Search Results
`Gemini II' cucumber (Cucumis sativus L.) fruits were stored for 2, 4, or 6 days at 5 and 6C in 1989 and for 5 days at SC or 10 days at 3C in 1990. Chilling injury (CI) symptoms were rated after 2 to 4 days at 25C. Cell wall polysaccharide concentrations in the peels and in injured and noninjured portions of the peels were determined only in 1990. High CO2 and low O2 delayed the onset of CI symptoms, but did not prevent symptom development. Chilling injury symptoms increased with longer exposure to chilling temperatures. Solubilization of cell wall polysaccharides was associated with development of CI symptoms. Variations in low methoxyl pectinates accounted for 70% of the variation in CI.
Abstract
Cell wall-degrading enzymes were extracted from the cell wall free space of mesocarp tissue from immature almonds [Prunus dulcis(Mill.)D.A. Webb, ‘Nonpareil’]. The activities of several of these enzymes were found to correlate with the development of gum ducts in this tissue. Polygalacturonase (EC 3.2.1.15) and 1,3-β-D-glucanase (EC 3.2.1.39) activities rose sharply at, or just prior to, the early schizogenous stage of duct initiation, while increases in α-galactosidase (EC 3.2.1.22), β-galactosidase (EC 3.2.1.23), α-arabinosidase (EC 3.2.1.55), and α-mannosidase (EC 3.2.1.24) activities were correlated with the later lysigenous stage of duct formation. Cell wall analysis of almond mesocarp tissue sampled the week preceding gum duct formation determined that the predominant noncellulosic sugars present in the mesocarp cell walls are arabinose, galactose, xylose, and glucose, with smaller amounts of rhamnose and mannose also present. The walls also contain a high percentage of galacturonic acid and trace amounts of glucuronic acid. Methylation analysis of the cell walls confirmed that many of the specific glycosidic linkages that are cleaved by the enzymes tested are present in the mesocarp cell walls immediately prior to gum duct formation.
Fruits of mid- (`O'Henry'), late (`Airtime'), and extra-late-season (`Autumn Gem') peach [Prunus persica (L.) Batsch] cultivars were examined for changes in cell wall structure and cytochemistry that accompany the onset of mealiness and leatheriness of the mesocarp due to chilling injury. The peaches were stored at 10C for up to 18 days or at SC for up to 29 days. Plastic-embedded sections were stained by the Schiff's-periodic acid reaction, Calcofluor white MR2, and Coriphosphine to demonstrate total insoluble carbohydrates, ß-1,4 glucans, and pectins, respectively. Mealiness was characterized by separation of mesocarp parenchyma cells leading to increased intercellular spaces and accumulation of pectic substances in the intercellular matrix. Little structural change was apparent in the cellulosic component of the cell walls of these fruits. In leathery peaches, the mesocarp parenchyma cells collapsed, intercellular space continued to increase, and pectin-positive staining in the intercellular matrix increased greatly. In addition, the component of the cell walls that stained positively for ß-1,4 glucans became thickened relative to freshly harvested or mealy fruit. At the ultrastructural level, dissolution of the middle lamella, cell separation, irregular thickening of the primary wall, and plasmolysis of the mesocarp parenchyma cells were seen as internal breakdown progressed.
Abstract
Eight-one percent of the harvest firmness in kiwifruit (Actinidia chinensis Planch.) was lost during the first 8 weeks of storage in air at 0°C. As softening proceeded, a solubilization of uronic acids and the neutral sugar residues usually associated with pectic polymers (galactose, arabinose, and rhamnose) was detected. No consistent changes were noted in cellulose or the neutral sugars usually associated with hemicelluloses. Starch degradation also occurred coincident with softening. The amount of cell wall components soluble in water following fruit homogenization and the proportion of ethanol-precipitable pectic neutral sugars in this fraction increased during the first 8 weeks of storage. Once the rate of softening slowed (8 to 20 weeks), an equilibrium situation apparently was established between the amounts of the sugars formed in the ethanol-precipitable (i.e., polymeric) and ethanol-soluble fractions, suggesting that digestion of wall components continues after their excision from the insoluble wall matrix. Controlled atmosphere (2% O2+ 5% CO2; CA) storage retarded flesh softening relative to that measured in fruit held at 0° in air. A comparison of the changes in the cell wall components of air-stored and CA-stored kiwifruit suggests that, in addition to cell wall degrading processes contributing to fruit softening, starch degradation (possibly causing cell turgor changes) also may be involved in low-temperature softening of kiwifruit. The losses in water-insoluble cell wall pectic neutral sugars and uronic acids in air and CA storage were similar during the first 8 weeks of storage. Once softening slowed in CA, small but consistent reductions in the amount of cell wall turnover were observed as compared to air storage.
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.
Cell wall composition and structure were examined in visually normal (N), granulated (G), and collapsed (VC) juice vesicles of `Marsh Seedless' grapefruit (Citrus paradisi) Macf.). According to gel-filtration data, VC appeared to be associated with a modification of water-soluble (WSP) and chelate-soluble (CSP) pectin molecular weight (Mr); small-Mr pectins increased, whereas large-J4. pectins decreased. The difference in M = of pectins did not appear to be mediated by polygalacturonases. Molecular weight of hemicelluloses did not differ. Granulated vesicles contained about two times more structural polysaccharides (pectins, hemicelhdose, and cellulose) than N vesicles, although hemicellulose and pectin M = modification were absent. Ion-exchange profiles of WSP, CSP, and hemicelhrlose fractions of VC and G vesicles were not different from those of N vesicles. Individual cells in vesicles with G and these vesicles themselves were much larger than those of N vesicles, whereas cells in VC were partially or completely collapsed.
Pawpaw fruit were harvested at the advent of the ripening process and were ripened at room temperature. Based on fruit firmness and respiration and ethylene production rates at harvest and during ripening, fruit were classified into one of four categories: preripening (no to very slight loss of firmness; preclimacteric), early ripening (some softening; increasing rates of ethylene and CO2 production), mid-ripening (soft; at or just past climacteric), and late ripening (very soft; postclimacteric). The activities of the cell-wall degrading enzymes polygalacturonase (PG), endo-(1→4)ß-D-glucanase (EGase), and endo-ß-1,4-mannanase (MAN) were low in the preripening and early ripening stages, increased dramatically by mid-ripening coincident with the respiratory and ethylene climacterics, and decreased at late ripening. However, pectin methylesterase (PME) activity per milligram protein was highest at the green stage when the fruit firmness was high and decreased as ripening progressed. Tissue prints indicated both EGase and MAN increased as ripening proceeded. The EGase activity was evident near the seeds and the surface of the fruit at preripening and eventually spread throughout, while MAN activity was evident near the fruit surface at preripening and was progressively expressed throughout the flesh as fruit ripened. The greatest decline in fruit firmness occurred between pre- and early ripening, before the peak activities of PG, EGase, and MAN, although MAN exhibited the greatest relative increase of the three enzymes in this period. The data suggest that PME may act first to demethylate polygalacturonate and may be followed by the action of the other enzymes resulting in cell wall disassembly and fruit softening in pawpaw.
A preliminary understanding of developmental processes among divergent species is essential to evaluate the applicability of information from model species to plants of agricultural importance. In tomato (Lycopersicon esculentum Mill.), where the molecular biology associated with fruit ripening has been studied most extensively, tissue softening is due at least in part to the activity of proteins called expansins, in concert with enzymatic activities that modify the pectin and xyloglucan components of the cell wall. We evaluated the potential for the concerted action of expansins and other cell wall-modifying enzymes during ripening in a highly divergent fruit species, sour cherry (Prunus cerasus L.). We identified a family of four expansin genes that was strongly upregulated at the advent of ripening. Activation of these genes was accompanied by strong upregulation of gene(s) encoding potential pectin methylesterases, pectate lyase(s), and xyloglucan endotransglycosylase(s). Initiation of ripening and gene induction were also associated with a rapid decrease in cell wall weight. These results suggest that expansin and several other distinct activities could be involved in ripening-associated cell wall modification in cherries.
Abstract
The capacity of ‘Eldorado’ pears to ripen increased dramatically after 4 weeks of exposure to 0°C and was associated with the synthesis of ethylene by pear tissue. Endogenous levels of ACC and internal ethylene were low after harvest, but increased rapidly after 4 weeks at 0°. Exposure to 0° for 4 weeks also resulted in an increase in soluble polyuronide during subsequent ripening at 20°. In contrast, after 9 months at 0°, soluble polyuronide content showed little increase when pears were transferred to 20°, and fruit failed to soften normally even though ACC content, internal ethylene concentration, ethylene evolution, and respiration remained relatively high. The content of arabinose, galactose, and rhamnose residues in cell walls decreased substantially during the ripening period after 4 weeks or longer at 0°. These cell wall neutral sugars decreased during ripening, even after 9 months of storage at 0°, while firmness and soluble polyuronide showed little change after fruit were transferred to 20°. These data indicate that the failure of pears to soften normally at 20° after prolonged storage at 0° is not related to ethylene synthesis or to changes in cell wall noncellulosic neutral sugar content, but is probably associated with mechanisms of polyuronide solubilization. Chemical name used: 1-aminocyclopropane-1-carboxylic acid (ACC).