Constitutive expression of apple endo-POLYGALACTURONASE1 in fruit induces early maturation, alters skin structure and accelerates softening.

During fruit ripening, polygalacturonases (PGs) are key contributors to the softening process in many species. Apple is a crisp fruit that normally exhibits only minor changes to cell walls and limited fruit softening. Here, we explore the effects of PG overexpression during fruit development using transgenic apple lines overexpressing the ripening-related endo-POLYGALACTURONASE1 gene. MdPG1-overexpressing (PGox) fruit displayed early maturation/ripening with black seeds, conversion of starch to sugars and ethylene production occurring by 80 days after pollination (DAP). PGox fruit exhibited a striking, white-skinned phenotype that was evident from 60 DAP and most likely resulted from increased air spaces and separation of cells in the hypodermis due to degradation of the middle lamellae. Irregularities in the integrity of the epidermis and cuticle were also observed. By 120 DAP, PGox fruit cracked and showed lenticel-associated russeting. Increased cuticular permeability was associated with microcracks in the cuticle around lenticels and was correlated with reduced cortical firmness at all time points and extensive post-harvest water loss from the fruit, resulting in premature shrivelling. Transcriptomic analysis suggested that early maturation was associated with upregulation of genes involved in stress responses, and overexpression of MdPG1 also altered the expression of genes involved in cell wall metabolism (e.g. ß-galactosidase, MD15G1221000) and ethylene biosynthesis (e.g. ACC synthase, MD14G1111500). The results show that upregulation of PG not only has dramatic effects on the structure of the fruit outer cell layers, indirectly affecting water status and turgor, but also has unexpected consequences for fruit development.

Microcracking of strawberry fruit cuticles: mechanism and factors.

Microscopic cracks in the cuticle (microcracks) are the first symptom of the strawberry fruit disorder 'water soaking' in which the fruit surface appears watery, translucent, and pale. Water soaking severely impacts fruit quality. The objective was to investigate the factors and mechanisms of cuticular microcracking in strawberry. Fluorescence microscopy revealed numerous microcracks in the achene depressions, on the rims between depressions and at the bases of trichomes. Microcracks in the achene depressions and on the rims were either parallel or transversely oriented relative to a radius drawn from the rim to the point of attachment of the achene. In the achene depression, the frequency of microcracks with parallel orientation decreased from the calyx end of the fruit, towards the fruit tip, while the frequency of those with transverse orientation remained constant. Most microcracks occurred above the periclinal cell walls of the epidermal cells. The long axes of the epidermal cells were primarily parallel-oriented. Microcracking increased during fruit development. Cuticle mass per fruit remained constant as fruit surface area increased but cuticle thickness decreased. When fruit developed under high relative humidity (RH) conditions, the cuticle had more microcracks than under low RH conditions. Exposing the fruit surface to increasing RHs, increased microcracking, especially above 75% RH. Liquid-phase water on the fruit surface was markedly more effective in inducing microcracking than high vapor-phase water (high RH). The results demonstrate that a combination of surface area growth strain and water exposure is causal in inducing microcracking of the strawberry cuticle.

Lenticels are sites of initiation of microcracking and russeting in 'Apple' mango.

The mango cultivar 'Apple' is an important fruitcrop in Kenya, but it is highly susceptible to russeting. The objective was to establish whether lenticels predispose cv. 'Apple' mango to russeting. Fruit mass and surface area increased in a sigmoidal pattern with time. The frequency of lenticels per unit surface area decreased during development. The number of lenticels per fruit was constant. Lenticels were most frequent in the apex region and least common in the cheek and nak (ventral) regions. The cheek region also had lenticels with the largest core areas, whereas the lenticel core areas in the apex region were significantly smaller. Microscopy revealed stomata became covered over with wax deposits at 33 days after full bloom (DAFB). By 78 DAFB, periderm had formed beneath the pore. At 110 and 161 DAFB, cracks had developed and the periderm had extended tangentially and radially. The presence of lenticels increased the strain released upon excision of an epidermal segment, further strain releases occurred subsequently upon isolation of the cuticle and on extraction of the cuticular waxes. The number of lenticels per unit surface area was negatively correlated with the fruit surface area (r2 = 0.62 **), but not affected by fruit size. Mango cv. 'Apple' had fewer, larger lenticels and more russet, compared with 'Ngowe', 'Kitovu' or 'Tommy Atkins' mango. In cv. 'Apple', the lowest lenticel frequency, the largest lenticels and the most russeting occurred at a growing site at the highest altitude, with the highest rainfall and the lowest temperature. Moisture exposure of the fruit surface resulted in enlarged lenticels and more microcracking of the cuticle. Our results establish that russeting in 'Apple' mango is initiated at lenticels and is exacerbated if lenticels are exposed to moisture.

Time course of changes in the transcriptome during russet induction in apple fruit.

BACKGROUND: Russeting is a major problem in many fruit crops. Russeting is caused by environmental factors such as wounding or moisture exposure of the fruit surface. Despite extensive research, the molecular sequence that triggers russet initiation remains unclear. Here, we present high-resolution transcriptomic data by controlled russet induction at very early stages of fruit development. During Phase I, a patch of the fruit surface is exposed to surface moisture. For Phase II, moisture exposure is terminated, and the formerly exposed surface remains dry. We targeted differentially expressed transcripts as soon as 24 h after russet induction. RESULTS: During moisture exposure (Phase I) of 'Pinova' apple, transcripts associated with the cell cycle, cell wall, and cuticle synthesis (SHN3) decrease, while those related to abiotic stress increase. NAC35 and MYB17 were the earliest induced genes during Phase I. They are therefore linked to the initial processes of cuticle microcracking. After moisture removal (Phase II), the expression of genes related to meristematic activity increased (WOX4 within 24 h, MYB84 within 48 h). Genes related to lignin synthesis (MYB52) and suberin synthesis (MYB93, WRKY56) were upregulated within 3 d after moisture removal. WOX4 and AP2B3 are the earliest differentially expressed genes induced in Phase II. They are therefore linked to early events in periderm formation. The expression profiles were consistent between two different seasons and mirrored differences in russet susceptibility in a comparison of cultivars. Furthermore, expression profiles during Phase II of moisture induction were largely identical to those following wounding. CONCLUSIONS: The combination of a unique controlled russet induction technique with high-resolution transcriptomic data allowed for the very first time to analyse the formation of cuticular microcracks and periderm in apple fruit immediately after the onset of triggering factors. This data provides valuable insights into the spatial-temporal dynamics of russeting, including the synthesis of cuticles, dedifferentiation of cells, and impregnation of cell walls with suberin and lignin.

Neck shrivel in European plum is caused by cuticular microcracks, resulting from rapid lateral expansion of the neck late in development.

MAIN CONCLUSION: Susceptibility to neck shrivel in European plum is due to cuticular microcracking resulting from high surface area growth rates in the neck region, late in development. Susceptibility to the commercially important fruit disorder 'neck shrivel' differs among European plum cultivars. Radial cuticular microcracking occurs in the neck regions of susceptible cultivars, but not in non-susceptible ones, so would seem to be causal. However, the reason for the microcracking is unknown. The objective was to identify potential relationships between fruit growth pattern and microcracking incidence in the neck (proximal) and stylar (distal) ends of selected shrivel-susceptible and non-susceptible cultivars. Growth analysis revealed two allometric categories: The first category, the 'narrow-neck' cultivars, showed hypoallometric growth in the neck region (i.e., slower growth than in the region of maximum diameter) during early development (stages I + II). Later (during stage III) the neck region was 'filled out' by hyperallometric growth (i.e., faster than in the region of maximum diameter). The second category, the 'broad-neck' cultivars, had more symmetrical, allometric growth (all regions grew equally fast) throughout development. The narrow-neck cultivars exhibited extensive radial cuticular microcracking in the neck region, but little microcracking in the stylar region. In contrast, the broad-neck cultivars exhibited little microcracking overall, with no difference between the neck and stylar regions. Across all cultivars, a positive relationship was obtained for the level of microcracking in the neck region and the difference in allometric growth ratios between stage III and stages I + II. There were no similar relationships for the stylar region. The results demonstrate that accelerated stage III neck growth in the narrow-neck plum cultivars is associated with more microcracking and thus with more shrivel.

Growth strains cause vascular browning and cavities in ´Nicoter´ apples.

'Nicoter' apples (Malus × domestica Borkh.) occasionally develop a disorder referred to as vascular browning. Symptomatic fruit are perceived as being of low quality. The objective was to identify the mechanistic basis of this disorder. The frequency of symptomatic 'Nicoter' apples differed between growing sites and increased with delayed harvest. Typical symptoms are tissue browning and cavities in the ray parenchyma of the calyx region, and occasionally also of the stem end. Cavity size is positively correlated with the extent of tissue browning. Cavities were oriented radially in the direction of the bisecting line between the radii connecting the calyx/pedicel axis to the sepal and petal bundles. Microscopy revealed cell wall fragments in the cavities indicating physical rupture of cell walls. Immunolabelling of cell wall epitopes offered no evidence for separation of cells along cell walls. The growth pattern in 'Nicoter' is similar to that in its parents 'Gala' and 'Braeburn'. Allometric analyses revealed no differences in growth in fruit length among the three cultivars. However, the allometric analyses of growth in diameter revealed a marked increase in the distance between the surface of the calyx cavity and the vascular bundle in 'Nicoter', that was absent in 'Braeburn' and 'Gala'. This increase displaced the petal bundles in the ray parenchyma outwards and subjected the tissue between the petal and sepal bundles to tangential strain. Rupture of cells results in tissue browning and cavity formation. A timely harvest is a practicable countermeasure for decreasing the incidence of vascular browning.

Necked strawberries are especially susceptible to cracking.

Fruit cracking is a commercially important disorder that reduces both quantity and quality of strawberries (Fragaria × ananassa Duch.). The objective was to identify the physiological mechanism of cracking and the factors affecting cracking. Cracking is more common in necked than in normal-shaped fruit. Most macroscopic cracks ('macrocracks') occur in the seedless neck. Large fruit is more cracking susceptible than medium size or small fruit. Macrocrack orientation is predominantly latitudinal in the proximal region of the neck and longitudinal in the mid and distal regions of the neck. The neck region of necked fruit has a thicker cuticle than the body of necked or normal-shaped fruit. The vascular bundles in the neck (seedless) are orientated longitudinally, while those in the body (with seeds) are both longitudinal and radial. Epidermal cells in the neck region are elongated longitudinally, with those in the proximal region of the neck being more elongated than those in the mid or distal regions of the neck. Cuticular microcracking was more severe in necked fruit than in normal-shaped fruit. The orientations of the microcracks matched those of the macrocracks, i.e., latitudinal in the proximal neck and longitudinal in the mid and distal neck regions. Following artificial incisions (blade), gaping was significantly more pronounced in necked than in normal-shaped fruit. Incubation of fruit in deionized water induced macrocracks in about 75% of fruit. Necked fruit cracked more than normal-shaped fruit. Most macrocracks were oriented latitudinally in the proximal neck and longitudinally in the distal neck regions. The results indicate cracking results from excessive growth strains which are further increased by surface water uptake.

Detached, wetted strawberries take up substantial water in the calyx region.

In strawberry, surface disorders like 'water soaking', 'cracking' and 'shrivel' impair fruit quality of this high value crop. Water movement through the fruit surface is implicated a role in these disorders. The objective was to identify the pathways of water uptake and water loss (transpiration) and to identify factors affecting these flows. Water movement was quantified gravimetrically in detached fruit. Cumulative transpiration and uptake increased linearly with time. During ripening, fruit osmotic potential and water potential became slightly more negative. Rates of transpiration and water uptake and their corresponding permeances were constant during early ripening but increased as the fruit turned red. The permeance for osmotic water uptake was more than 10-times that for transpiration. Sealing selected regions of the fruit surface with silicone rubber allowed identification of the petal and staminal abscission zones in the calyx region and cuticular microcracks of the calyx region and receptacle as high flux pathways particularly for water uptake (osmotic). These results were confirmed by acridine orange infiltration and fluorescence microscopy. Increasing the relative humidity (RH) decreased the rate of transpiration, while increasing temperature increased both transpiration and water uptake. There was no effect of storing fruit (2 °C, ~ 80% RH) for up to 10 days. Our results identify petal and staminal abscission zones and cuticular microcracks as high flux pathways for water uptake.

Calcium decreases cell wall swelling in sweet cherry fruit.

Swelling of epidermal cell walls decreases cell-to-cell adhesion and increases cracking susceptibility in sweet cherry. Ca is suggested to decrease cracking susceptibility by crosslinking of cell wall components and, possibly, by decreasing swelling. The objective is to test this hypothesis. The effect of Ca on swelling of anticlinal epidermal cell walls was quantified microscopically in vivo using excised skin sections and in vitro using extracted cell walls. After removal of turgor, cell wall thickness increased. Incubation in CaCl2 decreased cell wall thickness up to 3 mM CaCl2. At higher concentrations thickness remained constant. Decreased cell wall swelling in vivo also occurred with other salts of divalent and trivalent cations, but not with those of monovalent cations. Decreased swelling was due to the Ca cation, the anions had no effect. Ca also decreased swelling of cell walls that were already swollen. CaCl2 also decreased swelling of extracted cell walls in vitro. There was no effect on swelling pressure. The effect on swelling increased as the CaCl2 concentration increased. Chlorides of divalent and trivalent cations, but not those of monovalent cations decreased swelling in vitro. The decrease in swelling among the divalent cations was linearly related to the radius of the cation. The results indicate that Ca decreases cracking susceptibility by decreasing swelling.

Apple fruit periderms (russeting) induced by wounding or by moisture have the same histologies, chemistries and gene expressions.

Russeting is a cosmetic defect of some fruit skins. Russeting (botanically: induction of periderm formation) can result from various environmental factors including wounding and surface moisture. The objective was to compare periderms resulting from wounding with those from exposure to moisture in developing apple fruit. Wounding or moisture exposure both resulted in cuticular microcracking. Cross-sections revealed suberized hypodermal cell walls by 4 d, and the start of periderm formation by 8 d after wounding or moisture treatment. The expression of selected target genes was similar in wound and moisture induced periderms. Transcription factors involved in the regulation of suberin (MYB93) and lignin (MYB42) synthesis, genes involved in the synthesis (CYP86B1) and the transport (ABCG20) of suberin monomers and two uncharacterized transcription factors (NAC038 and NAC058) were all upregulated in induced periderm samples. Genes involved in cutin (GPAT6, SHN3) and wax synthesis (KCS10, WSD1, CER6) and transport of cutin monomers and wax components (ABCG11) were all downregulated. Levels of typical suberin monomers (ω-hydroxy-C20, -C22 and -C24 acids) and total suberin were high in the periderms, but low in the cuticle. Periderms were induced only when wounding occurred during early fruit development (32 and 66 days after full bloom (DAFB)) but not later (93 DAFB). Wound and moisture induced periderms are very similar morphologically, histologically, compositionally and molecularly.

Calcium ions decrease water-soaking in strawberries.

Water soaking is a common disorder of field-grown strawberries (Fragaria × ananassa Duch.). It develops when ripe fruit is exposed to rain. Here we investigate the effects of Ca on water soaking. Fruit was incubated in solutions of various Ca salts and the extent of water soaking quantified using a simple rating scheme. Exposure to CaCl2 (10 mM) decreased water soaking and anthocyanin leakage but had no effect on water uptake. The decrease in water soaking due to CaCl2 was not limited to a single cultivar but occurred in all cultivars examined. Incubating fruit in a chelating agent (EGTA) increased water soaking compared to the water control. Calcium salts of different acids varied in their effects on water soaking. Only CaCl2 reduced water soaking significantly. The chlorides of different cations, also varied in their effects on water soaking. Those of the monovalent cations had no effects on water soaking, while those of the divalent cations (CaCl2, BaCl2 and SrCl2) and of the trivalent cations (FeCl3 and AlCl3) were all effective in decreasing water soaking. Overall, CaCl2 decreased microcracking of the strawberry cuticle as compared to deionized water. Furthermore, CaCl2 also reduced the leakage of anthocyanins from flesh discs, irrespective of the osmotic potential of the incubation solution. Our results indicate that CaCl2 reduced water soaking by decreasing cuticular microcracking, by decreasing leakage of plasma membranes and, possibly, by increasing the crosslinking of cell wall constituents.

Xylogenesis and phloemogenesis in the flesh of sweet cherry fruit are limited to early-stage development.

Water inflows into sweet cherry fruit occur via the xylem and the phloem vasculatures of the pedicel. The rates of these inflows are subject to marked changes during fruit development. The objective was to establish if, and when, xylogenesis and phloemogenesis occur in the fruit flesh (mesocarp) during fruit development. Fruit were cut in half and the median and the lateral bundles inspected by light microscopy. Fruit mass increased with time in a double sigmoid pattern. Xylogenesis and phloemogenesis were both limited to early fruit development (stage I). There were no consistent changes in the areas of either xylem or phloem after stage I until maturity (i.e., during stages II and III). The cross-sectional areas of xylem and of phloem in a bundle were both linearly related to total bundle area. Most of the increases (stage I) in bundle area (62%, r2 = 0.99***) were accounted for by increases in phloem area and about 35% (r2 = 0.97***) by increases in xylem area. A small percentage of the xylem area increase (about 4% of the total area of the bundle; r2 = 0.48***) was contributed by the appearance of intercellular spaces within the xylem. Our results suggest, that new xylem and phloem tissues are differentiated only during early development.

Sweet cherry flesh cells burst in non-random clusters along minor veins.

MAIN CONCLUSION: Sweet cherry flesh cells burst when exposed to water but they do so in clusters indicating heterogeneity with respect to osmotic concentration, which depends on proximity to a minor vein. Water plays a key role in cracking in sweet cherry fruit. Magnetic resonance imaging has previously indicated preferential partitioning of water along veins. A more negative osmotic potential along veins seems the likely explanation. Here we establish if cell bursting in mature sweet cherry fruit is also associated with the veins. Cell bursting was identified by a novel light microscope technique involving exposure of a cut fruit surface to water or to sucrose solutions. Upon exposure to water there was no bursting of skin cells but for cells of the flesh (mesocarp) bursting increased with time. When the cut surface was exposed to sucrose solutions of decreasing osmotic concentrations (increasing water potentials) the incidence of cell bursting increased from hypertonic (no bursting), to isotonic, to hypotonic. Cell bursting in the outer mesocarp occurred primarily in the vicinity of minor veins that in the inner mesocarp was primarily between radial veins. The median distance between a minor vein and a bursting cell (mean diameter 0.129 mm) was about 0.318 mm that between a radial vein and a bursting cell was about 0.497 mm. In contrast, the distance between adjacent minor veins averaged 2.57 mm, that between adjacent radial veins averaged 0.83 mm. Cell bursting tends to occur in clusters. Mapping of cell bursting indicates (1) that a seemingly uniform population of mesocarp cells actually represents a heterogeneous population with regard to their cell osmotic potentials and (2) cell bursting afflicts clusters of neighbouring cells in the vicinities of minor veins.

Direct Evidence for a Radial Gradient in Age of the Apple Fruit Cuticle.

The pattern of cuticle deposition plays an important role in managing strain buildup in fruit cuticles. Cuticular strain is the primary trigger for numerous fruit-surface disorders in many fruit crop species. Recent evidence indicates a strain gradient may exist within the apple fruit cuticle. The outer layers of the cuticle are more strained and thus more susceptible to microcracking than the inner layers. A radial gradient in cuticle age is the most likely explanation. Our study aimed to establish whether (or not) deposition of new cutin in a developing apple fruit occurs on the inner surface of the cuticle, i.e., immediately abutting the outward-facing epidermal cell wall. Developing apples were fed with 13C oleic acid through the skin. Following a 14-d period for incorporation, the fruit was harvested and the cuticular membranes (CMs) isolated enzymatically. The CMs were then ablated to varying extents from the inner or the outer surfaces, using a cold atmospheric pressure plasma (CAPP). Afterwards, the ablated CMs were dewaxed and the 13C contents were determined by mass spectrometry. The incorporation of 13C in the cutin fraction was higher than in the wax fraction. The 13C content was highest in non-ablated, dewaxed CM (DCM) and decreased as ablation depth from the inner surface increased. There was no change in 13C content when ablation was carried out from the outer surface. As fruit development proceeded, more 13C label was found towards the middle of the DCM. These results offered direct evidence for deposition of cutin being on the inner surface of the cuticle, resulting in a radial gradient in cuticular age-the most recent deposition (youngest) being on the inner cuticle surface (abutting the epidermal cell wall) and the earliest deposition (oldest) being on the outer surface (abutting the atmosphere).

Low cuticle deposition rate in 'Apple' mango increases elastic strain, weakens the cuticle and increases russet.

Russeting compromises appearance and downgrades the market value of many fruitcrops, including of the mango cv. 'Apple'. The objective was to identify the mechanistic basis of 'Apple' mango's high susceptibility to russeting. We focused on fruit growth, cuticle deposition, stress/strain relaxation analysis and the mechanical properties of the cuticle. The non-susceptible mango cv. 'Tommy Atkins' served for comparison. Compared with 'Tommy Atkins', fruit of 'Apple' had a lower mass, a smaller surface area and a lower growth rate. There were little differences between the epidermal and hypodermal cells of 'Apple' and 'Tommy Atkins' including cell size, cell orientation and cell number. Lenticel density decreased during development, being lower in 'Apple' than in 'Tommy Atkins'. The mean lenticel area increased during development but was consistently greater in 'Apple' than in 'Tommy Atkins'. The deposition rate of the cuticular membrane was initially rapid but later slowed till it matched the area expansion rate, thereafter mass per unit area was effectively constant. The cuticle of 'Apple' is thinner than that of 'Tommy Atkins'. Cumulative strain increased sigmoidally with fruit growth. Strains released stepwise on excision and isolation (εexc+iso), and on wax extraction (εextr) were higher in 'Apple' than in 'Tommy Atkins'. Membrane stiffness increased during development being consistently lower in 'Apple' than in 'Tommy Atkins'. Membrane fracture force (Fmax) was low and constant in developing 'Apple' but increased in 'Tommy Atkin'. Membrane strain at fracture (εmax) decreased linearly during development but was lower in 'Apple' than in 'Tommy Atkins'. Frequency of membrane failure associated with lenticels increased during development and was consistently higher in 'Apple' than in 'Tommy Atkins'. The lower rate of cuticular deposition, the higher strain releases on excision, isolation and wax extraction and the weaker cuticle account for the high russet susceptibility of 'Apple' mango.

Water Soaking Disorder in Strawberries: Triggers, Factors, and Mechanisms.

Water soaking is an important surface disorder of strawberries that limits unprotected field production. The objective was to identify the mechanism(s) of water soaking. Symptomatic fruit show pale, deliquescent patches of skin. This damage extends into the flesh. Numerous cuticular microcracks occurred in water-soaked areas. Water soaking occurred only if the skin was exposed to liquid water. Water soaking was more rapid when the cuticle had been abraded. Water soaking, anthocyanin leakage, and water uptake all increased with incubation time. There was a lag phase for water soaking and anthocyanin leakage, but not for water uptake. Susceptibility to water soaking increased with fruit ripening and mass. Incubation in isotonic PEG 6000 increased cuticular microcracking but decreased water soaking and water uptake. Incubation in hypotonic fruit juice (natural and artificial) increased water soaking incidence and severity but reduced water uptake. Incubation in dilute citric and malic acids increased plasma membrane permeability as indexed by anthocyanin leakage and increased water soaking. Thus, water soaking involves cuticular microcracking, localized water uptake, bursting of cells, and the release of organic acids into the apoplast. The damage propagates from cell to cell.

Xylem, phloem and transpiration flows in developing European plums.

Neck shrivel is a quality disorder of European plum (Prunus × domestica L.). It has been suggested that backflow in the xylem (from fruit to tree) could contribute to the incidence of neck shrivel in plum. The objective was to quantify rates of xylem, phloem and of transpiration flow in developing plum fruit. Using linear variable displacement transducers, changes in fruit volume were recorded 1) in un-treated control fruit, 2) in fruit that had their pedicels steam-girdled (phloem interrupted, xylem still functional) and 3) in detached fruit, left in the canopy (xylem and phloem interrupted). Xylem flow rates were occasionally negative in the early hours after sunrise, indicating xylem sap backflow from fruit to tree. Later in the day, xylem flows were positive and generally higher in daytime and lower at night. Significant phloem flow occurred in daytime, but ceased after sunset. During stage II (but not during stage III), the rates of xylem flow and transpiration were variable and closely related to atmospheric vapor pressure deficit. The relative contribution of xylem inflow to total sap inflow averaged 79% during stage II, decreasing to 25% during stage III. In contrast, phloem sap inflow averaged 21% of total sap inflow during stage II, increasing to 75% in stage III. Our results indicate that xylem backflow occurs early in the day. However, xylem backflow rates are considered too low to significantly contribute to the incidence of neck shrivel.

Penetration of sweet cherry skin by 45Ca-salts: pathways and factors.

Calcium is beneficial to sweet cherry physiology. The objective was to investigate factors affecting uptake of Ca into mature sweet cherry fruit through their skins. Penetration of 45Ca-salts was monitored using whole fruit or excised fruit skins mounted in diffusion cells. Penetration of 45CaCl2 into intact fruit and through excised skins increased with time. Sealing the pedicel/fruit junction decreased penetration, but sealing the stylar scar had no effect. There was little difference in permeances of the fruit skin to 45CaCl2, 45Ca(NO3)2, 45Ca-formate, 45Ca-acetate, 45Ca-lactate or 45Ca-propionate. Only 45Ca-heptagluconate penetrated at a slower rate. Increasing temperature markedly increased Ca-penetration. Penetration was most rapid at 35 °C, intermediate at 22 °C and slowest at 12 °C. Increasing relative humidity (RH) from 0, 28, 75 to 100% increased penetration of 45CaCl2, but penetration of 45Ca-formate was restricted to 100% RH. Increasing the RH from 50 to 100% at 96 h after droplet application had no effect on penetration of 45CaCl2, but increased penetration of 45Ca-formate. The results reveal that: (1) the fruit/pedicel junction is a site of preferential Ca-uptake and (2) Ca-penetration is limited by the mobility of the Ca ion in the dried-down droplet residue when the point of deliquescence of the applied salt exceeds the ambient RH.

Strawberry fruit skins are far more permeable to osmotic water uptake than to transpirational water loss.

Water movements through the fruit skin play critical roles in many disorders of strawberry (Fragaria × ananassa Duch.) such as water soaking, cracking and shriveling. The objective was to identify the mechanisms of fruit water loss (dry skin, transpiration) and water uptake (wet skin, osmosis). Fruits were held above dried silica gel or incubated in deionized water. Water movements were quantified gravimetrically. Transpiration and osmotic uptake increased linearly with time. Abrading the thin cuticle (0.62 g m-2) increased rates of transpiration 2.6-fold, the rates of osmotic uptake 7.9-fold. The osmotic potential of the expressed juice was nearly the same for green and for white fruit but decreased in red fruit stages. Fruit turgor was low throughout development, except for green fruit. There was no relationship between the rates of water movement and fruit osmotic potential. The skin permeance for transpiration and for osmotic uptake were both high (relative to other fruit species) but were two orders of magnitude greater for osmotic uptake than for transpiration. Incubating fruit in isotonic solutions of osmolytes of different sizes resulted in increases in fruit mass that depended on the osmolyte. The rate of osmotic uptake decreased asymptotically as molecular size of the osmolyte increased. When transpiration and osmotic uptake experiments were conducted sequentially on the same fruit, the rates of transpiration were higher for fruit previously incubated in water. Fluorescence microscopy revealed considerable microcracking in a fruit previously incubated in water. Our findings indicate that the high permeance for osmotic uptake is accounted for by an extremely thin cuticle and by viscous water flow through microcracks and along polar pathways.

Cutin Synthesis in Developing, Field-Grown Apple Fruit Examined by External Feeding of Labelled Precursors.

An intact skin is essential in high-quality apples. Ongoing deposition of cuticular material during fruit development may decrease microcracking. Our objective was to establish a system for quantifying cutin and wax deposition in developing apple fruit. Oleic acid (13C and 14C labelled) and palmitic acid (14C labelled) were fed to developing apples and the amounts incorporated in the cutin and wax fractions were quantified. The incorporation of 14C oleic acid (C18) was significantly higher than that of 14C palmitic acid (C16) and the incorporation in the cutin fraction exceeded that in the wax fraction. The amount of precursor incorporated in the cutin increased asymptotically with time, but the amount in the wax fraction remained about constant. Increasing the concentration of the precursor applied generally increased incorporation. Incorporation in the cutin fraction was high during early development (43 days after full bloom) and decreased towards maturity. Incorporation was higher from a dilute donor solution (infinite dose feeding) than from a donor solution subjected to drying (finite dose feeding) or from perfusion of the precursor by injection. Feeding the skin of a developing apple with oleic acid resulted in significant incorporation in the cutin fraction under both laboratory and field conditions.