Germination and Early Seedling Development in Quercus ilex Recalcitrant and Non-dormant Seeds: Targeted Transcriptional, Hormonal, and Sugar Analysis

Seed germination and early seedling development have been studied in the recalcitrant species Quercus ilex using targeted transcriptional, hormonal, and sugar analysis. Embryos and seedlings were collected at eight morphologically defined developmental stages, S0–S7. A typical triphasic water uptake curve was observed throughout development, accompanied by a decrease in sucrose and an increase in glucose and fructose. Low levels of abscisic acid (ABA) and high levels of gibberellins (GAs) were observed in mature seeds. Post-germination, indole-3-acetic acid (IAA), increased, whereas GA remained high, a pattern commonly observed during growth and development. The abundance of transcripts from ABA-related genes was positively correlated with the changes in the content of the phytohormone. Transcripts of the drought-related genes Dhn3 and GolS were more abundant at S0, then decreased in parallel with increasing water content. Transcripts for Gapdh, and Nadh6 were abundant at S0, supporting the occurrence of an active metabolism in recalcitrant seeds at the time of shedding. The importance of ROS during germination is manifest in the high transcript levels for Sod and Gst, found in mature seeds. The results presented herein help distinguish recalcitrant (e.g., Q. ilex) seeds from their orthodox counterparts. Our results indicate that recalcitrance is established during seed development but not manifest until germination (S1–S3). Post-germination the patterns are quite similar for both orthodox and recalcitrant seeds.España Ministerio de Ciencia e Innovación AGL2009-12243-C02-0

Kaolin and salicylic acid alleviate summer stress in rainfed olive orchards by modulation of distinct physiological and biochemical responses

In a changing world, the search for new agronomic practices that help crops to maintain and/or increase yields and quality is a continuous challenge. We aim to evaluate kaolin (KL) and salicylic acid (SA) effectiveness as summer stress alleviating agents through physiological, biochemical and immunohistochemical analysis. Olive trees (Olea europaea L. cv. Cobrançosa) grown under rainfed conditions were sprayed with 5% KL and 100 μM SA, at the beginning of summer, during two consecutive years. KL enhanced relative water content (RWC), stomatal conductance (gs) net photosynthesis (A) and leaf indole-3-acetic acid (IAA) signal, and decreased leaf sclerophylly, secondary metabolites and non-structural carbohydrates accumulation and abscisic acid (ABA).The trees treated with SA showed changes on IAA and ABA dynamics, and an enhancement in RWC, gs, A, soluble proteins, and leaf P and Mg concentrations during the summer. Notably, KL and SA also allowed a faster restauration of the physiological functions during stress relief. In sum, KL and SA foliar sprays alleviated the negative effects induced by summer stress in olive trees performance, by modulation of distinct physiological and biochemical responses.info:eu-repo/semantics/publishedVersio

Induction of radiata pine somatic embryogenesis at high temperatures provokes a long-term decrease in dna methylation/hydroxymethylation and differential expression of stress-related genes

Based on the hypothesis that embryo development is a crucial stage for the formation of stable epigenetic marks that could modulate the behaviour of the resulting plants, in this study, radiata pine somatic embryogenesis was induced at high temperatures (23¿ C, eight weeks, control; 40¿ C, 4 h; 60¿ C, 5 min) and the global methylation and hydroxymethylation levels of emerging embryonal masses and somatic plants were analysed using LC-ESI-MS/ MS-MRM. In this context, the expression pattern of six genes previously described as stress-mediators was studied throughout the embryogenic process until plant level to assess whether the observed epigenetic changes could have provoked a sustained alteration of the transcriptome. Results indicated that the highest temperatures led to hypomethylation of both embryonal masses and somatic plants. Moreover, we detected for the first time in a pine species the presence of 5-hydroxymethylcytosine, and revealed its tissue specificity and potential involvement in heat-stress responses. Additionally, a heat shock protein-coding gene showed a down-regulation tendency along the process, with a special emphasis given to embryonal masses at first subculture and ex vitro somatic plants. Likewise, the transcripts of several proteins related with translation, oxidative stress response, and drought resilience were differentially expressed

Germination and Early Seedling Development in Quercus ilex Recalcitrant and Non-dormant Seeds: Targeted Transcriptional, Hormonal, and Sugar Analysis

Seed germination and early seedling development have been studied in the recalcitrant species Quercus ilex using targeted transcriptional, hormonal, and sugar analysis. Embryos and seedlings were collected at eight morphologically defined developmental stages, S0–S7. A typical triphasic water uptake curve was observed throughout development, accompanied by a decrease in sucrose and an increase in glucose and fructose. Low levels of abscisic acid (ABA) and high levels of gibberellins (GAs) were observed in mature seeds. Post-germination, indole-3-acetic acid (IAA), increased, whereas GA remained high, a pattern commonly observed during growth and development. The abundance of transcripts from ABA-related genes was positively correlated with the changes in the content of the phytohormone. Transcripts of the drought-related genes Dhn3 and GolS were more abundant at S0, then decreased in parallel with increasing water content. Transcripts for Gapdh, and Nadh6 were abundant at S0, supporting the occurrence of an active metabolism in recalcitrant seeds at the time of shedding. The importance of ROS during germination is manifest in the high transcript levels for Sod and Gst, found in mature seeds. The results presented herein help distinguish recalcitrant (e.g., Q. ilex) seeds from their orthodox counterparts. Our results indicate that recalcitrance is established during seed development but not manifest until germination (S1–S3). Post-germination the patterns are quite similar for both orthodox and recalcitrant seeds

Is the interplay between epigenetic markers related to the acclimation of Cork oak plants to high temperatures?

Trees necessarily experience changes in temperature, requiring efficient short-term strategies that become crucial in environmental change adaptability. DNA methylation and histone posttranslational modifications have been shown to play a key role in both epigenetic control and plant functional status under stress by controlling the functional state of chromatin and gene expression. Cork oak (Quercus suber L.) is a key stone of the Mediterranean region, growing at temperatures of 45°C. This species was subjected to a cumulative temperature increase from 25°C to 55°C under laboratory conditions in order to test the hypothesis that epigenetic code is related to heat stress tolerance. Electrolyte leakage increased after 35°C, but all plants survived to 55°C. DNA methylation and acetylated histone H3 (AcH3) levels were monitored by HPCE (high performance capillary electrophoresis), MS-RAPD (methylation-sensitive random-amplified polymorphic DNA) and Protein Gel Blot analysis and the spatial distribution of the modifications was assessed using a confocal microscope. DNA methylation analysed by HPCE revealed an increase at 55°C, while MS-RAPD results pointed to dynamic methylation-demethylation patterns over stress. Protein Gel Blot showed the abundance index of AcH3 decreasing from 25°C to 45°C. The immunohistochemical detection of 5-mC (5-methyl-2'-deoxycytidine) and AcH3 came upon the previous results. These results indicate that epigenetic mechanisms such as DNA methylation and histone H3 acetylation have opposite and particular dynamics that can be crucial for the stepwise establishment of this species into such high stress (55°C), allowing its acclimation and survival. This is the first report that assesses epigenetic regulation in order to investigate heat tolerance in forest trees.This work is supported by FEDER through COMPETE (Programa Operacional Factores de Competitividade) and by the FCT project PTDC/AGR-CFL/ 112996/2009. G. Pinto is hired under the programme Cie ˆncia 2008 (FCT, Portugal), co-funded by the Human Potential Operational Programme (National Strategic Reference Framework 2007–2013) and European Social Fund (EU). FCT supported the fellowship of M.C. Dias (SFRH/BPD/41700/2007). L. Valledor fellow was supported by a Marie Curie Action of the European Union (FP7-PEOPLE-IEF). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.publishe

Genome-wide association mapping identifies a new arsenate reductase enzyme critical for limiting arsenic accumulation in plants

Inorganic arsenic is a carcinogen, and its ingestion through foods such as rice presents a significant risk to human health. Plants chemically reduce arsenate to arsenite. Using genome-wide association (GWA) mapping of loci controlling natural variation in arsenic accumulation in Arabidopsis thaliana allowed us to identify the arsenate reductase required for this reduction, which we named High Arsenic Content 1 (HAC1). Complementation verified the identity of HAC1, and expression in Escherichia coli lacking a functional arsenate reductase confirmed the arsenate reductase activity of HAC1. The HAC1 protein accumulates in the epidermis, the outer cell layer of the root, and also in the pericycle cells surrounding the central vascular tissue. Plants lacking HAC1 lose their ability to efflux arsenite from roots, leading to both increased transport of arsenic into the central vascular tissue and on into the shoot. HAC1 therefore functions to reduce arsenate to arsenite in the outer cell layer of the root, facilitating efflux of arsenic as arsenite back into the soil to limit both its accumulation in the root and transport to the shoot. Arsenate reduction by HAC1 in the pericycle may play a role in limiting arsenic loading into the xylem. Loss of HAC1-encoded arsenic reduction leads to a significant increase in arsenic accumulation in shoots, causing an increased sensitivity to arsenate toxicity. We also confirmed the previous observation that the ACR2 arsenate reductase in A. thaliana plays no detectable role in arsenic metabolism. Furthermore, ACR2 does not interact epistatically with HAC1, since arsenic metabolism in the acr2 hac1 double mutant is disrupted in an identical manner to that described for the hac1 single mutant. Our identification of HAC1 and its associated natural variation provides an important new resource for the development of low arsenic-containing food such as rice

DNA methylation dynamics and MET1a-like gene expression changes during stress-induced pollen reprogramming to embryogenesis

This work was supported by Spanish Ministry of Science and Innovation (project BFU2011-23752). M.R.S. was a recipient of a postdoctoral Juan-de-la-Cierva grant (JCI-2007-123-1177). ÍNDICES DE IMPACTO DE LA REVISTA: Citas recibidas • Citas en Sopas: 66 (22-01-2023) • Citas en Web os Science: 63 (28-12-2023) JCR (Journal Impact Factor) • Año 2012 • Factor de impacto de la revista: 5.242 • Factor de impacto sin autocitas: 4.782 • Article influence score: 1.609 • Cuartil mayor: Q1 • Área: PLANT SCIENCES Cuartil: Q1 Posición en el área: 13/197 (Edicion: SCIE) SCImago Journal Rank • Año 2012 • Impacto SJR de la revista: 2.619 • Cuartil mayor: Q1 • Área: Plant Science Cuartil: Q1 Posición en el área: 15/417 • Área: Physiology Cuartil: Q1 Posición en el área: 15/185 Scopus CiteScore • Año 2012 • CiteScore de la revista: 8.5 • Área: Plant Science Percentil: 95 • Área: Physiology Percentil: 89 • Área: Medicine (all) Percentil: 90Stress-induced plant cell reprogramming involves changes in global genome organization, being the epigenetic modifications key factors in the regulation of genome flexibility. DNA methylation, accomplished by DNA methyltransferases, constitutes a prominent epigenetic modification of the chromatin fibre which is locked in a transcriptionally inactive conformation. Changes in DNA methylation accompany the reorganization of the nuclear architecture during plant cell differentiation and proliferation. After a stress treatment, in vitro-cultured microspores are reprogrammed and change their gametophytic developmental pathway towards embryogenesis, the process constituting a useful system of reprogramming in isolated cells for applied and basic research. Gene expression driven by developmental and stress cues often depends on DNA methylation; however, global DNA methylation and genome-wide expression patterns relationship is still poorly understood. In this work, the dynamics of DNA methylation patterns in relation to nuclear architecture and the expression of BnMET1a-like DNA methyltransferase genes have been analysed during pollen development and pollen reprogramming to embryogenesis in Brassica napus L. by a multidisciplinary approach. Results showed an epigenetic reprogramming after microspore embryogenesis induction which involved a decrease of global DNA methylation and its nuclear redistribution with the change of developmental programme and the activation of cell proliferation, while DNA methylation increases with pollen and embryo differentiation in a cell-type-specific manner. Changes in the presence, abundance, and distribution of BnMET1a-like transcripts highly correlated with variations in DNA methylation. Mature zygotic and pollen embryos presented analogous patterns of DNA methylation and MET1a-like expression, providing new evidence of the similarities between both developmental embryogenic programmes.Depto. de Genética, Fisiología y MicrobiologíaFac. de Ciencias BiológicasTRUEpu

Flower differentiation of azalea depends on genotype and not on the use of plant growth regulators

Flowering is a complex process which starts with the induction and development of the flower buds. For azalea (Rhododendron simsii hybrids), flower induction was hastened by the application of chlormequat and took place within 11 days after treatment. Subsequent flower bud differentiation was not altered by the application rate of the plant growth regulators (PGR) chlormequat and paclobutrazol, nor by temperature or light sum. There were however, large genotypic variations in flower bud differentiation rate. For all cultivars a linear phase until flower primordia were fully differentiated and the style started to enlarge (flower bud stage 7), was followed by a slower final development (to stage 8). The linear phase was fastest for the semi-early flowering cultivars (‘Mont Blanc’, ‘M. Marie’ and ‘Otto’), requiring only 46 or 48 days to reach flower bud stage 7 after the first PGR treatment. Two late flowering cultivars (‘Thesla’ and ‘Sachsenstern’) had the slowest differentiation, requiring 64 days to reach stage 7. The early flowering cultivars (‘H. Vogel’ sports) and two late flowering cultivars (‘Mw. G. Kint’ and ‘Tamira’) required 54 and 52 days, respectively, after the first PGR treatment to reach stage 7. To reach flower bud stage 8, a similar trend in velocity was seen, the semi-early flowering cultivars requiring the least amount of days (17 to 18 days), the late flowering cultivars ‘Thesla’ and ‘Sachsenstern’ requiring the highest amount of days (24) and the early flowering cultivars and the late flowering cultivars ‘Mw. G. Kint’ and ‘Tamira’ requiring an intermediate number of days (20 to 22 days)