Sorbitol is the major carbohydrate translocated into apple fruit where it is normally metabolized to fructose. In watercored apple fruit tissues, however, the intercellular spaces become flooded and sorbitol content is consistently higher than in nonwatercored apples, suggesting a defect in sugar alcohol metabolism or transport. Our previous results have identified and characterized two sorbitol transporters, MsSOT1 and MsSOT2, in apple fruit tissues. Sorbitol transporter gene expression has been implicated in development of watercore with MsSOT expression diminished or absent in certain watercored fruit tissues. To explore this further, we have investigated the relationships between watercore, fruit maturation, fruit composition, and MsSOT expression in a number of apple cultivars that differ in watercore susceptibility. We also compared transporter expression between affected (watercored) and healthy parts of the same fruit and between watercored and nonwatercored fruits throughout the maturation and ripening processes. The MsSOT expression was often dramatically reduced in fruit tissues exhibiting watercore. Thus, in susceptible cultivars, maturing (ripening) fruit parenchyma cells lose the ability to transport sorbitol, and this in turn leads to sorbitol accumulation in the apoplastic free space and subsequent flooding of these spaces. These results are consistent with a relationship between watercore and sorbitol transport and also with a genetic susceptibility to the disorder.
Wayne Loescher, Tad Johnson, Randolph Beaudry, and Sastry Jayanty
Shiow Y. Wang and Miklos Faust
The glycolipids, phospholipids, and sterols were determined in normal and watercore-affected apple (Malus domestica Borkh. cv. Delicious). Fruit with watercore contained higher amounts of glycolipids, phospholipids, and sterols. The ratios of unsaturated to saturated fatty acids and (18:3) to (18:1 + 18:2) were lower in watercore-affected tissue than in normal tissue. The ratio of free sterols to phospholipids was higher, whereas the ratio of phosphatidylcholine to phosphatidylethanolamine was lower in watercore-affected apple. Membrane lipids were altered in watercore-affected fruit.
Shiow Y. Wang and Miklos Faust
Ethylene biosynthesis and polyamine content were determined in normal and watercore-affected apple (Malus domestics Borkh. cv. Delicious). Fruit with watercore produced more ethylene and contained higher amounts of putrescine (PUT), spermidine (SPD), 1-aminocyclopropane-1-carboxylic acid (ACC), and 1-(malonylamino) cyclo-propane-1-carboxylic acid (MACC). The activities of ACC synthase and ethylene-forming enzyme (EFE) in watercore-affected fruit were also higher than in normal fruit. The EFE activity in severely affected flesh was inhibited, resulting in ACC accumulation and low ethylene production. S-adenosylmethionine (AdoMet) was maintained at a steady-state level even when C2 H4 and polyamides were actively synthesized in normal and affected fruit.
Hisashi Yamada, Hirokazu Ohmura, Chizuru Arai, and Makoto Terui
The influence of controlling temperature during apple (Malus domestica Borkh.) maturation on sugars, fruit maturity, and watercore occurrence was investigated in watercore-susceptible `Himekami' and `Fuji' apples. The incidence of watercore at 13/5 and 23/15C was greater than at 33/25C or ambient temperatures in `Himekami' apples in 1991 and was greater at 18/10C than at other temperatures in `Fuji' apples in 1990. In 1992, the extent of watercore increased as fruit temperature decreased from 28 to 14C and 23 to 9C in `Himekami' and `Fuji' apples, respectively. Watercore occurrence affected by fruit temperature was not related to fruit maturity, as judged by ethylene evolution. The effect of fruit temperature on sorbitol was relatively small compared with that on other sugars, and no relationship was found between watercore development and sugars. These results suggest that fruit temperature affects watercore expression independently of fruit maturity or sorbitol metabolism in the fruit in watercore-susceptible apple cultivars.
F. Roger Harker, Christopher B. Watkins, Paul L. Brookfield, Mellisa J. Miller, Suzanne Reid, Phillippa J. Jackson, Roderick L. Bieleski, and Tim Bartley
Preharvest development and postharvest disappearance of watercore in `Fuji' apples (Malus ×domestica Borkh.) from a northern (Hawke's Bay, latitude 39° south) and southern (Otago, latitude 45° south) region of New Zealand were compared. A new method for quantifying watercore was developed. A photocopy was taken of the symptoms after each fruit was cut in half through the equator, and then the area of affected flesh (photocopies black) was measured using morphometric methods and compared to the area of unaffected flesh (photocopies white). Watercore was more severe and developed earlier in the season in Otago than in Hawke's Bay. In Otago, a block-type watercore predominated, disorder symptoms initially appearing in the tissues located at the junction of two carpels, while in Hawke's Bay a radial-type of watercore predominated, initially appearing in the tissues surrounding the coreline vascular bundles. Regression analysis identified that orchard and harvest date accounted for most of the differences in watercore symptoms and that the initial appearance of low levels of watercore was the best predictor that fruit would start to develop commercially significant levels of watercore. Incorporation of background color, internal ethylene concentration, starch pattern index, and firmness only slightly improved the regression coefficient. Watercore disappeared from the flesh during storage of fruit from both regions. Fruit from early harvests had the least severe symptoms, and the highest rates of watercore disappearance during storage. In fruit with more severe symptoms at harvest, its disappearance during storage was associated with an increase in fruit volume and air space, which occurred despite continuing mass loss. We suggest that during storage, the extracellular fluid associated with watercore symptoms is absorbed into the cells, and thus drives the increase in fruit volume.
Zhifang Gao, Sastry Jayanty, Randolph Beaudry, and Wayne Loescher
In apple (Malus ×domestica Borkh.), where sorbitol is a primary photosynthetic product that is translocated throughout the plant, accumulation of sorbitol in sink cells appears to require an active carrier-mediated membrane transport step. Recent progress in isolation and characterization of genes for sorbitol transporters in sour cherry (Prunus cerasus L.) and mannitol transporters in celery (Apium graveolens L.) suggested that similar transporters may be present in apple tissues. A defect in these transporters could also explain the occurrence of the fruit disorder watercore, characterized by the accumulation of fluids and sorbitol in the apoplasmic free space. Our objectives therefore included isolation and characterization of genes for sorbitol transporters in apple tissues and comparisons of expression of transporter genes, especially in various sink tissues including watercored and non-watercored fruit tissues. We have isolated and characterized two sorbitol transporter genes, MdSOT1 and MdSOT2. Sequence analyses indicated that these are members of the major facilitator transporter superfamily that gives rise to highly hydrophobic integral membrane proteins. Heterologous expression and measurement of sorbitol uptake in yeast indicated that these are specific and with high affinities for sorbitol, with Kms for sorbitol of 1.0 and 7.8 mm for MdSOT1 and MdSOT2, respectively. Sorbitol transporter expression was evident in all sink tissues tested with the exception of watercore-affected fruit tissues. Sorbitol accumulation in apple sink tissues thus involves an apoplasmic active membrane transport step and watercore results from a defect in that process.
Incidence of scald in nontreated and DPA (2000 mg·liter-1)-treated `Delicious' apples (Malus domestics Borkh.) was assessed after 8.5 months in 1.5% or 0.7% O 2 plus 1.5% CO2 at 0.2C, with and without C2H4 scrubbing. Incidence of scald was high in non-DPA fruit held in 1.5% O2, and DPA treatment reduced scald in fruit held in 1.5% or 0.7% O2. Scald control was better with 0.7% O2 and no DPA `treatment than with 1.5% O 2 and a DPA dip. Ethylene scrubbing had no effect on scald in fruit held in 0.7% or 1.5% 02. Susceptibility of fruit to scald-and flesh browning exhibited seasonal variation, which was related to the differences in fruit maturity and the amount of watercore at harvest, respectively. Chemical name used: diphenylamine (DPA).
Kevin M. Keener, Richard L. Stroshine, and John A. Nyenhuis
A 5.40-MHz NMR system was used for measuring the self-diffusion coefficient of water (Dw) and the spin-spin relaxation constant (T2) in apple (Malus ×domestica Borkh.) tissue. The pulsed field gradient spin echo (PFGSE) technique was used to measure Dw, and the Carr-Purcell-Meiboom-Gill (CPMG) technique was used to measure T2. T2 and Dw values were compared for apples with differing amounts of soluble solids concentration (SSC) and with and without internal defects, such as bruising, watercore, and internal browning. `Granny Smith', `Golden Delicious', and `Delicious' apples were tested. In `Golden Delicious', Dw highly correlated with apple tissue SSC (P < 0.002, r 2 = 0.68). This indicates that Dw could potentially be used for sorting `Golden Delicious” apples based on SSC, but the coefficient of determination needs to be improved before it would be commercially viable. There were no measurable differences in Dw among healthy apple tissue and tissue affected by either watercore or internal browning. T2 values showed no relationship between healthy apple tissue and bruised tissue in `Golden Delicious' and `Granny Smith'. However, in `Delicious' tissue, T2 values were statistically different between healthy and bruised tissue (P < 0.02). Further comparisons in `Delicious' between watercore and healthy apple tissue showed no differences. But, there were statistical differences found between T2 in healthy apple tissue and tissue with internal browning (P < 0.01). These results indicate that T2 could potentially be used for separating `Delicious' apples with internal browning or with bruising from healthy apples. Titratable acids and pH were correlated for `Golden Delicious' (P < 0.08). This correlation is significant because one may be able to noninvasively measure pH in `Golden Delicious' apples using NMR, which could then be correlated to titratable acids.
In a 3-year study, the effectiveness of 0.7% and 1.5% O2 to attenuate scald was evaluated on three strains of `Delicious' apples (Malus ×domestica Borkh.)—`Harrold Red', `Starking', and `Starkrimson'—harvested five times at weekly intervals from a wide range of orchards. Scald susceptibility of fruit held in air, 1.5% O2 + 1.0% CO2, and 0.7% O2 + 1.0% CO2 at 0 °C decreased sharply as the fruit matured on the tree (starch index 1.0 to 2.5 on a 0 to 9 scale). Later harvests (starch index >2.8) further reduced scald but the fruit had more watercore-induced breakdown and were 3 N softer than fruit picked at a less advanced maturity. Early picked `Starkrimson' (starch index <2.0) scalded more than `Starking' and `Harrold Red' in air, 1.5% O2, and 0.7% O2 storage at 0 °C, and 0.7% O2 was less effective than 1.5% O2 in scald control. While 0.7% O2 storage effectively reduced scald (less than 10%) for 8 months in `Starking' and `Harrold Red' picked over a wide range of maturity (starch index 0.7 to 4.3), it did not adequately control scald (up to 45%) in early picked `Starkrimson' (starch index <2.0). Storage in 0.7% also reduced watercore-induced breakdown in `Starkrimson' (starch index >3.0) and did not result in skin purpling or alcoholic taste in `Harrold Red', `Starking', and `Starkrimson'.
A.H.D. Francesconi, C.B. Watkins, A.N. Lakso, J.P. Nyrop, J. Barnard, and S.S. Denning
Fruit maturity, quality, calcium concentration and economic value of `Starkrimson Delicious' (Malus domestica Borkh.) apples, under a range of crop levels and European red mite [Panonychus ulmi (Koch)] cumulative mite-days (CMD), were best explained by local surface regression models involving CMD and crop load. Fruit from trees with low CMD and a light crop (125 fruit/tree, about 20 t/ha) were the most mature at harvest. Those fruit had higher ethylene concentrations, starch pattern indices, soluble solids concentrations, and watercore incidence at harvest than fruit from trees with low CMD and a normal crop (300 fruit/tree, about 40 t/ha), or with high CMD at any crop level. Those fruit also had higher incidences of watercore and internal breakdown after 4 months of cold storage. Calcium concentrations in fruit increased as crop load and CMD increased. Whole-canopy net CO2 exchange rate per fruit related better to fruit quality and calcium concentrations than either crop load or CMD alone, but was always a much worse predictor than local surface regressions. Low CMD and normally cropped trees had the highest crop value; lightly cropped trees had an intermediate crop value; while high CMD and normally cropped trees had the lowest crop economic value. Crop load should be considered when defining action thresholds for mites, and harvest schedules for apples should reflect crop load and mite populations on apple trees.