Food reserves of Scots pine (Pinus sylvestris L.)

Summary The amounts of starch, soluble sugars, triacylglycerols, diacylglycerols and free fatty acids were studied in Scots pine (Pinus sylvestris L.) during an annual cycle in current-year needles and in 1-, 2- and 3-year-old needles collected shortly after bud break. Determination of the compounds was performed using specific enzymatic assays, capillary gas chromatography and thin layer chromatography. Newly emerging needles contained relatively large amounts of starch, but only trace amounts of fat. During autumn and winter, fat content rose, while starch content decreased; amounts of both these reserve materials were very high the next spring shortly before bud break and decreased again during shoot elongation. Concentration of intermediates in triacylglycerol biosynthesis (diacylglycerols and free fatty acids), were low in summer and high in winter. The same pattern was observed for fructose and glucose (the predominant soluble sugars), galactose/arabinose and raffinose/melibiose. In contrast, sucrose concentrations were highest in spring and in autumn. Mature needles of different ages collected in May showed significant differences only in their triacylglycerol and starch content. Concentration changes of reserve materials are discussed in relation to season, mobilization and translocation processes, dormancy, frost resistance and the possibility of carbohydrate-fat interconversions.     

Soluble carbohydrates of Pinus sylvestris L. sapwood and heartwood

Summary The amounts of glucose, fructose, sucrose, arabinose/galactose, raffinose/stachyose and starch were investigated in the outer sapwood, innermost sapwood, transition zone and heartwood of four stems of Pinus sylvestris L. The samples were taken in October and the determination of the compounds was done enzymatically. It was not possible to distinguish arabinose from galactose and raffinose from stachyose. The amounts of glucose, fructose and sucrose were greatest in the outer sapwood and decreased gradually towards the innermost sapwood and the heartwood. In the outermost heartwood glucose, fructose and sucrose were only present in trace amounts. Raffinose/stachyose showed highest concentrations in the outer sapwood and decreased towards the heartwood. In contrast, the concentrations of arabinose/galactose increased towards the heartwood and the greatest amount was found in the inner heartwood. When identified by thin-layer chromatography (TLC), arabinose was found to be present in greater amounts than galactose. The amount of starch decreased markedly towards heartwood. However, the amounts of sugars in all the studied stems was very variable. The changes in the amounts of carbohydrates in the different zones of the stems and the possible relationships of these phenomena with heartwood formation are discussed.     

Seasonal variation of neutral lipids in Pinus sylvestris L. sapwood and heartwood

Summary The lipid levels and fatty acid composition of three fractions (free fatty acids, triacylglycerols and sterol/triterpenoid esters) extracted from the sapwood and heartwood of three stems of Pinus sylvestris were determined to investigate both seasonal changes in sapwood and possible metabolic changes related to heartwood formation. Seasonal changes were observed only in the amount of the free fatty acids in the sapwood: the level of free fatty acids was greatest at the beginning and end of the growing season. In the January and March samples the amount of the free fatty acid fraction in the sapwood was very small. The amount of the other fractions remained at the same level throughout the study. Marked seasonal changes in the fatty acid composition occurred only in the free fatty acid fraction of the sapwood: the saturation grade increased during the winter.     

Lipids and sterols of Pinus sylvestris L. sapwood and heartwood

Summary The lipid and sterol content and composition of three lipid fractions (free fatty acids/ sterols, triacylglycerols and sterol/triterpenoid esters) extracted from three stem discs of Pinus sylvestris were assessed to investigate metabolic changes related to heartwood formation. The wood was separated into (1) cambial zone, (2) outer sapwood, (3) inner sapwood, (4) transition zone, (5) outer heartwood and 6) inner heart-wood. The fractions were separated by thin-layer chromatography (TLC) and analysed by gas-liquid chromatography (GLC). The amount of fatty acids of sapwood triacylglycerols was about 1.5% (dry wt.) but a large reduction occurred in the transition zone. In contrast, noticeable amounts of free fatty acids were present only in the heart-wood. The most important fatty acids in the sapwood fractions were 16:0, 18:0, 18:1, 18:2 (the dominant fatty acid in all fractions), 18:3 and 20:3. Together 18:1 and 18:2 formed about 70% of the total triacylglycerol fatty acids. Of the sterol/ triterpenoid esters, 18:2 and 18:3 were predominant. The fatty acid composition of all fractions changed in the transition zone. The sterols found were sitosterol, stigmastanol, campesterol and campestanol. The amount of sterol esters increased towards the heartwood, and the amount of free sterols was lowest in the inner sapwood. Sitosterol was the dominant sterol in both free sterols and sterol esters.     

Lipids and lipolytic enzymes in the trunkwood ofRobinia pseudoacacia L. during heartwood formation

The radial distribution of membrane and storage lipids was determined in the trunkwood ofRobinia pseudoacacia L. The trees were felled in November at the time of heartwood formation and fluctuations in the amount and composition of phospholipids, free sterols, steryl esters, diand triacylglycerols, and free fatty acids were investigated across the sapwood-heartwood boundary. The individual compounds were identified and quantified by thin layer chromatography, enzymatic and colorimetric assays, and by capillary gas chromatography. Phospholipids show a significant decrease towards the boundary area, and in the heartwood only trace amounts can be detected. The same pattern is observed for free sterols in the sapwood; in the heartwood, however, they reach maximum values with increasing depth of the trunk. Steryl esters exhibit a complementary behaviour by accumulating at the periphery of the heartwood. No concentration changes are found in the total amounts of diacylglycerols and free fatty acids. In contrast, the triacylglycerol concentration declines steadily across the trunk. With regard to qualitative composition, free fatty acids and the fatty acid moieties of the esterified constituents vary in their chain length from 14 to 24 carbon atoms and have up to three double bonds. A radial gradient in the ratio saturated/unsaturated fatty acids can be observed: except for the phospholipid fraction the relative amounts of unsaturated fatty acids increase in centripetal direction. Seven phospholipids were identified: phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, diphosphatidylglycerol, and phosphatidic acid, which constitutes the major proportion. In the sterol group, sitosterol is the most abundant component. The composition of the esterified sterols remains constant across the trunk cross-section, whereas the relative frequencies of individual free sterols change markedly.     

Variation in the amount of triacylglycerols and steryl esters in the outer sapwood of Pinus sylvestris L.

Variation in the amount of triacylglycerols and steryl esters was analysed enzymatically in the outer sapwood of Scots pine (Pinus sylvestris L.). Increment borings were taken at breast height from 40 stems of different diameter. Wood samples from outer sapwood (10 mm from the cambium) were extracted with acetone. Triacylglycerols and steryl esters were separated on TLC and the levels of glycerol and sterol were analysed enzymatically. The average amount of triacylglycerols and steryl esters was approximately 25 and 0.83 mg/g dry weight, respectively. However, variation between the stems studied was fairly large. The sapwood of young and small-diameter stems was not found to store larger amounts of either triacylglycerols or steryl esters than the sapwood of old stems. Neither was there any relationship between the amount of triacylglycerols and steryl esters in the outer sapwood. The possible role of these compounds in heartwood formation is discussed.     

Soluble carbohydrates in the nutrition of house longhorn beetle larvae, Hylotrupes bajulus (L.) (Col., Cerambycidae): from living sapwood to faeces

Abstract: Water-soluble carbohydrates from (a) living sapwood of Scots pine ( Pinus sylvestris L); (b) dry sapwood blocks from Scots pine (EUROPEAN STANDARD EN 46, 1988); and (c) faeces from Hylotrupes bajulus (L) larvae feeding from these blocks were analysed. Living sapwood contains high amounts of sucrose, glucose, fructose and starch. These carbohydrates seem to be hydrolysed and metabolized during the production of the sapwood blocks. The faeces contain carbohydrates which are not present either in living sapwood or in the sapwood blocks: xylose and an oligosaccharide of uncertain composition. This oligosaccharide is the dominant water-soluble carbohydrate in the faeces. In contrast to living sapwood and sapwood blocks, the faeces contain considerable amounts of soluble β-glucans. The results show that the composition and the amounts of carbohydrates differ significantly from that in the sapwood blocks (fodder for the larvae). The α-glucans (starch) present in the wood are not significantly utilized by the larvae. Regarding soluble carbohydrates, diet wood has only little in common with the living wood of the standing tree.     

Studies on Sugars and Starch in Light- and Dark-Germinated Seeds of Scots Pine (Pinus silvestris)

Stachyose, raffinose and sucrose have been tentatively identified in ungerminated Scots pine seeds with paper chromatography and TLC. After the onset of the germination process even glucose and fructose were detected. No free galactose has been found. The occurrence and distribution of starch has been investigated with histochemical technique. The same substances have been quantitatively studied during the first 24 hours of the germination in light and darkness. This was done with an enzymatic technique alone or in combination with TLC. The evidence of a light influence on the transformations of starch, glucose and fructose has been discussed in relation to the phytochrome mechanism.     

Resistance of Scots pine wood to Brown-rot fungi after long-term forest fertilization

The susceptibility of Scots pine (Pinus sylvestris L.) sap- and heartwood against the wood decaying brown-rot fungus (Coniophora puteana) was investigated after long-term forest fertilization at three different sites in central Finland. Different wood properties: wood extractives, wood chemistry, and wood anatomy were used to explain sap- and heartwood decay. Scots pine sapwood was more susceptible to decay than its heartwood. In one site, sapwood seemed to be more resistant to wood decay after forest fertilization whereas the susceptibility of heartwood increased. Significant changes in the sapwood chemistry were found between treatment and sites, however, no relationship between wood chemistry and wood decay was observed in the factor analysis. The results of this study show that there was an inconsistent relationship between decay susceptibility and fertilization and the measured physical and chemical attributes of the wood were not consistently correlated with the decay rate.     

Studies on the Carbohydrates of Horse Radish,Cochlearis Armoracia L

In the root of horse radish, sucrose, fructose, glucose and two unknown fructose-oligosac-charides were found in the form of free sugars, in the decreasing order just given, and large amounts of starch, hot 50% methanol-soluble polysaccharide, water-soluble polysaccharide, pectin, hemicellulose, cellulose and a small amount of lignin were found as polysaccharides. As the constructive sugar of these polysaccharides, the following, i.e., glucose, fructose, galactose, arabinose, galacturonic acid and xylose were detected.     

Estimating missing sapwood rings in three European gymnosperm species by the heartwood age rule

Precise dating of the year of felling is one intended outcome of dendrochronology. However, occasionally some or all sapwood rings might be missing, either due to deterioration or because they were carved off, or for some other reason. Consequently, while heartwood is preserved, sapwood might be fully or partially missing. In such cases, the year of felling must be estimated by adding a suitable number of sapwood rings. A heartwood age rule (HAR) has been advocated for Scots pine and adapted to European larch and Cembra pine, implying a linear relationship between sapwood ring count and the square root of heartwood ring count, largely irrespective of position in the stem. The same rule applied to all observations of a species, irrespective of silviculture, location or fertility of the growth site. Scots pine had twice or thrice as many sapwood rings as Cembra pine, which had 10% more rings than larch. The magnitude of model residuals was proportional to estimated sapwood ring count. Relative residuals were roughly normally distributed. To be applicable in Bayesian modelling in dendrochronology analyses, detailed information on model errors has been provided.     

Formation of heartwood substances in the stemwood of Robinia pseudoacacia L. II. Distribution of nonstructural carbohydrates and wood extractives across the trunk

Summary The distributions of reserve carbohydrates and of three dominant heartwood extractives were determined in the trunkwood of Robinia pseudoacacia L. The trees were cut at different times of the year (September, November, January, and April). With the exception of the tree felled in January, all trunks exhibited highest contents of nonstructural storage carbohydrates (glucose, fructose, sucrose, and starch) in the youngest, outermost sapwood zone. With increasing depth of the trunk, the levels of carbohydrates decreased. At the sapwood-heartwood transition zone, only trace amounts of nonstructural carbohydrates were present. The heartwood itself contained no storage material. The wood zones of different ages of the trees cut in September, November, and January exhibited glucose/fructose ratios of approximately 1. In April, however, there was a shift to glucose. In the youngest sapwood the amounts of soluble sugars were higher in the earlythan in the latewood. Older zones of the sapwood and the sap-wood-heartwood transition zone showed the opposite behaviour. Three main wood extractives of Robinia were characterized and quantified: the flavanonol dihydrorobinetin (DHR), the flavonol robinetin (ROB) and a hydroxycinnamic acid derivative (HCA). Only DHR was present — in very low amounts — in the younger sapwood of all trunks investigated. Higher amounts (>1 mol/g dry weight) of this compound and the HCA were present in the sapwood-heartwood transition zone. DHR augmented within the heartwood up to a more or less constant level. HCA increased towards the heartwood and decreased again in the inner heartwood parts. ROB appeared in the innermost parts of the sapwood-heartwood transition zone and reached maximum values in older parts of the heart-wood. The results indicate that starch is hydrolyzed at the sapwood-heartwood boundary and thus represents a primary major source of hydroxycinnamic acid and flavonoid synthesis.Dedicated to Prof. Meinhart H. Zenk on the occasion of his 60th birthday     

Fatty acids,starch and biomass of Scots pine needles and roots in open-air ozone exposure

Scots pine (Pinus sylvestris L.) seedlings were fumigated with 1.2–1.5 x ambient ozone (cumulative exposure) over 2 seasons in an open-air experiment. Starch and fatty acid concentrations were analyzed in needle and root tissue in the summer, autumn and early winter. Seedling growth was determined by measuring the height of the stem and the total shoot and root biomass. Significant decreases in growth were found in exposed seedlings, even though visible symptoms were lacking. Almost significant reductions in needle and root starch concentrations were found. In the ozone treated foliage, significant increases in myristic acid (140) were detected, but the major fatty acids remained unchanged. Fatty acid ratios showed that the degree of unsaturation decreased in treated needles in the summer. In the roots of ozone treated seedlings, changes in fatty acids were different from those in the foliage. Decreases of the main root fatty acids (160, 180, 181, 18:2, 183) were detected in the summer. These results show that Scots pine is susceptible to enhanced levels of ozone. If the tropospheric ozone levels continue to increase it may have deleterious effects on Scots pine forests in Finland.     

Concentration gradients of free sterols,steryl esters and lipid phosphorus in the trunkwood of Scot's pine (Pinus sylvestris L.)

The amounts of free sterols, steryl esters and lipid phosphorus were determined in the sapwood and heartwood of mature, and in the outer and inner sapwood of young Pinus sylvestris trees. In the mature trees (up to 70 years old) the heartwood contains significantly higher amounts of free sterols than the sapwood. No radial gradient can be demonstrated in the amounts of steryl esters. Lipids extracted from the sapwood contain higher amounts of phosphorus than those from the heartwood. Stems of young Pinus sylvestris trees (up to 13 years old) show in the inner sapwood higher amounts of both free sterols and steryl esters than the peripheral younger wood zone. The inner sapwood of the young stems shows slightly higher amounts of lipid phosphorus than the outer sapwood. The results indicate that Pinus sylvestris accumulates both free sterols and steryl esters in the stems at a very early stage of the life cycle. Sterol accumulation in the innermost parts of the stems seems not to depend on heartwood formation.     

The incorporation of [14C]palmitic acid into the lipids of mouse brain in vivo

Abstract— The half-life of free [¹⁴C]palmitic acid injected intracerebrally into C57BL/10J mice was less than 5 min. The rapid disappearance of radioactivity as palmitic acid was accompanied by increases in the radioactivity of the phosphatidic acids and the diacyl-glycerols. The peak specific radioactivity of the diacylglycerols occurred at about 6-8 min after injection. The triacylglycerols, phosphatidyl ethanolamines and phosphatidyl cholines exhibited increasing amounts of radioactivity during the first 40 min. At 160 min after injection, the distribution of radioactivity was similar to the pattern observed at 12 h. The biosynthetic pathway through the phosphatidic acids and the diacylglycerols to triacylglycerols, phosphatidyl ethanolamines and phosphatidyl cholines is apparently the major pathway in vivo for the esterification of free fatty acids in the brain.     

Seasonal changes of pinosylvin distribution in the sapwood/heartwood boundary of Pinus sylvestris

Changes in pinosylvin concentration and distribution across the sapwood/heartwood boundary were studied on Scots pine (Pinus sylvestris L.) tree stems to detect seasonal activity in heartwood formation. Pinosylvin concentrations were measured with FT-(NIR) Raman spectroscopy for a total of 96 trees equally distributed on 16 different sampling occasions. In another experiment, cores were sampled every month from six individual Scots pine trees from June to October and analysed for pinosylvin. There was a great between-tree variation in concentration and spatial distribution of pinosylvin. No seasonal trend in the distribution pattern or concentration of pinosylvin was found. Therefore, the results indicate that there is no specific period during the year when heartwood is formed. Received: 12 February 1998 / Accepted: 3 May 1999     

Lipid and fatty acid composition of the fat body during the female reproductive cycle of Labidura riparia (Insecta Dermaptera)

During the reproductive cycle of the female Labidura riparia, cytological observations show cyclical modifications of lipid droplets in the periovarian adipocyte. Fat body lipids and their constitutive fatty acids are analyzed. The lipids are predominantly triacylglycerols, which increase after adult ecdysis during vitellogenic and non-vitellogenic periods. Small amounts of diacylglycerols and phospholipids are found. Diacylglycerols increase during vitellogenesis and decrease during the non-vitellogenic period. Cytological modifications of lipid droplets are probably related to diacylglycerol fluctuations. Gas-liquid chromatography of fatty acid methyl esters shows oleic acid to be the predominant fatty acid in total lipids and triacylglycerols; unsaturated acids are approximately twice as abundant as saturated acids all along the reproductive cycle. Fatty acid composition of diacylglycerols and phospholipids differs from triacylglycerols and total lipids composition. Palmitic, stearic, oleic and linoleic acids represent the major fatty acids; their relative amounts vary during the different periods of the reproductive cycle. The correlations between fat body lipid changes and ovarian development were discussed and compared with observations made on other insect species. Accepted: 23 April 1997     

Influence of selected sugars and temperature on fatty acids composition in Candida lipolytica

Summary An investigation was made of the effects of temperature and of mono- and disaccharides on lipids, biomass, odd-chain and unsaturated fatty acids production of Candida lipolytica. With different sugars as carbon source at 30°C, the order for biomass production was: fructose > glucose > sucrose > lactose > galactose, while lipids production/g biomass decreased as follows: lactose, sucrose, galactose, fructose and glucose. On the other hand, the odd-chain fatty acids contents decreased in the following order: fructose, lactose, glucose, sucrose and galactose. Lowering the temperature of cultivation to 15°C, biomass, lipids and unsaturated fatty acids decreased. However, a notable decrease in the content of odd-chain fatty acids was detected.     

Effects of Fertilization and Irrigation on the Seasonal Changes of Carbohydrate Reserves in Different Age-Classes of Needle on 20-Year-Old Scots Pine Trees (Pinus silvestris)

The effects of fertilization, irrigation or both on the seasonal changes of starch and soluble carbohydrates (glucose, fructose, myo-inositol, pinitol and sucrose) in needles of 20-year-old Scots pine trees (Pinus silvestris L.) were studied during three consecutive years. The starch content of the mature needles increased during spring and early summer to about 25% of dry weight. Neither fertilization nor irrigation affected the general pattern of starch accumulation during the spring. The starch reserves were mobilized when the shoot started to grow. Starch content decreased more rapidly in needles from fertilized than in those from unfertilized trees. The current needles from the control trees accumulated starch while they were still growing. The current needles of the fertilized trees did so to a lesser extent. The amount of starch was closely correlated to the air temperature and to the growth rate. Large amounts were found at low temperatures and low growth rates. The concentrations of soluble carbohydrates showed the well-known seasonal variation, with the highest value during the winter. The levels of sugars were nearly similar, irrespective of fertilization. An exception was sucrose, which was found in small quantities in needles from fertilized plots. Small amounts of sucrose were also found in growing current needles. The results are discussed in relation to growth limitation by assimilate availability and indicate that the ‘sink demand’ is the limiting factor.