Abstract
Differences in storage root initiation among sweet potato (Ipomoea batatas Lam.) plants of the same cultivar were generated by treating them with different amounts of a slow-release fertilizer (19N-3P-10K). Activity of cell-wall-bound invertase (cwb invertase) was monitored during storage root initiation to determine whether differences in its activity could be associated with differences in storage root formation. Cwb invertase activity was increased in roots of plants receiving the lower amount of fertilizer and producing the greater number of storage roots. NAA enhanced cwb invertase activity in excised storage roots.
To compare to two types of Citrus fruit rind [i.e., soft type (satsuma mandarin, Citrus unshiu Marc.) and firm type (Hassaku, C. Hassaku Hort. Tanaka)], rind firmness and contents of cell wall polysaccharides were measured from August to January. In August, firmness was measured by a puncture test and found to be ≈3000g in both species. Firmness of satsuma mandarin decreased drastically with time from August to September and decreased slightly thereafter. In contrast, Hassaku firmness increased slightly from August to September, decreased from September to November, and fluctuated. Hassaku firmness, therefore, was significantly higher than satsuma mandarin firmness after September. We measured sugar content in each fraction after fractionalizing cell wall polysaccharides. In flavedo tissue, sugar content in cellulose fraction was the highest, followed by hot-water and EDTA fraction; hemicellulose fraction was the lowest. Although both species were almost the same in sugar content in cellulose and EDTA fraction in August, satsuma mandarin was significantly higher than Hassaku in January. These data showed that changing of rind firmness in citrus was related to the sugar content of cellulose and EDTA fraction in flavedo tissue. In albedo tissue, sugar content in the cellulose fraction was the highest, followed by hemicellulose and hot-water fraction, and EDTA fraction was the lowest. However the extent of seasonal fluctuation in albedo tissue was smaller than that of flavedo tissue, not having any relation to the changing of the firmness.
The softening of fruit dramatically reduces its market value, especially when this occurs on the tree before ripening. The causes of fruit softening, before ripening, were examined through anatomical and phytochemical comparative analyses between normal fruit, fruit softened on the tree, and stored fruit. The typical morphological changes that occurred with the fruit included early senescence and decreased firmness. The decrease in firmness of softening fruit was due to smaller cell sizes but larger intercellular spaces. The water and free sugar content of the fruit flesh, as well as the weight and sugar content of the cell walls, were significantly lower in softening fruit. Conversely, uronic acid levels and β-galactosidase activity were slightly higher in the softening compared to normal fruit, but the latter was lower than in stored fruit. The results indicated that reduced cell size and cell wall mass were major changes occurring during fruit softening on the tree before ripening, suggesting a difference in the softening mechanisms in ripening and stored fruit.
Changes in rind firmness and cell wall polysaccharide composition were measured in fruit with a) a soft rind, (`Satsuma' mandarin, Citrus unshiu Marc., cv. Aoshima), and b) a firm rind (hassaku, C. hassaku Hort. ex Tanaka), from August to January of the following year. Rind firmness was similar in both species in August, but hassaku had significantly firmer rind than did mandarin from September to January. Both flavedo and albedo tissues were extracted, and the extracts were hydrolyzed and fractionated to yield four fractions: (hot water, EDTA, hemicellulose, and cellulose). In flavedo tissue, sugar concentration was highest in the cellulose fraction, and lowest in the hemicellulose fraction. The concentration in all fractions decreased as the fruit developed and matured. Although the sugar concentration in the cellulose and EDTA fractions of both species was similar in August, it was significantly higher in both fractions in hassaku than in mandarin in January. The sugar concentration of each fraction from albedo tissue was in the order: cellulose > hemicellulose > hot water > EDTA. The range of variation in cell wall sugars in albedo tissue was smaller than that in flavedo tissue. Chemical name used: ethylenediaminetetraacetic acid (EDTA).
Harvested nectarine fruit [Prunus persica (L.) Batsch `Flavortop'] were held for 5 days at 20 °C, or stored at 0 °C either immediately (control), or after 2 days at 20 °C (delayed-cooling). Observations were conducted after removal from storage for 1, 3, or 5 weeks and a shelf life of 5 additional days at 20 °C. After 5 weeks storage, 87% of control fruit developed woolliness (mealiness in texture accompanied by dry tasting fruit as a result of reduced juice content), while only 7% of delayed-cooling fruit showed signs of woolliness. Firmness of fruit in the delayed-cooling treatment was less at the beginning of ripening than control fruit, but after shelf life in both treatments, fruit reached the same final softness. Expressible juice was lower in woolly fruit (46%) than in healthy fruit (65%). Along with woolliness, viscosity of the resuspended alcohol insoluble residue (cell wall material) of expressed juice increased, implying accumulation of large molecular-weight polymers. The high performance liquid chromatography profile confirmed there were more large pectin polymers (2000 to 76 Ku) in the cell wall components of juice from woolly fruit and a lower arabinose content in these polymers reflected greater side chain removal from pectins in the juice of woolly fruit. Accumulation of larger sized pectin polymers along with high viscosity correlated with lower polygalacturonase activity in woolly fruit. Degradation of soluble pectin released into the juice of woolly fruit may have been impeded by repressed polygalacturonase activity.
Activity of ß-galactosidase II (EC 3.2.1.23), which can hydrolyze ß-galactan from tomato cell walls, increased markedly during ripening of `Roma' and `Rutgers' tomatoes (Lycopersicon esculentum Mill.). Activity of two other ß-galactosidase isozymes, incapable of galactan hydrolysis, was present in green fruit and remained unchanged throughout ripening. ß-Galactosidase II activity was not detectable in green fruit of either cultivar, appearing first at the breaker stage of `Roma' fruit and not until the pink stage of `Rutgers' fruit. Consistent with this, galactose loss from Na2CO3-soluble pectin (NSP) was detectable at an earlier stage in `Roma' vs. `Rutgers' fruit. A greater decline in NSP galactose was evident in `Roma' fruit compared to `Rutgers' fruit, in keeping with the higher levels and longer period of ß-galactosidase II expression in the former. Significant galactose loss from trans -1,2-diaminocyclohexane-N,N,N',N' -tetraacetic acid-soluble pectin, in contrast, was not seen until the last stage of ripening. These results indicate that the long-reported, net galactosyl loss from the cell walls of ripening tomatoes correlates with ß-galactosidase II activity. Nonetheless, the observation that softening commenced before ß-galactosidase II activity or galactose loss was detectable suggests some other basis for the earliest stages of ripening-related fruit softening in tomato.
Tomato fruit (Solanum lycopersicon L.) can develop mealiness and enhanced softening when exposed to chilling temperatures during storage, but the involvement of cell wall-associated enzymes in chilling injury development is not well understood. To study this aspect of injury development, we have exposed breaker stage tomato cv. Trust fruit to a chilling temperature of 3 °C for 0, 7, 14, and 21 days followed by storage at 20 °C for 12 days. Ethylene production was not affected by storage except after 21 days, where production was greater at 20 °C. Exposure of fruit to chilling temperatures delayed the ripening-related color change (chroma and hue) and initially increased compression values, but percentage of extractable juice was not affected consistently. Increased polygalacturonase activity during ripening was reduced by about 50% after 7 days at 3 °C, and further inhibited with increasing storage periods. In contrast, the activities of pectin methylesterase and α-galactosidase were not significantly affected by the cold treatments. β-Galactosidase activity was greater in all chilled fruit compared with fruit ripened at harvest, whereas endo-β-1,4-glucanase activity was lower after 21 days at 3 °C. These results will be compared with equivalent changes in the activities of cell wall enzymes that are associated with wooliness development in chilling-injured peach fruit.
After 6 months of growth in 200,400, and 500 mm NaCl, cultured cells of Distichlis spicata showed a decreased cell volume (size) despite maintenance of turgor pressure sometimes 2-fold higher than that of the control. Tensile strength, as measured by a nitrogen gas decompression technique, showed empirically that the walls of NaCl-stressed cells were weaker than those of nonstressed cells. Breaking pressures of the walls of control cells were ≈68 ± 4 bars, while that of the walls of cells grown in 500 mm NaCl (-25 bars) were 14 ± 2 bars. The relative amount of cellulose per cell remained about constant despite salt stress. However, glucuronoarabinoxylans were more readily extractable, presumably because of a decrease in cross-linkage with phenol substances. Therefore, we suggest that cellulose microfibrils are not the only determinants that confer tensile strength to the primary cell wall, but rather subtle changes in the matrix polysaccharides are likely responsible for this event.
The `Stony Hard' gene of peach conferred a unique ability to manipulate softening and textural properties of the fruit by controlling the concentration and duration of exposure to ethylene. Fruit ripened in ethylene-free air softened very slowly. Exposure of fruit to 1 ppm ethylene continuously for 48 h, or discontinuously at 100 ppm over the same time period, significantly accelerated softening—to a normal texture. Exposure of fruit to 100 ppm ethylene continuously for 48 h induced softening to the same level, but to a mealy texture. We have prepared cell walls and conducted sequential chemical extractions from fruit exposed to the ethylene treatments above. Galacturonic acid content of chelator soluble pectin fractions decreased for mealy fruit, compared to fruit with normal texture, indicating that selective pectin degradation was associated with mealiness. Other differences in polysaccharide sugar composition and apparent molecular size associated with slow, accelerated, and abnormal softening in peach fruit will be addressed.
`McIlhenny Select' (easy detachment) and `Hard Pick' are two lines of tabasco pepper (Capsicum frutescens L.) that differ in the fruit detachment characteristics. Cellulase (Cx) and polygalacturonase (PG) activity, extracted from the fruit abscission zone, correlated inversely with the force needed to separate the fruit from the pedicel. A trend of higher Cx and PG is associated with the lower detachment force in the McIlhenny Select line. Differences in the fruit cell wall protein profile between both lines occurred during ripening. Two bands of 23 kDa and 40 kDa were higher in `McIlhenny Select'. A band of approximately 30 kDa was higher in `Hard Pick', while a band of ≈70 kDa increased in both lines. Isolation and characterization of these bands as well as Cx and PG is needed to understand the factors affecting fruit detachment in tabasco pepper.