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Hanling Yu and Claude Willemot

We examined the relationship between reduced galactolipid content in tomato fruit at 4C and chilling injury. Galactolipid biosynthesis from 14C-acetate was compared in pericarp discs of cold-tolerant `New York 280' (`NY') and -sensitive `Early Cherry' (`EC') at 4C and 20C. Labeled lipids were separated by 2D-TLC. Labeled monogalactosyldiglyceride (MGDG) molecular species were hydrolyzed using a position-specific lipase; the fatty acids released were hydrogenated and separated according to chain length by reverse-phase TLC. At 4C, the relative amount of radioactivity was reduced in MGDG and enhanced in phosphatidylcholine (PC) in both cultivars, in comparison with labeling at 20C. In discs from fruit chilled for 6 h, labeling was similar in `NY' and `EC'. In fruit held at 4C for 8 days, labeling of MGDG was reduced and that in PC was enhanced to a greater extent in chilling-sensitive `EC' than in `NY'. The proportion of the MGDG label in eukaryotic species (i.e., the ratio in C18/C16 fatty acids in position sn-2), was less in `EC' at 4C than at 20C, even for fruit held at 4C for only 6 h. The ratio was little affected in `NY'. The data indicate that biosynthesis of eukaryotic MGDG was inhibited in tomato fruit at chilling injury-inducing temperatures.

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Bruce D. Whitaker

Abbreviations: ASG, acylated steryl glycoside; DGDG, digalactosyldiacylglycerol; FS, free sterols; GL, galactolipids; GlyL, glycolipid; MGDG, monoga-lactosyldiacylglycerol; NL, neutral lipid; PA, phosphatidic acid; PC, phosphatidylcholine; PE

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Shiow Y. Wang and Miklos Faust

Composition changes in galactolipids, phospholipids, and sterols in apple shoots (Malus domestica Borkh. cv. Red Delicious) from August to April were determined. The predominant fatty acids in the membrane lipids of apple shoots were palmitic acid (C16:0), linoleic acid (C18:2), and linolenic acid (C18:3). The major galactolipid components in apple shoots were monogalactosyl diglyceride (MGDG) and digalactosyl diglyceride (DGDG). The amount of MGDG and DGDG increased from autumn to spring. Galactolipids contained highly unsaturated fatty adds, mainly linoleic (18:2) and linolenic (18:3) acid. The major individual phospholipids were phosphatidylcholine (PC) and phosphatidylethaeolamine (PE). β -Sitosterol and sitosteryl ester were the predominant sterols. The phloem contained higher amounts of galactolipids, phospholipids, and sterols than did the xylem tissue. There was a significant increase in the content of galactolipids and phospholipids and onsaturation of their fatty acids during cold acclimation. A decrease in the ratio of free sterols to phospholipids also occurred in apple shoots toward cold winter months. Composition changes in galactolipids, phospholipids, and sterols that were associated with growth cessation, defoliation and cold acclimation from fall to winter, were mostly reversed following deacclimation in spring.

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G.A. Picchioni, A.E. Watada, W.S. Conway, and B.D. Whitaker

Postharvest Ca infiltration delays senescence and improves storage quality of apple fruit, but the consequences on membrane lipid composition have received little evaluation. We studied changes in galactolipids (mono- and digalactosyl-diacylglycerol; MGDG and DGDG) and sterol conjugates (sterol glycosides and acylated sterol glycosides; SG and ASG) in `Golden Delicious' cortical tissue. Fruit were pressure-infiltrated with CaCl, at harvest (0, 2, or 4% w/v), stored for 6 months at 0C, and evaluated during subsequent exposure to 20C. MGDG, SG and ASG concentrations were greater in Ca-infiltrated fruit (CIF) than in control fruit. A 35-37% increase in ASG occurred during the first 7 days at 20C in CIF, when ASG decreased by 19% in control fruit. Ca infiltration may delay degradation of plastid membranes and increase sterol conjugation during apple fruit ripening.

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Shiow Y. Wang and Miklos Faust

The changes of membrane lipids in apple (Malus domestics Borkh. cv. Delicious) auxillary and terminal buds from August to April were determined. The predominant lipids were monogalactosyl diglyceride (MGDG), digalactosyl diglyceride (DGDG), phosphatidylcholine (PC), and phosphatidylethanolamine (PE). An increase in membrane polar lipids was associated with budbreak and bud growth from August to April. Linolenic acid was the predominant fatty acid in MGDG, DGDG, and PC, while linoleic acid was predominant in PE. Phosphatidylglycerol (PG) and phosphatidylinositol (PI) contained a high amount of palmitic acid. The ratio of (18:2 + 18:3) to 18:1 fatty acids in galactolipids in apple buds increased from August to April. ß-Sitosterol and sitosteryl ester were the predominant sterols in apple buds. An increase in sitosterol, a decrease in sitosteryl ester, and a decline in the ratio of free sterols to phospholipids occurred during budbreak in spring. A decrease in sitosterol was associated with bud expansion in spring.

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Bruce D. Whitaker

MG tomato fruit were stored for four or 12 days at chilling (2C) or nonchilling (15C) temperature. Fruits stored 12 days at 15C ripened to the turning stage, whereas fruits at 2C did not ripen. Lipids of microsomes and plastids from pericarp tissue were analyzed at harvest and after four or 12 days of storage. After 12 days at either 15C or 2C, the ratio of phospholipid (PL) to protein in microsomes declined, with a concomitant increase in the ratio of total membrane sterols (TMS) to PL. The TMS/PL ratio also increased in crude plastids. In both microsomes and plastids, free sterols (FS) increased more at 2C than at 15C, and thus accounted for a larger percentage of the TMS. The ratio of stigmasterol to sitosterol in steryl lipids, particularly in FS, increased more at 15C than at 2C. The unsaturation index of fatty acids in PL and galactolipids generally increased slightly during storage at both 15C and 2C. The ratio of phosphatidylethanolamine to P-choline increased in both membrane fractions at both temperatures. In plastids, the ratio of mono- to digalactosyldiacylglycerol declined substantially at 2C but not at 15C.

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Bruce D. Whitaker

Plastids and microsomal membranes were isolated from pericarp tissue of mature green and red-ripe tell pepper fruit harvested from greenhouse and field grown plants. The lipid composition of these membrane fractions changed far more with ripening of field grown than greenhouse grown fruit. Also, the phospholipid (PL), free sterol (FS), steryl glycoside (SG) and acylated steryl glycoside (ASG) content of microsomes and plastids from both green and red fruit were very different under the two growing conditions. Total steryl lipids (TSL = FS + SG + ASG), and the TSL/PL ratio, increased in microsomes and decreased in plastids with ripening. These changes were much greater in field grown fruit. The ASG/SG ratio decreased with ripening in both membrane fractions, under both growing conditions. Ripening and growth conditions affected the phospholipid and sterol composition in plastids much more than in microsomes. Lipid changes associated with the chloroplast – chromoplast transformation were similar in field and greenhouse grown fruit, including an increase in the galactolipid/PL ratio. Future studies will assess how differences in membrane lipid composition affect postharvest storage life of the fruit.

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Bruce D. Whitaker

Altered metabolism of membrane lipids has been proposed as a mechanism for the beneficial effects of postharvest calcium treatment on apple quality. A previous study showed that after transfer of apples stored 6 months at 0C to 20C, calcium-treated fruit exhibited slower loss of galactolipid and altered levels of sterol conjugates. The present study of lipids in “control” fruit was conducted as a prelude to further in-depth analyses of the effects of postharvest calcium and heat treatments on lipid metabolism in apples during and after cold storage. Neutral lipid, glycolipid (GL), and phospholipid (PL) fractions were obtained by column chromatography followed by TLC separation of GL and PL classes. The major GL were steryl glycosides (SG), acylated steryl glycosides (ASG), cerebrosides (CB), and mono- and digalactosyl diacylglycerols. Phosphatidylcholine (PC) > P-ethanolamine (PE) > P-irositol (PI) were the major PL. The fatty acids of PC and PE were quite similar, whereas those of PI were more saturated. CB included only 2-hydroxy fatty acids. Among the steryl lipids, free sterols > SG > ASG, with beta-sitosterol >90% of the total sterol in each.

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Bruce D. Whitaker, Joshua D. Klein, and William S. Conway

Postharvest heat treatment of apples maintains fruit firmness and reduces decay during storage. Four days at 38C are beneficial, but 1 or 2 days are detrimental. The cellular basis of these effects may involve changes in cell wall and membrane lipid metabolism. Lipids from hypodermal tissue of `Golden Delicious' apples were analyzed after 0, 1, 2, or 4 days at 38C. Major lipids included phospholipids (PL), free sterols (FS), steryl glycosides (SG), and cerebrosides (CB). Galactolipids (GL) were minor components. PL content fell ?10% after 1 day at 38C, was unchanged after 2 days, and began to rise again after 4 days. PL class composition did not change with heating, but fatty-acid unsaturation declined throughout. FS and CB content and composition changed little, whereas SG content cropped by ≈20% over 4 days. GL fell ≈50% during 1 day at 38C, with no change at days 2 or 4. A burst of PL catabolism followed by recovery of synthesis may in part explain the different effects of 1-, 2-, or 4-day heat treatments. GL loss (in plastids) may be related to the effect of heat on fruit color (yellowing).

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G.A. Picchioni, A.E. Watada, W.S. Conway, B.D. Whitaker, and C.E. Sams

Postharvest CaCl2 pressure infiltration improves firmness and storage quality of apples but is still in the experimental stages. Its effectiveness could be increased if we had a better understanding of how Ca affects the tissue at the cellular level. `Golden Delicious' fruit were harvested from a commercial orchard and were pressure-infiltrated with CaCl2 (0%, 2%, or 4% w/v), stored for 6 months at 0C, and then for 7 days at 20C. Between harvest and the end of storage at 20C, the net breakdown of galactolipids and phospholipids decreased with increasing CaCl2 in infiltration solutions. During 0C storage, CaCl2-infiltrated fruit maintained greater concentrations of conjugated sterol lipids, and these lipid classes are thought to be closely associated with the plasma membrane. As membrane lipid alterations are viewed as a central factor in the senescence of fruits, Ca (from postharvest infiltration) may serve a major role in regulating fruit quality losses through its interactions with cell membranes.