6 Lignin-protein complex in cell walls of Pinus elliottii: Amino acid constituents

Cell walls of Pinus elliottii callus contain ca 12 % protein. Klason lignin prepared from the walls contained 9 % protein and represented 4.5 % of the wall. The lignin fraction was increased to 22 % of the wall weight by reacting washed cell-wall tissue with coniferyl alcohol and H₂O₂, a reaction catalysed by peroxidase that remained bound to the wall. The augmented lignin preparation yielded 10 % protein. The acid hydrolysate of whole wall tissue included five amino acids at a concentration higher than hydroxyproline. The hydrolysates of both natural and augmented lignin preparations yielded distributions of amino acids in which the concentration of hydroxyproline was higher than that of all other amino acids. The results suggest that polymerizing lignin links covalently with cell-wall glycoprotein, and that the bonds may be formed preferentially with hydroxyproline.     

Polyisoprenols in Pinus sylvestris needles

A polyisoprenoid fraction was isolated from the needles of Pinus sylvestris and comprised about 1% of the dry weight. MS, IR, GLC and HPLC analyses showed that the fraction consisted of polyisoprenylacetates, with 10–19 isoprene units predominantly in the cis configuration.     

Analysis of 3-indole carboxylic acid in Pinus sylvestris needles

3-Indole carboxylic acid (ICA) has been characterized as an endogenous constituent of Pinus sylvestris needles. Quantitative estimates of 3-indole acetic acid (IAA) and ICA, corrected for both sample losses and the conversion of IAA to ICA occurring during purification, indicate that Pinus needles contain 24.5 ± 6.5 ng IAA/g and 2.3 ± 0.4 ng ICA/g.     

Oxidative dimerization of ferulic acid by extracts from Sorghum

Extracts from leaves and first internodes of Sorghum bicolor catalyze the conversion of ferulic acid to a β-β-coupled dimer, the dilactone dimer of ferulic acid. The dilactone is then hydrolyzed, probably non-enzymatically, to a blue fluorescing compound, tentatively identified as a β-β-coupled dimer with at least one lactone ring opened to form a carboxylic acid. Both the initial enzymatic and the subsequent non-enzymatic steps are greater at pH 8 than pH 6. The most active preparation is a crude particulate fraction from leaves obtained by centrifugation at 37000 g; white light increases the amount of dimer formed.     

A cutin acid in Pinus sylvestris microspores

Nonacosan-10-ol (0.7%) and the cutin acid, 9,16-dihydroxyhexadecanoic acid (0.3%) are present in Pinus sylvestris microspores. The pollen coat hence has some features in common with leaf cuticles.     

Regioselectivity of ferulic acid polymerization catalyzed by oxidases

Enzymatic oxidation of ferulic acid catalyzed by oxidases (laccase and peroxidase) was carried out. Ferulic acid was shown to be subjected to oxidative processes leading to the formation of oligomeric and polymeric structures. The polymer formation takes place due to the formation of C_Ar-C_Ar- and C_Ar-O-C_Ar-bonds, as well as due to reactions of opening of the propane chain double bond. Different dynamic conditions of the enzymatic reactions were used to study the effects of conditions on the biosynthesis in vitro of some dehydropolymers: the method of dropwise mixing (endwise polymers) and a single addition (bulk polymers). The chemical structures of the resulting compounds were examined by the methods of IR, ¹H NMR, and ¹³C NMR spectroscopy. Differences in the quantitative ratio of structural fragments in a polymer cause changes in its thermal characteristics.     

Variability in the essential oils of wood and foliage of Pinus aristata and Pinus longaeva

Variations in the monoterpenoid composition of the essential oil of wood, and of the mono- and sesquiterpenoids of foliage collected from more than 200 specimens of pines of subsection Balfourianae (P. aristata, P. longaeva and P. balfouriana) substantiate the previously reported morphological and chemical separation of P. aristata and P. longaeva and suggest a restricted intermediacy in northern Arizona and eastern California. On the basis of wood essential oil the limited population of P. aristata in northern Arizona, while exhibiting to a slight degree morphological intermediacy, is found to be practically identical to the main populations of P. aristata in Colorado and New Mexico. On the basis of foliage oil, however, the same population is clearly intermediate, but closer to P. aristata. The detailed chemical data suggest a distinct chemical status for the Arizona population, with selection for the present-day chemophenotypes being the dominant factor in their evolution. Analysis of monoterpenoids from P. balfouriana × longaeva hybrids indicates that incomplete selective elimination of 3-carene-producing P. longaeva chemophenotypes rather than gene exchange with P. balfouriana is responsible for the presence of abnormal P. aristata-like 3-carene-producing trees in eastern California. The latter trees could have resulted from limited gene exchange between P. aristata and P. longaeva by routes no longer available to them lying to the south of the Grand Canyon.     

Lipids of Pinus halepensis pollen

The total lipids of Pinus halepensis pollen were separated into individual classes of neutral and polar lipids and the components of each class were identified and determined quantitatively. Free fatty acids, waxes and triacylglycerols were found as the main constituents of neutral lipids and phosphatidylcholine and phosphatidylethanolamine of polar lipids. Glycerylether derivatives were detected in neutral and polar lipid fractions. Free and esterified volatile fatty acids were also found in pollen and its neutral lipid fraction.     

Correlations between baltic amber and Pinus resins

The composition of the acid fractions of four pine resins has been studied before and after an ‘ageing’ process. Correlations are established between Baltic amber and ‘aged’ Pinus halepensis resin.     

Variability of the wood monoterpenoids from Pinus aristata

Striking differences in turpentine composition between California-Nevada-Utah populations and Colorado-New Mexico-Arizona populations of the Bristlecone pine (Pinus aristata) were demonstrated by GLC analyses of the wood extracts. These differences are in line with Bailey's taxonomic division of Pinus aristata sensu lato into western Pinus longaeva and eastern Pinus aristata sensu stricto.     

Resin acids of Pinus resinosa needles

The principal resin acids in the needles of Pinus resinosa are the labdane diterpenes, the new 8,13-epoxy-14-labden-19-oic acid (epimanoyl oxide acid), 8,13β-epoxy-14-labden-19-oic acid (manoyl oxide acid), 8(17),E-12,14-labdatrien-19-oic acid (communic acid) and 15-oxo-8(17)-labden-19-oic acid (imbricataloic acid). A survey of needles from representative populations of P. resinosa showed a limited variability in resin acid composition consistent with the uniformity of other traits. The composition of needle resin acids for putative P. nigra x resinosa hybrids strongly suggests the improbability of P. resinosa as the pollen parent.     

Constituent acids of Pinus radiata stem cutin

Stem cutin from P. radiata seedlings grown under winter and summer environmental conditions comprised n-alkanoic, (C₁₀–C₂₆), α, ω-alkanedioic (C₁₄–C₂₂), ω-hydroxyalkanoic (C₁₂–C₂₄), hydroxy-α, ω-alkanedioic and polyhydroxyalkanoic acids. 9-Hydroxyheptadecane-1, 17-dioic, 9-hydroxyoctadecene-1, 18-dioic, 9-hydroxynonadecane-1, 19-dioic, and 10, 17-dihydroxyheptadecanoic acids are newly-identified constituents of gymnosperm cutin. Cutin grown under winter temperatures and photoperiod contained twice the amount of 9, 16-dihydroxyhexadecanoic acid than that in summer-grown cutin, suggesting that the winter-grown cutin was formed from a highly cross-linked polymer, and that summer-grown cutin contained more linear polyester portions in the polymer.     

Lupin peroxidases. II. Binding of acidic isoperoxidases to cell walls

Extracellular acidic isoperoxidases (EC 1.11.1.7), isolated from both the cell walls and intercellular spaces of lupin ( Lupinus albus L. cv. multolupa) hypocotyls, bound to water-insoluble pectins of wall fragments also isolated from the hypocotyls. The binding was sáturable by increasing the isoenzyme concentration in the assay medium and it was dependent on the pH; neutral pH (6.0–7.0) favoured release, while acidic pH (4.0–5.0) favoured the attachment to the cell wall. Binding of acidic isoperoxidases to wall fractions was correlated with the in vitro acid-induced growth of hypocotyl segments, and both were modulated in the same direction by the Ca²⁺/H⁺ ratio in the incubation media, although the two responses were clearly separated when the Ca²⁺/H⁺ ratio varied. Binding of acidic isoperoxidases of cell walls could operate as a fine control of the activity of these cell wall enzymes, although its physiological role in the cell wall stiffening remains unclear. Some aspects of Ca²⁺ on the control of peroxidase activity at this level are also discussed.     

Formation of coniferyl alcohol from ferulic acid by the white rot fungus Trametes

The white rot fungus, Trametes sp., was cultivated in a medium containing ferulic acid, glucose and ethanol under aerobic conditions in submerged culture. The ferulic acid was transformed into coniferyl alcohol, coniferylaldehyde, dihydroconiferyl alcohol, vanillic acid, vanillyl alcohol, 2-methoxyhydroquinone and 2-methoxyquinone during 48–120 hr of cultivation. The amount of coniferyl alcohol in the culture reached a maximum after 90 hr with ca 40% of the initial amount of ferulic acid. Cinnamic acid, p-methoxycinnamic acid, 3,4-dimethoxycinnamic acid, p -coumaric acid and sinapic acid were also transformed into the corresponding alcohols, benzoic acids and benzyl alcohols in the fungus culture.     

Characterization of ribosomes from the seed of Pinus lambertiana

Ribosomes isolated from seeds of the sugar pine, Pinus lambertiana, have been characterized: The ribosome has a sedimentation coefficient (s_20,w⁰) of 78·2 S and contains 41 % RNA and 58 % protein. On dialysis against buffer containing 0·5-1 mM MgCl₂, the ribosome was reversibly transformed into an intermediate form (60 S). Further removal of Mg²⁺ causes the intermediate ribosome to dissociate into subunits (30 S and 40 S). Treatment of the intermediate ribosome with p-chloromercuribenzoic acid caused the dissociation of the particle into subunits. Incubating the 80 S ribosome with the sulfhydryl reagent caused a rapid transformation of the particle into an intermediate type particle. These results suggest that sulfhydryl groups are involved not only in associating the subunits but also in maintaining the compact structure of the ribosomes. The ribosome contains three ribosomal RNA components of 28 S, 18 S and 5 S. The base compositions of the three ribosomal RNA components are different.     

Epicuticular wax of Pinus radiata needles

Epicuticular wax isolated from the cotyledons and primary needles of 10-week-old Pinus radiata seedlings is similar in composition and contains 86% neutral compounds, viz. alkyl esters (25%, C₂₄–C₆₄), nonacosan-10-ol (52%), heptacosane-5,10-diol (2%), nonacosane-4,10-diol, nonacosane-5,10-diol, and nonacosane-10,13-diol (total 12%) and estolides, MW ca 800 (2%), MW ca 1100 (6%), and MW ca 1500 (1%). The acidic fraction (14%) contains n-acids (78%, C₁₂–C₃₂) and diterpene acids (22%, mainly abieta-8,11,13-trien-18-oic, with lesser amounts of pimara-8(14),15-dien-18-oic, isopimara-7,15-dien-18-oic and hydroxylated aromatic, diene and mono-ene acids). Wax isolated from primary needles of 1-yr-old seedlings had a similar neutral fraction composition, but the acidic fraction contained predominantly the diterpene acid mixture, with only trace amounts of n-acids. The wax from 1-yr-old secondary, needles from P. radiata forest trees aged 5 yr and 40 yr contained an acid fraction (12% 5 yr, 17% 40 yr trees) comprising the diterpene acid mixture, with trace amounts of n-acids together with ω-hydroxy acids (C₁₂, C₁₄ and C₁₆). The neutral fraction from both young and old trees had a similar composition containing alkyl esters (7%, C₂₄–C₆₆), estolides (90%, MW 566-ca 1500), nonacosan-10-ol (2%) and the heptacosane and nonacosane diols (1%). During growth and maturation of P. radiata, the nonacosan-10-ol content of the needle wax decreases while the proportion of estolides and diterpene acids increases, the latter probably being located around the stomatal pore.     

Monoterpene composition of cortical oleoresin from different clones of Pinus sylvestris

Monoterpene composition of cortical oleoresin was determined in a number of 9-yr-old grafts of Scots pine (Pinus sylvestris), growing in a clone trial at two different levels of mineral nutrient supply. Variation between clones was considerable for some of the monoterpene constituents, indicating genetic control. In one of the clones, δ³-carene, α-terpinene and terpinolene were totally absent. The monoterpene composition was not significantly affected by the annual application of macro-nutrients.     

Major resin acids of Pinus nigra needles

Labdane diterpene acids were found to be the major resin acid components in Pinus nigra needles of various seed sources. The major constituents have been identified as 4-epiimbricataloic acid, manoyl oxide 19-oic acid, 4-epicommunic acid, and 15-monomethyl pinifolate. A GC method was developed to analytically differentiate pinifolic acid from its monomethyl ester in an admixture of both compounds. A minor resin acid was identified as 18-acetoxy-8(17)-labden-15-oic acid. 10-Nonacosanol and isoabienol were identified as major constituents of the needle and cortex extractives, respectively.     

Gas chromatography-mass spectrometry study of sterols from Pinus elliotti tissues

A comparative study of the sterol components of slash pine (Pinus elliottii) callus tissue cultures, seeds, and seedlings was carried out using GC-MS techniques. Cholesterol, desmosterol, campesterol, stigmasterol, sitosterol and cycloeucalenol were identified in all tissues while lophenol and 24-methylenelophenol were identified in only the seed and seedlings. 24-Ethylidenelophenol was detected in trace concentrations in only the seedlings. Sitosterol was the predominant sterol component, i.e. 80·8, 38·1 and 47·8% of the tissue culture, seed and seedling sterols, respectively.