Alkaloid variability in Leucojum aestivum from wild populations

Leucojum aestivum (summer snowflake) is a plant species used for the extraction of galanthamine, an acetylcholinesterase inhibitor for the treatment of Alzheimer's disease. Extracts from bulbs collected from 18 Bulgarian populations and from shoot-clumps obtained in vitro from 8 different populations showed variations in their alkaloid composition. Nineteen alkaloids were detected in the studied samples by GC-MS. Typically, the alkaloid fractions of L. aestivum bulbs were dominated by galanthamine type compounds, but lycorine, haemanthamine and homolycorine type alkaloids were also found as dominant compounds in some of the samples. Extracts from the shoot-clumps obtained in vitro were found to contain galanthamine or lycorine as main alkaloids. The galanthamine content ranged from 28 to 2104 microg/g dry weight in the bulbs, and from traces to 454 microg/g dry weight in the shoot-clumps.     

Lipids of the tropical seagrass Thallassia hemprichii

The component sterols, alcohols, hydrocarbons, monocarboxylic, α,ω-dicarboxylic and α- and ω-hydroxy acids from the leaves and roots of the tropical seagrass Thallassia hemprichii are reported. The leaves contained significant concentrations of cholest-5-en-3β-ol, a sterol not normally detected in either higher plants or seagrasses. The lower abundance of polyunsaturated fatty acids found in both the leaves and roots compared to other seagrass species may be a result of the warmer waters from which this species was collected. Solvent-extractable, long-chain (> C₂₂)α,ω-diacids, α- and ω-hydroxy and monocarboxylic acids were also isolated from the leaves. The distribution pattern of these lipids should enable these components along with other distinctive components to be used as chemical markers for this seagrass.     

Epicuticular wax composition in relation to aphid infestation and resistance in red raspberry (Rubus idaeus L.)

Epicuticular waxes from the aphid-resistant red raspberry (Rubus idaeus) cultivar Autumn Bliss and the aphid-susceptible cultivar Malling Jewel were collected from the newly emerging crown leaves, and also from the group of four more mature leaves immediately below the crown. Resistance and susceptibility status of the leaves to infestation by the large raspberry aphid, Amphorophora idaei, were determined by bioassay with the insect just prior to collection of the wax. Analysis showed the waxes to consist of a complex mixture of free fatty acids; free primary alcohols and their acetates; secondary alcohols; ketones; terpenoids including squalene, phytosterols, tocopherol and amyrins; alkanes and long chain alkyl and terpenyl esters. Compositional differences which may relate to A. idaei-resistance status were noticeably higher levels of sterols, particularly cycloartenol, together with the presence of branched alkanes, and an absence of C₂₉ ketones and the symmetrical C₂₉ secondary alcohol in wax from the resistant cultivar Bliss. There were also differences between the cultivars in the distribution of individual amyrins and tocopherols and in the chain length distribution for homologues of fatty acids, primary alcohols and alkanes, and these may also be related to resistance to A. idaei. Emerging leaves had lower levels of primary alcohols and terpenes, but higher levels of long-chain alkyl esters, and in general, more compounds of shorter chain-length than the more mature leaves. During bioassay A. idaei displayed a preference to settle on the more mature leaves. This may be due to greater wax coverage and higher levels of the compounds of shorter chain length found in the newly emerged younger leaves at the crown of the plant.     

Chromosome variations in regenerants of Arabidopsis thaliana derived from 2- and 6-week-old callus detected using flow cytometry and FISH analyses

Shoot organogenesis was induced from 2- and 6-week-old callus derived from the leaves of Arabidopsis thaliana ecotype Columbia (2n = 10). Regenerated plants were evaluated for chromosomal variations by means of flow cytometry and fluorescent in situ hybridization (FISH). Flow cytometric measurements revealed the occurrence of diploid, tetraploid, and octoploid plants among the regenerants of 2-week-old calli, whereas only diploid and tetraploid plants were regenerated from the 6-week-old calli. Chromosome counting showed that plants developed from the 2-week-old calli exhibited mixoploidy and a high frequency of aneuploid cells. These plants were infertile and displayed altered morphology. FISH with 5S and 25S rDNA probes allowed to detect some structural chromosomal rearrangements in regenerated plants. Along with cells which exhibited correct localisation of rDNA loci, also cells bearing chromosomal translocations, deletions or duplications were found. The type of structural aberrations varied between diploid and tetraploid regenerants.     

Epicuticular waxes of two sorghum varieties

The epicuticular waxes of the two sorghum varieties Alliance A and SD 102 have been analyzed, after separation of the leaf blades from the sheaths. The major constituents were found to be free fatty acids but small amounts of esters, aldehydes, alcohols, n-alkanes and sterols were also detected. The typical chain lengths of aldehydes, free alcohols and free fatty acids were C₂₈ and C₃₀.     

Sterols and fatty acids from three species of mangrove

The hydrolysis of steryl esters on thin-layer chromatographic plates by porcine pancreatic lipase is described. The sterols and fatty acids produced were separated on the same plate, recovered, and analysed by gas-liquid chromatography for their compositions. Synthetic cholesteryl esters containing various saturated and unsaturated fatty acids and synthetic steryl oleates with various sterols were lipolysed along with steryl esters of Acanthus ilicifolius, Bruguiera gymnorhiza and Rhizophora mucronata mangrove leaves. The major sterol was sitosterol which was accompanied by cholesterol, campesterol, stigmasterol and 28-isofucosterol. In addition, stigmast-7-en-3β-ol was present in R. mucronata leaves. The component fatty acids found in all three species were 16:0, 18:0, 18:1, 18:2 and 18:3. The relative proportions of the sterols and fatty acids were significantly different from the chemotaxonomic standpoint. The results obtained by carrying out plate lipolysis for 45 min at 40° compared well with those produced by conventional chemical hydrolysis.     

Free and ester-bound triterpene alcohols and sterols in cellular subfractions of Calendula officinalis flowers

In the chromoplast fraction and in the chromoplast-free fraction, obtained from Calendula officinalis ligulate flowers, the contents of individual free and ester-bound triterpene alcohols and sterols as well as the fatty acid components of the ester form were determined. It was shown that all sterols and triterpene monols in both forms occur in the two subtractions investigated, whereas all diols are localized only in the chromoplast fraction. The compositions of the fatty acids esterifying monols and sterols were similar to those esterifying diols in the chromoplasts. However, the fatty acids esterifying extra-chromoplast monols and sterols were different. This result indicates that triterpene monol esters are substrates for the biosynthesis of 3-monoesters of diols.     

Antioxidant activity and phenol content of Crithmum maritimum L. leaves

Sea fennel (Crithmum maritimum L.) is an edible halophyte with various economical interests because of its high secondary metabolite content. However, little is known about water-soluble compounds in that species. Here, we have studied major solutes in C. maritimum leaves. Among these solutes, carbohydrates (sucrose, glucose) were the most abundant, followed by organic acids (malate and quinate) and a phenolic compound never described in a halophyte before: chlorogenic acid (CGA). Total phenols and chlorogenic acid contents were followed throughout one year, as well as antioxidant activity, in two populations of C. maritimum growing in contrasting habitats: sand hills and cliffs. Sea fennel leaves appeared to be rich in phenolic compounds, particularly in chlorogenic acid. On that point, differences between the two populations were found, sand hill plants accumulating more CGA than those growing on cliffs. Moreover, the former presented a higher radical-scavenging activity, and the two observations were positively correlated. These results indicate that sea fennel can be considered as a valuable source of antioxidant products, especially of chlorogenic acid.     

Neutral lipids of leaves and stems of Trifolium repens

The neutral lipids of white clover leaves and stems have been separated into wax esters, free fatty acids, free fatty alcohols, free sterols, triglycerides and hydrocarbons. The wax esters were mainly of C₁₈ di- and tri-unsaturated fatty acids and C₃₀ fatty alcohol. Linolenic acid was the predominant free fatty acid and triacontanol was the principal free fatty alcohol. Of the hydrocarbons, C₂₉ and C₃₁ were present in the largest amounts.     

Growth requirements ofArabidopsis thaliana crown galls

Crown galls induced onArabidopsis thaliana plants by octopine or nopaline strains ofAgrobacterium tumefaciens were grownin vitro on different media. Dark growth of all tumor tissues was strictly hormone-dependent. In contrast, hormonal autonomy was observed in the light where crown gall calli readily differentiated into teratomas and (sometimes fertile) plants. Differentiating tissues always grew more vigorously than subtended calli. The growth of transformed calli was stimulated by vitamins and partly inhibited by growth regulators in concentrations used for the maintenance of untransformed calli. Crown gall calli, teratomas and sometimes regenerated plants were shown to express lysopine or nopaline dehydrogenase activities.     

Foliar Fatty Acids and Sterols of Soybean Field Fumigated with SO(2)

Sixty-day-old soybean plants were exposed in the field to 78.7 parts per one-hundred million of SO₂ in an open-air fumigation system for 20 days. Leaves from the top one-fourth and bottom one-fourth of the plants were analyzed for chlorophyll, free fatty acids, fatty acid esters, polar lipid fatty acids, and sterols. Fumigated plants had a lower chlorophyll, free fatty acid, and polar lipid content, but a higher fatty acid ester content. Of the individual fatty acids, linoleic and linolenic acid increased with SO₂ fumigation while palmitic acid decreased. SO₂ fumigations had only a minor effect on leaf sterols. In general, the lower, more mature leaves showed a greater response to SO₂ exposure.     

Effectiveness of the fatty acid and sterol composition of seeds for the chemotaxonomy of Coffea subgenus Coffea

The chemotaxonomic relationships between Coffea (subgenus Coffea) species have been poorly studied to date and the compounds tested so far - chlorogenic acids, diterpenoids and purine alkaloids - did not enable the establishment of phylogenetic relationships analogous to those revealed by chloroplast and nuclear DNA studies. In the present study, the relationships between African Coffea species were assessed on the basis of their seed lipid composition. Fatty acids and sterols were determined in 59 genotypes belonging to 17 distinct Coffea species/origins. Principal Component Analysis of fatty acid and sterol data enabled easy identification of the few species for which one or several compounds could serve as a quantitative signature. Hierarchical Clustering classified the Coffea species in seven groups with both fatty acids and sterols. However, while groupings based on seed fatty acid composition showed remarkable ecological and geographical coherence, no phylogeographic explanation was found for the clusters retrieved from sterol data. When compared with previous phylogenetic studies, the groups deduced from seed fatty acid composition were remarkably congruent with the clades inferred from nuclear and plastid DNA sequences.     

Regulation of Sterol Content in Membranes by Subcellular Compartmentation of Steryl-Esters Accumulating in a Sterol-Overproducing Tobacco Mutant

The study of sterol overproduction in tissues of LAB 1-4 mutant tobacco (Nicotiana tabacum L. cv Xanthi) (P. Maillot-Vernier, H. Schaller, P. Benveniste, G. Belliard [1989] Biochem Biophys Res Commun 165: 125-130) over several generations showed that the overproduction phenotype is stable in calli, with a 10-fold stimulation of sterol content when compared with wild-type calli. However, leaves of LAB 1-4 plants obtained after two steps of self-fertilization were characterized by a mere 3-fold stimulation, whereas calli obtained from these plants retained a typical sterol-overproducing mutant phenotype (i.e. a 10-fold increase of sterol content). These results suggest that the expression of the LAB 1-4 phenotype is dependent on the differentiation state of cells. Most of the sterols accumulating in the mutant tissues were present as steryl-esters, which were minor species in wild-type tissues. Subcellular fractionation showed that in both mutant and wild-type tissues, free sterols were associated mainly with microsomal membranes. In contrast, the bulk of steryl-esters present in mutant tissues was found in the soluble fraction of cells. Numerous lipid droplets were detected in the hyaloplasm of LAB 1-4 cells by cytochemical and cytological techniques. After isolation, these lipid granules were shown to contain steryl-esters. These results show that the overproduced sterols of mutant tissues accumulate as steryl-esters in hyaloplasmic bodies. The esterification process thus allows regulation of the amount of free sterols in membranes by subcellular compartmentation.     

Lipid Composition of Citrus Leaves from Plants Resistant and Susceptible to Citrus Bacterial Canker

The contents and composition of lipids in citrus leaves in relation to their general resistance to infection by strains of Xanthomonas campestris pv. citri (Xcc) were determined. The composition and contents of total polar lipids and phospholipids and the degree of fatty acid unsaturation were significantly different between resistant and susceptible species. Leaves from resistant plants had less phospholipids, but more free sterols than those from susceptible plants. The predominant fatty acids in the phospholipids were palmitic (16:0), linoleic (18:2) and α-linolenic acid (18:3). The degree of fatty acid unsaturation was higher in susceptible plants than in resistant plants. Major phospholipids in citrus leaves were phosphatidylchloline (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), and phosphatidylinositol (PI). β-Sitosterol, campesterol and lanosterol were major sterols in the leaves of citrus plants with resistant species having a higher ratio of free sterols to total phospholipids than susceptible species. Differences in lipid metabolism may contribute to differences in Xcc-resistance of citrus leaves.     

Plant cells express telomerase activity upon transfer to callus culture, without extensively changing telomere lengths

Changes in telomere lengths and telomerase activity in tobacco cells were studied during dedifferentiation and differentiation; leaf tissues were used to initiate callus cultures, which were then induced to regenerate plants. While no significant changes in the range of telomere lengths were observed in response to dedifferentiation and differentiation, there was a conspicuous increase in telomerase activity in calli compared to the source leaves, where the activity was hardly detectable. In leaves of regenerated plants, the telomerase activity fell to almost the same level as in the original plant, showing on the average 0.04% of the level in callus. The process was then repeated using the regenerants as the source material. In the second round of dedifferentiation and differentiation, telomerase activity showed a similar increase in calli derived from regenerated plants and a drop in plants regenerated from these calli. Telomere lengths remained unchanged both in calli and in leaves of regenerants. The conservation of telomere lengths over repeated rounds of dedifferentiation and differentiation, which are associated with dramatic changes in cell division rate and corresponding variation in telomerase activity may reflect the function of a regulatory mechanism in plant cells which controls telomerase action to compensate for replicative loss of telomeric DNA. Received: 31 July 1998 / Accepted: 21 September 1998     

Occurrence of Plant-Uncoupling Mitochondrial Protein (PUMP) in Diverse Organs and Tissues of Several Plants

The presence of plant-uncoupling mitochondrial protein (PUMP), previously described by Vercesi et al. (1995), was screened in mitochondria of various organs or tissues of several plant species. This was done functionally, by monitoring purine nucleotide-sensitive linoleic acid-induced uncoupling, or by Western blots. The following findings were established: (1) PUMP was found in most of the higher plants tested; (2) since ATP inhibition of linoleic acid-induced membrane potential decrease varied, PUMP content might differ in different plant tissues, as observed with mitochondria from maize roots, maize seeds, spinach leaves, wheat shoots, carrot roots, cauliflower, broccoli, maize shoots, turnip root, and potato calli. Western blots also indicated PUMP presence in oat shoots, carnation petals, onion bulbs, red beet root, green cabbage, and Sedum leaves. (3) PUMP was not detected in mushrooms. We conclude that PUMP is likely present in the mitochondria of organs and tissues of all higher plants.     

Manipulating membrane Fatty Acid compositions of whole plants with tween-Fatty Acid esters

This paper describes a method for manipulating plant membrane fatty acid compositions without altering growth temperature or other conditions. Tween-fatty acid esters carrying specific fatty acids were synthesized and applied to various organs of plants growing axenically in glass jars. Treated plants incorporated large amounts of exogenous fatty acids into all acylated membrane lipids detected. Fatty acids were taken up by both roots and leaves. Fatty acids applied to roots were found in leaves, while fatty acids applied to leaves appeared in both leaves higher on the plant and in roots, indicating translocation (probably in the phloem). Foliar application was most effective; up to 20% of membrane fatty acids of leaves above the treated leaf and up to 40% of root membrane fatty acids were exogenously derived. Plants which took up exogenous fatty acids changed their patterns of fatty acid synthesis such that ratios of saturated to unsaturated fatty acids remained essentially unaltered. Fatty acid uptake was most extensively studied in soybean (Glycine max [L.] Merr.), but was also observed in other species, including maize (Zea mays L.), mung beans (Vigna radiata L.), peas (Pisum sativum L.), petunia (Petunia hybrida L.) and tomato (Lycopersicon esculentum Mill.). Potential applications of this system include studying internal transport of fatty acids, regulation of fatty acid and membrane synthesis, and influences of membrane fatty acid composition on plant physiology.     

Free and Conjugated Indoleacetic Acid (IAA) Contents in Transgenic Tobacco Plants Expressing the iaaM and iaaH IAA Biosynthesis Genes from Agrobacterium tumefaciens

The Agrobacterium tumefaciens T-DNA gene iaaM was introduced by leaf-disc transformation into transgenic tobacco (Nicotiana tabacum) plants expressing the iaaH gene. Regenerated calli were screened for the presence of indole-3-acetamide (IAM), by gas chromatography-multiple ion monitoring-mass spectrometry, and IAM-containing calli were further analyzed for free and conjugated indoleacetic acid (IAA). It was found that transgenic calli on average contained twice as much free IAA and three times more conjugated IAA than calli from wild-type plants. About 40% of the transformed calli could be regenerated to plants. The distribution of free and conjugated IAA was measured in transformed plants with a normal phenotype and compared with equivalent wild-type plants. The IAA content of transgenic plants was only slightly increased, whereas IAA-conjugate levels were enhanced significantly. These data suggest that conjugation of IAA may serve as a regulatory mechanism, contributing to maintenance of steady-state IAA pool sizes during tobacco growth and development.     

Fatty acid composition of lipids from differentiated tissues and cell cultures of Euonymus europaeus

The fatty acid patterns of Euonymus europaeus callus cultures and cell suspension cultures were analysed at the beginning of stationary growth phase and compared with those from the respective differentiated tissues. The lipid and fatty acid patterns in cell cultures differed remarkably from those in the tissues of the mother plant. No glycerol triacetate was detected in the callus cultures derived from differentiated tissues whereas in seeds this lipid compound amounts to 29%. In addition to fatty acids normally occurring in differentiated tissues, lipids in cultured cells also contained short-chain (C₁₂–C₁₄) as well as very long-chain fatty acids (C₂₀–C₂₄). In tissue culture cells the major fatty acids were found to be saturated, whereas in the mother cells unsaturated fatty acids were predominant. Palmitic acid is the most abundant fatty acid in most of the cultures. Lauric, myristic and palmitic acid amount to 50% in lipids of cell suspension cultures.     

Regulation of growth,nutritive, phytochemical and antioxidant potential of cultivated <Emphasis Type="Italic">Drimiopsis maculata</Emphasis> in response to biostimulant (vermicompost leachate,VCL) application

The effect of vermicompost leachate (VCL, low-cost biostimulant) on the growth, elemental (macro and micro-nutrients) and phytochemical content as well as the antioxidant potential of Drimiopsis maculata was evaluated. Three dilutions (1:5; 1:10 and 1:20) of VCL were tested and the cultivation lasted for 3 months. In addition to the recorded growth parameters, dried and ground plant materials (leaves and bulbs) were evaluated for nutrients, phenolic acids and antioxidant capacity. Vermicompost leachate application enhanced the growth of D. maculata, particularly, the leaves (VCL 1:10) and bulbs (VCL 1:20) which were significantly bigger than the controls. Apart from the concentration of phosphorus which was significantly lower in the leaves of VCL (1:20)-treated plants, the quantity of all four macro-nutrients analysed were similar with and without VCL. Similar observations were also demonstrated in the majority of quantified micro-nutrients in D. maculata. Relative to the control, VCL-treated plants had higher concentrations of the 10 phenolic acids quantified in the leaves. However, the majority of the quantified phenolic acids were not significantly enhanced in bulbs. Antioxidant activity of D. maculata extracts was generally higher in leaves than in the bulbs. The leaf extract from VCL (1:10 and 1:20)-treated plants exhibited lower oxygen radical absorbance capacity (ORAC) when compared to the control. However, bulbs from VCL (1:5) treatment had significantly higher ORAC than the control. From a conservational perspective, the current findings provided insight on viable approaches useful for mitigating challenges associated with over-harvesting of highly utilized but slow-growing plant species.