Building a Barrier: The Influence of Different Wax Fractions on the Water Transpiration Barrier of Leaf Cuticles

Waxes are critical in limiting non-stomatal water loss in higher terrestrial plants by making up the limiting barrier for water diffusion across cuticles. Using a differential extraction protocol, we investigated the influence of various wax fractions on the cuticular transpiration barrier. Triterpenoids (TRPs) and very long-chain aliphatics (VLCAs) were selectively extracted from isolated adaxial leaf cuticles using methanol (MeOH) followed by chloroform (TCM). The water permeabilities of the native and the solvent-treated cuticles were measured gravimetrically. Seven plant species (Camellia sinensis, Ficus elastica, Hedera helix, Ilex aquifolium, Nerium oleander, Vinca minor, and Zamioculcas zamiifolia) with highly varying wax compositions ranging from nearly pure VLCA- to TRP-dominated waxes were selected. After TRP removal with MeOH, water permeability did not or only slightly increase. The subsequent VLCA extraction with TCM led to increases in cuticular water permeabilities by up to two orders of magnitude. These effects were consistent across all species investigated, providing direct evidence that the cuticular transpiration barrier is mainly composed of VLCA. In contrast, TRPs play no or only a minor role in controlling water loss.

Variation in Petal and Leaf Wax Deposition Affects Cuticular Transpiration in Cut Lily Flowers

The vase life of cut flowers is largely affected by post-harvest water loss. Cuticular wax is the primary barrier to uncontrolled water loss for aerial plant organs. Studies on leaf cuticular transpiration have been widely conducted; however, little is known about cuticular transpiration in flowers. Here, the cuticular transpiration rate and wax composition of three lily cultivars were determined. The minimum water conductance of tepal cuticles was higher at the green bud than open flower stage. Lily cuticular transpiration exhibited cultivar- and organ-specific differences, where transpiration from the tepals was higher than leaves and was higher in the ‘Huang Tianba’ than ‘Tiber’ cultivar. The overall wax coverage of the tepals was higher compared to that of the leaves. Very-long-chain aliphatics were the main wax constituents and were dominated by n-alkanes with carbon (C) chain lengths of C27 and C29, and C29 and C31 in the tepal and leaf waxes, respectively. Primary alcohols and fatty acids as well as small amounts of alkyl esters, ketones, and branched or unsaturated n-alkanes were also detected in both tepal and leaf waxes, depending on the cultivar and organ. In addition, the chain-length distributions were similar between compound classes within cultivars, whereas the predominant C-chain lengths were substantially different between organs. This suggests that the less effective transpiration barrier provided by the tepal waxes may result from the shorter C-chain aliphatics in the tepal cuticle, compared to those in the leaf cuticle. These findings provide further insights to support the exploration of potential techniques for extending the shelf life of cut flowers based on cuticular transpiration barrier properties.

Leaf morphology, wax composition, and residual (cuticular) transpiration of four poplar clones

Key message We identified two poplar clones of the same species as highly comparable, yet clones of two further species of the same genus to be distinctly different regarding multiple morphological and ecophysiological traits. Abstract Leaf morphology, wax composition, and residual (cuticular) transpiration of four poplar clones (two clones of the hybrid species P. × canescens, P. trichocarpa, and P. euphratica) were monitored from the beginning to end of the growing season 2020. A pronounced epicuticular wax coverage was found only with P. euphratica. As the most prominent substance classes of cuticular wax primary alcohols, alkanes and esters were identified with P. × canescens and P. trichocarpa, whereas esters and alkanes were completely lacking in P. euphratica. Wax amounts were slightly decreasing during the season and significantly lower wax amounts were found for newly formed leaves in summer compared to leaves of the same age formed in spring. Residual (cuticular) transpiration was about five to tenfold lower for P. × canescens compared with the two other poplar species. Interestingly, with three of the four investigated species, newly formed leaves in summer had lower wax coverages and lower rates of residual (cuticular) transpiration compared to leaves of exactly the same age formed in spring. Our findings were especially surprising with P. euphratica, representing the only one of the four investigated poplar species naturally growing in very dry and hot climates in Central Asia. Instead of developing very low rates of residual (cuticular) transpiration, it seems to be of major advantage for P. euphratica to develop a pronounced epicuticular wax bloom efficiently reflecting light.

Leaf Cuticular Transpiration Barrier Organization in Tea Tree Under Normal Growth Conditions

The cuticle plays a major role in restricting nonstomatal water transpiration in plants. There is therefore a long-standing interest to understand the structure and function of the plant cuticle. Although many efforts have been devoted, it remains controversial to what degree the various cuticular parameters contribute to the water transpiration barrier. In this study, eight tea germplasms were grown under normal conditions; cuticle thickness, wax coverage, and compositions were analyzed from the epicuticular waxes and the intracuticular waxes of both leaf surfaces. The cuticular transpiration rates were measured from the individual leaf surface as well as the intracuticular wax layer. Epicuticular wax resistances were also calculated from both leaf surfaces. The correlation analysis between the cuticular transpiration rates (or resistances) and various cuticle parameters was conducted. We found that the abaxial cuticular transpiration rates accounted for 64–78% of total cuticular transpiration and were the dominant factor in the variations for the total cuticular transpiration. On the adaxial surface, the major cuticular transpiration barrier was located on the intracuticular waxes; however, on the abaxial surface, the major cuticular transpiration barrier was located on the epicuticular waxes. Cuticle thickness was not a factor affecting cuticular transpiration. However, the abaxial epicuticular wax coverage was found to be significantly and positively correlated with the abaxial epicuticular resistance. Correlation analysis suggested that the very-long-chain aliphatic compounds and glycol esters play major roles in the cuticular transpiration barrier in tea trees grown under normal conditions. Our results provided novel insights about the complex structure–functional relationships in the tea cuticle.

Diversity of Leaf Cuticular Transpiration and Growth Traits in Field-Grown Wheat and Aegilops Genetic Resources

Plants are subjected to unregulated water loss from their surface by cuticular transpiration. Therefore, specific morphophysiological changes may occur during leaf development to eliminate water loss. This study aimed to examine the cuticular transpiration of 23 winter wheat genotypes and their wild-growing predecessors of the genus Aegilops, which were divided into three groups to demonstrate their diversity. The genotypes were sown in autumn and grown in regular field trials at the Research Institute of Plant Production in Piešťany, Slovakia. Cuticular transpiration and growth parameters were analyzed in the postanthesis growth stage. Gravimetric measurement of residual water loss was performed on detached leaves with a precisely measured leaf area. The lowest nonproductive transpiration values were observed in modern wheat genotypes, while higher cuticular transpiration was observed in a group of landraces. Aegilops species generally showed the highest cuticular transpiration with increased water loss, but the total water loss per plot was low due to the low leaf area of the wild wheat relatives. Some of the growth parameters showed a good correlation with cuticular transpiration (e.g., dry mass per plant), but direct relationships between leaf traits and cuticular transpiration were not observed. This study identified a high diversity in cuticular resistance to water loss in wheat and Aegilops accessions of different origins. The potential of identifying and exploiting genetic resources with favorable cuticular transpiration in crop breeding is discussed.

Plasticity of the Cuticular Transpiration Barrier in Response to Water Shortage and Resupply in Camellia sinensis: A Role of Cuticular Waxes

The cuticle is regarded as a non-living tissue; it remains unknown whether the cuticle could be reversibly modified and what are the potential mechanisms. In this study, three tea germplasms (Wuniuzao, 0202-10, and 0306A) were subjected to water deprivation followed by rehydration. The epicuticular waxes and intracuticular waxes from both leaf surfaces were quantified from the mature 5th leaf. Cuticular transpiration rates were then measured from leaf drying curves, and the correlations between cuticular transpiration rates and cuticular wax coverage were analyzed. We found that the cuticular transpiration barriers were reinforced by drought and reversed by rehydration treatment; the initial weak cuticular transpiration barriers were preferentially reinforced by drought stress, while the original major cuticular transpiration barriers were either strengthened or unaltered. Correlation analysis suggests that cuticle modifications could be realized by selective deposition of specific wax compounds into individual cuticular compartments through multiple mechanisms, including in vivo wax synthesis or transport, dynamic phase separation between epicuticular waxes and the intracuticular waxes, in vitro polymerization, and retro transportation into epidermal cell wall or protoplast for further transformation. Our data suggest that modifications of a limited set of specific wax components from individual cuticular compartments are sufficient to alter cuticular transpiration barrier properties.

Variability and Plasticity in Cuticular Transpiration and Leaf Permeability Allow Differentiation of Eucalyptus Clones at an Early Age

Background and Objectives. Water stress is a major constraining factor of Eucalyptus plantations’ growth. Within a genetic improvement program, the selection of genotypes that improve drought resistance would help to improve productivity and to expand plantations. Leaf characteristics, among others, are important factors to consider when evaluating drought resistance evaluation, as well as the clone’s ability to modify leaf properties (e.g., stomatal density (d) and size, relative water content at the time of stomatal closure (RWCc), cuticular transpiration (Ec), specific leaf area (SLA)) according to growing conditions. Therefore, this study aimed at analyzing these properties in nursery plants of nine high-productivity Eucalyptus clones. Material and Methods: Five Eucalyptus globulus Labill. clones and four hybrids clones (Eucalyptus urophylla S.T. Blake × Eucalyptus grandis W. Hill ex Maiden, 12€; Eucalyptus urograndis × E. globulus, HE; Eucalyptus dunnii Maiden–E. grandis × E. globulus, HG; Eucalyptus saligna Sm. × Eucalyptus maidenii F. Muell., HI) were studied. Several parameters relating to the aforementioned leaf traits were evaluated for 2.5 years. Results: Significant differences in stomatal d and size, RWCc, Ec, and SLA among clones (p < 0.001) and according to the dates (p < 0.001) were obtained. Each clone varied seasonally the characteristics of its new developing leaves to acclimatize to the growth conditions. The pore opening surface potential (i.e., the stomatal d × size) did not affect transpiration rates with full open stomata, so the water transpired under these conditions might depend on other leaf factors. The clones HE, HG, and 12€ were the ones that differed the most from the drought resistant E. globulus control clone (C14). Those three clones showed lower leaf epidermis impermeability (HE, HG, 12€), higher SLA (12€, HG), and lower stomatal control under moderate water stress (HE, HG) not being, therefore, good candidates to be selected for drought resistance, at least for these measured traits. Conclusions: These parameters can be incorporated into genetic selection and breeding programs, especially Ec, SLA, RWCc, and stomatal control under moderate water stress.

Stomatal anatomy and closing ability is affected by supplementary light intensity in rose (Rosa hybrida L.)

Increasing the light level in protected cultivation of ornamental crops via supplementary lighting is critical to enhance both production and external quality especially during the periods of low light availability. Despite wide applications the effects of light intensities were not previously addressed on water loss pathways. In this study rose plants were cultivated at 100, 200 or 400 μmol/(m2·s) photosynthetic photon flux density (PPFD). The stomatal responsiveness to desiccation, stomatal anatomical features and cuticular transpiration were determined. Plant biomass as well as photosynthesis response to light and CO2 were also assessed. Increasing growth PPFD led to a considerable increase in plant biomass (85 and 57% for 100 to 200 and 200 to 400 μmol/(m2·s) respectively). Photosynthesis was marginally affected by increasing growth PPFD from 100 to 200 μmol/(m2·s) while a further rise to 400 μmol/(m2·s) considerably increased photosynthetic rate at high light intensities. Higher PPFD during cultivation generally led to larger stomata with bigger pores. A PPFD increase from 100 to 200 μmol/(m2·s) had a small negative effect on stomatal closing ability whereas a further rise to 400 μmol/(m2·s) had a substantial stimulatory effect. Cultivation at a PPFD higher than 100 μmol/(m2·s) led to lower rates of cuticular transpiration. In conclusion, high growth PPFD (&gt; 200 μmol/(m2·s)) enchanced both photosynthetic and stomatal anatomical traits. High light intensity (&gt; 200 μmol/(m2·s)) also led to a better control of water loss due to more responsive stomata and decreased cuticular permeability.