Terpenoid and morphological variability of Pinus quadrifolia and the natural hybridization with Pinus monophylla in the San Jacinto Mountains in California

Monoterpenoids from wood of Pinus quadrifolia and Pinus monophylla from south of the San Jacinto Mountains in southern California were analysed by gas-liquid chromatography (GLC). The number of needles per fascicle, the number of resin canals, and the number of abaxial and adaxial stomatal rows in the needles were determined. Percentages of mycrene, α-pinene, and to a minor extent of camphene, β-pinene, limonene, and β-phellandrene, the above mentioned morphological characteristics and previously obtained data were all used for identification and characterisation of species intermediacy in the San Jacinto area. It was concluded that many P. quadrifolia trees in the mixed Buck Ridge stand and some trees in two pure stands of the same area were hybrids and that sympatric and to a lesser extent allopatric introgression of P. monophylla into P. quadrifolia takes place in the San Jacinto region. Introgression of P. quadrifolia into P. monophylla could not be demonstrated.     

On the question of natural hybridization between Pinus discolor and Pinus cembrides

Sixty Pinus cembroides and P. discolor trees from two mixed stands and eight pure stands were analysed for the composition of their wood monoterpenes, number of abaxial and adaxial stomatal rows and the number of needles per fascicle. While by far the majority of the characters analysed showed highly significant differences between the two species, only the sum of thujene, sabinene, γ-terpenene, terpinolene and p-cymene percentages, the sum of myrcene and cis-ocimene percentages, the number of abaxial stomatal rows, and the number of needles per fascicle could be used to evaluate the potential intermediacy of trees from the common sites. The results indicated the absence of any hybridization in the Sierra de la Catarina Mountains of Chihuathua (Mexico) and either a possible minute introgression of P. cembroides into P. discolor in the San Miguelito Mountains of San Luis Potosi (Mexico) or a slightly different genetic composition of the trees from this site. The results strongly support the proposal of Bailey and Hawksworth to regard P. discolor as a distinct species.     

Monoterpenoid and morphological differentiation within Pinus cembroides

Wood of Pinus cembroides from 232 trees sampled in 23 localities throughout its range in the United States and Mexico has been analysed for monoterpenes. With a few exceptions the trees of the continental populations from the state of Hidalgo northward produced α-pinene as the main monoterpene. The majority of trees of the southern populations (subsp. orizabensis) from the state of Puebla and adjoining regions of the neighboring Veracruz and Tlaxcala produced 3-carene in appreciable amounts. The population from Baja California (subsp. lagunae) produced monoterpenes of the sabinene family (thujene, sabinene, γ-terpinene, terpinolene and $\text{p-}\text{cymene}$) as the main constituents. The variations in morphological characters, including number of needles per fascicle, number of stomatal rows in needles, number of resin canals in needles, as well as other characters of the internal needle morphology, needle length and thickness, needle retention, thickness of seed shells, bark and cone characteristics, rate of growth and cotyledon number either paralleled or did not contradict the chemical variations. The evolution of the southern populations (subsp. orizabensis) into a separate group most likely resulted from geographic separation and climatic differences, while the evolutionary separation of the Baja California trees (subsp. lagunae) was connected with northwestern movement of the California coastal region from Miocene onward and the formation of Sierra Madre Occidental resulting in separation of this population from the mainland populations by the gap of the Gulf of California and the coastal mountain ranges.     

Variability in essential oils and needle resin canals of Pinus longaeva from Eastern California and Western Nevada in relation to other members of subsection Balfourianae

Mono- and sesquiterpenoids from wood and leaves of Pinus longaeva trees collected in various localities of eastern California and western Nevada were quantitatively analyzed by GLC. Concurrently the number and size of resin canals in foliage of the same trees and as well as Pinus aristata trees from Arizona, Colorado and New Mexico were determined. While some monoterpenes (mainly α-pinene and 3-carene) suggest that P. longaeva trees from the Panamint Mountains of California are P. aristata-like, the total chemical data and morphological data indicate that Panamint trees should be grouped with P. longaeva. A chemical latitudinal gradient is evidenced by the appearance of trees producing 3-carene in large amounts along the line joining the Panamint, Inyo and White Mountains of California. Pinus rzedowskii, well separated geographically and morphologically from the accepted members of subsection Balfourianae, is shown to be sufficiently distinct chemically, as to leave its suggested membership in the subsection an open question.     

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.     

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.     

Differences between Pinus pinea and Pinus pinaster as bioindicators of polycyclic aromatic hydrocarbons

The potential of Univariate and Multivariate Analysis and specifically Principal Components Analysis (PCA), has been employed to assess the performance of pine needles as bioindicators of polycyclic aromatic hydrocarbons (PAHs) and particularly emphasize the crucial importance of making a distinction between different pine species if more than one is sampled. Four sampling sessions were done in 29 sites and needles of two common pine species (Pinus pinaster Ait. in 19 sites and Pinus pinea L. in 12) were collected and analysed using gas chromatography-mass spectrometry (GC-MS) and PCA. The results obtained indicated significant differences between species, attributed to their different morphology. The mean total PAH concentration of the P. pinaster needles are over two times higher than P. pinea's. This difference is lower when the results are presented in lipid weight, but still statistically significant. Samples from the two sites with adjacent trees reinforce these conclusions, showing significant differences in terms of PAH mean concentration and aromatic ring patterns.     

Non-linear response of stomata in Pinus koraiensis to tree age and elevation

Knowledge on variations in stomata is useful in reflecting leaf physiological characteristics of CO₂ uptake and water transpiration, and predicting the responses of plants to future climate change. Stomatal density and number of stomatal rows (current-year, 1- and 2-year-old needles) in relation to tree age (ranging from 25 to 320 years old), elevation (ranging from 738 to 1,380 m a.s.l.), and sun exposure (sun and shade exposure) were investigated in Pinus koraiensis trees. Stomatal density and number of stomatal rows in relation to tree age and elevation showed a humped curve with the maximum values at intermediate levels of tree age (210 years old) and elevation (1,050 m a.s.l.), respectively. Needle age but not sun exposure significantly affected the stomatal density across tree ages and elevations. Our results suggest that variations in stomatal density of Pinus koraiensis needles are related to ontogenetic growth and environmental factors.     

Variation in needle terpenoids among Pinus sylvestris L. (Pinaceae) provenances from Turkey

Variation of terpenes and resin acids in needles of young Scots pine (Pinus sylvestris L.) seedlings from nine different provenances in Turkey was investigated. The provenances represent 1200-km West to East and 400-km South to North transects. Seven monoterpenes and two sesquiterpenes were reported in the needles of pines studied. Generally, the kinds of terpenes were similar but the relative amount of some compounds differed among the origins. The major components of the monoterpene fraction in Turkish sources were α-pinene (84.8%), β-pinene (4.1%) and limonene (3.0%), corresponding to 91.9% of the crude needle extract. In a PCA-analysis, 3-carene, myrcene and terpinolene in seedlings from Turkish provenances were quite low and thus, they were clearly different from a Northern European Scots pine provenance from Finland. In the resin acid fraction, abietic acid (62.4%) and dehydroabietic acid (16.1%) were the most abundant constituents in the needles of the Scots pine from Turkish provenances.     

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.     

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.     

Role of acyclic compounds in monoterpene biosynthesis in Pinus pinaster

The biosynthesis of monoterpene hydrocarbons was studied in maritime pine needles by incorporation of ¹⁴CO₂. It was shown that the acyclic terpenes β-myrcene and trans-β-ocimene, act as transitory compounds before the biosynthesis of cyclic monoterpenes such as α- and β-pinene. The mechanisms of biosynthesis are genetically controlled.     

Light-dependent monoterpene synthesis in Pinus radiata cultures

Callus cultures of Pinus radiata that synthesized monoterpenes de novo and which were stable for at least 1 year have been established. The products differed from those of parent plants in that α-pinene (87–100%) rather than β-pinene was the main component. The best lines accumulated monoterpenes (ca 2 × 10^?3% wt/wet wt)in yields 40–20% of that in the parent stem and needles. The composition of the extractable oil depended on the light regime. After culture in total darkness toluene and acetone accumulated. These compounds also occurred (at low levels) in dark-grown seedlings and in seeds of P. radiata and a route for their formation from α-pinene is suggested. Cell-free extracts of the culture lines converted [¹⁴C] IPP into geraniol, nerol and α- and β-pinenes in up to 46% total yield. These are the most active crude extracts for monoterpene biosynthesis that have been reported from either tissue cultures or higher plants.     

Terpene composition of Pinus pinaster seedlings and plants

-The production of essential oils in Pinus pinaster increases with the age of the seedlings, being higher in plants grown under continuous illumination. In the seedlings, nearly all the terpene is α- and β-pinene, the relative proportions of which are completely reversed between the 6th and 10th days of growth, regardless of the illumination period. Another reversion takes place after 60–65 days, the proportion found in the adult pine being very constant with α-pinene as the main component. The amount of oil in seedlings from high resin yielding parent trees was 4-fold higher than that in seedlings of normal (wild type) seeds. This finding is interesting because it can be employed for the preselection at the early stage of seedling of seeds to be used in forestry to obtain plantations of a high resin yield.     

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.     

Changes in diversity and storage function of ectomycorrhiza and soil organoprofile dynamics after introduction of beech into Scots pine forests

Diversity and storage function of mycorrhiza as well as soil organoprofile formation were investigated in a chronosequence of a pure Scots pine (Pinus sylvestris L.) stand, of Scots pine stands that were underplanted with beech (Fagus sylvatica L.) and in three pure beech stands of different age. Mycorrhiza diversity was higher in the pure beech stands compared to the pure pine stand. Beech and pine trees in the mixed stands had similar dominant mycorrhiza morphotypes. However, trees in two of the three pure beech stands were mycorrhized with other types. Mycorrhizal abundance and nutrient amounts of mycorrhizae associated with beech trees were higher in the mixed and in the pure beech stands compared to pine mycorrhizae indicating that nutrient uptake was higher in older beech than in older pine trees. Humus quality varied from pine to beech stands. Plant litter storage in the humus layer was highest in the youngest mixed stand and lowest in the oldest beech stand. Humus forms changed from moder grass-type in the pure Scots pine stand to mor-like moder and moder rich in fine humus with increasing age of beeches in the mixed stands. The older beech stands were characterised by oligomull and mull-like moder as the dominating humus forms. The ecologically favourable humus forms, i.e., nutrient rich humus forms in the older beech stands correlate well with the higher mycorrhizal diversity and abundance as well as the higher nutrient storage of their mycorrhizae in these stands. The results are also discussed with regard to the 'base-pump effect' of beech trees.     

Monolignol and dilignol glycosides from Pinus contorta leaves

Four monolignol glucosides have been isolated and identified from the needles of Pinus contorta. Chavicol 4-O-α-L-arabinofuranosyl-(1 →     

Sesquiterpene lactones from dicoria canescens

Dicoria canescens from Baja California, Mexico afforded the known monoterpene glucoside, (?)-cis-chrysanthenol O-β-d-glucopyranoside, ivalbatin, a xanthanolide and two new sesquiterpene lactones, 4-dihydroivalbatin (dicorin) and 11β, 13-tetrahydrovalbatin (dihydrocorin).     

Relationship between stomatal frequencies,yield components and morphological characters in collections of winter wheat cultivars

Genotypes on the Belgian national list as well as about 120 genotypes of winter wheat from the world collection were examined for the relationships of their stomatal frequencies in the flag leaf with yield, yield components and other morphological characters. Stomata were in rows, with a higher density in adaxial leaf surface. Cultivars differed significantly in both stomatal density and number of rows of stomata per unit of leaf width. Yield and morphological characters in general showed no significant correlation with stomatal frequencies. Densities in all series examined ranged from 46 to 78 stomata per mm² in the adaxial leaf epidermis and from 32 to 93 in the abaxial one.