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Figure 1.
Phenotypes of (a) plants and (b) leaves of five Dianthus plants: D. caryophyllus, D. gratianopolitanus, D. spiculifolius (WT), D. spiculifolius (HW), and D. carthusianorum. WT: wild type; HW: high wax content. Bar = 1 cm (b).
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Figure 2.
Scanning electron microscopy (SEM) images of cuticular wax crystals on the adaxial leaf surface of five Dianthus plants obtained by three leaf sample fixation methods. (a) Cuticular wax morphology of leaf samples fixed with glutaraldehyde (2.5%) and then dehydrated with alcohol. (b) Cuticular wax morphology of leaf samples dried at 40 °C. (c) Cuticular wax morphology of leaf samples air-dried at room temperature. Red arrows indicate rodlet-shaped wax crystals, and red arrowheads indicate irregular platelet-shaped wax crystals. Bar = 1 µm. WT, wild type; HW, high wax content.
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Figure 3.
Cryo-SEM images of cuticular wax crystals on the adaxial leaf surface of five Dianthus plants. (a) D. caryophyllus, (b) D. gratianopolitanus, (c) D. spiculifolius (wild-type, WT), (d) D. spiculifolius (high wax mutant, HW), (e) D. carthusianorum. Red arrows indicate tubular wax crystals, and red arrowhead indicates irregular platelet-shaped wax crystals. Bar = 1 µm. WT, wild type; HW, high wax content.
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Figure 4.
Comparison of cuticle permeability, non-stomatal water loss and chlorophyll leaching rates in leaves of five Dianthus plants. (a) Five Dianthus leaves were immersed for 24 h in 0.05% toluidine blue-O and then rinsed with water. (b) The rate of non-stomatal water loss from Dianthus leaves at 38 °C for 240 min. Water loss rate was expressed as a percentage of fresh weight at each individual time point versus initial fresh weight. Error bars represent standard error (SE) (n = 3). (c) The rate of chlorophyll leaching from Dianthus leaves within 24 h. Chlorophyll leaching rate was expressed as a percentage of extracted chlorophyll content at each individual time point versus chlorophyll content at 48 h after initial immersion. Error bars represent SE (n = 3). WT, wild type; HW, high wax content.
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Figure 5.
Main components of cuticular wax on the leaves of five Dianthus plants and their proportion to total wax abundance. (a) D. caryophyllus, (b) D. gratianopolitanus, (c) D. spiculifolius (WT, wild type), (d) D. spiculifolius (HW, high wax content), (e) D. carthusianorum.
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Classification Name Chemical formula Wax load (µg/cm2) Ds (HW) SE (n = 3) Ds (WT) SE (n = 3) Dcart SE (n = 3) Dg SE (n = 3) Dcary SE (n = 3) Fatty acids Oleic acid C18H34O2 0.0117 0.0033 0.0038 0.0013 0.0045 0.0009 0.0100 0.0062 0.0112 0.0080 Erucic acid C22H42O2 0.0011 0.0002 0.0006 0.0002 0.0010 0.0001 0.0033 0.0021 0.0028 0.0019 Alkanes Heptadecane, 2-methyl- C18H38 0.0007 0.0002 0.0004 0.0004 0.0009 0.0007 0.0036 0.0020 0.0026 0.0011 Eicosane C20H42 0.0404 0.0404 0.0314 0.0075 0.1133 0.0500 0.5224 0.1524 0.4323 0.1792 Heneicosane C21H44 0.0273 0.0061 0.0138 0.0033 0.0356 0.0118 0.2248 0.0643 0.2821 0.2118 Heptacosane C27H56 1.3036 0.3190 1.0488 0.2382 0.7294 0.1948 2.5350 0.2712 1.4806 0.1268 2-methyloctacosane C29H60 0.0053 0.0014 0.0012 0.0004 0.0009 0.0000 0.0292 0.0227 0.0043 0.0021 Hentriacontane C31H64 2.0727 0.4401 0.9618 0.2272 0.9219 0.0980 1.4263 0.1561 1.6730 0.2989 Dotriacontane C32H66 0 0 0.0011 0.0010 0.0013 0.0006 0 0 0.0032 0.0041 Hexatriacontane C36H74 0.0043 0.0022 0.0031 0.0018 0.0045 0.0019 0.0095 0.0048 0.0311 0.0216 Aldehydes 13-Octadecenal, (Z)- C18H34O 0.0139 0.0058 0.0019 0.0004 0.0017 0.0006 0.0243 0.0193 0.0044 0.0029 Octadecanal C18H36O 0.0100 0.0043 0.0033 0.0004 0.0027 0.0003 0.0061 0.0046 0.0056 0.0027 Eicosanal C20H40O 2.0066 0.2791 0.6801 0.1167 0.5929 0.1364 0.9309 0.2463 1.1186 0.2585 Alcohols 1-Octacosanol C28H58O 3.9916 0.2895 4.3199 0.7506 2.2719 0.4369 0.0000 0.0000 0.0000 0.0000 Ketones 2-Nonadecanone C19H38O 0.0418 0.0085 0.0091 0.0022 0.0160 0.0039 0.0900 0.0633 0.0308 0.0389 Tricosane-2,4-dione C23H44O2 0.2934 0.1054 0.0033 0.0007 0.0092 0.0040 0.9492 0.1375 0.3463 0.1357 Esters Myristic acid isobutyl ester C18H36O2 0.0012 0.0004 0 0 0 0 0.0291 0.0205 0 0 Hexadecanoic acid, 15-methyl-, methyl ester C18H36O2 0.0048 0.0020 0.0009 0.0002 0.0016 0.0003 0.0015 0.0009 0.0058 0.0021 Octadecanoic acid, phenylmethyl ester C25H42O2 0.0047 0.0011 0.0018 0.0004 0.0147 0.0056 0.0572 0.0291 0.0332 0.0337 Total wax 9.8352 1.5093 7.0865 1.3530 4.7240 0.9468 6.8525 1.2031 5.4679 1.3302 Table 1.
Quantification of multiple cuticular wax components identified from the leaves of five Dianthus plants.
Figures
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Tables
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