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Water conductance of the cuticular membrane (CM) of mature sweet cherry fruit (Prunus avium L. cv. Sam) was investigated by monitoring water loss from segments of the outer pericarp excised from the cheek of the fruit. Segments consisted of epidermis, hypodermis and several cell layers of the mesocarp. Segments were mounted in stainless-steel diffusion cells with the mesocarp surface in contact with water, while the outer cuticular surface was exposed to dry silica (22 +/- 1 degrees C). Conductance was calculated by dividing the amount of water transpired per unit area and time by the difference in water vapour concentration across the segment. Conductance values had a log normal distribution with a median of 1.15 x 10(-4) m s(-1) (n=357). Transpiration increased linearly with time. Conductance remained constant and was not affected by metabolic inhibitors (1 mM NaN3 or 0.1 mM carbonylcyanide m-chlorophenylhydrazone) or thickness of segments (range 0.8-2.8 mm). Storing fruit (up to 42 d, 1 degrees C) used as a source of segments had no consistent effect on conductance. Conductance of the CM increased from cheek (1.16 +/- 0.10 x 10(-4) m s(-1)) to ventral suture (1.32 +/- 0.07 x 10(-4) m s(-1)) and to stylar end (2.53 +/- 0.17 x 10(-4) m s(-1)). There was a positive relationship (r2=0.066**; n=108) between conductance and stomatal density. From this relationship the cuticular conductance of a hypothetical astomatous CM was estimated to be 0.97 +/- 0.09 x 10(-4) m s(-1). Removal of epicuticular wax by stripping with cellulose acetate or extracting epicuticular plus cuticular wax by dipping in CHCl3/methanol increased conductance 3.6- and 48.6-fold, respectively. Water fluxes increased with increasing temperature (range 10-39 degrees C) and energies of activation, calculated for the temperature range from 10 to 30 degrees C, were 64.8 +/- 5.8 and 22.2 +/- 5.0 kJ mol(-1) for flux and vapour-concentration-based conductance, respectively.

Changes in surface area, deposition and elastic strain of the cuticular membrane (CM) were monitored during development of sweet cherry (Prunus avium L.) fruit. Fruit mass and surface area ('Sam') increased in a sigmoidal pattern between 16 and 85 days after full bloom (DAFB) with maximum rates of 0.35 g day(-1) and 0.62 cm(2) day(-1), respectively. Rates of total area strain, namely the sum of elastic plus plastic strain, were highest in cheek and stem cavity regions followed by stylar and suture regions. Rates of total uniaxial strain were higher in transverse, namely perpendicular to the stem/stylar axis, than in longitudinal direction, namely parallel to the stem/stylar axis. On a whole fruit basis CM mass remained essentially constant during fruit development. Mass of CM, dewaxed CM and wax per unit surface area decreased during development, particularly between 43 and 71 DAFB. There was no change in wax content of isolated CM. Up to 43 DAFB the surface area of isolated CM was similar to the area prior to excision indicating little elastic strain, but markedly decreased thereafter. Calculating elastic and plastic components of total strain of the CM revealed, that initial deformation up to 22 to 43 DAFB was mostly plastic. Thereafter, elastic strain was evident and both, elastic and plastic deformation, increased linearly with an increase in total strain. There was no consistent difference in the relative contribution of elastic strain to total strain between transverse and longitudinal directions, but both total and elastic strain were larger in the transverse direction. Abrading the CM had only little effect on fruit turgor. However, turgor decreased when the exocarp was cut indicating that the exocarp provided a significant structural shell of a mature sweet cherry fruit ('Regina'). Our data demonstrate, that (1) surface area expansion in sweet cherry fruit causes elastic and plastic strain of the CM, and (2) the onset of elastic strain coincided with the cessation of CM formation.

The results support the view that the cessation of CM deposition during early sweet cherry fruit development is accounted for by a downregulation of genes involved in CM deposition. Genes that merit further investigation include PaWINA, PaWINB, PaLipase, PaLTPG1, PaATT1, PaLCR, PaGPAT4/8, PaLACS2, PaLACS1 and PaCER1.

The effect of surface water on the frequency of microcracks in the cuticular membrane (CM) of exocarp segments (ES) of developing sweet cherry fruit (Prunus avium L.) was studied. Strain of CM and ES on the fruit surface was preserved by mounting a stainless steel washer on the fruit surface in the cheek region using an ethyl-cyanacrylate adhesive. ES were excised by tangentially cutting underneath the washer. Frequency of microcracks in the CM of ES was determined following infi ltration for 10 minutes with a 0.1% acridine orange solution by fl uorescence microscopy before and after exposure to deionized water (generally 48 hours). Exposing the surface of ES of mature 'Burlat' sweet cherry fruit to water resulted in a rapid increase in microcracks in the CM that approached an asymptote at about 30 microcracks/cm 2 within 24 hours. There was no change in microcracks in the CM when the surface of the ES remained dry. Incubating ES in polyethylene glycol solution that was isotonic to fruit juice extracted from the same batch of fruit resulted in a greater increase in frequency of microcracks as compared to incubation in deionized water. The waterinduced increase in microcracks was closely related to strain of the CM across different developmental stages within a cultivar [between 45 and 94 days after full bloom (DAFB); r 2 = 0.96, P ≤ 0.001, n = 9] or across different cultivars at maturity (r 2 = 0.92, P ≤ 0.0022, n = 6). Incubating ES of developing fruit in enzyme solution containing pectinase and cellulase such that the outer surface remained dry resulted in complete rupture and failure of the ES. Time to rupture and percentage of ruptured ES were closely related to the strain of the CM (r 2 = 0.92, P ≤ 0.001, n = 9 and r 2 = 0.68, P ≤ 0.0063, n = 9, respectively). Removal of epicuticular wax had no effect on frequency of water-induced microcracks. Also, temperature had no effect on frequency of water-induced microcracks, but frequency of microcracks increased exponentially when exposing the outer surface of ES to relative humidities above 75%. At 100% humidity the increase in frequency of microcracks did not differ from that induced by liquid water. Local wetting the surface of intact fruit in the pedicel cavity or stylar end region resulted in formation of macroscopically visible cracks despite of a net water loss of fruit. Uniaxiale tensile tests using dry and fully hydrated CM strips isolated from mature 'Sam' sweet cherry fruit established that hydration increased fracture strain, but decreased fracture stress and moduli of elasticity. Our data demonstrate that exposure of the fruit surface to liquid water or high concentrations of water vapor resulted in formation of microcracks in the CM.

Frequency and distribution of microcracks in the cuticular membrane (CM) were monitored in cheek, suture, pedicel cavity and stylar regions of developing sweet cherry (Prunus avium L.) fruit using fluorescence microscopy following infiltration with a fluorescence tracer (1 to 2 min in 0.1% w/v acridine orange containing 50 mm citric acid and 0.1% Silwet L-77, pH 6.5). These microcracks were limited to the cuticle, did not extend into the pericarp and were only detected by microscopy. Fruit mass and surface area increased in a sigmoidal pattern with time between 16 days after full bloom (DAFB) and maturity. The increase in frequency of fruit with microcracks paralleled the increase in fruit mass. During early development (up to 43 DAFB) the CM of `Sam' fruit remained intact. However, by 57 DAFB essentially all `Sam' fruit had microcracks in the pedicel cavity and ≈25% in the suture region with little change thereafter. At maturity percentage of `Sam' fruit with microcracks in cheek, suture, pedicel cavity and stylar end region averaged 23%, 25%, 100%, and 63%, respectively. Similar data were obtained for `Hedelfinger' (70% and 100% for cheek and pedicel cavity, respectively), `Kordia' (80% and 100%) and `Van' (100% and 100%). Generally, microcracks were most severe in pedicel cavity and stylar end region. Most of the first detectable microcracks formed above periclinal walls of epidermal cells perpendicular to their longest axis (72% and 92% in cheek and stylar regions, respectively). The other microcracks formed above the anticlinal walls were mostly oriented in the direction of the underlying cell wall. There was no difference in projected surface area, length/width ratio or orientation among epidermal cells below, adjacent to or distant from the first detectable microcracks in the CM. However, as length of microcracks increased the projected surface area of cells underlying cracks increased suggesting strain induced upon cracking of the CM. Permeability of excised exocarp segments in osmotic water uptake was positively correlated with number of stomata and number of microcracks in the CM. From our results we suggest that strain of the epidermal system during stage III of fruit growth is a factor in “microcracking” of the CM that may predispose fruit to subsequent rain-induced cracking.

The effect of four applications of gibberellin A4+7 [GA4+7 (10 mg·L−1 at 10-day intervals beginning with petal fall)] on water-induced russeting, formation of microcracks. and on fruit growth and deposition of the cuticular membrane (CM) was studied in developing ‘Golden Delicious’ fruit (Malus ×domestica Borkh.). Submerging developing apple fruit in deionized water for 48 h induced russeting in untreated control but not in GA4+7-treated fruit. Immersing in water during early fruit development, 19 days after full bloom (19 DAFB), resulted in more russeting than immersions occurring later (139 DAFB). Water on the outer surface of epidermal segments increased the frequency of microscopic cracks in untreated controls but to a lesser degree in GA4+7-treated fruit. The effect of GA4+7 on water-induced russeting and formation of microcracks was larger during early as compared with later stages of fruit development. Fruit treated with GA4+7 consistently had fewer microcracks as compared with non-treated control fruit. GA4+7 had no effect on amounts or rates of cutin or wax deposition, strain, or mechanical properties of the CM as compared with the non-treated control. Thus, the decrease in russeting and formation of microcracks in the cuticle of GA4+7-treated fruit must be accounted for effects on underlying epi- and hypodermal tissues.