Pore size regulates operating stomatal conductance, while stomatal densities drive the partitioning of conductance between leaf sides.

BACKGROUND AND AIMS: Leaf gas exchange is influenced by stomatal size, density, distribution between the leaf adaxial and abaxial sides, as well as by pore dimensions. This study aims to quantify which of these traits mainly underlie genetic differences in operating stomatal conductance (gs) and addresses possible links between anatomical traits and regulation of pore width. METHODS: Stomatal responsiveness to desiccation, gs-related anatomical traits of each leaf side and estimated gs (based on these traits) were determined for 54 introgression lines (ILs) generated by introgressing segments of Solanum pennelli into the S. lycopersicum 'M82'. A quantitative trait locus (QTL) analysis for stomatal traits was also performed. KEY RESULTS: A wide genetic variation in stomatal responsiveness to desiccation was observed, a large part of which was explained by stomatal length. Operating gs ranged over a factor of five between ILs. The pore area per stomatal area varied 8-fold among ILs (2-16 %), and was the main determinant of differences in operating gs between ILs. Operating gs was primarily positioned on the abaxial surface (60-83 %), due to higher abaxial stomatal density and, secondarily, to larger abaxial pore area. An analysis revealed 64 QTLs for stomatal traits in the ILs, most of which were in the direction of S. pennellii. CONCLUSIONS: The data indicate that operating and maximum gs of non-stressed leaves maintained under stable conditions deviate considerably (by 45-91 %), because stomatal size inadequately reflects operating pore area (R(2) = 0·46). Furthermore, it was found that variation between ILs in both stomatal sensitivity to desiccation and operating gs is associated with features of individual stoma. In contrast, genotypic variation in gs partitioning depends on the distribution of stomata between the leaf adaxial and abaxial epidermis.

Threshold response of stomatal closing ability to leaf abscisic acid concentration during growth.

Leaf abscisic acid concentration ([ABA]) during growth influences morpho-physiological traits associated with the plant's ability to cope with stress. A dose-response curve between [ABA] during growth and the leaf's ability to regulate water loss during desiccation or rehydrate upon re-watering was obtained. Rosa hybrida plants were grown at two relative air humidities (RHs, 60% or 90%) under different soil water potentials (-0.01, -0.06, or -0.08MPa) or upon grafting onto the rootstock of a cultivar sustaining [ABA] at elevated RH. Measurements included [ABA], stomatal anatomical features, stomatal responsiveness to desiccation, and the ability of leaves, desiccated to varying degrees, to recover their weight (rehydrate) following re-watering. Transpiration efficiency (plant mass per transpired water) was also determined. Soil water deficit resulted in a lower transpiration rate and higher transpiration efficiency at both RHs. The lowest [ABA] was observed in well-watered plants grown at high RH. [ABA] was increased by soil water deficit or grafting, at both RHs. The growth environment-induced changes in stomatal size were mediated by [ABA]. When [ABA] was increased from the level of (well-watered) high RH-grown plants to the value of (well-watered) plants grown at moderate RH, stomatal responsiveness was proportionally improved. A further increase in [ABA] did not affect stomatal responsiveness to desiccation. [ABA] was positively related to the ability of dehydrated leaves to rehydrate. The data indicate a growth [ABA]-related threshold for stomatal sensitivity to desiccation, which was not apparent either for stomatal size or for recovery (rehydration) upon re-watering.

Foliar abscisic acid content underlies genotypic variation in stomatal responsiveness after growth at high relative air humidity.

BACKGROUND AND AIMS: Stomata formed at high relative air humidity (RH) respond less to abscisic acid (ABA), an effect that varies widely between cultivars. This study tested the hypotheses that this genotypic variation in stomatal responsiveness originates from differential impairment in intermediates of the ABA signalling pathway during closure and differences in leaf ABA concentration during growth. METHODS: Stomatal anatomical features and stomatal responsiveness to desiccation, feeding with ABA, three transduction elements of its signalling pathway (H2O2, NO, Ca(2+)) and elicitors of these elements were determined in four rose cultivars grown at moderate (60 %) and high (90 %) RH. Leaf ABA concentration was assessed throughout the photoperiod and following mild desiccation (10 % leaf weight loss). KEY RESULTS: Stomatal responsiveness to desiccation and ABA feeding was little affected by high RH in two cultivars, whereas it was considerably attenuated in two other cultivars (thus termed sensitive). Leaf ABA concentration was lower in plants grown at high RH, an effect that was more pronounced in the sensitive cultivars. Mild desiccation triggered an increase in leaf ABA concentration and equalized differences between leaves grown at moderate and high RH. High RH impaired stomatal responses to all transduction elements, but cultivar differences were not observed. CONCLUSIONS: High RH resulted in decreased leaf ABA concentration during growth as a result of lack of water deficit, since desiccation induced ABA accumulation. Sensitive cultivars underwent a larger decrease in leaf ABA concentration rather than having a higher ABA concentration threshold for inducing stomatal functioning. However, cultivar differences in stomatal closure following ABA feeding were not apparent in response to H2O2 and downstream elements, indicating that signalling events prior to H2O2 generation are involved in the observed genotypic variation.

Additional Blue LED during Cultivation Induces Cold Tolerance in Tomato Fruit but Only to an Optimum.

Tomato is a chilling-sensitive fruit. The aim of this study is to examine the role of preharvest blue LED lighting (BL) to induce cold tolerance in 'Foundation' tomatoes. Blue and red supplemental LED light was applied to achieve either 0, 12 or 24% additional BL (0B, 12B and 24B). Mature green (MG) or red (R) tomatoes were harvested and cold stored at 4 °C for 0, 5, 10, 15 and 20 d, and then stored for 20 d at 20 °C (shelf life). Chilling injury (CI) indices, color and firmness, hydrogen peroxide, malondialdehyde, ascorbic acid and catalase activity were characterized. At harvest, R tomatoes cultivated at 12B were firmer and showed less coloration compared to fruit of other treatments. These fruits also showed higher loss of red color during cold storage and lower CI symptoms during shelf-life. MG tomatoes cultivated at 12B showed delayed coloring (non-chilled) and decreased weight loss (long cold stored) during shelf life compared to fruit in the other treatments. No effects of light treatments, both for MG and R tomatoes, were observed for the selected antioxidant capacity indicators. Improved cold tolerance for R tomatoes cultivated at 12B points to lycopene having higher scavenging activity at lower concentrations to mitigate chilling injury.

Stomatal behavior following mid- or long-term exposure to high relative air humidity: A review.

High relative air humidity (RH ≥ 85%) is frequent in controlled environments, and not uncommon in nature. In this review, we examine the high RH effects on plants with a special focus on stomatal characters. All aspects of stomatal physiology are attenuated by elevated RH during leaf expansion (long-term) in C3 species. These include impaired opening and closing response, as well as weak diel oscillations. Consequently, the high RH-grown plants are not only vulnerable to biotic and abiotic stress, but also undergo a deregulation between CO2 uptake and water loss. Stomatal behavior of a single leaf is determined by the local microclimate during expansion, and may be different than the remaining leaves of the same plant. No effect of high RH is apparent in C4 and CAM species, while the same is expected for species with hydropassive stomatal closure. Formation of bigger stomata with larger pores is a universal response to high RH during leaf expansion, whereas the effect on stomatal density appears to be species- and leaf side-specific. Compelling evidence suggests that ABA mediates the high RH-induced stomatal malfunction, as well as the stomatal size increase. Although high RH stimulates leaf ethylene evolution, it remains elusive whether or not this contributes to stomatal malfunction. Most species lose stomatal function following mid-term (4-7 d) exposure to high RH following leaf expansion. Consequently, the regulatory role of ambient humidity on stomatal functionality is not limited to the period of leaf expansion, but holds throughout the leaf life span.

Adding Blue to Red Supplemental Light Increases Biomass and Yield of Greenhouse-Grown Tomatoes, but Only to an Optimum.

Greenhouse crop production in northern countries often relies heavily on supplemental lighting for year-round yield and product quality. Among the different spectra used in supplemental lighting, red is often considered the most efficient, but plants do not develop normally when grown solely under monochromatic red light ("red light syndrome"). Addition of blue light has been shown to aid normal development, and typical lighting spectra in greenhouse production include a mixture of red and blue light. However, it is unclear whether sunlight, as part of the light available to plants in the greenhouse, may be sufficient as a source of blue light. In a greenhouse high-wire tomato (Solanum lycopersicum), we varied the percentage of blue supplemental light (in a red background) as 0, 6, 12, and 24%, while keeping total photosynthetically active radiation constant. Light was supplied as a mixture of overhead (99 µmol m-2 s-1) and intracanopy (48 µmol m-2 s-1) LEDs, together with sunlight. Averaged over the whole experiment (111 days), sunlight comprised 58% of total light incident onto the crop. Total biomass, yield and number of fruits increased with the addition of blue light to an optimum, suggesting that both low (0%) and high (24%) blue light intensities were suboptimal for growth. Stem and internode lengths, as well as leaf area, decreased with increases in blue light percentage. While photosynthetic capacity increased linearly with increases in blue light percentage, photosynthesis in the low blue light treatment (0%) was not low enough to suggest the occurrence of the red light syndrome. Decreased biomass at low (0%) blue light was likely caused by decreased photosynthetic light use efficiency. Conversely, decreased biomass at high (24%) blue light was likely caused by reductions in canopy light interception. We conclude that while it is not strictly necessary to add blue light to greenhouse supplemental red light to obtain a functional crop, adding some (6-12%) blue light is advantageous for growth and yield while adding 24% blue light is suboptimal for growth.

Improving stomatal functioning at elevated growth air humidity: A review.

Plants grown at high relative air humidity (RH≥85%) are prone to lethal wilting upon transfer to conditions of high evaporative demand. The reduced survival of these plants is related to (i) increased cuticular permeability, (ii) changed anatomical features (i.e., longer pore length and higher stomatal density), (iii) reduced rehydration ability, (iv) impaired water potential sensitivity to leaf dehydration and, most importantly, (v) compromised stomatal closing ability. This review presents a critical analysis of the strategies which stimulate stomatal functioning during plant development at high RH. These include (a) breeding for tolerant cultivars, (b) interventions with respect to the belowground environment (i.e., water deficit, increased salinity, nutrient culture and grafting) as well as (c) manipulation of the aerial environment [i.e., increased proportion of blue light, increased air movement, temporal temperature rise, and spraying with abscisic acid (ABA)]. Root hypoxia, mechanical disturbance, as well as spraying with compounds mimicking ABA, lessening its inactivation or stimulating its within-leaf redistribution are also expected to improve stomatal functioning of leaves expanded in humid air. Available evidence leaves little doubt that genotypic and phenotypic differences in stomatal functioning following cultivation at high RH are realized through the intermediacy of ABA.

Smaller stomata require less severe leaf drying to close: a case study in Rosa hydrida.

Stomata formed at high relative air humidity (RH) close less as leaf dries; an effect that varies depending on the genotype. We here quantified the contribution of each stomatal response characteristic to the higher water loss of high RH-grown plants, and assessed the relationship between response characteristics and intraspecific variation in stomatal size. Stomatal size (length multiplied by width), density and responsiveness to desiccation, as well as pore dimensions were analyzed in ten rose cultivars grown at moderate (60%) or high (85%) RH. Leaf morphological components and transpiration at growth conditions were also assessed. High growth RH resulted in thinner (11%) leaves with larger area. A strong positive genetic correlation of daytime and nighttime transpiration at either RH was observed. Stomatal size determined pore area (r=0.7) and varied by a factor of two, as a result of proportional changes in length and width. Size and density of stomata were not related. Following desiccation, high RH resulted in a significantly lower (6-19%) decline of transpiration in three cultivars, whereas the relative water content (RWC) of high RH-expanded leaflets was lower (29-297%) in seven cultivars. The lower RWC of these leaflets was caused by (a) higher (33-72%) stable transpiration and/or (b) lower (12-143%) RWC at which this stable transpiration occurred, depending on the cultivar. Stomatal size was significantly correlated with both characteristics (r=0.5 and -0.7, respectively). These results indicate that stomatal size explains much of the intraspecific variation in the regulation of transpiration upon water deprivation on rose.