Different abscisic acid-deficient mutants show unique morphological and hydraulic responses to high air humidity.

High relative humidity (RH) perturbs plant growth, stomatal functioning and abscisic acid (ABA) homeostasis, but the role of ABA in this physiological regulation is equivocal. To determine the role(s) of ABA in plant responses to high RH, wild-type (WT) tomato and barley plants and their respective ABA-deficient mutants flacca and Az34 (which are mutated in the same locus of the ABA biosynthesis pathway) were grown in contrasting RHs (60% and 90%) to measure biomass partitioning, stomatal traits and water relations. Surprisingly, growth RH did not affect foliar ABA levels in either species. While Az34 showed similar stomatal size and density as WT plants, flacca had larger and more abundant stomata. High RH increased stomatal size in tomato, but decreased it in barley, and decreased stomatal density in tomato, but not in barley. Altered stomatal responses in ABA-deficient plants to high RH had little effect on tomato photosynthesis, but Az34 barley showed lower photosynthesis. ABA deficiency decreased relative shoot growth rate (RGRSHOOT ) in both species, yet this was counteracted by high RH increasing leaf water status in tomato, but not in barley. High RH increased RGRSHOOT in flacca, but not in WT tomatoes, while having no effect on RGRSHOOT in barley, but affecting barley net assimilation rate, leaf area ratio (LAR) and specific leaf area in an ABA-dependent manner. ABA-RH interaction affected leaf development in tomato only. Thus, different crop species show variable responses to both high RH and ABA deficiency, making it difficult to generalise on the role of ABA in growth regulation at contrasting RHs.

Blue Light Improves Stomatal Function and Dark-Induced Closure of Rose Leaves (Rosa x hybrida) Developed at High Air Humidity.

Plants developed under constant high (>85%) relative air humidity (RH) have larger stomata that are unable to close completely in response to closing stimuli. Roses (Rosa x hybrida) developed in high RH have previously been shown to have high water loss during leaf dehydration and reduced dark-induced closure resulting in a shorter postharvest life. In this study, the effect of B-light on stomatal function under high RH conditions was investigated. The ability of rose leaves developed under continuous high (90%) or moderate (60%) RH to close their stomata in response to darkness and leaf dehydration assay was studied. Moreover, the level and regulation of ABA in light and darkness in relation to B-light was measured. Our results show that increased B-light proportion improved stomatal function and dark-induced stomatal closure under high RH conditions and that was associated with increased [ABA] in general and a dynamic ABA peak during darkness. Furthermore, increased B-light during the day was associated with the presence of high ß-glucosidase activity during night. This indicates that B-light is important as a signal to activate the ß-glucosidase enzyme and release ABA during night. Altogether, the improved stomatal function and reduced transpiration in combination with increased [ABA] indicate that preharvest B-light plays an important role in governing stomatal functionality and ABA homeostasis under high RH and can be a useful method to improve postharvest water balance of roses.

Elevated air humidity increases UV mediated leaf and DNA damage in pea (Pisum sativum) due to reduced flavonoid content and antioxidant power.

Growth in high relative air humidity (RH, >85%) affects plant morphology and causes diminished response to stomatal closing signals. Many greenhouses are prone to high RH conditions, which may negatively affect production and post-harvest quality. UV radiation induces stomatal closure in several species, and facilitates disease control. We hypothesised that UV exposure may trigger stomatal closure in pea plants (Pisum sativum) grown in high RH, thereby restoring stomatal function. The effects of UV exposure were tested on plants grown in moderate (60%) or high (90%) RH. UV exposure occurred at night, according to a disease control protocol. Lower stomatal conductance rates were found in UV-exposed plants, though UV exposure did not improve the rate of response to closing stimuli or desiccation tolerance. UV-exposed plants showed leaf curling, chlorosis, necrosis, and DNA damage measured by the presence of cyclobutane pyrimidine dimers (CPD), all of which were significantly greater in high RH plants. These plants also had lower total flavonoid content than moderate RH plants, and UV-exposed plants had less than controls. Plants exposed to UV had a higher content of cuticular layer uronic compounds than control plants. However, high RH plants had a higher relative amount of cuticular waxes, but decreased proteins and uronic compounds. Plants grown in high RH had reduced foliar antioxidant power compared to moderate RH. These results indicate that high RH plants were more susceptible to UV-induced damage than moderate RH plants due to reduced flavonoid content and oxidative stress defence.

Daily changes in VPD during leaf development in high air humidity increase the stomatal responsiveness to darkness and dry air.

Previous studies have shown that plants developed under high relative air humidity (RH>85%) develop malfunctioning stomata and therefor have increased transpiration and reduced desiccation tolerance when transferred to lower RH conditions and darkness. In this study, plants developed at high RH were exposed to daily VPD fluctuations created by changes in temperature and/or RH to evaluate the potential improvements in stomatal functioning. Daily periods with an 11°C temperature increase and consequently a VPD increase (vpd: 0.36-2.37KPa) reduced the stomatal apertures and improved the stomatal functionality and desiccation tolerance of the rosette plant Arabidopsis thaliana. A similar experiment was performed with only a 4°C temperature increase and/or a RH decrease on tomato. The results showed that a daily change in VPD (vpd: 0.36-1.43KPa) also resulted in improved stomatal responsiveness and decreased water usage during growth. In tomato, the most effective treatment to increase the stomatal responsiveness to darkness as a signal for closure was daily changes in RH without a temperature increase.

Thermoperiodic Control of Floral Induction Involves Modulation of the Diurnal FLOWERING LOCUS T Expression Pattern.

Thermoperiodism is defined as the ability to discriminate between day temperature (DT) and night temperature (NT). Our aim was to shed light on the mechanistic basis of thermoperiodic floral induction with acceleration under lower DT than NT compared with other DT-NT combinations at the same average daily temperature (ADT), a response exploited in temperate area greenhouses. Arabidopsis thaliana floral pathway mutants and a lhy circadian clock mutant as well as the expression of floral integrators and LHY (LATE ELONGATED HYPOCOTYL) were studied under different DT-NT combinations, all at the same ADT. We show that acceleration of floral induction under lower DT than NT is linked to increased FT expression early during the day and generally increased LFY expression preceding visible flower buds, compared with higher DT than NT or equal DT and NT. Consistent with FLOWERING LOCUS T (FT) action through LEAFY (LFY), time to floral transition in ft-1 and lfy-1 was similar under all treatments, in contrast to the situation for soc1-1, which behaved like the wild type (WT). The lhy-21 mutants did not discriminate between opposite DT-NT combinations, whereas LHY expression in the WT differed in these temperature regimes. This might suggest that LHY plays a role in thermoperiodic control of floral induction. We conclude that thermoperiodic control of floral transition is associated with modulation of the diurnal expression patterns of FT, with timing of temperature alteration being important rather than ADT.

Elevated air movement enhances stomatal sensitivity to abscisic acid in leaves developed at high relative air humidity.

High relative air humidity (RH ≥ 85%) during growth leads to stomata malfunctioning, resulting in water stress when plants are transferred to conditions of high evaporative demand. In this study, we hypothesized that an elevated air movement (MOV) 24 h per day, during the whole period of leaf development would increase abscisic acid concentration ([ABA]) enhancing stomatal functioning. Pot rose 'Toril' was grown at moderate (61%) or high (92%) RH combined with a continuous low (0.08 m s(-1)) or high (0.92 m s(-1)) MOV. High MOV reduced stomatal pore length and aperture in plants developed at high RH. Moreover, stomatal function improved when high MOV-treated plants were subjected to leaflet desiccation and ABA feeding. Endogenous concentration of ABA and its metabolites in the leaves was reduced by 35% in high RH, but contrary to our hypothesis this concentration was not significantly affected by high MOV. Interestingly, in detached leaflets grown at high RH, high MOV increased stomatal sensitivity to ABA since the amount of exogenous ABA required to decrease the transpiration rate was significantly reduced. This is the first study to show that high MOV increases stomatal functionality in leaves developed at high RH by reducing the stomatal pore length and aperture and enhancing stomatal sensitivity to ABA rather than increasing leaf [ABA].

Ethylene is involved in high air humidity promoted stomatal opening of tomato (Lycopersicon esculentum) leaves.

High relative air humidity (RH) promotes stomatal opening in tomato leaves. This study examined the role of the plant hormones abscisic acid (ABA) and ethylene in high RH induced stomatal opening. Plants were grown in high (90%) and moderate (60%) RH or transferred from moderate to high RH. ABA levels were only slightly, but significantly decreased during darkness by increasing RH. However, a significantly higher ethylene evolution was found in high RH compared with moderate RH. Ethephon increased conductance and stomatal aperture in moderate RH. Treatment with amino-ethoxyvinylglycine (AVG) suppressed stomatal opening when plants were transferred from moderate to high RH. Similarly, blocking the ethylene receptor or using an ethylene-insensitive mutant (NR) reduced the response to high RH. These results demonstrate that both ethylene production and sensitivity play a role in high RH-induced stomatal opening in tomato leaves. The increased conductance found when plants were transferred to high RH could be counteracted by exogenous ABA spray. The ABA deficient mutant 'Flacca' produced high levels of ethylene irrespective of the RH and the difference in water loss and conductance between high and moderate grown 'Flacca' plants was attenuated compared with WT. The results indicate that both ABA and ethylene play a role in air humidity control of stomatal movement in tomato.

ABA induces H2O2 production in guard cells, but does not close the stomata on Vicia faba leaves developed at high air humidity.

Plants developed under constant high (> 85%) relative air humidity (RH) have larger stomata that are unable to close completely. One of the hypotheses for the less responsive stomata is that the plants have reduced sensitivity to abscisic acid (ABA). Both ABA and darkness are signals for stomatal closure and induce the production of the secondary messenger hydrogen peroxide (H2O2). In this study, the ability of Vicia faba plants developed in moderate or high RH to close the stomata in response to darkness, ABA and H2O2 was investigated. Moreover, the ability of the plants to produce H2O2 when treated with ABA or transferred to darkness was also assessed. Our results show that the ABA concentration in moderate RH is not increased during darkness even though the stomata are closing. This indicates that stomatal closure in V. faba during darkness is independent of ABA production. ABA induced both H2O2 production and stomatal closure in stomata formed at moderate RH. H2O2 production, as a result of treatment with ABA, was also observed in stomata formed at high RH, though the closing response was considerably smaller as compared with moderate RH. In either RH, leaf ABA concentration was not affected by darkness. Similarly to ABA treatment, darkness elicited both H2O2 production and stomatal closure following plant cultivation at moderate RH. Contrary to this, neither H2O2 production nor stomatal closure took place when stomata were formed at high RH. These results suggest that the reduced stomatal response in plants developed in continuous high RH is caused by one or more factors downstream of H2O2 in the signaling pathway toward stomatal closure.

Overexpression of the AtSHI gene in poinsettia, Euphorbia pulcherrima, results in compact plants.

Euphorbia pulcherrima, poinsettia, is a non-food and non-feed vegetatively propagated ornamental plant. Appropriate plant height is one of the most important traits in poinsettia production and is commonly achieved by application of chemical growth retardants. To produce compact poinsettia plants with desirable height and reduce the utilization of growth retardants, the Arabidopsis SHORT INTERNODE (AtSHI) gene controlled by the cauliflower mosaic virus 35S promoter was introduced into poinsettia by Agrobacterium-mediated transformation. Three independent transgenic lines were produced and stable integration of transgene was verified by PCR and Southern blot analysis. Reduced plant height (21-52%) and internode lengths (31-49%) were obtained in the transgenic lines compared to control plants. This correlates positively with the AtSHI transcript levels, with the highest levels in the most dwarfed transgenic line (TL1). The indole-3-acetic acid (IAA) content appeared lower (11-31% reduction) in the transgenic lines compared to the wild type (WT) controls, with the lowest level (31% reduction) in TL1. Total internode numbers, bract numbers and bract area were significantly reduced in all transgenic lines in comparison with the WT controls. Only TL1 showed significantly lower plant diameter, total leaf area and total dry weight, whereas none of the AtSHI expressing lines showed altered timing of flower initiation, cyathia abscission or bract necrosis. This study demonstrated that introduction of the AtSHI gene into poinsettia by genetic engineering can be an effective approach in controlling plant height without negatively affecting flowering time. This can help to reduce or avoid the use of toxic growth retardants of environmental and human health concern. This is the first report that AtSHI gene was overexpressed in poinsettia and transgenic poinsettia plants with compact growth were produced.

High relative air humidity and continuous light reduce stomata functionality by affecting the ABA regulation in rose leaves.

Plants developed under high (90%) relative air humidity (RH) have previously been shown to have large, malfunctioning stomata, which results in high water loss during desiccation and reduced dark induced closure. Stomatal movement is to a large extent regulated by abscisic acid (ABA). It has therefore been proposed that low ABA levels contribute to the development of malfunctioning stomata. In this study, we investigated the regulation of ABA content in rose leaves, through hormone analysis and ß-glucosidase quantification. Compared with high RH, rose plants developed in moderate RH (60%) and 20 h photoperiod contained higher levels of ABA and ß-glucosidase activity. Also, the amount of ABA increased during darkness simultaneously as the ABA-glucose ester (GE) levels decreased. In contrast, plants developed under high RH with 20 h photoperiod showed no increase in ABA levels during darkness, and had low ß-glucosidase activity converting ABA-GE to ABA. Continuous lighting (24 h) resulted in low levels of ß-glucosidase activity irrespective of RH, indicating that a dark period is essential to activate ß-glucosidase. Our results provide new insight into the regulation of ABA under different humidities and photoperiods, and clearly show that ß-glucosidase is a key enzyme regulating the ABA pool in rose plants.

A high proportion of blue light increases the photosynthesis capacity and leaf formation rate of Rosa × hybrida but does not affect time to flower opening.

Alterations in light quality affect plant morphogenesis and photosynthetic responses but the effects vary significantly between species. Roses exhibit an irradiance-dependent flowering control but knowledge on light quality responses is scarce. In this study we analyzed, the responses in morphology, photosynthesis and flowering of Rosa × hybrida to different blue (B) light proportions provided by light-emitting diodes (LED, high B 20%) and high pressure sodium (HPS, low B 5%) lamps. There was a strong morphological and growth effect of the light sources but no significant difference in total dry matter production and flowering. HPS-grown plants had significantly higher leaf area and plant height, yet a higher dry weight proportion was allocated to leaves than stems under LED. LED plants showed 20% higher photosynthetic capacity (Amax ) and higher levels of soluble carbohydrates. The increase in Amax correlated with an increase in leaf mass per unit leaf area, higher stomata conductance and CO2 exchange, total chlorophyll (Chl) content per area and Chl a/b ratio. LED-grown leaves also displayed a more sun-type leaf anatomy with more and longer palisade cells and a higher stomata frequency. Although floral initiation occurred at a higher leaf number in LED, the time to open flowers was the same under both light conditions. Thereby the study shows that a higher portion of B light is efficient in increasing photosynthesis performance per unit leaf area, enhancing growth and morphological changes in roses but does not affect the total Dry Matter (DM) production or time to open flower.

UV responses of Lolium perenne raised along a latitudinal gradient across Europe: a filtration study.

Lolium perenne (cv. AberDart) was grown at 14 locations along a latitudinal gradient across Europe (37-68°N) to study the impact of ultraviolet radiation (UV) and climate on aboveground growth and foliar UV-B absorbing compounds. At each location, plants were grown outdoors for 5 weeks in a replicated UV-B filtration experiment consisting of open, UV-B transparent (cellulose diacetate) and UV-B opaque (polyester) environments. Fourier transform-infrared spectroscopy was used to compare plant metabolite profiles in relation to treatment and location. UV radiation and climatic parameters were determined for each location from online sources and the data were assessed using a combination of anova and multiple regression analyses. Most of the variation in growth between the locations was attributable to the combination of climatic parameters, with minimum temperature identified as an important growth constraint. However, no single environmental parameter could consistently account for the variability in plant growth. Concentrations of foliar UV-B absorbing compounds showed a positive trend with solar UV across the latitudinal gradient; however, this relationship was not consistent in all treatments. The most striking experimental outcome from this study was the effect of presence or absence of filtration frames on UV-absorbing compounds. Overall, the study demonstrates the value of an European approach in studying the impacts of natural UV across a large latitudinal gradient. We have shown the feasibility of coordinated UV filtration at multiple sites but have also highlighted the need for open controls and careful interpretation of plant responses.

Day and night temperature responses in Arabidopsis: effects on gibberellin and auxin content, cell size, morphology and flowering time.

The effect of 16 different day (DT) and night (NT) temperature combinations (DT and NT 12, 17, 22 and 27 degrees C) on rosette leaf growth, flower stem elongation and flowering time in Arabidopsis thaliana Ler was investigated. Final leaf length decreased with increasing NT due to a combination of reduced elongation period and reduced elongation rate. Final stem length increased with increasing DT due to increased elongation rate, and decreased with increasing NT due to a decrease in elongation period. Under NT 27 degrees C, however, stem elongation rate increased greatly, resulting in the same final stem length as under NT 12 degrees C. The transition to flowering was accelerated by increasing NT. A linear regression analysis was performed to clarify the relationship between final leaf length, final stem length and flowering time with DIF (DT minus NT) and/or ADT (average daily temperature). For all three variables, the effect of DIF depended on ADT and vice versa. The relationship of final stem length with DIF also depended on the temperature range. Increased cell volume in flower stems developing at DT/NT 22/12 degrees C gave rise to longer and thicker stems compared with stems developing at DT/NT 12/22 degrees C. GC-MS analysis (gas chromatography-mass spectrometry) showed that the endogenous level of IAA was 56 % higher in stems grown under DT/NT 22/12 degrees C compared with DT/NT 12/22 degrees C. Of the 12 gibberellins analysed, however, only the level of non-bioactive GA29 was affected by the temperature treatment.