Conversion of Phosphatidyl Choline to Phosphatidic Acid in Freeze Injured Rye and Wheat Cultivars

The effect of exposure to freezing temperature (?15°C) on leaf phospholipid composition of hardened rye (Secale cereale L.) and hardened wheat cultivars (‘Miranovskaja 808’, ‘Bezostaja 1’, ‘Short Mexican’ and ‘Penjamo 62’), which differ in their resistance to frost, was investigated. Hardening took place under natural conditions. All the seedlings attained an equal level of linolenic acid in their leaves during hardening. Exposure to freezing temperatures resulted in a loss of phosphatidyl choline and accumulation of phosphatidic acid in the leaves. The ratio of phosphatidic acid to phosphatidyl choline, but not the level of poly-unsaturated fatty acids in the leaves, was related to their ability to survive at low temperatures. As freezing injury is caused by the formation of ice crystals in both extra- and intracellular space, it is probable that the plasma membranes of the investigated cultivars differed with respect to their water permeability. It is concluded that the plants, depending on the degree of their resistance to cold, produce an unknown substance of lipidic nature upon exposure to cold, with the aid of which they adjust the transitional state of their membranes to the prevailing temperature and, at the same time, facilitate the efflux of water from the cells.     

Lipids of Cuscuta reflexa and changes in lipids of its host plants after infection

The lipid level (fresh weight basis) of Cuscuta reflexa Roxb. was related to the lipid content of the host plants Meilicago saliva L., Helianthus annuus L., Pisum sativum L. and Lantana camara L. Parasitizing by the dodder significantly increased the total lipid level of the hosts. The increase was mainly due to enhancement in the neutral lipid fraction.
The level of phospholipid in the parasite was always higher than in its hosts. Phospholidyl choline and phosphatidyl ethanolamine constituted about 65% of the total phospholipid of Cuscuta. This was followed by phosphatidyl inositol (ca 20%) and phosphatidyl glycerol (ca 12%). Phosphatidic acid constituted only ca 3% of the phospholipids of Cuscuta. Although the total phospholipid levels of various host plants were not affected as a result of the infection by Cuscuta, a significant decrease occurred in the levels of phosphatidyl eholine and phosphatidyl ethanolamine as well as marked increases in phosphatidyl inositol and phosphatidic acid. The infected tissue showed an increase in phospholipase D activity as compared with the controls. The results have been discussed in relation to changes in permeability of the infected tissue.     

Turnover of Phospholipids in Normal and Phosphorus-deficient Spirodela

When ³²P₁ was supplied as a 15-minute pulse to normal Spirodela oligorrhiza plants, the first phospholipid to become fully labeled was phosphatidic acid. Phosphatidyl glycerol reached maximum labeling before the other major phospholipids. In phosphorus-deficient plants, however, phosphatidyl glycerol became labeled much more slowly than either phosphatidyl choline or phosphatidyl ethanolamine, and also the proportion of phosphatidyl glycerol present was smaller. Thus, phosphatidyl glycerol synthesis is sensitive to phosphorus deficiency. Since most of the phosphatidyl glycerol present in Spirodela was localized in the chloroplast, this effect appeared to be specifically one on chloroplast composition. The phosphorus-deficient chloroplast had a 60% lower phospholipid content and a normal phospholipid pattern, but the phospholipid which was present was apparently cycling much less rapidly. Zeatin, which ameliorates the visual symptoms of phosphorus deficiency, also reduces the effect of phosphorus deficiency on phospholipid synthesis.     

Phospholipid changes in wheat and barley leaves under water stress

Total phospholipid content of leaves of wheat and barley increased and phospholipid components changed under water stress. Notable among these were the absence of phosphatidyl serine in barley varieties, decrease in phosphatidyl glycerol content in a less drought-tolerant variety of wheat (S-308) and barley (BG-25), and appearance of phosphatidic acid in both crops. The phospholipid content and its components did not return to normal upon release of the stress by subsequent irrigations. Such observations are indicative of water stress effected alterations in membranes.     

Effect of prolactin on the phospholipid composition and cholesterol level in liver microsomes of the carp during acclimation to different temperatures

The phospholipid composition, content of cholesterol and its esters in the carp (Cyprinus carpio L.) liver microsomes depend on the environmental temperature. The free cholesterol amount and cholesterol/phospholipids ratio in microsomes decrease after the lowering of temperature from 20 to 5 degrees C. The temperature elevation to 30 degrees C results in an increase of the cholesterol ester content. The relative proportions of phosphatidyl choline, phosphatidyl ethanolamine, sphingomyelin, phosphatidyl inositol, phosphatidyl serine, phosphatidic acid increase with a significant decrease of the unidentified phospholipids amount at 30 degrees C. Prolactin affects the cholesterol content and phospholipid composition of liver microsomes. The hormone has a more pronounced effect at subextremal temperatures (5 and 30 degrees C). The actions of prolactin and temperature on the cholesterol content are similar. The hormone influence on the membrane phospholipid composition is opposite to the effect of the temperature acclimation. The possible role of prolactin in the temperature adaptation of the membrane lipids metabolism in poikilotherms is discussed.     

Effect of Low Temperature upon Lipid and Fatty Acid Composition of Roots and Leaves of Winter Rape Plants

When winter rape plants were transferred from favourable temperature conditions (25/20°C day/night temperature) to 5°C, the frost resistance of the leaves was increased whereas the frost tolerance of the roots remained unaffected. This permitted an analysis of the changes in lipid and fatty acid composition both as related to functioning of the plant at low temperature alone (roots) and as related to adaptation to freezing and functioning at low temperature (leaves). — Transfer of the plants to 5°C lead to an increase in the level of linolenic acid in roots and leaves. This increase was most evident in the phosphatidyl choline and ethanolamine fractions of the leaves, and in the neutral lipids and in an unidentified phospholipid from the roots. It was concluded that upon transfer of the plants to 5°C a general and non-specific increase in linolenic acid level contributed to functioning of the rape plants at low temperature; and that parallel but minor increases in linolenic acid level of digalactosyl diglyceride, phosphatidyl inositol and the unknown phospholipid in roots and leaves could only contribute to low-temperature functioning in specific membrane enzyme locations. Combined adaptation of the leaves to freezing tolerance and low-temperature functioning was correlated with a higher level of phosphatidyl choline and ethanolamine, predominantly esterified with linolenic acid.     

Inhibition of chilling-induced photooxidative damage to leaves of Cucumis sativus L. by treatment with amino alcohols

The effect of pretreatment of cucumber (Cucumis sativus L.) roots with choline chloride or ethanolamine on leaf phospholipid composition and light-induced leaf damage during chilling was studied. Photooxidative chlorophyll degradation was similarly inhibited by both amino alcohols. The decrease of the chlorophyll a/chlorophyll b ratio and the increase of polyunsaturated-fatty-acid degradation during chilling in the light were equally inhibited by pretreatment with choline chloride or ethanolamine. Treatment with choline chloride and ethanolamine caused, respectively, 43% and 26% increases in the total phospholipid contents of the leaves. After treatment with choline chloride, the phosphatidylcholine content was higher than the content of phosphatidylethanolamine; the reverse was true after treatment with ethanolamine. The chlorophyll concentration increased less than the phospholipid concentration, resulting in a decreased chlorophyll/phospholipid ratio of treated leaves. During chilling in the light, degradation of phosphatidylcholine, ethanolamine and phosphatidyl glycerol occurred. Phosphatidyl glycerol was less sensitive than phosphatidylcholine and ethanolamine. The degradation was equally inhibited by pretreatment with either amino alcohol. Possible connections between the phospholipid content of leaf membranes and the inhibition of chilling-induced photooxidative leaf damage are discussed.Abbreviations CC choline chloride - Chl chlorophyll - EA ethanolamine - PC phosphatidyl choline - PE phosphatidyl ethanolamine - PG phosphatidyl glycerol     

Hypervitaminosis A and Liver Phospholipids

Effect of daily oral feeding of 33 mg retinol for nine days on the liver phospholipids of rats has been studied. As early as two days after feeding retinol an increase in the amounts of liver triglycerides, proteins, phospholipids, and cholesterol was noted which kept increasing and reached the peak concentration 6 days after daily retinol feeding and thereafter a decrease in their amounts was noted. Hepatic phospholipid fractions viz. phosphatidyl choline, phosphatidyl ethanolamine, phosphatidic acid and polyglycerol phosphatide, phosphatidyl inositol, phosphatidyl serine, sphingomyelin, lysophosphatidyl ethanolamine and lysophos- phatidyl choline showed the same pattern. Labelling of these phospholipids with NaH₂³²PO₄ in rats fed daily 33 mg of retinol for a period of two days also exhibited the pattern which was observed in their amounts two days after daily feeding of retinol. The results suggest a close relationship between the metabolism of hepatic triglycerides and phospholipids of rats fed excessive amounts of retinol.     

Fatty acid composition of glycerolipids from potato leaves

A method for rapid isolation of glyco- and phospholipids from potato leaves by a two-fold separation in a thin layer of silica gel is described. Using gas-liquid chromatography, the fatty acid compositions of monogalactosyldiglyceride, digalactosyldiglyceride, sulfolipid, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl glycerol, phosphatidyl inositol, diphosphatidyl glycerol, phosphatidic acid and non-identified lipid from potato leaves were determined. The monogalactosyl diglyceride was found to contain up to 25% of 7,10,13-hexadecatrienic acid. Trans-3-hexadecenic acid as well as phosphatidyl glycerol is a constituent component of phosphatidic acid, diphosphatidyl glycerol and the non-identified lipid.     

Lipid composition in leaves of cucumber genotypes as affected by different temperature regimes and grafting

The lipid composition of leaves has been investigated in different genotypes of cucumber ( Cucumis sativus L.), which differ in temperature requirement for cultivation. In addition the effects of hardening by low but non-chilling temperature, soil heating and grafting (on the chilling-resistant C. ficifolia L.) on lipid composition have been studied. Content and composition of phospholipids and sterols were determined as well as phospholipid/sterol ratio, and fatty acid composition of total lipids and the different phospholipids.
The effects of genetic differentiation and of the various culture treatments on lipid composition of the leaves were very different. Genetic differentiation was evident as higher levels of Iinolenic acid in several phospholipids in the more cold-tolerant cultivars. Hardening the plants by low temperature resulted in a higher phospholipid level (especially phosphatidyl choline), more unsaturated phospholipid, and lowering of the sterol/phiospholipid ratio, all properties which may contribute to a higher membrane fluidity and lower growth temperature limit. Soil healing reduced the phospholipid level of the leaves slightly, and a higher content of 3- trans -hexadece-noic acid in phosphatidyl glycerol was observed. Grafting cucumber on the cold-resistant rootstock of C. ficifolia also raised the level of trans -hexadecenoic acid in phosphatidyl glycerol. The role of this fatty acid in the functioning of the chloroplast is discussed.     

Phospholipids inhibit proteolysis of protein kinase Cα by mM calcium-requiring calpain

The alpha isoform of protein kinase C (PKC) is rapidly hydrolyzed by mM Ca²⁺-requiring calpain (calcium-activated neutral proteinase) under cell-free conditions (Shea et al, 1994, FEBS Lett. 350: 223). In the present study, we demonstrate that this hydrolysis is inhibited by phosphatidyl serine, diacylglycerol, phosphatidyl choline, phosphatidyl inositol, and phosphatidic acid. With the exception of phosphatidic acid, these phospholipids did not directly inhibit calpain activity as evidenced by degradation of [¹⁴C]azocasein, suggesting that the nature of inhibition of calpain-mediated PKC degradation is due to an effect of phospholipids on PKC conformation. These findings suggest that m calpain-mediated PKC hydrolysis may be specifically minimized at the plasma membrane, and leave open the possibility that such a mechanism exists in situ. In addition, the unique inhibition of calpain activity by phosphatidic acid suggests the existence of a specific mechanism by which this phospholipid regulates PKC activity.Special issue dedicated to Dr. Leon S. Wolfe.     

Complex lipids of Rhodomicrobium vannielii

Eight components, seven of which contained phosphorus, were found in the phospholipid fraction of Rhodomicrobium vannielii. The major components were lipoamino acid (o-ornithine ester of phosphatidyl glycerol, 46.5%) and phosphatidyl choline (26.5%). The other six components were phosphatidyl glycerol (9.7%), bisphosphatidic acid (6.7%), phosphatidyl ethanolamine (4.5%), phosphatidic acid (1.8%), lysophosphatidyl glycerol-o-ornithine ester (3.2%), and N,N-ornithine amide of unidentified fatty acid (0.95%). Total phospholipid accounted for 4.2% of cell dry weight. The major fatty acid was vaccenic acid, C_18:1, which accounted for approximately 90% of the total fatty acids of the complex lipid fraction. The other four fatty acids were C_16:0 (6.25%), C_18:0 (3.8%), C_14:0 (0.7%), and C_16:1 (0.35%). The sulfolipid content was 0.01% of the cell dry weight or 0.14 μmoles per g of dried cells, assuming that its fatty acid component is vaccenic acid. No steroids were detected.     

Composition of Phospholipids and Fatty Acids cf Mitochondria of Chilling-Sensitive and Chilling-Tolerant Corns

The phospholipids of the mitochondria of chiUing-sensitive (Zhang Dang No. 9) and chilling-tolerant (HD103×Ferumac) corn shoots are composed of phosphatidyl choline, phosphatidyl ethaaolamine, phosphatidic acid, phosphatidyl inositol, diphosphatidyl glycerol, lysophosphatidyl choline and so on. The weight per cent composition of various phospholipid components in there two varieties of corn are not apparently different. The fatty acid composition of the mitochondria of both corn shoots arc similar except the palmitic acid and linoleic acid.     

The Effects of Acetylcholine on the Turnover of Phosphatidic Acid and Phosphoinositide in Sympathetic Ganglia, and in Various Parts of the Central Nervous System in Vitro

The effect of acetylcholine on the incorporation of P³² into the individual phosphatides in slices of various structures of the nervous system has been studied. There was a marked stimulation of P³² incorporation into phosphoinositide and phosphatidic acid, but not into phosphatidyl choline and phosphatidyl ethanolamine, in the cat stellate and celiac ganglia in vitro. Acetylcholine stimulated P³² incorporation into certain phosphatides, primarily phosphoinositide and phosphatidic acid, in several structures of the cat and guinea pig brain; there was little or no effect of acetylcholine on phosphatide turnover in the inferior corpora quadrigsemina and cerebellar cortex. The suggestion is made that the phospholipid effect can best be explained as being concerned with the active transport of sodium ions out of the cell across the postsynaptic membrane of cholinergic neurons in response to acetylcholine.     

Membrane phospholipids of normal and crown gall callus cultures

The relative proportions of different phospholipid species were determined in membrane-enriched fractions from normal and crown-gall callus cultures. In general the pattern of phospholipids was similar, but the tumour cultures contained relatively more phosphatidyl choline and less phosphatidic acid and phosphatidyl inositol than the corresponding normal cultures. These differences are probably not caused by phospholipase-D activity during homogenization.     

Effects of prolactin and androgens on seminal vesicular lipids of castrated mature bonnet monkeys Macaca radiata

Effects of prolactin (PRL), bromocriptine (Br), testosterone propionate (TP), dihydrotestosterone (DHT) and the combination of these androgens with PRL/Br on the total lipid, total cholesterol, total glyceride glycerols, total phospholipid and their fractions in seminal vesicles of castrated mature monkeys were studied. Glyceride glycerols formed the major portion (50%) of total lipids in normal monkeys. Cholesterol and phospholipids were of equal share (25%). Esterified cholesterol formed major share (75%) of total cholesterol. Diacyl glycerol was the major (60%) glyceride glycerol and phosphatidyl choline and ethanolamine were the major phospholipid classes. Except triacyl glycerol castration markedly decreased all the lipid classes. PRL restored normal free and esterified cholesterol and phosphatidyl inositol but Br invariably decreased all the lipid classes. TP/DHT treatment stimulated the free and esterified cholesterol more than the control; it restored the normal glyceride glycerols. Phosphatidyl inositol, choline and ethanolamine were stimulated by androgens and other phospholipid classes were brought to normal. Addition of PRL + TP/DHT markedly increased esterified cholesterol, phosphatidyl inositol, choline, ethanolamine and phosphatidic acid. In all these aspects, Br counteracted the effects of androgens and PRL.     

Gibberellin modulation of phosphatidyl-choline turnover in wheat aleurone tissue

Phosphatidyl choline (PC) is synthesised in wheat (Triticum aestivum L. cv. Flanders) aleurone tissue during early germination when new endomembranes are being formed. Although gibberellic acid does not ostensibly affect PC levels, it inhibits the incorporation of choline and differentially and specifically modulates the turnover of the N-methyl and methylene carbons of the choline headgroup of PC. Gibberellic acid has no effect on turnover of the phosphate moiety of either PC or the other major phosphatides. The possible biological importance of the findings is discussed.Abbreviations ER endoplasmic reticulum - GA gibberellin - GA₃ gibberellic acid - PA phosphatidic acid - PC phosphatidyl choline - PE phosphatidyl ethanolamine - PG phosphatidyl glycerol - PI phosphatidyl inositol - t_1/2 half-life     

The manipulation of polar head group composition of phospholipids in the wheat Miranovskaja 808 affects frost tolerance

Caryopses of the frost-resistant cultivar of the wheat Triticum aestivum L., Miranovskaja 808, were germinated and grown in the presence of various concentrations of choline chloride. Changes in the composition of leaf total phospholipids and leaf total fatty acids at two extreme temperatures (25°C and 2°C) as well as changes in frost resistance were followed. A choline chloride concentration-dependent accumulation of phosphatidyl choline was observed in the leaves. Seedlings grown at 2°C accumulated more phosphatidyl choline at each choline chloride concentration than those grown at 25°C. There was an inverse relationship between the contents of phosphatidyl choline and phosphatidic acid in the leaves. Neither the temperature nor choline chloride seemed to affect fatty-acid composition. Modification of polar-head group composition of phospholipids affected frost tolerance: Seedlings grown in the presence of 15 mM choline chloride at 25°C exhibited a freezing resistance equal to that of hardened controls. The data indicate that the polar-head group composition of membrane phospholipids in plants can be easily manipulated and point to the importance of phosphatidyl choline in cold adaptation processes.     

Effect of Temperature and BASF 13 338 on the Lipid Composition and Respiration of Wheat Roots

The fatty acid composition of wheat seedling roots changed in response to temperature. As temperature declined, the level of linolenic acid increased and the level of linoleic acid decreased. The distribution of phospholipid classes was not influenced by temperature. Phosphatidyl choline and phosphatidyl ethanolamine were the predominant phospholipids isolated and comprised 85% of the total lipid phosphorus. Smaller quantities of phosphatidyl glycerol, phosphatidyl inositol, phosphatidic acid, and phosphatidyl serine were isolated. The fatty acid composition of phosphatidyl choline and phosphatidyl ethanolamine were the same and temperature affected the fatty acid composition of both phospholipids in the same manner.Growth in the presence of the substituted pyridazinone, BASF 13 338 (4-chloro-5-dimethylamino-2-phenyl-3(2H)pyridazinone), reduced the level of linolenic acid and increased the level of linoleic acid in the phosphatidyl choline, phosphatidyl ethanolamine, and total polar lipid fractions. BASF 13 338 did not affect the levels of palmitate, stearate, and oleate or the distribution of phospholipid classes.Respiration rates of wheat root tips were measured over a range of temperatures. The respiration rate declined as the temperature decreased. Neither the temperature at which the tissue was grown nor BASF 13 338 treatment influenced the ability of root tips to respire at any temperature from 4 to 30 C. The results indicated that the relative proportion of linolenic acid to linoleic acid did not influence the plants ability to grow and respire over the range of temperatures tested.     

Effect of freezing and cold storage on phospholipids in developing soybean cotyledons

Freezing of plant tissue adversely affects lipid composition. Immature soybean cotyledons (Glycine max L. Merr.) var. “Harosoy 63” were frozen with liquid N₂, dry ice, or stored in a freezer (−20 C) before lipid extraction. The effects of freezing temperature, thawing rate, and cold storage on the lipid composition of frozen tissue revealed significantly higher levels of phosphatidic acid, and diminished levels of phosphatidylcholine, phosphatidylethanolamine, and N-acylphosphatidylethanolamine from the control. Regardless of freezing temperature, phosphatidic acid levels increased from 4.7 mole% to nearly 50 mole% of the total phospholipid when frozen tissues were stored 10 days at −20 C. During the same period, N-acylphosphatidylethanolamine decreased from 54.1 mole% to 6.6 mole% phospholipid. At least 8 mole% of the phosphatidic acid increase occurred during slow thawing of the frozen tissues. In autoclaved samples, phosphatidic acid, phosphatidylcholine, phosphatidylethanolamine, and N-acylphosphatidylethanolamine levels were not different from the control. Labeling of the lipid-glycerol with ³H, and fatty acids with ¹⁴C, demonstrated the degradation product was primarily phosphatidic acid. Apparently enzymic destruction of the phospholipids occurred during freezing, cold storage, and thawing.