The electrical impedance spectroscopy of Scots pine needles during cold acclimation

The electrical impedance spectroscopy (EIS) was applied to current-year needles of Scots pine ( Pinus sylvestris L.) in an 8-year provenance field trial in central Finland during frost hardening. The EIS analysis of the needles using a Model-A equivalent circuit indicated a sequence of events in the needles during their cold acclimation. Some of the EIS-parameters referred to maturation phenomena occurring during the pre-hardening phase at the end of the growing season, and some parameters displayed a clear coincidence with the frost hardening itself. Significant differences between provenances were found in several of the Model-A parameters. Extracellular resistance (r_e) and β -coefficient decreased in all provenances in the pre-hardening phase in August and until mid-September. In the same phase, both the intracellular resistance (r_i) and the cell membrane time constant (τ_m) first increased and then decreased. According to τ_m, r_e and β there was a clear gradation between provenances in the pre-hardening phase. From the end of September significant differences were found in the intracellular resistance between provenances, corresponding with the differences in their hardening pattern. The dry weight (DW) content of needles increased during the study period but no clear differences were found between the provenances.     

Composition and positional distribution of fatty acids in polar lipids from Chlorella ellipsoidea differing in chilling susceptibility and frost hardiness

The composition and positional distribution of fatty acids in the polar lipids from 4 strains of Chlorella differing in chilling susceptibility and frost hardiness were analyzed by enzymatic hydrolysis and gas-liquid chromatography. Analysis of the polar lipids from chilling-sensitive, chilling-resistant and chilling-sensitive revertant strains of Chlorella ellipsoidea IAM C-102 showed that the sum of palmitic and trans -3-hexadecenoic acid in phosphatidylglycerol (PG) is about 60% for the sensitive strains and 53% for the resistant strain. The sum of dipalmitoyl and 1-palmitoyl-2-( trans -3-hexadecenoyl) PG as estimated from the positional distribution of their fatty acids, is about 10% in the case of each of the three strains. The contents of unsaturated fatty acids in phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were higher in the resistant than in the sensitive strain. This suggests that unsaturation of fatty acids in not only PG but also PC and PE is involved in chilling sensitivity of Chlorella . On the other hand, lipid changes during the development of frost hardiness of C. ellipsoidea IAM C-27, a frost hardy strain, were examined. The results showed that fatty acids in most lipid classes are unsaturated in the hardening process but their degree of unsaturation is not greatly different from that of the chilling-resistant strain, suggesting that not only unsaturation of fatty acids in lipids but also other factors are necessary for the development of frost hardiness.     

Effect of nitrogen supply on frost resistance, nitrogen metabolism and carbohydrate content in white clover (Trifolium repens)

Effects of mineral nitrogen (2, 4, 6 and 8 m M NH₄NO₃) and nodulation with Rhizobium on frost hardiness in seedlings of white clover ( Trifolium repens ) have been studied. Seedlings of a population from Bodø (67°N lat.) were grown in Leonard jars under controlled conditions in a phytotron. For induction of frost hardening, plants were first exposed to 12 h photoperiod conditions for 2 weeks at 18°C, then for 2 weeks at 6°C and finally for 2 weeks at 0.5°C. Frost hardiness after treatments at 6 and 0.5°C was significantly enhanced by increasing nitrogen supply and was positively correlated with total nitrogen content of the stolons. Frost hardiness of nodulated plants correlated to the tissue nitrogen concentration. Content of soluble proteins in stolons decreased during hardening at 6°C but did not change during treatment at 0.5°C. There were minor changes in total amount of free amino acids during hardening. Both absolute and relative amounts of proline and arginine increased, and those of asparagine decreased during hardening. Absolute amounts of all free amino acids increased with increasing nitrogen supply, but the changes during hardening were similar in all treatments. There was a significant increase in the content of soluble carbohydrates during hardening. However, this increase was inversely related to nitrogen supply.     

Effects of hardening and freezing stress on membrane lipids and CO2 fixation of Ceratodon purpureus protonemata

Membrane lipids and steady-state CO₂ fixation rates were studied in moss protonemata in order to evaluate separately the effects of growth temperature, freezing stress and the achievement of frost hardiness. Protonemata of Ceratodon purpureus (Hedw.) Brid, were grown at 20 and 4°C and parts of both materials were then hardened. The low growth temperature increased the content and unsaturation level of membrane lipids significantly. This did not, however, cause a noticeable increase in the frost hardiness. Nor was the achievement of frost hardiness in this material accompanied by further changes in the amount or unsaturtion level of any membrane lipid class. Cytoplasmic membranes were abundant in both unhardened and hardened materials grown at 4°C, which agreed with the high phospholipid content of these protonemata. The only significant difference in membrane lipids between unhardened and hardened materials was a 50% lower level of trans 16:1 fatty acid in the phosphatidylglycerol fraction of hardened protonemata.
In hardened protonemata monogalactosyldiacylglycerol (MGDG) was the membrane lipid most liable to decrease during the freeze-thaw cycle. The loss of MGDG was accompanied by partial inhibition of CO₂ fixation. Provided the content of phospholipids remained unchanged (freeze-thaw cycle with – 10°C in hardened protonemata), this inhibition was mostly reversible. If loss of the phospholipids also had occurred during the freeze-thaw cycle, as was the case in unhardened material at or below -10°C, CO₂ fixation was severely and nearly irreversibly inhibited after thawing.     

Vegetative growth and frost hardiness of cloudberry (Rubus chamaemorus) as affected by temperature and photoperiod

The effect of photoperiod and temperature on growth and induction and development of frost hardiness in cloudberry ( Rubus chamaemorus L.) was examined in two experiments. The photoperiods were 8, 12 or 24 h and the temperatures were 18, 15, 12, 9, 4, 3, –3 or –4°C depending on the experiment. The level of hardiness was expressed as LT₆₆ or LT₅₀ (the lethal temperature for 66 or 50% of the plant material) for percentage of bud break and for the degree of coloring by triphenyltetrazolium chloride for rhizomes. The vegetative growth was clearly affected by daylength; petiole elongation, leaf growth, shoot dry weight and number of shoots per plant were all reduced under short days compared with long days. However, the photoperiod had no significant effect on hardening of buds or rhizomes. Hardening increased with successively decreasing temperatures. To get the maximum hardiness, plants had to be exposed to freezing temperatures.     

Accumulation of Free Fatty Acids during Hardening of Chlorella ellipsoidea

Chlorella ellipsoidea Gerneck (IAM C-27) was synchronously grown, and cells at an intermediate stage in the ripening phase of the cell cycle were hardened at 3°C for 48 hours. A nonpolar lipid which increased greatly during hardening was analyzed by gas-liquid chromatography. Palmitic, oleic, linoleic, and linolenic acids were the main components of the lipid. Electron micrographs revealed the appearance of lipid bodies in hardened cells. When formation of free fatty acids and lipid bodies was inhibited with cycloheximide, oligomycin, and 3-(3,4-dichlorophenyl)-1,1-dimethylurea, the development of a high level of hardiness was always inhibited. However, the converse results were not always realized. Cells hardened in the dark in the absence of glucose developed a measurable hardiness in spite of their failure to form free fatty acids. The appearance of lipid bodies was invariably accompanied by the formation of the fatty acids. In pulse-labeling with [¹⁴C]NaHCO₃ for 4 minutes at zero time and at the 12th hour of hardening, initial incorporation rates of ¹⁴C into total lipids of whole cells and the cellular membrane fraction were significantly higher than that into free fatty acids. These results suggest that, although fatty acids are inserted into membrane lipids during hardening, the accumulation of free fatty acids and the appearance of lipid bodies per se are not involved in the development of frost hardiness.     

Changes in fatty acid composition of winter wheat during frost hardening

Twelve day old winter wheat seedlings (cvs Kharkov, frost hardy and Champlein, less hardy) accumulated linolenic acid at the expense of linoleic acid during controlled hardening. The change was most pronounced in the roots, where it was not specific to the phospholipid fraction. It was less marked in the leaves, but occurred there mainly in the phospholipids. The lack of differences between fatty acid profiles of the two cultivars rules out the explanation of varietal differences in frost hardiness in winter wheat on the basis of major changes in fatty acid unsaturation.     

Temperature-dependent changes in phospholipid and fatty acid composition and membrane lipid fluidity of Yersinia enterocolitica

Temperature-dependent compositional changes of phospholipids and their fatty acids were analysed in Yersinia enterocolitica grown at 5°, 25° and 37°C. The relative amounts of the four phospholipids, phosphatidylethanolamine (75–78%), phosphatidylglycerol (10–11%), cardiolipin (<7%) and lysophosphatidylethanolamine (<5%), were essentially the same at all growth temperatures. The degree of fatty acid unsaturation of the four phospholipids increased with decrease in growth temperature, mainly due to an increase of C_16:1 and C_18:1 and a corresponding decrease of C_16;0, C_18:0 and cyclo C_17:0. An electron spin resonance spectroscopic study of the membrane lipids showed that membrane lipid fluidity was enhanced by decreasing the growth temperatures. The changes in fatty acid composition of phospholipids in response to varied temperatures were consistent with the temperature-dependent changes in the membrane lipid fluidity of Y. enterocolitica , and were similar to those reported for other bacteria.     

Specific features of source-sink relations in alloplasmic hybrid of winter wheat with alien cytoplasm of goatgrass with emphasis on resistance to low temperature stress

We studied the influence of alien cytoplasm of spring goatgrass Aegilops ovata L. on some physiological parameters in winter wheat (Triticum aestivum L.), Mironovskaya 808, under normal conditions and in the case of modified source-sink relations. Measurements of relative rates of plant dry matter growth and its distribution among organs, CO₂ exchange (photosynthesis upon light saturation and dark respiration), content of sugars (sucrose + glucose + fructose) and their ratio in leaves, frost hardiness, and indices of membrane stability and damage of leaves by frost have shown that, on average, alloplasmic hybrid differed from the initial cultivar by almost all parameters. Reduced frost hardiness, increased index of leaf damage by frost, lowered leaf content of sugars, and reduced sucrose/(glucose + fructose) ratio in the alloplasmic hybrid were combined with higher roots/leaves ratio, relative rate of dry matter growth, and photosynthesis and respiration rates. The alloplasmic hybrid was more tolerant to decreased source strength in source-sink relations as compared to the initial cultivar.     

Effects of Hormones on Morphogenesis and Cold Resistance in Berseem Clover (Trifolium alexandrinum L.)

Plants of berseem clover (Trifolium alexandrinum L.) cv. Taborwere raised under conditions inhibiting the acquisition of coldhardiness (non-hardened) or inducing cold hardiness (hardened).All non-hardened plants developed an elongated shoot and exhibitedconsiderable frost sensitivity, as measured by the extent ofthe reduction in yield of variable chlorophyll fluorescenceafter exposure to sub-zero temperature. Hardened plants developeda shorter shoot, with fewer leaves and a greater percentageof dry matter in the root system. These parameters were associatedwith a marked increase in frost resistance. Exogenous applicationof ABA to plants effected similar morphological modificationsin both hardening and non-hardening temperature regimes; plantsdeveloped a shorter primary shoot axis and leaves exhibiteda marked increase in frost hardiness. In berseem clover ABAcan thus substitute, at least partially, for the low temperaturetreatment required to induce cold hardiness. Spraying plantsraised under hardening conditions with gibberellic acid reversedthe effects of the hardening treatment, since they developedan elongated shoot and exhibited frost sensitivity comparableto non-treated plants grown under non-hardening conditions.It is concluded that these endogenous hormones are directlyinvolved in triggering changes in morphogenesis which accompanyphysiological and metabolic events associated with the inductionof plant cold hardiness. Key words: Frost resistance, morphogenesis, abscisic acid, giberellic acid, Trifolium alexandrinum     

Chemical and Biophysical Changes in the Plasma Membrane during Cold Acclimation of Mulberry Bark Cells (Morus bombycis Koidz. cv Goroji)

The lipid and protein composition of the plasma membrane isolated from mulberry (Morus bombycis Koidz.) bark cells was analyzed throughout the cold acclimation period under natural and controlled environment conditions. There was a significant increase in phospholipids and unsaturation of their fatty acids during cold acclimation. The ratio of sterols to phospholipids decreased with hardiness, primarily due to the large increase in phospholipids. The fluidity of the plasma membrane, as determined by fluorescent polarization technique, increased with hardiness. Electrophoresis of plasma membrane proteins including glycoproteins revealed change in banding pattern during the early fall to winter period. Some of the protein changes could be related to growth cessation and defoliation. However, minor changes in proteins also occurred during the most active period of hardening. Changes in glycoproteins were coincident both with changes in growth stages and with the development of cold hardiness.     

Altered Composition of Cerebral Microvessel Membrane Phosphoglycerides from Senescent Mouse

Abstract— Phosphoglyceride and fatty acid composition was determined in the cellular membranes of isolated cerebral microvessels and brain parenchymal cells (neurons and glia) taken from 10-, 20-, and 27–30-month-old C57BL6/NNIA mice. Lipids were extracted from each fraction and the fatty acid profiles of ethanolamine, cho-line, serine, and inositol phosphoglycerides analyzed by gas chromatography. The results suggest that membrane phosphoglycerides from cerebral microvessels are significantly more affected by the aging process than are those of the brain parenchyma. Relative percentage for fatty acids in cerebral microvessels indicate an overall decline in membrane unsaturation with a concomitant elevation in the level of saturation. The decline in unsaturation is reflected primarily in the loss of precursor fatty acids for arachidonic (18:2n-6 and 20:3n-6) and docosahexaenoic (20:5n-3 and 22:5n-3) acids. Levels of arachidonic (20:4n-6) and docosahexaenoic (22:6n-3) acids in each phos-phoglyceride remained unchanged with age; however, mol% for ethanolamine plasmalogen, a major source of these fatty acids, was significantly reduced in 27–30-month-old mice. Conversely, mol% for choline phospho-glyceride increased with age. The age-related changes in fatty acid profile for microvessel membrane phosphoglycerides are reflected by increased saturation/unsaturation ratios and decreased unsaturation indices. These parameters were not affected by aging in parenchymal membranes.     

Simultaneous Inhibition of Linolenic Acid Synthesis in Winter Wheat Roots and Frost Hardening by BASF 13-338, a Derivative of Pyridazinone

Treatment of 12-day-old winter wheat (Triticum aestivum) plants with BASF 13-338 {4-chloro-5 (dimethylamino)-2-phenyl-3(2H)-pyridazinone} 36 hours before frost hardening simultaneously and completely inhibits accumulation of linolenic acid in the roots during the hardening period and the acquisition of frost resistance. Increased unsaturation of fatty acids is therefore probably an important part of the mechanism of cold adaptation in winter wheat.     

Lipids,amino acids,sugars, hardiness and growth of Festuca arundinacea

Tall fescue (Festuca arundinacea Schreb. cv S.170) plants were grown in environments differing only in temperature: 6/4, 16/14 or 21/19°. The content of total and individual sugars and amino acids in leaf laminae and roots did not relate closely to the hardiness of the organ. The unsaturation of lipid fatty acids alone was clearly unrelated to hardiness because the difference in unsaturation was greater in the neutral lipids, glycolipids and phospholipids from roots than from leaves but only the latter differed in hardiness. Total amounts of lipids could have been related to hardiness but phospholipids and glycolipids in the roots were not. At least some of these changes may be related to adaptation of growth to temperature.     

水杨酸对大叶黄杨茎抗寒性和电阻抗图谱参数的影响

在抗寒锻炼前,对当年生大叶黄杨(Euonymus japonicus)扦插苗喷施不同浓度水杨酸,用电阻抗图谱(EIS)法和电导(EL)法估测茎的抗寒性,以探明抗寒锻炼期间水杨酸对大叶黄杨抗寒性和电阻抗图谱参数的影响,找到适合不经冷冻处理估测大叶黄杨茎抗寒性和经冷冻处理后测定其抗寒性的EIS参数。结果表明,水杨酸处理能够提高大叶黄杨茎的抗寒性,最适浓度为5.0mmol.L-1;不经冷冻处理茎的EIS参数电阻率r和r1、胞外电阻率re、胞内电阻率ri、弛豫时间τ、弛豫时间分布系数ψ与EL法测定的抗寒性有较高的相关性(r=0.70~0.87),说明不经冷冻处理样本用以上参数估测大叶黄杨茎抗寒性是可行的,r1为最佳参数;冷冻处理后茎的re、τ、ri求得的抗寒性与EL法测定的抗寒性有较高的相关性(r=0.85~0.94),说明冷冻处理后re、τ、ri可以作为测定大叶黄杨茎抗寒性的参数,re为最佳参数。     

Seasonal Changes of Fatty Acid Compositions in Overwintering Larvae of Rice Stem Borer,Chilo suppressalis (Lepidoptera: Pyralidae)

Cold acclimation and overwintering state can affect fatty acid compositions of insects. To determine compositional change of fatty acids during nondiapause and diapause stages, an experiment was conducted to investigate fatty acid constituents from whole body of C. suppressalis larvae. Five most abundant fatty acids were found to be palmitoleic (35–58%), palmitic (18–44%), oleic (14–23%), stearic (0.5–2.5%) and linoleic acid (0.4–2%). However, linolenic, erucic, lauric and myristic acid were found at lower level. Saturated fatty acids significantly decreased and conversely unsaturated fatty acids increased from August (pre-diapause) to October (initiation of diapause). The increase in seasonal cold hardiness during cold acclimation, exposed at −15°C for 24 h, was related to degree of fatty acid unsaturation. The elevation of palmitoleic acid content at low temperature resulted in an increase in the overall degree of unsaturation in the whole body. These results indicated the importance of unsaturated fatty acids composition to prepare larvae entering diapause phase.     

Fatty acid and lipid compositions of Conidiobolus

The fatty acid composition of the total lipids from two Conidiobolus species was studied by gas—liquid chromatography. The major fatty acids of C. lamprauges were palmitic acid (C_16:0), oleic acid (C_18:1), linolenic acid (C_18:3), and arachidonic acid (C_20:4). For C. eurymitus , myristic acid (C_14:0), C_16:0, and linoleic acid (C_18:2) were the most abundant acids. The fatty acid composition of C. eurymitus was quite different from that of the Conidiobolus species as mentioned in other reports. The lipid composition of the total lipids of C. lamprauges and C. eurymitus was also studied by thinchrography on quartz rods. Triglycerides and phospholipids were the major components in the two Conidiobolus species.     

The Effect of Temperature on the Fatty Acid Composition of Tetrahymena pyriformis WH-14

SYNOPSIS. A reduction in the growth temperature of Tetrahymena pyriformis strain WH-14 from 35 C to 15 C resulted in distinct alterations in the fatty acid composition of the glycerophospholipids. The proportion of normal saturated acids declined from 26 to 19%; palmitoleic acid increased by 6%, and the composition of the polyunsaturated fatty acids increased in 18:2 Δ^6,11(n) and decreased in 18:2 Δ^9,12(n) and 18:3 Δ^6,9,12(n). The unsaturation index (the average number of double bonds/100 molecules) did not change with a shift in temperature.
Two biosynthetic pathways exist in Tetrahymena for the formation of unsaturated fatty acids. The observed changes in fatty acid composition that accompany a lowering of the environmental temperature can be accounted for by a reduction in the accumulation of products of the fatty acid pathway leading to the formation of γ-linolenic acid [16:0(n) → 18:0(n) → 18:1 Δ⁹(n) → 18:2 Δ^9,12(n) → 18:3 Δ^6,9,12(n)] and an increase in the components of the pathway leading to the formation of 18:2 Δ^6,11(n) [16:0(n) → 16:1 Δ⁹(n) → 18:1 Δ¹¹(n) → 18:2 Δ^6,11(n)]. The data suggest that the regulatory mechanism in Tetrahymena differs from that found in some bacteria where a simple substitution of unsaturated fatty acids for saturated fatty acids occurs at low culture temperatures.     

Effect of non-target cells on the sensitivity of the PCR for Escherichia coli O157 : H7

The fatty acid composition from mycelia of Streptomyces hygroscopicus strains was studied. A significant proportion of C_{18 : 2} was found in cultures. High levels of C_{16 : 0}, iso-C_{16 : 0} and C_{18 : 1} were also detected in all S. hygroscopicus strains. The different representatives of S. hygroscopicus had almost the same proportion of unsaturated fatty acids. Certain shifts in the amount of iso, anteiso and straight-chain fatty acids in some cultures were revealed. This might be explained by the adaptation capability of strains belonging to one species to form a variety of available fatty acids determined by particular cell membrane composition favouring certain antibiotic biosynthesis.     

LIPID CLASS DISTRIBUTION OF HIGHLY UNSATURATED LONG CHAIN FATTY ACIDS IN MARINE DINOFLAGELLATES

The very long chain highly unsaturated C₂₈ fatty acids, octacosaheptaenoic [28:7(n-6)] and octacosaoctaenoic acid [28:8(n-3)], were found to be associated with phospholipids, obtained by fractionation of total lipid extracts into distinct lipid classes, in 4 and 6, respectively, of 16 examined dinoflagellates. An interfraction comparison of fatty acids associated with phospholipids and glycolipids has also shown that the phospholipid fractions contained the majority (over 75% in 12 of 16 strains) of docosahexaenoic acid [22:6(n-3)] and traces of tetracosanoic acid (24:0). By contrast, the highly unsaturated C₁₈ fatty acids octadecatetraenoic [18:4(n-3)] and octadecapentaenoic acid [18:5(n-3)] were primarily recovered from a chloroplast-associated glycolipid fraction comprised of monogalactosyldiacylglycerol, digalactosyldiacylglycerol, and sulfoquinovosyldiacylglycerol. In 12 of 16 strains, an interfraction comparison showed that over 90% of 18:5(n-3) was found to be associated with glycolipids. These findings indicate that the C₂₈ fatty acids are located and probably synthesized in the cytoplasm or in an organelle other than the chloroplast, possibly with 22:6(n-3) and 24:0 as precursors, whereas the C₁₈ fatty acids 18:4(n-3) and 18:5(n-3) are glycolipid constituents apparently synthesized within the chloroplast. The function(s) of these C₂₈ fatty acids as components of phospholipids in cellular membranes is currently unknown.