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PubMed2002

Removal of cadmium and mercury ions from aqueous solution by sorption on treated Pinus pinaster bark: kinetics and isotherms.

Formaldehyde pretreated Pinus pinaster bark was used to sorb Cd2+ and Hg2+ from aqueous solutions. The sorption kinetics showed hyperbolic dependence of the proportion of cation adsorbed on time, and the sorption isotherms were satisfactorily fitted by Freundlich equations, with k and n values showing Hg2+ to be more efficiently sorbed than Cd2+. Except for low cation concentrations, for which sorption was practically total at all initial pH > or = 6, sorption increased in this range, in keeping with a mechanism based on ion exchange with the hydroxyl protons of ring B of the procyanidin units of the tannins in the bark.

Absorption↗

PubMed2002

Operational factors and nutrient effects on activated sludge treatment of Pinus radiata kraft mill wastewater.

The biodegradability of Pinus radiata bleached kraft mill wastewater by an activated sludge treatment during a period of 280 days was evaluated. The effect of varying hydraulic retention time (HRT) in the range of 48 to 4.5 h and nitrogen (N) and phosphorus (P) addition on removal of biological oxygen demand (BOD5), chemical oxygen demand (COD), suspended solids (TSS and VSS), total phenolic compounds, tannin and lignin and reduction of toxicity was investigated. Removal of BOD5 was higher than 90% when HRT varied from 16 to 6 h, but decreased when HRT was less than 6 h. Similar performance was observed for COD removal, which was about 60% when HRT was varied from 16 to 6 h. Removal of total phenolic compounds and tannin and lignin was seriously affected by HRT. N and P addition to maintaining a ratio of 100:5:0.3 provided optimal BOD5, COD and suspended solids removal when HRT varied from 16 to 7 h, and no toxicity (using Daphnia) was detected in the treated effluent. When HRT was less than 6 h, the system showed destabilisation and pH, COD, BOD5 and suspended solids removal decreased.

Bioreactors↗

PubMed2003

Slow pyrolysis of wood barks from Pinus brutia Ten. and product compositions.

Biomass in the form of pine bark (Pinus brutia Ten.) was pyrolysed in an externally heated fixed-bed reactor. The effects of temperature and heating rate on the yields and compositions of the products were investigated. Pyrolysis runs were performed using reactor temperatures between 300 and 500 degrees C with heating rates of 7 and 40 degrees Cmin(-1). The product yields were significantly influenced by the process conditions. The bio-oil obtained at 450 degrees C, at which the liquid product yield was maximum, was analysed. It was characterized by Fourier transform infrared spectroscopy. In addition, the solid and liquid products were analysed to determine their elemental composition and calorific value. Chemical fractionation of bio-oil showed that only low quantities of hydrocarbons were present, while oxygenated and polar fractions dominated. The empirical formula of the bio-oil with heating value of 31.03 MJkg(-1) was established as CH(1.43)O(0.332)N(0.0013).

Hot Temperature↗

PubMed2003

Gravitational stress on germinating Pinus pinea seeds.

In the germination of lipid-rich seeds, the glyoxylate cycle plays a control role in that, bypassing the two decarboxylative steps of the Krebs cycle; it allows the net synthesis of carbohydrates from lipids. The activity of isocitrate lyase, the key enzyme of the glyoxylate cycle, is an indicator of the state of seed germination: stage of germination, growth of embryo, activation and progress of protein synthesis, depletion of lipidic supplies. In order to investigate the effects of gravity on seed germination, we carried out a study on the time pattern of germination of Pinus pinea seeds that were subjected to a hypergravitational stress (1000 g for 64 h at 4 degrees C), either in a dry or in a wet environment, before to be placed in germination plates. During the whole time of germination, we monitored the state of embryo growth and the most representative enzymes of the main metabolic pathways. In treated wet seeds, we observed an average germination of only 20% with a slowdown of the enzyme activities assayed and a noticeable degradation of lipidic reserves with respect to the controls. These differences in germination are not found for dry seeds.

Citric Acid Cycle↗

PubMed2003

Molecular identification of white morphotype strains of endophytic fungi from Pinus tabulaeformis.

Sterile mycelia isolated from Pinus tabulaeformis were grouped into white morphotype strains based on cultural characteristics. Eighteen of the isolates were randomly selected and identified to various taxonomic levels based on nuclear ribosomal DNA (nrDNA) sequence analysis. The 5.8S gene and flanking internal transcribed spacer (ITS1 and ITS2) regions of nrDNA were amplified and sequenced. Phylogenetic analysis of the 5.8S gene sequences indicated that the white morphotype strains were Ascomycota. Further identification was achieved by means of sequence similarity comparison and phylogenetic analysis of the ITS regions. Results showed that strains WMS9 and WMS10 were Lophodermium species (Rhytismataceae), while strains WMS11, WMS13 and WMS18 were species of Rhytismataceae. Strains WMS2, WMS3, WMS4, WMS5 and WMS6 were identified to Rosellinia, strain WMS1 to Entoleuca, and strain WMS14 to Nemania (Xylariaceae). Strains WMS7, WMS8, WMS12, WMS15, WMS16 and WMS17 were xylariaceous species. The potential of using DNA sequence analysis in the identification of endophytic fungi is discussed.

Ascomycota↗

PubMed2003

Local variations in microfungal populations on Pinus sylvestris needles.

We studied the fungal colonization of Pinus sylvestris needles in two neighbouring sites, comparing stands of isolated and grouped trees. We observed large variations among the proportions of needles bearing fruit bodies of Cyclaneusma minus, Lophodermium pinastri, Verticicladium trifidum and black lines characteristic of L. pinastri colonization. Variations between sites and within trees were greater than that between stands or between trees. The frequency of L. pinastri colonization was negatively correlated with C. minus fruit body frequency, while the frequency of V. trifidum conidiophores was positively correlated with L. pinastri colonization frequency without fruiting, and negatively correlated with C. minus apothecia frequency. Although L. pinastri black lines and C. minus apothecia were nearly randomly associated on individual needles in each sample, the two fungi occupied different segments when they occupied the same needle. These patterns at needle and sample scales do not explain the negative correlation between the frequencies of these two species observed at larger scales. In each sample, frequency of V. trifidum conidiophores was highest on needles colonized by L. pinastri without fruiting. On individual needles, V. trifidum conidiophores developed on segments colonized by L. pinastri without fruiting, but not on segments bearing fruit bodies of L. pinastri or C. minus. These patterns at needle and sample scales were consistent with the correlations between frequencies observed at larger scales. These results were compared to variations observed with stand age and climate in others studies. The observed variations might result from both microclimate variations and fungal interactions.

Ascomycota↗

PubMed2004

Leptographium bistatum sp. nov., a new species with a Sporothrix synanamorph from Pinus radiata in Korea.

Radiata pine (Pinus radiata) lumber from Australia, Chile, and New Zealand is imported into Korea where it represents one of the most important sources of timber. Blue stain of this timber is a serious problem for which an integrated control strategy is being developed. One of the fungi that has been isolated from stained radiata pine sapwood in Korea is a Leptographium sp. that has a distinct Sporothrix synanamorph. The aim of this study was to classify this fungus. Morphological comparisons showed that this fungus is distinct from all other species of Leptographium and especially L. elegans and L. francke-grosmanniae that also have Sporothrix synanamorphs. Comparisons of sequence data for the ITS2 and part of the 28S rDNA genes as well as the beta-tubulin gene also confirmed that this fungus represents an undescribed taxon, described here as Leptographium bistatum sp. nov.

Ascomycota↗

PubMed2004

Identification by gas chromatography-mass spectrometry of the volatile organic compounds emitted from the wood-rotting fungi Serpula lacrymans and Coniophora puteana, and from Pinus sylvestris timber.

Volatile organic compounds (VOCs) emitted by two wood-rotting basidiomycete fungi, Serpula lacrymans (dry rot fungus) and Coniophora puteana (cellar fungus), and the timber of Pinus sylvestris (Scots pine), were identified. Several volatile collection techniques were employed including dichloromethane solvent extraction, solid-phase microextraction (SPME) and thermal desorption of VOCs entrained on Tenax GR. In addition, a new method of solid sample injection (SSI) is described which utilises a low injector temperature and an all-glass deactivated injector liner designed to minimise both the formation of pyrolysis products and analyte degradation. All the volatile compounds collected were analysed using electron impact capillary gas chromatography-mass spectrometry (GC-MS) on HP-5, HP-Innowax and beta-cyclodextrin columns. SSI and Tenax thermal desorption were found to be the most effective extraction methods. A total of 19 VOCs were observed from S. lacrymans grown on glass slides and pine, 15 from C. puteana grown on glass slides and 12 from P. sylvestris timber. S. lacrymans was found to emit, in low abundance, six unique VOCs, of which 2-methylbutanal was the greatest. The major volatile compound emitted by S. lacrymans was 1-octen-3-ol, which was also found in lower abundance from C. puteana. Six VOCs, including diethylene glycol and 4-methyl methylbenzoate, were found to be unique to C. puteana, all in medium abundance: From P. sylvestris, the major volatiles identified were S-alpha-pinene and 3-carene.

Basidiomycota↗

PubMed2004

Geographic isolation of Diplodia scrobiculata and its association with native Pinus radiata.

Diplodia pinea (syn. Sphaeropsis sapinea) is a well-known latent pathogen of Pinus spp. with a worldwide distribution. As such, this fungus is native where pines are endemic in the northern hemisphere and it has been introduced into all countries of the Southern Hemisphere where pines are exotic. The newly described D. scrobiculata (formerly known as the B morphotype of D. pinea) is thought to have a much more limited distribution. D. scrobiculata was first reported as an endophyte and weak pathogen of P. banksiana, where it was found to coexist with D. pinea. Diplodia scrobiculata is now known to have a much broader distribution in Northern America and Europe. In this study, seven Simple Sequence Repeat (SSR) markers were used to evaluate genetic diversity and gene flow between populations of D. scrobiculata. Results indicate a strong geographic isolation between populations of D. scrobiculata from different regions in North America, with unique alleles fixed in the different populations. The data fits the isolation by distance model indicating limited dispersal. Geographic isolation in combination with isolation by distance suggests prolonged reproductive isolation. Intensive collections of endophytes from native P. radiata in California have yielded only D. scrobiculata and not the significantly more pathogenic D. pinea. SSR analysis of three populations of D. scrobiculata from native P. radiata identified many shared alleles among the populations and moderate to high gene flow between them. The three Californian populations are distant and distinct from populations of D. scrobiculata from elsewhere. Under stress conditions, P. radiata is known to be very susceptible to D. pinea in plantations in the Southern Hemisphere. Native P. radiata is currently experiencing severe stress due to pitch canker caused by Fusarium circinatum. Such stress would provide ideal conditions for an associated outbreak of D. pinea. Thus, it is critical to prevent the movement of D. pinea into the last remaining native stands of P. radiata.

Ascomycota↗

PubMed2003

Antioxidative properties of the essential oil from Pinus mugo.

The essential oil from Pinus mugo (PMEO) was tested on its antioxidative capacity. For this purpose, several biochemical test systems were chosen (e.g., the Fenton System, the xanthine oxidase assay, or the copper-induced oxidation of low-density lipoprotein (LDL)). The results show that there is moderate or weak antioxidative activity when tested in aqueous environments, like in the Fenton system, xanthine oxidase induced superoxide radical formation, or in the HOCl driven fragmentation of 1-aminocyclopropane-1-carboxylic acid (ACC). In contrast, when tested in more lipophilic environments (e.g., the ACC-cleavage by activated neutrophils in whole blood) the PMEO exhibits good antioxidative activity. PMEO does also show good antioxidative capacity in another lipophilic test system (i.e., the copper induced oxidation of LDL). Some components of PMEO (i.e., Delta(3)-carene, camphene, alpha-pinene, (+)-limonene and terpinolene) were also tested. As the PMEO, they showed weak or no antioxidant activity in aqueous environments, but some of them were effective antioxidants regarding ACC-cleavage by activated neutrophils in whole blood or copper-induced LDL-oxidation. Terpinolene, a minor component of PMEO, exhibited remarkable protection against LDL-oxidation.

Antioxidants↗

PubMed2004

Phenolic extractives from the bark of Pinus sylvestris L. and their effects on inflammatory mediators nitric oxide and prostaglandin E2.

The anti-inflammatory properties of phenolic pine (Pinus sylvestris L.) bark extract were studied. The pine bark extract was fractionated by liquid-liquid extractions and semipreparative high-performance liquid chromatography to reveal the most potent constituents. The phenolic compositions of three pine bark samples obtained, a crude extract, a chloroform fraction, and a semipreparative fraction, were analyzed using high-performance liquid chromatography with UV diode array detection and/or electrospray ionization mass spectrometry. In addition, eight compounds were isolated and identified by NMR and MS techniques. In total 28 phenolic compounds were identified. The effects of the three pine bark samples on the synthesis of two proinflammatory mediators, nitric oxide and prostaglandin E(2), were measured. It was shown that pine bark contains compounds that inhibit the production of these proinflammatory mediators.

Chloroform↗

PubMed2005

Procyanidin fractions from pine (Pinus pinaster) bark: radical scavenging power in solution, antioxidant activity in emulsion, and antiproliferative effect in melanoma cells.

Pine (Pinus pinaster) bark is a rich source of procyanidin oligomers. From a total polyphenolic extract, we have generated fractions of different procyanidin composition. The mixtures, devoid of gallate esters, were active as free radical scavengers against ABTS(*+), DPPH, and HNTTM. Pine bark fractions were tested for antioxidant activity in solution (hydrogen donation and electron transfer) and emulsion (inhibition of lipid peroxidation) and compared with their galloylated counterparts from grape origin. While galloylation clearly influenced the free radical scavenging efficiency in solution, it did not seem to play a determinant role in protection against lipid peroxidation in emulsion. The fractions were very mild inhibitors of cell proliferation. Because gallate esters appear to interfere with crucial cell functions, gallate free pine procyanidins may be the innocuous chemopreventative agents of choice for many applications in food and skin protection.

Antioxidants↗

PubMed2005

Efficient preparation of catechin thio conjugates by one step extraction/depolymerization of pine (Pinus pinaster) bark procyanidins.

The skin penetrating antioxidant cysteamine derivative of (-)-epicatechin as well as other thio conjugates were efficiently obtained with high yields from pine (Pinus pinaster) bark by simultaneous one pot extraction and depolymerization using water and cysteamine hydrochloride. The influence of the concentration of bark, acid, and cysteamine, as well as the reaction time on the total conversion, was studied. The total conversion into the epicatechin and catechin conjugates was as high as 47 g/kg pine bark with 1666 g cysteamine/kg bark and 28 g/kg with 166 g cysteamine/kg bark. A fast cleanup step by absorption/desorption on XAD-16 greatly facilitated further purification of the active major component. At a pilot scale, 4beta-(2-aminoethylthio)epicatechin (1) (conversion 263 g, purity 35% by reversed phase high-performance liquid chromatography/weight) was obtained from 17 kg of pine bark after simultaneous extraction/depolymerization followed by cleanup with the polymeric resin in approximately 10 h. The results show that pine (P. pinaster) bark is a suitable source of flavanols for the preparation of active thio derivatives. Conditions are given for the fast and efficient preparation of the conjugates.

Catechin↗

PubMed2002

Potential antitumor-promoting diterpenes from the cones of Pinus luchuensis.

A new nor-labdane-type diterpene, 15-nor-labda-8(17),12E-dien-13,19-dienoic acid (1), along with five known diterpenes, 15-nor-14-oxolabda-8(17),12E-dien-19-oic acid (2), trans-communic acid (3), sandaracopimaric acid (4), dehydroabietic acid (5), and abieta-8,11,13-triene-15,18-diol (6), was isolated from the cones of Pinus luchuensis. The structure of 1 was established by chemical and spectroscopic methods. Among these isolates, compounds 2, 4, and 6 showed potent inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter 12-O-tetradecanoylphorbol 13-acetate.

Antigens, Viral↗

PubMed2001

Regulation of loblolly pine (Pinus taeda L.) arginase in developing seedling tissue during germination and post-germinative growth.

After seed germination, hydrolysis of storage proteins provides a nitrogen source for the developing seedling. In conifers the majority of these reserves are located in the living haploid megagametophyte tissue. In the developing loblolly pine (Pinus taeda L.) seedling an influx of free amino acids from the megagametophyte accompanies germination and early seedling growth. The major component of this amino acid pool is arginine, which is transported rapidly and efficiently to the seedling without prior conversion. This arginine accounts for nearly half of the total nitrogen entering the cotyledons and is likely a defining factor in early seedling nitrogen metabolism. In the seedling, the enzyme arginase is responsible for liberating nitrogen, in the form of ornithine and urea, from free arginine supplied by the megagametophyte. In this report we investigate how the seedling uses arginase to cope with the large arginine influx. As part of this work we have cloned an arginase cDNA from a loblolly pine expression library. Analysis of enzyme activity data, accumulation of arginase protein and mRNA abundance indicates that increased arginase activity after seed germination is due to de novo synthesis of the enzyme. Our results suggest that arginase is primarily regulated at the RNA level during loblolly pine seed germination and post-germinative growth.

Amino Acid Sequence↗

PubMed2001

Pine weevil (Hylobius abietis) antifeedants from lodgepole pine (Pinus contorta).

Pine weevils (Hylobius abietis) fed less on bark of lodgepole pine (Pinus contorta) than on bark of Scots pine (P. sylvestris). Two pine weevil antifeedants, ethyl trans-cinnamate and ethyl 2,3-dibromo-3-phenyl-propanoate, were isolated from bark of lodgepole pine. These two compounds significantly reduced pine weevil feeding in a laboratory bioassay. In field assays, the second compound significantly decreased pine weevil damage on planted seedlings. Ethyl 2,3-dibromo-3-phenylpropanoate has not previously been reported as a natural product.

Animals↗

PubMed2002

Biochemical responses of Pinus pinaster trees to fire-induced trunk girdling and crown scorch: secondary metabolites and pigments as needle chemical indicators.

The biochemical responses to varying levels of localized heat damage to stems and crowns were studied in Pinus pinaster trees in Galicia, Spain. The objective was to ascertain the utility of secondary metabolites (total polyphenols. condensed tannins, astringency, free and esterified phenolic acids) and pigments (chlorophylls and carotenoids) as chemical indicators of localized fire damage. The study involved three levels of girdling by trunk heating (0%, 60%, and 75%), three levels of crown scorching (0%, 50%, and 75%), and all combinations of those trunk and crown treatment levels. Secondary metabolites and pigments were analyzed in needles before, during, and up to 8 months after fire treatments. High levels of polyphenols, condensed tannin, and pigments occurred in situations where the sole treatment was 75% crown scorching. Low levels of hydroxycinnamic acids were the result when the treatment was 75% trunk girdling. These responses occurred in the first two months after fire. Thereafter, normal values were observed. Low values of chlorophyll a/b ratio and high levels of free protocatechuic and esterified syringic acids were found to be long-term indicators of trunk girdling damage combined with crown scorching.

Analysis of Variance↗

PubMed2003

The production of transgenic Scots pine (Pinus sylvestris L.) via the application of transformed pollen in controlled crossings.

The study demonstrates the production of a transgenic Scots pine (Pinus sylvestris L.) seedling through the application of transformed pollen in controlled crossings. The pollen lots were transformed by particle bombardment, resulting in transient transformation frequencies varying from 15 to 49% of the germinated pollen grains, and bombarded pollen was used to pollinate megasporangiate strobili. Progeny was screened by histochemical, GUS assays, and selected seedlings were further analysed by PCR. PCR amplification revealed the presence of both the nptII and gusA genes in one seedling (23/237). Results were confirmed by Southern blot analysis. The morphology and growth of this transgenic seedling was normal. Although the transformation frequency of recovered plants was very low (1/14999), the present protocol suggests that production of transgenic Scots pine is possible without the use of any tissue culture methods or the involvement of marker genes, for selection of transformants.

Crosses, Genetic↗