Contrasting effects of urban trees on air quality: From the aerodynamic effects in streets to impacts of biogenic emissions in cities.

Urban trees are often not considered in air-quality models although they can significantly impact the concentrations of pollutants. Gas and particles can deposit on leaf surfaces, lowering their concentrations, but the tree crown aerodynamic effect is antagonist, limiting the dispersion of pollutants in streets. Furthermore, trees emit Biogenic Volatile Organic Compounds (BVOCs) that react with other compounds to form ozone and secondary organic aerosols. This study aims to quantify the impacts of these three tree effects (dry deposition, aerodynamic effect and BVOC emissions) on air quality from the regional to the street scale over Paris city. Each tree effect is added in the model chain CHIMERE/MUNICH/SSH-aerosol. The tree location and characteristics are determined using the Paris tree inventory, combined with allometric equations. The air-quality simulations are performed over June and July 2022. The results show that the aerodynamic tree effect increases the concentrations of gas and particles emitted in streets, such as NOx (+4.6 % on average in streets with trees and up to +37 % for NO2). This effect increases with the tree Leaf Area Index and it is more important in streets with high traffic, suggesting to limit the planting of trees with large crowns on high-traffic streets. The effect of dry deposition of gas and particles on leaves is very limited, reducing the concentrations of O3 concentrations by -0.6 % on average and at most -2.5 %. Tree biogenic emissions largely increase the isoprene and monoterpene concentrations, bringing the simulated concentrations closer to observations. Over the two-week sensitivity analysis, biogenic emissions induce an increase of O3, organic particles and PM2.5 street concentrations by respectively +1.1, +2.4 and + 0.5 % on average over all streets. This concentration increase may reach locally +3.5, +12.3 and + 2.9 % respectively for O3, organic particles and PM2.5, suggesting to prefer the plantation of low-emitting VOC species in cities.

In-Depth Proteomic Analysis of the Secondary Dormancy Induction by Hypoxia or High Temperature in Barley Grains.

In barley, incubation of primary dormant (D1) grains on water under conditions that do not allow germination, i.e. 30°C in air and 15°C or 30°C in 5% O2, induces a secondary dormancy (D2) expressed as a loss of the ability to germinate at 15°C in air. The aim of this study was to compare the proteome of barley embryos isolated from D1 grains and D2 ones after induction of D2 at 30°C or in hypoxia at 15°C or 30°C. Total soluble proteins were analyzed by 2DE gel-based proteomics, allowing the selection of 130 differentially accumulated proteins (DAPs) among 1,575 detected spots. According to the protein abundance profiles, the DAPs were grouped into six abundance-based similarity clusters. Induction of D2 is mainly characterized by a down-accumulation of proteins belonging to cluster 3 (storage proteins, proteases, alpha-amylase inhibitors and histone deacetylase HD2) and an up-accumulation of proteins belonging to cluster 4 (1-Cys peroxiredoxin, lipoxygenase2 and caleosin). The correlation-based network analysis for each cluster highlighted central protein hub. In addition, most of genes encoding DAPs display high co-expression degree with 19 transcription factors. Finally, this work points out that similar molecular events accompany the modulation of dormancy cycling by both temperature and oxygen, including post-translational, transcriptional and epigenetic regulation.

Potential involvement of proline and flavonols in plant responses to ozone.

Ozone is considered to be a major phytotoxic pollutant. It is an oxidizing molecule with harmful effects that can affect human health and vegetation. Due to its phytotoxicity, it constitutes a threat to food security in a context of climate change. Proline accumulation is induced in response to numerous stresses and is assumed to be involved in plant antioxidant defense. We therefore addressed the question of the putative involvement of proline in plant ozone responses by analyzing the responses of two Arabidopsis mutants (obtained in the Col-0 genetic background) altered in proline metabolism and different ecotypes with various degrees of ozone sensitivity, to controlled ozone treatments. Among the mutants, the p5cs1 mutant plants accumulated less proline than the double prodh1xprodh2 (p1p2) mutants. Ozone treatments did not induce accumulation of proline in Col-0 nor in the mutant plants. However, the variation of the photosynthetic parameter Fv/Fm in the p1p2 mutant suggests a positive effect of proline. Proline accumulation induced by ozone was only observed in the most ozone-sensitive ecotypes, Cvi-0 and Ler. Contrary to our expectations, proline accumulation could not be correlated with variations in protein oxidation (carbonylation). On the other hand, flavonols content, measured here, using non-destructive methods, reflected exactly the genotypes ranking according to ozone sensitivity.

A New Role for Plastid Thioredoxins in Seed Physiology in Relation to Hormone Regulation.

In Arabidopsis seeds, ROS have been shown to be enabling actors of cellular signaling pathways promoting germination, but their accumulation under stress conditions or during aging leads to a decrease in the ability to germinate. Previous biochemical work revealed that a specific class of plastid thioredoxins (Trxs), the y-type Trxs, can fulfill antioxidant functions. Among the ten plastidial Trx isoforms identified in Arabidopsis, Trx y1 mRNA is the most abundant in dry seeds. We hypothesized that Trx y1 and Trx y2 would play an important role in seed physiology as antioxidants. Using reverse genetics, we found important changes in the corresponding Arabidopsis mutant seeds. They display remarkable traits such as increased longevity and higher and faster germination in conditions of reduced water availability or oxidative stress. These phenotypes suggest that Trxs y do not play an antioxidant role in seeds, as further evidenced by no changes in global ROS contents and protein redox status found in the corresponding mutant seeds. Instead, we provide evidence that marker genes of ABA and GAs pathways are perturbed in mutant seeds, together with their sensitivity to specific hormone inhibitors. Altogether, our results suggest that Trxs y function in Arabidopsis seeds is not linked to their previously identified antioxidant roles and reveal a new role for plastid Trxs linked to hormone regulation.

Evaluation of trace metal accumulation in six vegetable crops intercropped with phytostabilizing plant species, in a French urban wasteland.

The extensive development of agriculture in urban and peri-urban wastelands polluted with several trace elements (TE) poses risks to human health through contaminated food products. The objective was to explore the accumulation of TE in the various parts of vegetable crop plants (tomato, French bean, radish, potato, spinach, and leek) intercropped with phytostabilizing plant species (ryegrass and white clover, respectively). Field studies were conducted in a multicontaminated French urban wasteland with Cd, Cu, Pb and Zn, and an alkaline soil pH. Analyses of the respective non-edible parts of monocultured vegetable crops showed accumulation of all TE, mostly Zn, then Pb and Cu, and finally Cd. The corresponding TE accumulation factors (soil to plant) were all below 0.25. In the edible parts, average concentrations for TE were above the limit values, according to European and Chinese standards. TE contents in the phytostabilizing species chosen were in the same orders of magnitude and the same ranking as described for vegetable crops and most accumulation was in the roots. Unexpectedly, the presence of the phytostabilizing plants had a very strong positive impact on the soil to plant accumulation factor. Moreover, the edible plant parts were poorly impacted by the co-cropping with phytostabilizing plants.

Chilling temperature remodels phospholipidome of Zea mays seeds during imbibition.

Global warming is a major agricultural issue in the Northern hemisphere where higher temperatures are expected to be associated with restricted water availability. In Europe, for maize, earlier and further northward sowings are forecasted in order to avoid water deficit periods in the crop life cycle. However these conditions may compromise seed germination and stand establishment since they will take place at cold temperatures. It is urgent to better understand the molecular bases of response of germinating maize seeds to cold in order to design genotypes adapted to these novel agricultural practices. Here we have performed a global phospholipidomic study to profile changes in membrane reorganisation during seed imbibition at 10 °C of cold-tolerant and -sensitive maize hybrids. Using a Multiple Reaction Monitoring (MRM-MS/MS) method coupled with HPLC we have identified 80 distinct phospholipids. We show that seed sensitivity to cold temperatures during imbibition relies on the accumulation of saturated or poorly unsaturated fatty acids, whatever the phospholipid class. In contrast seeds of cold-tolerant hybrid accumulated polyunsaturated chains which was associated with lower electrolyte leakage during imbibition at 10 °C. The expression of fatty acid desaturase genes provides a molecular model of maize seed sensitivity to imbibitional chilling damage.

5' to 3' mRNA Decay Contributes to the Regulation of Arabidopsis Seed Germination by Dormancy.

The regulation of plant gene expression, necessary for development and adaptive responses, relies not only on RNA transcription but also on messenger RNA (mRNA) fate. To understand whether seed germination relies on the degradation of specific subsets of mRNA, we investigated whether the 5' to 3' RNA decay machinery participated in the regulation of this process. Arabidopsis (Arabidopsis thaliana) seeds of exoribonuclease4 (xrn4) and varicose (vcs) mutants displayed distinct dormancy phenotypes. Transcriptome analysis of xrn4-5 and vcs-8 mutant seeds allowed us to identify genes that are likely to play a role in the control of germination. Study of 5' untranslated region features of these transcripts revealed that specific motifs, secondary energy, and GC content could play a role in their degradation by XRN4 and VCS, and Gene Ontology clustering revealed novel actors of seed dormancy and germination. Several specific transcripts identified as being putative targets of XRN4 and VCS in seeds (PECTIN LYASE-LIKE, ASPARTYL PROTEASE, DWD-HYPERSENSITIVE-TO-ABA3, and YELLOW STRIPE-LIKE5) were further studied by reverse genetics, and their functional roles in the germination process were confirmed by mutant analysis. These findings suggest that completion of germination and its regulation by dormancy also depend on the degradation of specific subsets of mRNA.

Fluctuation of Arabidopsis seed dormancy with relative humidity and temperature during dry storage.

The changes in germination potential of freshly harvested seeds of Arabidopsis thaliana stored in various combinations of temperature and relative humidity were investigated over 63 weeks of storage. Seeds of the wild type Col-0 and of two mutants displaying low and high levels of dormancy, cat2-1 and mtr4-1, respectively, were stored at harvest in 24 different environments including a combination of eight relative humidities, from 1 to 85%, and four temperatures (10, 15, 20, and 25 °C). These mutations did not influence behaviour of seeds during storage. Primary dormant seeds did not germinate in darkness at 25 °C but acquired the potential to germinate at this temperature within 7 weeks when stored in relative humidities close to 50% across all temperatures. Sorption isotherms and Arrhenius plots demonstrated that the seed moisture content of 0.06 g H2O/g dry weight was a critical value below which dormancy release was associated with reactions of negative activation energy and above which dormancy release increased with temperature. Longer storage times when relative humidity did not exceed 75-85% led to decreased germination at 25 °C, corresponding to the induction of secondary dormancy. Dormancy release and induction of secondary dormancy in the dry state were associated with induction or repression of key genes related to abscisic acid and gibberellins biosynthesis and signalling pathways. In high relative humidity, prolonged storage of seeds induced ageing and progressive loss of viability, but this was not related to the initial level of dormancy.

Germination Potential of Dormant and Nondormant Arabidopsis Seeds Is Driven by Distinct Recruitment of Messenger RNAs to Polysomes.

Dormancy is a complex evolutionary trait that temporally prevents seed germination, thus allowing seedling growth at a favorable season. High-throughput analyses of transcriptomes have led to significant progress in understanding the molecular regulation of this process, but the role of posttranscriptional mechanisms has received little attention. In this work, we have studied the dynamics of messenger RNA association with polysomes and compared the transcriptome with the translatome in dormant and nondormant seeds of Arabidopsis (Arabidopsis thaliana) during their imbibition at 25 °C in darkness, a temperature preventing germination of dormant seeds only. DNA microarray analysis revealed that 4,670 and 7,028 transcripts were differentially abundant in dormant and nondormant seeds in the transcriptome and the translatome, respectively. We show that there is no correlation between transcriptome and translatome and that germination regulation is also largely translational, implying a selective and dynamic recruitment of messenger RNAs to polysomes in both dormant and nondormant seeds. The study of 5' untranslated region features revealed that GC content and the number of upstream open reading frames could play a role in selective translation occurring during germination. Gene Ontology clustering showed that the functions of polysome-associated transcripts differed between dormant and nondormant seeds and revealed actors in seed dormancy and germination. In conclusion, our results demonstrate the essential role of selective polysome loading in this biological process.

Translatome profiling in dormant and nondormant sunflower (Helianthus annuus) seeds highlights post-transcriptional regulation of germination.

Seed dormancy, which blocks germination in apparently favourable conditions, is a key regulatory control point of plant population establishment. As germination requires de novo translation, its regulation by dormancy is likely to be related to the association of individual transcripts to polysomes. Here, the polysome-associated mRNAs, that is, the translatome, were fractionated and characterized with microarrays in dormant and nondormant sunflower (Helianthus annuus) embryos during their imbibition at 10°C, a temperature preventing germination of dormant embryos. Profiling of mRNAs in polysomal complexes revealed that the translatome differs between germinating and nongerminating embryos. Association of transcripts with polysomes reached a maximum after 15 h of imbibition; at this time-point 194 polysome-associated transcripts were specifically found in nondormant embryos and 47 in dormant embryos only. The proteins corresponding to the polysomal mRNAs in nondormant embryos appeared to be very pertinent for germination and were involved mainly in transport, regulation of transcription or cell wall modifications. This work demonstrates that seed germination results from a timely regulated and selective recruitment of mRNAs to polysomes, thus opening novel fields of investigation for the understanding of this developmental process.

Inhibition of germination of dormant barley (Hordeum vulgare L.) grains by blue light as related to oxygen and hormonal regulation.

Germination of primary dormant barley grains is promoted by darkness and temperatures below 20 °C, but is strongly inhibited by blue light. Exposure under blue light at 10 °C for periods longer than five days, results in a progressive inability to germinate in the dark, considered as secondary dormancy. We demonstrate that the inhibitory effect of blue light is reinforced in hypoxia. The inhibitory effect of blue light is associated with an increase in embryo abscisic acid (ABA) content (by 3.5- to 3.8-fold) and embryo sensitivity to both ABA and hypoxia. Analysis of expression of ABA metabolism genes shows that increase in ABA mainly results in a strong increase in HvNCED1 and HvNCED2 expression, and a slight decrease in HvABA8'OH-1. Among the gibberellins (GA) metabolism genes examined, blue light decreases the expression of HvGA3ox2, involved in GA synthesis, increases that of GA2ox3 and GA2ox5, involved in GA catabolism, and reduces the GA signalling evaluated by the HvExpA11 expression. Expression of secondary dormancy is associated with maintenance of high embryo ABA content and a low HvExpA11 expression. The partial reversion of the inhibitory effect of blue light by green light also suggests that cryptochrome might be involved in this hormonal regulation.

Arabidopsis lipocalins AtCHL and AtTIL have distinct but overlapping functions essential for lipid protection and seed longevity.

Lipocalins are a group of multifunctional proteins, recognized as carriers of small lipophilic molecules, which have been characterized in bacteria and animals. Two true lipocalins have been recently identified in plants, the temperature-induced lipocalin (TIL) and the chloroplastic lipocalin (CHL), the expression of which is induced by various abiotic stresses. Each lipocalin appeared to be specialized in the responses to specific stress conditions in Arabidopsis thaliana, with AtTIL and AtCHL playing a protective role against heat and high light, respectively. The double mutant AtCHL KO × AtTIL KO deficient in both lipocalins was more sensitive to temperature, drought and light stresses than the single mutants, exhibiting intense lipid peroxidation. AtCHL deficiency dramatically enhanced the photosensitivity of mutants (vte1, npq1) affected in lipid protection mechanisms (tocopherols, zeaxanthin), confirming the role of lipocalins in the prevention of lipid peroxidation. Seeds of the AtCHL KO × AtTIL KO double mutant were very sensitive to natural and artificial ageing, and again this phenomenon was associated with the oxidation of polyunsaturated lipids. The presented results show that the Arabidopsis lipocalins AtTIL and AtCHL have overlapping functions in lipid protection which are essential for stress resistance and survival.

Induction of secondary dormancy by hypoxia in barley grains and its hormonal regulation.

In barley, primary dormant grains did not germinate at 30 °C in air and at 15 °C in an atmosphere containing less than 10% O2, while they germinated easily at 15 °C in air. O2 tension in embryos measured with microsensors was 15.8% at 15 °C but only 0.3% at 30 °C. Incubation of grains at 30 °C is known to induce secondary dormancy in barley, and it was shown here that secondary dormancy was also induced by a 3 d treatment in O2 tensions lower than 10% at 15 °C. After such treatments, the grains lost their ability to germinate subsequently at 15 °C in air. During seed treatment in 5% O2, embryo abscisic acid (ABA) content decreased more slowly than in air and was not altered after transfer into air. Hypoxia did not alter the expression of ABA metabolism genes after 1 d, and induction of HvNCED2 occurred only after 3 d in hypoxia. Embryo sensitivity to ABA was similar in both primary and hypoxia-induced secondary dormant grains. Gibberellic acid (GA) metabolism genes were highly regulated and regulated earlier by the hypoxia treatment, with major changes in HvGA2ox3, HvGA3ox2 and HvGA20ox1 expression after 1 d, resulting in reduced GA signalling. Although a high temperature has an indirect effect on O2 availability, the data showed that it did not affect expression of prolyl-4-hydroxylases and that induction of secondary dormancy by hypoxia at 15 °C or by high temperature in air involved separate signalling pathways. Induction by hypoxia at 15 °C appears to be more regulated by GA and less by ABA than the induction by high temperature.

Water content: a key factor of the induction of secondary dormancy in barley grains as related to ABA metabolism.

Primary dormant barley (Hordeum vulgare) grains germinate at 10-15°C but not at 30°C, and there exist a positive correlation between embryo ABA content after 24 h on water and the depth of dormancy. Incubation at 30°C results in a progressive loss of the ability to germinate at 15°C. This induction of a secondary dormancy is optimal after 3 days and requires an embryo water content higher than 0.50 g H2O g⁻¹ DW, this corresponding with activation of the cell cycle. There exists no correlation between ABA content after 3 days at 30°C and the induction of secondary dormancy. However, at high water content (1.60-1.87 g H2O g⁻¹ DW), secondary dormancy is associated with an high embryo ABA content after transfer to 15°C, resulting from an increase in HvNCED1 and HvNCED2 expression and a decrease in HvABA8'OH-1. Such changes are not observed at 0.45 g H2O g⁻¹ DW. Incubation at 30°C also results in an increase in expression of genes involved in GA catabolism (HvGA2ox1, HvGA2ox3 and HvGA2ox5) and synthesis (HvGA3ox2, HvGA20ox1 and HvGA20ox3). The HvGA3ox2/HvGA2ox3 transcript ratio remains low (0.27-0.37) at 30°C and after transfer to 15°C in secondary dormant seeds, but it is higher than two when secondary dormancy is not induced. Changes in HvExpA11 expression indicate that GA signaling decreases when a secondary dormancy is expressed. Our results clearly indicate that expression of genes involved in ABA and GA metabolism differs in primary and secondary dormancies and furthermore, their expression is related to embryo water content.

Role of reactive oxygen species in the regulation of Arabidopsis seed dormancy.

Freshly harvested seeds of Arabidopsis thaliana, Columbia (Col) accession were dormant when imbibed at 25°C in the dark. Their dormancy was alleviated by continuous light during imbibition or by 5 weeks of storage at 20°C (after-ripening). We investigated the possible role of reactive oxygen species (ROS) in the regulation of Col seed dormancy. After 24 h of imbibition at 25°C, non-dormant seeds produced more ROS than dormant seeds, and their catalase activity was lower. In situ ROS localization revealed that germination was associated with an accumulation of superoxide and hydrogen peroxide in the radicle. ROS production was temporally and spatially regulated: ROS were first localized within the cytoplasm upon imbibition of non-dormant seeds, then in the nucleus and finally in the cell wall, which suggests that ROS play different roles during germination. Imbibition of dormant and non-dormant seeds in the presence of ROS scavengers or donors, which inhibited or stimulated germination, respectively, confirmed the role of ROS in germination. Freshly harvested seeds of the mutants defective in catalase (cat2-1) and vitamin E (vte1-1) did not display dormancy; however, seeds of the NADPH oxidase mutants (rbohD) were deeply dormant. Expression of a set of genes related to dormancy upon imbibition in the cat2-1 and vet1-1 seeds revealed that their non-dormant phenotype was probably not related to ABA or gibberellin metabolism, but suggested that ROS could trigger germination through gibberellin signaling activation.

Crosstalk between reactive oxygen species and hormonal signalling pathways regulates grain dormancy in barley.

Seed dormancy, defined as the inability to germinate under favourable conditions, is controlled by abscisic acid (ABA) and gibberellins (GAs). Phytohormone signalling interacts with reactive oxygen species (ROS) signalling regarding diverse aspects of plant physiology and is assumed to be important in dormancy alleviation. Using dormant barley grains that do not germinate at 30 °C in darkness, we analysed ROS content and ROS-processing systems, ABA content and metabolism, GA-responsive genes and genes involved in GA metabolism in response to hydrogen peroxide (H2O2) treatment. During after-ripening, the ROS content in the embryo was not affected, while the antioxidant glutathione (GSH) was gradually converted to glutathione disulphide (GSSG). ABA treatment up-regulated catalase activity through transcriptional activation of HvCAT2. Exogenous H2O2 partially alleviated dormancy although it was associated with a small increase in embryonic ABA content related to a slight induction of HvNCED transcripts. H2O2 treatment did not affect ABA sensitivity but up-regulated the expression of HvExpA11 (GA-induced gene), inhibited the expression of HvGA2ox3 involved in GA catabolism and enhanced the expression of HvGA20ox1 implicated in GA synthesis. In barley, H2O2 could be implicated in dormancy alleviation through activation of GA signalling and synthesis rather than repression of ABA signalling.

Involvement of ABA in induction of secondary dormancy in barley (Hordeum vulgare L.) seeds.

At harvest, barley seeds are dormant because their germination is difficult above 20 degrees C. Incubation of primary dormant seeds at 30 degrees C, a temperature at which they do not germinate, results in a loss of their ability to germinate at 20 degrees C. This phenomenon which corresponds to an induction of a secondary dormancy is already observed after a pre-treatment at 30 degrees C as short as 4-6 h, and is optimal after 24-48 h. It is associated with maintenance of a high level of embryo ABA content during seed incubation at 30 degrees C, and after seed transfer at 20 degrees C, while ABA content decreases rapidly in embryos of primary dormant seeds placed directly at 20 degrees C. Induction of secondary dormancy also results in an increase in embryo responsiveness to ABA at 20 degrees C. Application of ABA during seed treatment at 30 degrees C has no significant additive effect on the further germination at 20 degrees C. In contrast, incubation of primary dormant seeds at 20 degrees C for 48 and 72 h in the presence of ABA inhibits further germination on water similarly to 24-48 h incubation at 30 degrees C. However fluridone, an inhibitor of ABA synthesis, applied during incubation of the grains at 30 degrees C has only a slight effect on ABA content and secondary dormancy. Expression of genes involved in ABA metabolism (HvABA8'OH-1, HvNCED1 and HvNCED2) was studied in relation to the expression of primary and secondary dormancies. The results presented suggest a specific role for HvNCED1 and HvNCED2 in regulation of ABA synthesis in secondary seed dormancy.

Regulation of cell cycle activity in the embryo of barley seeds during germination as related to grain hydration.

Various studies indicate that cell division is a post-germination phenomenon, with radicle protrusion occurring by cell elongation, while others demonstrate that induction of the cell cycle occurs in osmo-conditioned seeds prior to radicle growth. The aim of the present work was to investigate the occurrence of the cell cycle during germination as related to grain hydration, using: (i) a flow cytometry technique to estimate the percentage of cell nuclei in G(1) and G(2) phases of the cell cycle; and (ii) reverse transcription-PCR (RT-PCR) in order to characterize the expression of the genes encoding cyclin-dependent kinases (CDKA1, CDKB1, and CDKD1) and cyclins (CYCA3, CYCB1, and CYCD4), the main genes involved in the cell cycle and its regulation. Radicle tips of embryos were isolated from seeds placed for various times on water at 30 degrees C and from grains partially hydrated at moisture contents ranging from 11% to 51% fresh weight (FW), which prevent radicle elongation. Abscisic acid (ABA) contents of the embryos during seed germination at 30 degrees C and after 48 h of partial hydration were also measured. In dry embryos, cells are mostly arrested in the G(1) phase of the cell cycle (82%), the remaining cells being in the G(2) phase, and the ABA content of the embryo was 432.7 ng g(-1) dry weight (DW). Seed imbibition was associated with a sharp decrease in ABA content as early as 5 h, while the cell cycle reactivation was a late process taking place approximately 4-6 h prior to radicle protrusion. Hydration of seeds resulted in a decrease in embryo ABA content, but it remained at a high level (207-273 ng g(-1) DW) even after 48 h at 0.41-0.51 g H2O g(-1) FW. The cell population of the radicle tips in the G(2) phase of the cell cycle, i.e. 4C nuclei, increased from 9% up to 34% at a moisture content of 51% FW. In dry seeds, CDKA1 and CDKD1 mRNAs were present at low levels, but transcripts of CDKB1, CYCA3, CYCB1, and CYCD4 were not detected. Radicle protrusion was associated with a higher expression of CDKA1, CDKB1, CYCA3, and CYCB1. Blockage of germination of partially hydrated grains resulted in a reduction in the expression of CDKA1 and CDKB1, and of CYCA3 and CYCB1, and in a reinforcement of that of CDKD1 and CYCD4. Patterns of gene expression show differential sensitivity of the genes studied to hydration of the grain. They will be discussed with regard to embryo ABA content and embryo sensitivity to ABA.

Identification of transcripts potentially involved in barley seed germination and dormancy using cDNA-AFLP.

Freshly harvested barley seeds are considered as dormant since they do not germinate at temperatures above 20 degrees C. This dormancy is broken during dry storage. Molecular regulation of dormancy was investigated using cDNA-AFLP to identify transcripts differentially expressed in dormant and non-dormant embryos. Transcript patterns in embryos from dry dormant and non-dormant seeds and from both seeds imbibed for 5 h at 30 degrees C, a temperature at which dormancy is expressed, were compared. Thirty-nine Transcript-Derived Fragments (TDF) that were reproducibly differentially expressed among treatments were identified, and 25 of these were cloned and sequenced. Among these, eight transcripts were observed to be differentially expressed during after-ripening, seven of which decline, probably due to post-maturation degradation. HV13B, TDF identified as having homology to fructose-6-phosphate-2-kinase/fructose-2,6-biphosphatase, may have a role in the maintenance of dormancy in barley and probably in other cereals. During the first 5 h of imbibition, there was expression of 24 TDF which was apparently independent of dormancy, revealing putative epigenetic regulation. This was typified by HV44A, a SET domain protein. Seven TDF differentially expressed, and especially HV12D, HV42B, and HV32B, in dormant and non-dormant seeds were potential signalling elements. HV12D had homology with an ARIADNE gene which could be implicated in ABA signalling.