Molecular and physiological responses to salt stress in salinity-sensitive and tolerant Hibiscus rosa-sinensis cultivars.

Ornamental plants are used to decorate urban and peri-urban areas, and during their cultivation or utilisation, they can be exposed to abiotic stress. Salinity is an abiotic stress factor that limits plant growth and reduces the ornamental value of sensitive species. In this study, transcriptomic analysis was conducted to identify genes associated with tolerance or sensitivity to salinity in two hibiscus (Hibiscus rosa-sinensis L.) cultivars, 'Porto' and 'Sunny wind'. The physiological and biochemical parameters of plants exposed to 50, 100, or 200 mM NaCl and water (control) were monitored. Salinity treatments were applied for six weeks. After four weeks, differences between cultivars were clearly evident and 'Porto' was more tolerant than 'Sunny wind'. The tolerant cultivar showed lower electrolyte leakage and ABA concentrations, and higher proline content in the leaves. Accumulation of Na in different organs was lower in the flower organs of 'Porto'. At the molecular level, several differential expressed genes were observed between the cultivars and flower organs. Among the highly expressed DEGs, coat protein, alcohol dehydrogenase, and AP2/EREBP transcription factor ERF-1. Among the downregulated genes, GH3 and NCED were the most interesting. The differential expression of these genes may explain the salt stress tolerance of 'Porto'.

Red versus green leaves: transcriptomic comparison of foliar senescence between two Prunus cerasifera genotypes.

The final stage of leaf ontogenesis is represented by senescence, a highly regulated process driven by a sequential cellular breakdown involving, as the first step, chloroplast dismantling with consequent reduction of photosynthetic efficiency. Different processes, such as pigment accumulation, could protect the vulnerable photosynthetic apparatus of senescent leaves. Although several studies have produced transcriptomic data on foliar senescence, just few works have attempted to explain differences in red and green leaves throughout ontogenesis. In this work, a transcriptomic approach was used on green and red leaves of Prunus cerasifera to unveil molecular differences from leaf maturity to senescence. Our analysis revealed a higher gene regulation in red leaves compared to green ones, during leaf transition. Most of the observed DEGs were shared and involved in transcription factor activities, senescing processes and cell wall remodelling. Significant differences were detected in cellular functions: genes related to photosystem I and II were highly down-regulated in the green genotype, whereas transcripts involved in flavonoid biosynthesis, such as UDP glucose-flavonoid-3-O-glucosyltransferase (UFGT) were exclusively up-regulated in red leaves. In addition, cellular functions involved in stress response (glutathione-S-transferase, Pathogen-Related) and sugar metabolism, such as three threalose-6-phosphate synthases, were activated in senescent red leaves. In conclusion, data suggests that P. cerasifera red genotypes can regulate a set of genes and molecular mechanisms that cope with senescence, promoting more advantages during leaf ontogenesis than compared to the green ones.

UV-B Physiological Changes Under Conditions of Distress and Eustress in Sweet Basil.

UV-B radiation has been previously reported to induce protective or deleterious effects on plants depending on the UV-B irradiation doses. To elucidate how these contrasting events are physiologically coordinated, we exposed sweet basil plants to two UV-B doses: low (8.5 kJ m-2 day-1, 30 min exposure) and high (68 kJ m-2 day-1, 4 h exposure), with the plants given both doses once continuously in a single day. Physiological tests during and after both UV-B exposures were performed by comparing the stress-induced damage and adverse effects on photosynthetic activity, the concentration and composition of photosynthetic and non-photosynthetic pigments, and stress-related hormones biosynthesis in basil plants. Our results showed that upon receiving a high UV-B dose, a severe inactivation of oxygen evolving complex (OEC) activity at the PSII donor side and irreversible PSII photodamage caused primarily by limitation of the acceptor side occurred, which overloaded protective mechanisms and finally led to the death of the plants. In contrast, low UV-B levels did not induce any signs of UV-B stress injuries. The OEC partial limitation and the inactivation of the electron transport chain allowed the activation of photoprotective mechanisms, avoiding irreversible damage to PSII. Overall results indicate the importance of a specific response mechanisms regulating photoprotection vs irreversible photoinhibition in basil that were modulated depending on the UV-B doses.

When "thirsty" means "less able to activate the signalling wave trigged by a pulse of ozone": A case of study in two Mediterranean deciduous oak species with different drought sensitivity.

There is a lack of knowledge about the possibility that plants facing abiotic stressors, such as drought, have an altered perception of a pulse of O3 and incur in alterations of their signalling network. This poses some concerns as to whether defensive strategy to cope episodic O3 peaks in healthy plants may fail under stress. In this study, a set of saplings of two Mediterranean deciduous species, Quercus cerris and Q. pubescens, was subjected to water withholding (20% of daily evapotranspiration for 15 days) while another set was kept well-watered. Saplings were then subjected to a pulse of O3 (200 nl l-1 for 5 h) or maintained in filtered air. Q. pubescens had a more severe decline of photosynthesis and leaf PDΨw (about -65% and 5-fold lower than in well-watered ones) and events of cell death were observed under drought when compared to Q. cerris, which is supportive for a higher sensitivity to drought exhibited by this species. When O3 was applied after drought, patterns of signalling compounds were altered in both species. Only in Q. pubescens, the typical O3-induced accumulation of apoplastic reactive oxygen species, which is the first necessary step for the activation of signalling cascade, was completely lost. In Q. cerris the most frequent changes encompassed the weakening of peaks of key signalling molecules (ethylene and salicylic acid), whereas in Q. pubescens both delayed (salicylic and jasmonic acid) or weakened (ethylene and salicylic acid) peaks were observed. This is translated to a higher ability of Q. cerris to maintain a prompt activation of defensive reaction to counteract oxidative damage due to the pollutant. Our results reveal the complexity of the signalling network in plants facing multiple stresses and highlight the need to further investigate possible alteration of defensive mechanism of tree species to predict their behavior.

How Quercus ilex L. saplings face combined salt and ozone stress: a transcriptome analysis.

BACKGROUND: Similar to other urban trees, holm oaks (Quercus ilex L.) provide a physiological, ecological and social service in the urban environment, since they remove atmospheric pollution. However, the urban environment has several abiotic factors that negatively influence plant life, which are further exacerbated due to climate change, especially in the Mediterranean area. Among these abiotic factors, increased uptake of Na + and Cl - usually occurs in trees in the urban ecosystem; moreover, an excess of the tropospheric ozone concentration in Mediterranean cities further affects plant growth and survival. Here, we produced and annotated a de novo leaf transcriptome of Q. ilex as well as transcripts over- or under-expressed after a single episode of O3 (80 nl l-1, 5 h), a salt treatment (150 mM for 15 days) or a combination of these treatments, mimicking a situation that plants commonly face, especially in urban environments. RESULTS: Salinity dramatically changed the profile of expressed transcripts, while the short O3 pulse had less effect on the transcript profile. However, the short O3 pulse had a very strong effect in inducing over- or under-expression of some genes in plants coping with soil salinity. Many differentially regulated genes were related to stress sensing and signalling, cell wall remodelling, ROS sensing and scavenging, photosynthesis and to sugar and lipid metabolism. Most differentially expressed transcripts revealed here are in accordance with a previous report on Q. ilex at the physiological and biochemical levels, even though the expression profiles were overall more striking than those found at the biochemical and physiological levels. CONCLUSIONS: We produced for the first time a reference transcriptome for Q. ilex, and performed gene expression analysis for this species when subjected to salt, ozone and a combination of the two. The comparison of gene expression between the combined salt + ozone treatment and salt or ozone alone showed that even though many differentially expressed genes overlap all treatments, combined stress triggered a unique response in terms of gene expression modification. The obtained results represent a useful tool for studies aiming to investigate the effects of environmental stresses in urban-adapted tree species.

Multiple Consequences Induced by Epidermally-Located Anthocyanins in Young, Mature and Senescent Leaves of Prunus.

Anthocyanic morphs are generally less efficient in terms of carbon gain, but, in turn, are more photoprotected than anthocyanin-less ones. To date, mature leaves of different morphs or leaves at different developmental stages within the same species have generally been compared, whereas there is a lack of knowledge regarding different stages of development of red vs. green leaves. Leaves (1-, 7-, and 13-week-old) of red- (RLP) and green-leafed (GLP) Prunus in terms of photosynthetic rate, carbon metabolism and photoprotective mechanisms were compared to test whether anthocyanin-equipped leaves perform better than anthocyanin-less leaves and whether photoprotection is the primary role of epidermally-located anthocyanins, using for the first time a recently-developed parameter of chlorophyll fluorescence (qPd). GLP leaves had a higher photosynthetic rate in 1- and 7-week-old leaves, but RLP leaves performed better at an early stage of senescence and had a longer leaf lifespan. Anthocyanins contributed to leaf photoprotection throughout the leaf development, but were tightly coordinated with carotenoids. Besides photoprotecting, we propose that epidermal anthocyanins may be principally synthetized to maintain an efficient carbon-sink strength in young and senescent leaves, thus extending the RLP leaf lifespan.

Losing the Warning Signal: Drought Compromises the Cross-Talk of Signaling Molecules in Quercus ilex Exposed to Ozone.

Understanding the interactions between drought and acute ozone (O3) stress in terms of signaling molecules and cell death would improve the predictions of plant responses to climate change. The aim was to investigate whether drought stress influences the responses of plants to acute episodes of O3 exposure. In this study, the behavior of 84 Mediterranean evergreen Quercus ilex plants was evaluated in terms of cross-talk responses among signaling molecules. Half of the sample was subjected to drought (20% of the effective daily evapotranspiration, for 15 days) and was later exposed to an acute O3 exposure (200 nL L-1 for 5 h). First, our results indicate that in well-water conditions, O3 induced a signaling pathway specific to O3-sensitive behavior. Second, different trends and consequently different roles of phytohormones and signaling molecules (ethylene, ET; abscisic acid, ABA; salycilic acid, SA and jasmonic acid, JA) were observed in relation to water stress and O3. A spatial and functional correlation between these signaling molecules was observed in modulating O3-induced responses in well-watered plants. In contrast, in drought-stressed plants, these compounds were not involved either in O3-induced signaling mechanisms or in leaf senescence (a response observed in water-stressed plants before the O3-exposure). Third, these differences were ascribable to the fact that in drought conditions, most defense processes induced by O3 were compromised and/or altered. Our results highlight how Q. ilex plants suffering from water deprivation respond differently to an acute O3 episode compared to well-watered plants, and suggest new effect to be considered in plant responses to environmental changes. This poses the serious question as to whether or not multiple high-magnitude O3 events (as predicted) can change these cross-talk responses, thus opening it up possible further investigations.

The harsh life of an urban tree: the effect of a single pulse of ozone in salt-stressed Quercus ilex saplings.

Ozone (O3) and salinity are usually tested as combined factors on plant performance. However, the response to a single episode of O3 in plants already stressed by an excess of NaCl as occurs in the natural environment has never been investigated, but is important given that it is commonly experienced in Mediterranean areas. Three-year-old Quercus ilex L. (holm oak) saplings were exposed to salinity (150 mM NaCl, 15 days), and the effect on photosynthesis, hydric relations and ion partitioning was evaluated (Experiment I). In Experiment II, salt-treated saplings were exposed to 80 nl l-1 of O3 for 5 h, which is a realistic dose in a Mediterranean environment. Gas exchanges, chlorophyll fluorescence and antioxidant systems were characterized to test whether the salt-induced stomatal closure limited O3 uptake and stress or whether the pollutant represents an additional stressor for plants. Salt-dependent stomatal closure depressed the photosynthetic process (-71.6% of light-saturated rate of photosynthesis (A380)) and strongly enhanced the dissipation of energy via the xanthophyll cycle. However, salt-treated plants had higher values of net assimilation rate/stomatal conductance (A/gs) than the controls, which was attributable to a greater mesophyll conductance gm/gs and carboxylation efficiency (higher gm/maximal rate of Rubisco carboxylation (Vcmax)), thus suggesting no damage to chloroplasts. O3 did not exacerbate the effect of salinity on photosynthesis, however a general enhancement of the Halliwell-Asada cycle was necessary to counteract the O3-triggered oxidative stress. Despite the 79.4% gs reduction in salt-stressed plants, which strongly limited the O3 uptake, a single peak in the air pollutant led to an additional burden for the antioxidant system when plants had been previously subjected to salinity.

Seedling Establishment of Tall Fescue Exposed to Long-Term Starvation Stress.

In germinating seeds under unfavorable environmental conditions, the mobilization of stores in the cotyledons is delayed, which may result in a different modulation of carbohydrates balance and a decrease in seedling vigor. Tall fescue (Festuca arundinacea Schreb.) caryopses grown at 4°C in the dark for an extended period in complete absence of nutrients, showed an unexpected ability to survive. Seedlings grown at 4°C for 210 days were morphologically identical to seedlings grown at 23°C for 21 days. After 400 days, seedlings grown at 4°C were able to differentiate plastids to chloroplast in just few days once transferred to the light and 23°C. Tall fescue exposed to prolonged period at 4°C showed marked anatomical changes: cell wall thickening, undifferentiated plastids, more root hairs and less xylem lignification. Physiological modifications were also observed, in particular related to sugar content, GA and ABA levels and amylolytic enzymes pattern. The phytohormones profiles exhibited at 4 and 23°C were comparable when normalized to the respective physiological states. Both the onset and the completion of germination were linked to GA and ABA levels, as well as to the ratio between these two hormones. All plants showed a sharp decline in carbohydrate content, with a consequent onset of gradual sugar starvation. This explained the slowed then full arrest in growth under both treatment regimes. The analysis of amylolytic activity showed that Ca2+ played a central role in the stabilization of several isoforms. Overall, convergence of starvation and hormone signals meet in crosstalk to regulate germination, growth and development in tall fescue.

Spatial and temporal transcriptome changes occurring during flower opening and senescence of the ephemeral hibiscus flower, Hibiscus rosa-sinensis.

Flowers are complex systems whose vegetative and sexual structures initiate and die in a synchronous manner. The rapidity of this process varies widely in flowers, with some lasting for months while others such as Hibiscus rosa-sinensis survive for only a day. The genetic regulation underlying these differences is unclear. To identify key genes and pathways that coordinate floral organ senescence of ephemeral flowers, we identified transcripts in H. rosa-sinensis floral organs by 454 sequencing. During development, 2053 transcripts increased and 2135 decreased significantly in abundance. The senescence of the flower was associated with increased abundance of many hydrolytic genes, including aspartic and cysteine proteases, vacuolar processing enzymes, and nucleases. Pathway analysis suggested that transcripts altering significantly in abundance were enriched in functions related to cell wall-, aquaporin-, light/circadian clock-, autophagy-, and calcium-related genes. Finding enrichment in light/circadian clock-related genes fits well with the observation that hibiscus floral development is highly synchronized with light and the hypothesis that ageing/senescence of the flower is orchestrated by a molecular clock. Further study of these genes will provide novel insight into how the molecular clock is able to regulate the timing of programmed cell death in tissues.

Effect of cytokinins on delaying petunia flower senescence: a transcriptome study approach.

Flower senescence is a fascinating natural process that represents the final developmental stage in the life of a flower. Plant hormones play an important role in regulating the timing of flower senescence. Ethylene is a trigger and usually accelerates the senescence rate, while cytokinins are known to delay it. The aim of this work was to study the effect of 6-benzylaminopurine (BA) on petal senescence by transcript profile comparison after 3 or 6 h using a cross-species method by hybridizing petunia samples to a 4 × 44 K Agilent tomato array. The relative content of ethylene, abscisic acid, anthocyanins, total carotenoids and total phenols that determine the physiological behaviours of the petal tissue were measured. BA treatment prolonged the flower life and increased the concentrations of phenols and anthocyanins, while total carotenoids did not increase and were lower than the control. The ethylene biosynthetic and perception gene expressions were studied immediately after treatment until 24 h and all genes were repressed, while ethylene production was strongly induced after 4 days. The microarray analyses highlighted that BA strongly affected gene regulation after 3 h, but only 14% of genes remained differentially expressed after 6 h. The most affected pathways and genes were those related to stress, such as heat shock proteins, abscisic acid (ABA) catabolism and its signalling pathway, lipid metabolism and antioxidant defence systems. A gene annotation enrichment analysis using DAVID showed that the most important gene clusters were involved in energy generation and conservation processes. In addition to the ethylene pathway, cytokinins seem to be strongly involved the regulation of the ABA response in flower tissues.

Effect of salt stress in the regulation of anthocyanins and color of hibiscus flowers by digital image analysis.

The effect of salt stress (200 mM NaCl for 28 days) on physiological characteristics of Hibiscus rosa-sinensis, such as abscisic acid (ABA) content, electrolyte leakage, and photochemical efficiency in leaves, and its influence on biomass production, anthocyanin composition, and color expression of flowers were evaluated. Salinity significantly increased electrolyte leakage and ABA content in leaves and reduced the flower fresh weight. Chlorophyll fluorescence parameters were lower in salt stress condition, compared to control. Moreover, salt stress negatively affected the content of anthocyanins (mainly cyanidin-3-sophoroside), which resulted in a visually perceptible loss of color. The detailed anthocyanin composition monitored by HPLC-DAD-MS and the color variations by digital image analysis due to salt stress showed that the effect was more noticeable at the basal portion of petals. A forward stepwise multiple regression was performed for predicting the content of anthocyanins from appearance characteristics obtained by image analysis, reaching R-square values up to 0.90.

Signaling molecules and cell death in Melissa officinalis plants exposed to ozone.

KEY MESSAGE: The study focuses on the interaction between reactive oxygen species and hormones that regulate the programmed cell death in plants of Melissa officinalis exposed to ozone. Interaction between hormone and redox signaling pathways has been investigated in ozone-stressed (200 ppb, 5 h) lemon balm to verify if the response resembles the biotic defense reactions. In comparison to controls, plants exhibited foliar injury and the cell death was induced by (1) biphasic production of hydrogen peroxide and superoxide radical; (2) hormonal regulation of ozone-induced lesion formation with a significant production of ethylene, salicylic, jasmonic and abscisic acid; (3) ozone degradation to reactive oxygen species and their detoxification by some enzymatic (such as superoxide dismutase) and non-enzymatic antioxidant systems (such as ascorbic acid, glutathione and carotenoids), that worked in cooperation without providing a defense against free radicals (such as confirmed by the modification of the antioxidant properties of leaf tissue). This integrated view showed that reactive oxygen species interact with hormonal signaling pathway regulating cell death and the sensitivity of lemon balm to ozone.

Chlorophyll a fluorescence as a tool in evaluating the effects of ABA content and ethylene inhibitors on quality of flowering potted Bougainvillea.

Flowering potted plants during the postproduction stage are usually stored in inadequate environmental conditions. We evaluated the effect of the most common storage conditions and treatments on two Bougainvillea cultivars after harvest and during recovery. Flowering potted Bougainvillea plants were treated with 100 mL 2 mM amino-oxyacetic acid (AOA) or 500 ppb 1-methylcyclopropene (1-MCP) prior storage in dark at 14°C for simulating transport or storage conditions and, subsequently, transferred to growth chambers at 20°C in the light for one week for evaluating the recovery ability. The plant stress during the experiments was assessed by ethylene, ABA, and chlorophyll a fluorescence measurements. Ethylene production was affected by temperature rather than treatments. ABA concentration declined in leaves and flowers during storage and was not affected by treatments. Fluorescence parameters appear to be very useful for screening Bougainvillea cultivars resistant to prolonged storage periods.

Effects of abscisic acid on ethylene biosynthesis and perception in Hibiscus rosa-sinensis L. flower development.

The effect of the complex relationship between ethylene and abscisic acid (ABA) on flower development and senescence in Hibiscus rosa-sinensis L. was investigated. Ethylene biosynthetic (HrsACS and HrsACO) and receptor (HrsETR and HrsERS) genes were isolated and their expression evaluated in three different floral tissues (petals, style-stigma plus stamens, and ovaries) of detached buds and open flowers. This was achieved through treatment with 0.1 mM 1-aminocyclopropane-1-carboxylic acid (ACC) solution, 500 nl l(-1) methylcyclopropene (1-MCP), and 0.1 mM ABA solution. Treatment with ACC and 1-MCP confirmed that flower senescence in hibiscus is ethylene dependent, and treatment with exogenous ABA suggested that ABA may play a role in this process. The 1-MCP impeded petal in-rolling and decreased ABA content in detached open flowers after 9 h. This was preceded by an earlier and sequential increase in ABA content in 1-MCP-treated petals and style-stigma plus stamens between 1 h and 6 h. ACC treatment markedly accelerated flower senescence and increased ethylene production after 6 h and 9 h, particularly in style-stigma plus stamens. Ethylene evolution was positively correlated in these floral tissues with the induction of the gene expression of ethylene biosynthetic and receptor genes. Finally, ABA negatively affected the ethylene biosynthetic pathway and tissue sensitivity in all flower tissues. Transcript abundance of HrsACS, HrsACO, HrsETR, and HrsERS was reduced by exogenous ABA treatment. This research underlines the regulatory effect of ABA on the ethylene biosynthetic and perception machinery at a physiological and molecular level when inhibitors or promoters of senescence are exogenously applied.

Exogenous ABA accentuates the differences in root hydraulic properties between mycorrhizal and non mycorrhizal maize plants through regulation of PIP aquaporins.

The arbuscular mycorrhizal (AM) symbiosis has been shown to modulate the same physiological processes as the phytohormone abscisic acid (ABA) and to improve plant tolerance to water deficit. The aim of the present research was to evaluate the combined influence of AM symbiosis and exogenous ABA application on plant root hydraulic properties and on plasma-membrane intrinsic proteins (PIP) aquaporin gene expression and protein accumulation after both a drought and a recovery period. Results obtained showed that the application of exogenous ABA enhanced osmotic root hydraulic conductivity (L) in all plants, regardless of water conditions, and that AM plants showed lower L values than nonAM plants, a difference that was especially accentuated when plants were supplied with exogenous ABA. This effect was clearly correlated with the accumulation pattern of the different PIPs analyzed, since most showed reduced expression and protein levels in AM plants fed with ABA as compared to their nonAM counterparts. The possible involvement of plant PIP aquaporins in the differential regulation of L by ABA in AM and nonAM plants is further discussed.

Mycorrhizal and non-mycorrhizal Lactuca sativa plants exhibit contrasting responses to exogenous ABA during drought stress and recovery.

The arbuscular mycorrhizal (AM) symbiosis enhances plant tolerance to water deficit through the alteration of plant physiology and the expression of plant genes. These changes have been postulated to be caused (among others) by different contents of abscisic acid (ABA) between AM and non-AM plants. However, there are no studies dealing with the effects of exogenous ABA on the expression of stress-related genes and on the physiology of AM plants. The aim of the present study was to evaluate the influence of AM symbiosis and exogenous ABA application on plant development, physiology, and expression of several stress-related genes after both drought and a recovery period. Results show that the application of exogenous ABA had contrasting effects on AM and non-AM plants. Only AM plants fed with exogenous ABA maintained shoot biomass production unaltered by drought stress. The addition of exogenous ABA enhanced considerably the ABA content in shoots of non-AM plants, concomitantly with the expression of the stress marker genes Lsp5cs and Lslea and the gene Lsnced. By contrast, the addition of exogenous ABA decreased the content of ABA in shoots of AM plants and did not produce any further enhancement of the expression of these three genes. AM plants always exhibited higher values of root hydraulic conductivity and reduced transpiration rate under drought stress. From plants subjected to drought, only the AM plants recovered their root hydraulic conductivity completely after the 3 d recovery period. As a whole, the results indicate that AM plants regulate their ABA levels better and faster than non-AM plants, allowing a more adequate balance between leaf transpiration and root water movement during drought and recovery.

Plant responses to drought stress and exogenous ABA application are modulated differently by mycorrhization in tomato and an ABA-deficient mutant (sitiens).

The aims of the present study are to find out whether the effects of arbuscular mycorrhizal (AM) symbiosis on plant resistance to water deficit are mediated by the endogenous abscisic acid (ABA) content of the host plant and whether the exogenous ABA application modifies such effects. The ABA-deficient tomato mutant sitiens and its near-isogenic wild-type parental line were used. Plant development, physiology, and expression of plant genes expected to be modulated by AM symbiosis, drought, and ABA were studied. Results showed that only wild-type tomato plants responded positively to mycorrhizal inoculation, while AM symbiosis was not observed to have any effect on plant development in sitiens plants grown under well-watered conditions. The application of ABA to sitiens plants enhanced plant growth both under well-watered and drought stress conditions. In respect to sitiens plants subjected to drought stress, the addition of ABA had a cumulative effect in relation to that of inoculation with G. intraradices. Most of the genes analyzed in this study showed different regulation patterns in wild-type and sitiens plants, suggesting that their gene expression is modulated by the plant ABA phenotype. In the same way, the colonization of roots with the AM fungus G. intraradices differently regulated the expression of these genes in wild-type and in sitiens plants, which could explain the distinctive effect of the symbiosis on each plant ABA phenotype. This also suggests that the effects of the AM symbiosis on plant responses and resistance to water deficit are mediated by the plant ABA phenotype.

On the role of abscisic acid in seed dormancy of red rice.

Abscisic acid (ABA) is commonly assumed to be the primary effector of seed dormancy, but conclusive evidence for this role is lacking. This paper reports on the relationships occurring in red rice between ABA and seed dormancy. Content of free ABA in dry and imbibed caryopses, both dormant and after-ripened, the effects of inhibitors, and the ability of applied ABA to revert dormancy breakage were considered. The results indicate: (i) no direct correlation of ABA content with the dormancy status of the seed, either dry or imbibed; (ii) different sensitivity to ABA of non-dormant seed and seed that was forced to germinate by fluridone; and (iii) an inability of exogenous ABA to reinstate dormancy in fluridone-treated seed, even though applied at a pH which favoured high ABA accumulation. These considerations suggest that ABA is involved in regulating the first steps of germination, but unidentified developmental effectors that are specific to dormancy appear to stimulate ABA synthesis and to enforce the responsiveness to this phytohormone. These primary effectors appear physiologically to modulate dormancy and via ABA they effect the growth of the embryo. Therefore, it is suggested that ABA plays a key role in integrating the dormancy-specific developmental signals with the control of growth.

Light induces expression of a dehydrin-encoding gene during seedling de-etiolation in sunflower (Helianthus annuus L.).

The effects of light quality on the expression of a sunflower dehydrin-encoding gene, HaDhn1, were studied during seedling de-etiolation. Seeds were germinated in the dark and, after 5 days, seedlings were maintained well watered and de-etiolated under different lights for 3, 6, 12, and 24h. Exposure to white light stimulated HaDhn1 transcript accumulation in the cotyledons of these seedlings, contrary to seedlings grown in the dark. HaDhn1 transcripts increased also treating plantlets with monochromatic lights, especially red light. The increase of HaDhn1 transcripts is provoked by the formation of the active form of phytochrome. Further experiments, performed saturating active phytochrome by yellow light, in combination or not with blue light, showed that also cryptochrome can increase HaDhn1 transcripts accumulation after exposure to light. In situ analysis of HaDhn1 expression domains in cotyledons of light-treated seedlings showed a hybridisation signal spread in all mesophyll cells, especially in the basal portion and in the vascular tissue. In the distal portion of the cotyledons, less intense signal was observed. Western blot analysis indicated that HaDhn1 transcription is not followed by dehydrin-protein accumulation. The isolated putative promoter sequence of the HaDhn1 gene showed that different putative cis-elements recognisable by transcription factors occur in the isolated sequence, including a putative light-responsive G-box. On the whole, our results indicate that HaDhn1 gene expression is induced by light during de-etiolation, in absence of water stress.