Seasonal variations in trans-2-hexenal and linolenic acid in homogenates of Thea sinensis leaves

The seasonal variations in the amounts of C₆-volatile components cis-3-hexenal trans-2-hexenal n-hexanal) and their precursors (linoleic and linolenic acid) in homogenates of Thea sinensis leaves were quantitatively analyzed throughout the year. Formation of trans-2-hexenal began in the middle of April and reached a maximum during July. Towards autumn the aldehyde gradually decreased and, in winter (December to March), was virtually absent. The levels of cis-3-hexenol remained constant during May–December. cis-3-Hexenal showed a similar variation pattern to that of trans-2-hexenal. The major fatty acids in the leaves were palmitic, palmitoleic, oleic, linoleic and linolenic acid, and occurred in non-ionic lipids and phospholipid fractions. The amounts of linoleic and linolenic acid did not show any marked variation except for a big peak in October.     

Lipid degradation during manufacture of black tea

About 85% of the fatty acids liberated during the manufacture of black tea can be attributed to autolysis of 4 major polar lipid classes in tea leaf tissue, phosphatidylcholine, monogalactosyldiglyceride, digalactosyldiglyceride and phosphatidylethanolamine. Linolenic, linoleic and palmitic acids are the principal fatty acids released from these lipids and they all undergo further degradation. Linolenic acid (60% of fatty acids released) is derived mainly from galactolipids and thus the upper limit of release is dependent on the chloroplast maturity and content of the leaf tissues. Lipid breakdown is complete after 2 hr fermentation and, as there appears to be no accumulation of long chain fatty acid intermediates, it is probable that volatile production has also ceased at this time.     

Seasonal Changes in Activities of Enzymes Responsible for the Formation of C6-aldehydes and C6-alcohols in Tea Leaves, and the Effects of Environmental Temperatures on the Enzyme Activities

When tea leaves were homogenized and incubated, the volatileC₆-compounds hexanal, cis-3-hexenal, cis-3-hexenol and trans-2-hexenalwere formed much more by summer leaves than by winter leavesof tea plants (Camellia sinensis). The enzymes lipolytic acylhydrolase (LAH), lipoxygenase, fatty acid hydroperoxide lyase(HPO lyase) and alcohol dehydrogenase (ADH) and an isomerizationfactor were responsible for the sequential reactions of C₆-compoundformation from linoleic and linolenic acids in tea leaf lipids,and there were seasonal changes in their activities. The tealeaf enzymes were of 3 types: LAH and lipoxygenase, which hadhigh activities in summer leaves and low activities in winterleaves; ADH, which had low activity in summer leaves and highactivity in winter ones; and HPO lyase and the isomerizationfactor, which did not seem to have any effect on the rate ofC₆-compound formation throughout the year. Changes in enzymeactivities were induced by shifts in the environmental air temperaturerather than by the age of the leaves. The combined activitiesof these enzymes determined the amounts and compositions ofthe volatile C₆-compounds formed, which are the factors thatcontrol the quality of the raw leaves processed for green tea. (Received October 6, 1983; Accepted December 20, 1983)     

Formation of cis-3-hexenal,trans-2-hexenal and cis-3-hexenol in macerated Thea sinensis leaves

Thea sinensis; Theaceae; tea; cis-3-hexenal: leaf aldehyde; leaf alcohol; linolenic acid; biosynthesis of leaf alcohol.Linolenic acid and cis-3-hexenal were found in macerated leaves of Thea sinensis and this aldehyde may be produced from linolenic acid by an enzyme contained in macerated leaves in the presence of oxygen. This aldehyde was easily isomerized to trans-2-hexenal, and was converted to cis-3-hexenol by alcohol dehydrogenase. During maceration of freshly picked tea leaves, the amounts of trans-2-hexenal quickly increased and were influenced by maceration time, heating, oxygen and the pH. But in unpicked tea leaves the occurrence of trans-2-hexenal is extremely doubtful.     

The importance of linoleic acid and linolenic acid as precursors of hexanal and trans-2-hexenal in black tea

During tea fermentation, linoleic acid in the neutral fat fraction,and linolenic acid in both the neutral fat and phospholipidfractions from leaves decreased. The addition of linoleic orlinolenic acid to leaf macerates during fermentation resultedin an increase in hexanal or trans-2-hexenal in the volatilefraction. Tracer experiments showed the direct conversion oflinoleic-U-¹⁴C and linolenic-U-¹⁴C acids to labeled hexanaland trans-2-hexenal, respectively, which were identified as2,4-DNPH derivatives. Further conversion of hexanal and trans-2-hexenal into hexanoicand trans-2-hexenoic acids during tea fermentation was suggestedby the increases in these compounds after the addition of hexanaland trans-2-hexenal to leaf macerates. (Received December 21, 1971; )     

Comparison of fatty acid patterns in plant parts and respective callus cultures of Cucumis melo

Root, hypocotyl, cotyledon, stem and leaf of Cucumis melo var. utilissimus seedlings were used for callus induction. Comparison was made between these parts, between callus tissues originating from all the parts and between each part and its callus, with respect to the fatty acid composition of total lipids. In all the parts there was a greater proportion of unsaturated fatty acids, the predominant fatty acid in root, stem and leaf being linolenic acid whilst in the cotyledon linoleic predominated. In the hypocotyl these two acids were present in equal amounts. In callus cultures the proportion of saturated acids was greater and the predominant fatty acid was palmitic. The major unsaturated fatty acid in callus cultures was linolenic. The analysis showed that callus tissue and its respective plant part had different fatty acid patterns and that all the callus cultures had very similar patterns irrespective of their origin.     

Lipoxygenase-mediated cleavage of fatty acids to carbonyl fragments in tomato fruits

Homogenates of tomato fruits catalysed the enzymic conversion of linoleic and linolenic acids (but not oleic acid) to C₆ aldehydes in low (3–5%) molar yield. Hexanal was formed from linoleic acid; cis-3-hexenal and smaller amounts of trans-2-hexenal were formed from linolenic acid. With the fatty acids as substrates, the major products were fatty acid hydroperoxides (50–80% yield) and the ratio of 9- to 13-hydroperoxides as isolated from an incubation with linoleic acid was at least 95:5 in favour of the 9-hydroperoxide isomer. When the 9- and 13-hydroperoxides of linoleic acid were used as substrates with tomato homogenates, the 13-hydroperoxide was readily cleaved to hexanal in high molar yield (60%) but the 9-hydroperoxide isomer was not converted to cleavage products. Properties of the hydroperoxide cleavage system are described. The results indicate that the C₆ aldehydes are formed from C₁₈ polyunsaturated fatty acids in a sequential enzyme system involving lipoxygenase (which preferentially oxygenates at the 9-position) followed by a hydroperoxide cleavage system which is, however, specific for the 13-hydroperoxy isomers.     

The influence of linoleic acid and linolenic acid on adult moth emergence of Homona coffearia from meridic diets in vitro

The fatty acids linoleic and linolenic acids have been found to be critical dietary supplements for the successful adult emergence of Homona coffearia reared in meridic diets in vitro. When the levels of these fatty acids were sub-optimal, the moths were able to emerge only partially and a few that emerged fully were deformed with naked wings. In the presence of optimal amounts of these fatty acids, the Δ⁷ sterol, ergosterol, available in the brewer's yeast provided in the diets was found to be adequate for growth and adult emergence. The availability of linoleic acid, which appeared to be the more critical requirement, was sufficient for successful adult emergence, provided that the diet was supplemented with additional amounts of ergosterol. The above two fatty acids seemed to be the only critical dietary supplements offered by whole tea leaves incorporated in a previously described oligidic diet. The provision of a dietary source of a Δ⁷ sterol was found essential as this insect was unable to utilize cholesterol.     

Potato tubers exhibit both homolytic and heterolytic hydroperoxide fatty acid-cleaving activities

The action of a crude potato-tuber extract on 9- and 13-hydroperoxides of linoleic and linolenic acids was investigated. HPLC analysis revealed that 50% of the 9-hydroperoxide isomers and almost all the 13-hydroperoxide isomers were rapidly enzymically metabolized. No degradation of fatty acid hydroperoxides was observed with a thermally denatured enzymic extract. GC-MS identification of the volatiles formed by the reaction revealed that no volatiles were detected from the 9-hydroperoxide isomers, whereas 13-hydroperoxide of linolenic acid was cleaved into (Z)-3-hexenal, pentenols or dimers of pentene.     

EAG and behavioral responses of the wingless tea aphid Toxoptera aurantii (Homoptera: Aphididae) to tea plant volatiles

Electrophysiological and behavioral responses of the wingless tea aphid, Toxoptera aurantii (Boyer), to 14 synthetic volatiles identified from tea shoots, their partial (GLV mixture) and full (ACB mixture) blends, and fresh young tea leaves, buds, tender stems, adult tea leaves and tea aphid-damaged young leaves (ADYL) were studied by using an electroantennography (EAG) and a four-arm olfactometer. ACB elicited the largest EAG responses. Major volatile components, Z-3-hexen-1-ol, E-2-hexenal, n-hexanol, methyl salicylate and benzylalcohol, from the tea shoots were strongly EAG active. All the 4 tested tea shoot tissues also elicited significant EAG responses, with the young tea leaves being the strongest, followed by buds, tender stems and adult tea leaves. Surprisingly, ADYL elicited a weakly negative EAG response. In the olfactory assays, the fresh and tender tea leaves, as well as the individual major volatile components, e.g. Z-3-hexenyl acetate, methyl salicylate, E-2-hexen-1-ol and Z-3-hexen-1-ol, from the tender shoots (EAG-active) were all attractive. This result might indicate that the wingless tea aphids may use tea shoot volatiles as kairomone to find their optimal feeding sites, e.g. fresh tender tea shoots.     

蚜害茶树挥发物组分变化的聚类分析

舒茶早(Camellia sinensis cv. ‘shuchazao’)是皖西南产区一种茶树品种。为研究茶蚜取食为害诱导的茶树挥发物释放特征,运用气相色谱-质谱联用技术,比较了健康茶梢和蚜害茶梢的挥发物组成和相对含量。结果显示,健康茶梢挥发物种类(16种)和相对含量较少,而蚜害茶梢挥发物种类(24种)较为丰富且相对...     

Enzymic formation of carbonyls from linoleic acid in leaves of Phaseolus vulgaris

Homogenization of Phaseolus vulgaris leaves at acid pH results in the evolution of hexanal, cis-3- and trans-2-hexenal. With cell-free extracts of leaves, linoleic and linolenic acids are enzymically converted to their hydroperoxides (predominantly the 13-hydroperoxide isomers) and to hexanal or hexenal respectively. Activity was highest in young, dark-green leaves and was stimulated by Triton X-100. Oleic acid is not a substrate for these reactions. Both 9- and 13-hydroperoxides were cleaved to carbonyl fragments and are proposed as intermediates in the formation of volatile aldehydes and non-volatile ω-oxoacids in P. vulgaris leaves. Properties of the enzyme systems are described.     

Volatile Products of the Lipoxygenase Pathway Evolved from Phaseolus vulgaris (L.) Leaves Inoculated with Pseudomonas syringae pv phaseolicola

Activation of the "lipoxygenase pathway" in plants gives rise to a series of products derived from fatty acids. Analysis by gas chromatography-mass spectroscopy of volatile products produced by Phaseolus vulgaris (L.) cv Red Mexican leaves during a hypersensitive resistance response (HR) to the plant pathogenic bacterium Pseudomonas syringae pv phaseolicola showed evolution of several lipid-derived volatiles, including cis-3-hexenol and trans-2-hexenal, which arise from the 13-hydroperoxide of linolenic acid. These compounds were not produced in detectable amounts by buffer-inoculated leaves, nor did they evolve to such a high degree during comparable stages of the susceptible response. The absence of trans-2,cis-6-nonadienal, a product expected from 9-hydroperoxide of linolenic acid, suggests that lipid peroxidation during the HR proceeded primarily enzymically via bean lipoxygenase, which produces the 13-hydroperoxide, and not via autoxidative processes. The effects of trans-2-hexenal, cis-3-hexenol, and traumatic acid on P.s pv phaseolicola were investigaed. trans-2-Hexenal appeared to be highly bactericidal at low concentrations, whereas cis-3-hexenol was bactericidal only at much higher concentrations. Traumatic acid appeared to have no effect on P.s. pv. phaseolicola at the concentrations tested. These results demonstrate that during plant defense responses against microbial attack, several lipid-derived compounds are produced by the plant, some of which possess antimicrobial activity and conceivably are involved in plant disease resistance. The time of production of these substances, in amounts that would be expected to be antibacterial in vitro, correlated with a slowing down of the growth rate of bacteria in the leaves and was seen at a time before the accumulation of isoflavonoid phytoalexins in the host.     

Lipids in plant tissue cultures. VIII: Reversible changes in the composition of lipids and their constituent fatty acids in response to alternate shifts in the mode of carbon supply

When heterotrophic cell cultures of red goosefoot (Chenopodium rubrum) turned photoautotrophic, their contents of various glycolipids and phospholipids increased. The total lipids and the individual lipid classes, especially monogalactosyldiacylglycerols, became richer in linolenic and poorer in linoleic acids. When photoautotrophic cell cultures were rendered heterotrophic again a reversal of changes occurred; both the composition of lipids and the patterns of their constituent fatty acids became similar to those of the starting heterotrophic cultures.The results indicate that the biosynthesis of linolenic acid in photoautotrophic cell cultures involves mainly desaturation of linoleic acid and that chain extension of hexadecatrienoic acid is possibly another, though minor pathway. Monogalactosyldiacylglycerols are apparently the substrates preferred for linolenic acid biosynthesis, whereas various phospholipids are the substrates preferred for linoleic acid biosynthesis.During a growth period of 6 weeks, the levels of polyunsaturated fatty acids in the lipids from both heterotrophic and photoautotrophic cell cultures decrease with time, whereas the proportions of palmitic acid increase.     

Lipids in plant tissue cultures. VI. Effect of temperature on the lipids of Brassica napus and Tropaeolum majus cultures

The lipid contents of callus cultures of rape (Brassica napus) and nasturtium (Tropaeolum majus) increase in response to decreasing the temperature, though to different degree. Irrespective of the incubation temperature the lipids in cultures of both plants contain as predominant classes steryl glycosides, esterified steryl glycosides, sterols, steryl esters and fatty acids and, as minor constituents, various proportions of triacylglycerols, phospholipids and several unidentified fractions.The ratio of phospholipids to triacylglycerols as well as the ratio of diacylglycerophosphorylethanolamines to dicylglycerophosphorylcholines increase with time both in rape cultures incubated at 30°C and in those kept at 5°C.The lipids in rape and nasturtium cultures grown at 30°C contain smaller proportions of polyunsaturated fatty acids than the lipids in cultures incubated at 5°C. Erucic acid, the major constituent fatty acid of the seed lipids, in both plants, occurs only in trace amounts in the lipids of callus cultures. In contrast, linoleic and linolenic acids, which occur only in traces in the seed lipids of nasturtium, are major constituent fatty acids in the lipids of callus cultures derived from seedlings of this plant.The levels of constituent polyunsaturated fatty acids in the diacylglycerophosphorylethanolamines and the diacylglycerophosphorylcholines increase with time whereas in the triacylglycerols only linolenic acid is slightly increased.     

Effect of Growth Temperature on the Biosynthesis of Chloroplastic Galactosyldiacylglycerol Molecular Species in Brassica napus Leaves

Brassica napus leaves developed at low temperature display rapid in situ desaturation of monogalactosyldiacylglycerol (MGDG) fatty acids leading to the production of hexadecatrienoic/linolenic acid. This was shown by radioactivity-tracer experiments to occur via a sequence of desaturations proceeding from the initially synthesized palmitic/oleic acid molecular species to palmitic/linoleic acid, palmitoleic/linoleic acid, hexadecadienoic/linoleic acid, hexadecadienoic/linolenic acid, and finally to hexadecatrienoic/linolenic acid. The results suggest that there is increased activity in all five desaturation steps in leaves developed at low temperatures. Labeling data also indicate that there is another pool of MGDG which is more slowly desaturated before galactosylation to digalactosyldiacylglycerol (DGDG). Our data further suggest that relative rates of galactosylation of chloroplastic and cytosolic MGDG molecular species may regulate the final amounts of chloroplastic and cytosolic MGDG and DGDG in the leaf. We have proposed a model for chloroplastic biosynthesis and desaturation of galactosyldiacylglycerols in the leaves of Brassica napus, a 16:3 plant.     

Influence of cold temperatures on leaf lipids of Hibiscus rosa-sinensis

Triaclyglycerols (TAG) accounted for 15% of the leaf acyl lipids in a Hibiscus rosa-sinensis plant that survived the January 1986 freeze in Florida, U.S.A. This high TAG level suggested that the plant had cold hardened. The TAG concentration in leaves on new shoots from this plant was greater than that in new leaves on new shoots from five plants frozen to the roots during the freeze. However, five months later, the amounts of TAG in the six Hibiscus plants were nearly the same. The minor differences in TAG levels, however, related to the survival rate of these six plants during subsequent freezes. In addition to linoleic acid, two cyclopropene fatty acids, malvalic and sterculic, were major constituents of the leaf TAG. Hibiscus plants placed in environmental chambers under control and cold-hardening regimes optimized for Citrus showed TAG concentrations of 7% in control and 20% in hardened plants. A survey of neutral lipids in Hibiscus and other plants showed that plastoquinone A (B) and α-tocopherol decreased and plastoquinone C increased under cold-hardening conditions. Polyprenols, a major component of Hibiscus leaves under normal conditions, declined greatly under cold-hardening regimes.     

Flavour biogenesis. Partial purification and properties of a fatty acid hydroperoxide cleaving enzyme from fruits of cucumber

A membrane-bound enzyme, which catalyses the cleavage of fatty acid hydroperoxides to carbonyl fragments, has been partially purified from cucumber fruit. The isomeric 9- and 13-hydroperoxydienes (but not the hydroxydienes) derived from both linoleic and linolenic acids are cleaved by the enzyme but a mixture of 9- and 10-hydroperoxymonoenoic derivatives of oleic acid was not attacked. No evidence was obtained for free intermediates between fatty acid hydroperoxides and the cleavage products. Major volatile products were: cis-3-nonenal and hexanal (from 9- and 13-hydroperoxides of linoleic acid respectively) or cis-3,cis-6-nonadienal and cis-3-hexenal (from 9- and 13-hydroperoxides of linolenic acid). The increase in the ratio of cis-3- to trans-2-enal products with enzyme purification indicated that cis-3-enals are the immediate cleavage products and that the trans-2- forms are produced by subsequent isomerization.     

Lipids in developing and mature rice grain

The neutral fraction of nonstarch lipids in developing brown rice (Oryza sativa L., cv IR42) was accumulated up to 16 days after flowering (DAF), but phospholipids and glycolipids increased only up to 8 DAF. Fatty acids accumulated in nonstarch lipids until 12 DAF. However, the proportion of linolenic acid in the lipid fraction decreased and that of oleic acid increased during this period. Accumulation of fat-by-hydrolysis in the brown rice occurred until 20 DAF and followed closely that of starch. The proportion of linolenic acid decreased and that of linoleic acid increased until 16 DAF. The fatty acid composition of fat-by-hydrolysis and starch lipids were identical and fat-by-hydrolysis accounted for 48% by weight of starch lipids. Nonstarch lipids were mainly composed of triglycerides and were located in the bran and embryo of mature brown rice. Starch lipids were mainly composed of lysophosphatidyl choline, free fatty acids and lysophosphatidyl ethanolamine, and were located in the endosperm.     

Isolation of biochemical mutants using haploid mesophyll protosplasts of Hyoscyamus muticus

The question of whether membrane expansion, which is caused by anesthetics in animal systems, alters the lipid composition of plant cell membranes was investigated. We have measured the effects of several anesthetics on the relative amounts of the principal fatty acids from the polar lipids of barley (Hordeum vulgare L.) root membranes. Procaine, dibucaine, tetracaine, chloroform and, to a lesser degree, methanol increased the proportions of palmitic, stearic and oleic acids and decreased the proportions of linoleic and linolenic acids. Ethanol had no significant effect. Total amounts of the fatty acids from the polar lipids of roots in procaine solution decreased markedly so that all of the acids decreased in amount. The anesthetic was effective as soon as the roots were introduced to the solution and the changes progressed at constant rates for 6 h. Only the polar membrane lipids were altered; other lipids were not affected. Increased hydrostatic pressure of about 1.0 MPa largely prevented the anesthetic effects, including the decrease in the total amounts of the fatty acids. Hydrostatic pressure as high as 2 MPa had no effect per se on the membrane lipid composition. These results indicate that anesthetics cause expansion of the root membranes which results in the lipid changes. That a compositional change in the membrane lipids involves a conformational change such as expansion is an indication of the nature of the link between changes in the membrane lipids and changes in function of areas where hydrophilic ions permeate.Abbreviations 16:0 palmitic acid - 18:0 stearic acid - 18:1 oleic acid - 18:2 linoleic acid - 18:3 linolenic acid