteins in post-UV response in higher plants is currently limited. In Arabidopsis thaliana, an initial analysis of in vivo localization of ASF1A andASF1B indicates that both proteins are mainly expressed in proliferative tissues. In silico promoteranalysis identified ASF1A and ASF1B as potential targets of E2F transcription factors. Theseobservations were experimentally validated, both in vitro by electrophoretic mobility shift assays, and in vivo by chromatin immunoprecipitation assays and expression analysis using transgenic plants with altered levels of different E2F transcription factors. These data suggest that ASF1A and ASF1B are regulated during cell cycle progression through E2F transcription factors. In addition, we found that ASF1A and ASF1B are associated with the UV-B induced DNA damage response in A. thaliana. Transcript levels of ASF1A and ASF1B were increased following a UV-B-treatment. Consistent with a potential role in ultraviolet-B (UV-B) response, RNAi silenced plants of both genes showed increased sensitivity to UV-B compared to wild type plants. Finally, by coimmunoprecipitation analysis, we found that ASF1 physically interacts with N-terminal acetylated histones H3 and H4, and with acetyltransferases of the HAM subfamily, which are known to be involved in cell cycle control and DNA repair, among other functions. Together, here we provide evidence that ASF1A and ASF1B are regulated by cell cycle progression and are involved in DNA repair after UV-B irradiation.Fil: Lario, Luciana Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); ArgentinaFil: Gutierrez, Crisanto. Universidad Autónoma de Madrid; EspañaFil: Ramirez Parra, Elena. Universidad Politecnica de Madrid; EspañaFil: Spampinato, Claudia Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); ArgentinaFil: Casati, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); Argentin

Repression of Growth Regulating Factors by the MicroRNA396 Inhibits Cell Proliferation by UV-B Radiation in Arabidopsis Leaves

Because of their sessile lifestyle, plants are continuously exposed to solar UV-B radiation. Inhibition of leaf growth is one of the most consistent responses of plants upon exposure to UV-B radiation. In this work, we investigated the role of GROWTH-REGULATING FACTORs (GRFs) and of microRNA miR396 in UV-B–mediated inhibition of leaf growth in Arabidopsis thaliana plants. We demonstrate that miRNA396 is upregulated by UV-B radiation in proliferating tissues and that this induction is correlated with a decrease in GRF1, GRF2, and GRF3 transcripts. Induction of miR396 results in inhibition of cell proliferation, and this outcome is independent of the UV-B photoreceptor UV resistance locus 8, as well as ATM AND RAD3–RELATED and the mitogen-activated protein kinase MPK6, but is dependent on MPK3. Transgenic plants expressing an artificial target mimic directed against miR396 (MIM396) with a decrease in the endogenous microRNA activity or plants expressing miR396-resistant copies of several GRFs are less sensitive to this inhibition. Consequently, at intensities that can induce DNA damage in Arabidopsis plants, UV-B radiation limits leaf growth by inhibiting cell division in proliferating tissues, a process mediated by miR396 and GRFs.Fil: Casadevall, Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); ArgentinaFil: Rodriguez Virasoro, Ramiro Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Debernardi, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Palatnik, Javier Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Casati, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); Argentin

Analysis of E2FA protein in the response of arabidopsis thaliana plants to UV-B radiation

Plants use sunlight to direct and regulate essential processes. Among the components of solar radiation is UV-B radiation (280-315 nm) that at high intensities generates harmful effects on plants. Because of this, plants have developed multiple mechanisms of tolerance and adaptation to UV-B radiation. Among the effects caused by high doses of UV-B radiation in plants are damage to DNA, lipids and proteins. Two common plant phenotypes after UV-B exposure are inhibition of leaf growth and primary root elongation.The shape and architecture of plants are determined by processes that modulate growth and differentiation of organs, which control the number, size and type of cells that constitute them. One of the pathways involved in the regulation of cell division, growth and differentiation is the Retinoblastoma pathway, in which the Retinoblastoma protein (RBR), the E2F transcription factors and the DP dimerization proteins participate. This pathway regulates the G1/S cell cycle transition, one of the key stages of cell cycle control in eukaryotes. E2F transcription factors serve crucial and antagonistic roles in several pathways related to cell division, DNA repair, and differentiation. In particular, E2Fa activates transcription binding to DNA cooperatively with DP proteins through a specific recognition site that is found in the promoter region of several genes whose products are involved in cell cycle regulation or DNA replication.Based on this, the aim of this work is to understand the participation of E2F transcription factors, in particular E2Fa, in the response of Arabidopsis thaliana plants to ultraviolet-B radiation. In the laboratory, using mutant lines in the E2Fa gene (e2fa-1), we observed that the primary roots are less affected by a treatment with UV-B radiation than WT lines, when we analyzed their elongation and also at the cellular level. In addition, the primary roots of e2fa mutants showed significantly fewer dead meristematic cells after a UV-B treatment than WT plants. Regarding the aerial part of the plants, we also found that growth of the proliferating leaves of e2fa-1 lines is less affected by UV-B radiation than the WT leaves. Together, these results suggest that E2Fa regulates growth, development and programmed cell death in response to UV-B radiation.Fil: Sheridan, María Luján. 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; ArgentinaFil: 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; ArgentinaFil: 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; ArgentinaLVI Annual Meeting Argentine Society for Biochemistry and Molecular Biology; XV Annual Meeting Argentinean Society for General MicrobiologyArgentinaSociedad Argentina de Investigación en Bioquímica y Biología MolecularSociedad Argentina de Microbiología Genera