Victor R. Villanueva

Changes in polyamine concentration during seed germination

Abstract Phaseolus mungo seeds 0 to 10 days after germination contained putrescine, spermidine, spermine, cadaverine, agmatine and tyramine. The rate of biosynthesis of total polyamines, proteins and RNA in the developing seeds follows similar profiles, reaching maxima 3 hr from germination. Putrescine, cadaverine, spermidine, spermine and agmatine were the major amines found in Pisum sativum 0–7 days after germination. RNA and proteins seem to follow the same pattern as polyamines during the first 12 hr in the developing pea seeds. RNA reaches a peak at 15 hr and polyamines and proteins peak 24 hr after germination. A rise to total polyamine concentration was also observed in seeds of Tragopogon porrifolius, Zea mays and Triticum aestivum 2–12 hr after germination.

Automated analysis of common basic amino acids, mono-, di-, and polyamines, phenolicamines, and indoleamines in crude biological samples

A fully automated, fast, and sensitive method for the separation of common basic amino acids and mono-, di-, and polyamines as well as phenolic- and indoleamines is described. Picomole level determination of hydroxytryptophan, tryptophan, histidine, lysine, ethanol amine, arginine, noradrenaline, diaminopropane, putrescine, histamine, cadaverine, dopamine, hexamethylenediamine, agmatine, tyramine, phenethylamine, serotonin, 5,6-dihydroxytryptamine, 5-methoxytryptamine, tryptamine, spermidine, and spermine is carried out by ion-exchange column chromatography on a single sample in 170 min of total analysis. This method is well suited for crude extracts without preliminary purification, thus reducing preparative losses. The reproducibility of the method has been studied and the percentage recovery of the different compounds after column chromatography is reported. Its application to crude samples from different biological sources such as microorganisms, vegetables, platelets, and urine is presented. This method could serve as a powerful tool for the analysis of these amino compounds in which there is currently a considerable interest.

Are polyamines involved in the induction and regulation of the Crassulacean acid metabolism

Leaves of plants with Crassulacean acid metabolism (CAM) were analyzed for variation in the content of polyamines in connection with the metabolism of malic acid in the dark and in the light, and with the induction of full-CAM activity. Under conditions (long days) resulting in extremely low CAM activity, young leaves of K. blossfeldiana have very low content in the polyamine-precursor arginine and in putrescine. The content in these two substances was increased dramatically by full-CAM induction with short days. During the course of the night/day cycle two peaks of putrescine content were observed in leaves of Kalanchoe blossfeldiana Poelln. Tom Thumb performing full-CAM operation: a large increase occurs toward the end of the day and the first half of the night, and its kinetics corresponds to the increase in the rate of malic acid synthesis; another peak, very sharp, appears during the first hours of the day, concomitant with the time of release of malic acid from the vacuole into the cytoplasm. In the case of Bryophyllum daigremontianum Berger similar variations were observed for the content in spermidine. These results support the hypothesis that polyamines could be involved in countering the tendency toward acidification of the cytoplasm at those moments of CAM operation at which the local concentration of malic acid is increased (i.e., during active synthesis in the dark and during the efflux from the vacuole in the light).

Effect of acid rain on polyamines in Picea

Abstract A comparative study on the content and evolution of putrescine, spermidine and spermine in healthy and diseased Picea trees living in a polluted (acid rain) area (Vosges, France) showed marked differences according to the physiological state of trees. Diseased trees contained higher levels of these compounds as compared to healthy trees. These results indicate that amine synthesis is one of the metabolic responses to acid pollution.

Inhibition of polyamine biosynthesis and seed germination in Picea abies

Abstract Control of seed germination in Picea abies by inhibiting polyamine biosynthesis was studied using cycloheximide, cyclohexylamine, dicyclohexylamine, 1,4-diaminobutanone, l - C -allylglycine and MGBG. All compounds tested showed an inhibitory action on the biosynthesis of at least one of the three polyamines viz. putrescine, spermidine and spermine. Inhibition of polyamine biosynthesis slowed down the germination process but only when both polyamines, spermidine and spermine, were simultaneously inhibited.

Biosynthesis of polyamines in Euglena gracilis

Abstract In Euglena gracilis Z the biosynthesis of spermidine and spermine closely resembles the pathways occurring in mammalian tissues and in most microorganisms. l -Ornithine and not l -arginine, as is the case in most plants, is the main precursor of putrescine, and S-adenosylmethionine donates the propylamino moiety for the biosynthesis of spermidine and spermine. Cell-free extracts of Euglena synthesized sym-norspermidine and sym-norspermine from 1,3-diaminopropane and labelled S-adenosylmenthionine. The synthases for the biosynthesis of these two polyamines have a pH optimum of 7.6, like that of spermidine and spermine synthases. Ion exchange chromatography showed two peaks corresponding to the retention times of 2,4-diaminobutyric acid and 1,3-diaminopropane, lower homologues of ornithine and putrescine, respectively. Experiments with dl -2,4-diaminobutyric acid-[4-14C] did not result in significant incorporation of the label into 1,3-diaminopropane.

Development of a multi-component analysis system : Application and preliminary results of a comparative study of cellular metabolism in healthy and damaged Picea trees from polluted areas

Abstract A multi-component analysis system is described, based on electrophoresis (for high-molecular-weight analysis), automatic column chromatography (for low-molecular-weight analysis) and flow injection analysis (for total functional analysis). The whole system is interconnected with the LABNET network. In this system chromatography is carried out with the help of automatic analysers (mostly laboratory-made) using high-performance column chromatography on ion exchangers adapted for the specific analysis of essential metabolites grouped by “family” (sugars, amino acids, polyamines, etc.). The most important feature is that all the different “family” analyses are run on a single sample without purification. This system has been partially applied to a comparative analytical study of cellular metabolism between healthy and damaged Picea trees in polluted areas (acid rain). This makes it possible to detect metabolic disfunctioning caused by pollution and to deduce further research goals. The first results on sugar and amine content showed marked differences between healthy and damaged trees that can be correlated with their physiological state. The results are presented and discussed in the light of the biochemical knowledge of the biological role of the different metabolites with particular emphasis on the precursor—product relationship.

Arrest of cell division blocks the utilization of polyamines in synchronized cultures of photoautotrophically grown Euglena

Abstract A trimodal change in the cellular levels of three major polyamines: spermidine, N,N′ -bis(3-aminopropyl)-1, 3-propanediamine (BAP) and 3,3′-diaminodipropylamine (DAD) was observed during two successive cell cycles in synchronously dividing cultures of the algal flagellate, Euglena gracilis Z photoautotrophically grown in a 24-h light-dark cycle. The intracellular levels of these three polyamines decreased as cells divided and then were enhanced as cells exited the G 1 phase and proceeded through the S and G 2 phases. Spermidine, BAP and DAD concentrations increased about 2.5-fold during the S phase. Putrescine and 1,3-diaminopropane levels did not vary significantly. One peak of polyamine synthesis occurred in the G 1 phase prior to DNA synthesis, followed by a second more important peak during the S-G 2 phases before cell division; both peaks were observed during the light period. A third minor peak was observed during the pre-G 1 (or G 0 ) phase in the dark period after mitosis had been completed. In contrast, when the cells attained the “stationary” phase of growth, there was no significant increase in the content of polyamines during the light period although spermidine and BAP increased slightly twice during the dark period (putrescine and 1,3-diaminopropane and DAD levels remained almost constant). To ascertain whether the synthesis of polyamines was merely a direct effect of the photoperiod, parallel experiments with synchronous cultures were carried out in the presence and absence of 3-(3,4-dichlorophenyl)-1, 1-dimethyl urea, a photosynthetic inhibitor. Although a slight decrease in the concentration of polyamines was observed, the three maxima of polyamines synthesis were observed as in normal cultures. These results clearly suggest that polyamine biosynthesis is closely related to DNA replication and cell division in Euglena cells.

Triterpénes et stérols de Lenzites trabea

Abstract Triterpenoid acids and sterols from the mycelium of the woodrotting fungus Lenzites trabea have been investigated. In addition to trametenolic and 15α-hydroxytrametenolic acids the following compounds were identified: eburicoic and sulfurenic acids, 24-methylenelanosterol, ergosterol and fungisterol. Triterpenoid composition of normal cultures was compared to that of submerged cultures. The main effects of submerged cultures is the increase of trametenolic and eburicoic acids and the decrease of 15α-hydroxytrametenolic acid.

Polyamines in turions and young plants of hydrocharis morsus-ranae and utricularia intermedia

Abstract Spermidine is the most abundant polyamine in dormant turions of Hydrocharis morsus-ranae and Utricularia intermedia , and it is also the dominant polyamine in sprouts of U. intermedia . The putrescine level is high in young leaves of H. morsus-ranae . Cadaverine and homospermidine occur respectively in vernalized turions of H. morsus-ranae and of U. intermedia .