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Inheritance of allozyme variants in bishop pine (Pinus muricuta D. Don).

Isozyme phenotypes are described for 45 structural loci and 1 modifier locus in bishop pine (Pinus muricata D. Don,) and segregation data are presented for a subset of 31 polymorphic loci from 19 enzyme systems. All polymorphic loci had alleles that segregated within single-locus Mendelian expectations, although one pair of alleles at each of three loci showed significant segregation distortion. The consistency of resolution and segregation at many loci in bishop pine makes electrophoretic analyses feasible for many purposes in this species.

Electrophoresis, Polyacrylamide Gel↗

A study on the nutritive value of pollen from the Chinese Masson Pine (Pinus massoniana) and its effect on fecal characteristics in rats.

A digestion experiment with growing rats was conducted to study the effect of native and broken pollen of Chinese Masson Pine (Pinus massoniana) on fecal composition and digestibilities of dry matter, crude protein and crude ash. For that issue each 10 of 30 animals were fed a semisynthetic diet containing either no pollen or and addition of 6% native or broken pollen for 17 days at restricted amounts. In a second experiment each 6 of 12 growing rats received a semisynthetic diet containing either no or 5% broken pollen. At day 4, 11 and 18 samples of the feces were collected and analyzed for mesophilic aerobic bacteria. The pollen contained large amounts of cell wall constituents (26-30% lignin and 10-15% cellulose, 2-13% hemicellulose) with higher values found in native pollen. The contents of crude protein, crude fat, total lipids, available carbohydrates (starch and sugars), crude ash, gross energy and metabolizable energy were 13%, 2-10%, 8-10%, 17-18%, 3.1-3.5%, 21-22 kJ/g and 5.7-9.0 kJ/g. The addition of native pollen to the diet increased the total amount of feces by 71% as compared to the control level. The fecal contents of dry matter and of crude protein and crude ash in fecal dry matter changed by -5, +4 and -5 percentage units. The apparent digestibilites of dry matter and crude protein decreased by 3 and 5 percentage units, respectively, while the apparent digestibility of crude ash remained unchanged. Broken pollen acted in the same direction, however the effects were about 20% less pronounced as compared to native pollen. The pollen feeding reduced the fecal germ contents of Proteus mirabilis and Escherichia coli, while the amounts of alpha-hemolysing streptococci was increased. In total, the observed effects of an addition of native broken or pollen to the diet seemed to be based mainly on the increased intake of cell wall constituents.

Animals↗

Effect of dose rate and exposure time on the stimulation effect of tube growth of Pinus silvestris pollen.

The stimulating effect of ionizing radiation in respect to dose rate and exposure time was studied using the tube growth of Pinus silvestris pollen. Stimulation was registered with a small dose (50 rad) supplied at low dose rates (0.5; 1.0; 3.0 and 5.0 rad/sec) and with higher doses (300, 800 and 1400 rad) supplied at higher dose rates (10; 40 and 50 rad/sec). This suggests that only the exposure time is of importance for radiation-induced stimulation provided that the exposure time does not exceed 100 sec.

Dose-Response Relationship, Radiation↗

Delayed conversion of squalene to sterols during development of Pinus pinea seeds.

During germination of seeds of the gymnosperm, Pinus pinea, radioactivity from [2-14C]-mevalonate proceeded principally through the anaerobic reactions leading to squalene in the first 24 hr in both the haploid endosperm and the diploid embryo, and only with succeeding time (3-9 days) in both cases was label transferred to sterols in oxygen-requiring steps. The rates of turnover must be real and independent in the two tissues, since no consequential interchange of labelled lipids occurred between the endosperm and the embryo. Similar delayed conversion of squalene to sterols has been observed previously during germination of seeds of the angiosperm, Pisum sativum.

Female↗

Genetic analysis of growth curves for a woody perennial species, Pinus taeda L.

Inheritance of growth curves is critical for understanding evolutionary change and formulating efficient breeding plans, yet has received limited attention. Growth curves, like other characters that change in concert with development, often have higher heritability than age-specific traits. This study compared genetic parameters of height-growth curves with those of age-specific heights for a conifer, Pinus taeda L. Growth curves were fitted with: (1) a linear regression model, and (2) a non-linear model based on Richards' function using two sources of height data: two six-parent diallel tests assessed at age 2 to 10 years and two tests from a nested mating design with 222 parents assessed at 1 to 25 years. Additive genetic control of growth-curve parameters was moderate (h(2) = 0.06 to 0.26) and slightly lower than that for age-specific heights. Additive variance exceeded dominance variance for rate and shape parameters, but not for the asymptote. Genetic correlations among growth-curve parameters were high. Early selection on height was as efficient as selection on growth-curve parameters.

Journal Article↗

Seed gene flow and fine-scale structure in a Mediterranean pine ( Pinus pinaster Ait.) using nuclear microsatellite markers.

The Mediterranean populations of maritime pine ( Pinus pinaster Ait.) are typically small and have a scattered distribution, being threatened by human activities and forest fires. In the framework of the genetic-resources conservation program of this species, a native multi-age stand located in a Mediterranean area (central Spain) was studied using three highly polymorphic nuclear microsatellites (SSRs). Spatial autocorrelation analysis was conducted using Moran's index in order to detect fine-scale structure in both natural regeneration and mature trees. The spatial pattern of seed flow based on dispersed progeny was studied using a highly reliable subset of parent-offspring matches obtained by means of parentage analysis and simulation-based calculation of statistical confidence. Maritime pine showed a fine-scale structure at the seedling stage. In natural regeneration, the autocorrelograms indicated a patch size of approximately 10 m. The fine-scale structure seems to be produced by a restricted seed gene flow. In fact, there was an excess of parent-offspring matches in a radius of 15 m from the parent trees. Pines with a heavy seed, such as P. pinaster, are expected to have a short dispersal distance, thus producing a fine-scale structure. However, the fine-scale structure did not persist in the mature trees. Within-population genetic structure in Mediterranean pines may be affected by a number of post-dispersal events (e.g. mortality due to the severity of the Mediterranean climate and animal-mediated secondary dispersal during the summer period). Thus, great alteration in the pattern produced by the initial seed rain and differences in genetic structure between tree cohorts are expected.

Journal Article↗

Considerations of correlated fertility between genders on genetic diversity: the Pinus densiflora seed orchard as a model.

The correlation between 99 clone female and male fertilities in a first generation seed orchard of Pinus densiflora was studied over 6 years. The effective number of the parent ( N(p)) and the variance effective population number [ N(e)((v))] were used to assess the impact of total (Psi(T)), female (psi(f)) and male (psi(m)) fertility variation. A theoretical framework was developed to account for female and male fertility correlations as well as the impact of possible pollen contamination. Total fertility variation was described by the sibling coefficient (Psi(T): the probability that two genes randomly chosen from the gamete gene pool originate from the same parent), which was further subdivided into psi(f) and psi(m). These parameters were compared under various conditions including the total seed harvest, imposing on equal seed harvest among the orchard's clones and two contamination scenarios ( M = 0 and 20%). Fertility variations among females, males and clones were observed within and among years. Sibling coefficients (Psi(T)) were lower, but the effective number of parent ( N(p)) and variance effective population number ( N(e)((v))) were higher in years with moderate female and good male strobilus production. N(p) for female and male reproductive outputs varied from 49 to 82 and from 57 to 93, respectively. N(p) was higher for males than females. When the crop of the 6 years was pooled, N(p) for female, male and the clone were 73, 87 and 85, respectively. The impact of female-male fertility correlation for conditions with no-, positive- and negative-correlations were assessed and their impact on Psi(T), N(p) and N(e)((v)) was also evaluated. It was demonstrated that the practice of equal seed harvesting from every clone, or the mixing of seeds from several years, would substantially improve the genetic diversity and the genetic representation of the seed orchard population when a positive correlation between gender fertilities was observed. The relevance of these results to supplemental-mass-pollination was discussed under two cases where equal- and un-equal amounts of pollen from clones were included in the pollen mixes.

Journal Article↗

Chloroplast phylogeography and evolution of highly polymorphic microsatellites in lodgepole pine ( Pinus contorta).

We employed a novel set of six highly polymophic chloroplastic simple sequence repeat (cpSSR) loci to investigate the phylogeography of lodgepole pine ( Pinus contorta Dougl. Ex. Loud.), and to examine aspects of the evolutionary process operating on these repetitive DNA sequences. Chloroplast haplotypes of 500 trees, sampled throughout the range of lodgepole pine, were determined. We found a marked association of genetic distance with physical distance within the scale of 0 to 1,000 km, but no association beyond that range. Likewise, geographic clustering was observed only among recent clades in a dendrogram. These phylogeographic patterns are consistant with a rapid rangewide expansion ("big-bang") followed by recent, local population differentiation ("galaxy formation"). In support of this expansion, coalescent simulations of the genealogical process gave a long-term effective population size in the low thousands, and a time to common ancestry of about 1,500 generations (12,000 years), consistent with a post-Pleistocene population expansion as documented by previous pollen-sediment analyses. Two lines of evidence (mapping mutational events onto a phylogeny, and evaluation of observed versus expected gene diversity) suggest that five of the cpSSR loci evolve primarily by a stepwise model of evolution of single repeat changes (but with a small proportion of changes involving two or more repeats), and the coalescent simulations point to a mutation rate of about 10(-3).

Journal Article↗

Identification of QTLs influencing wood property traits in loblolly pine ( Pinus taeda L.). II. Chemical wood properties.

Chemical wood property traits were analyzed for the presence of quantitative trait loci (QTLs) in a three-generation outbred pedigree of loblolly pine ( Pinus taeda L.). These traits were assayed using pyrolysis molecular beam mass spectrometry and include mass spectrum peak intensities associated with carbohydrates, alpha-cellulose and hemicellulose sugars, and lignin. Models for projection to latent structures (PLS) were used to also estimate the chemical composition of cell walls (i.e., alpha-cellulose, galactan and lignin) from mass spectrum data using multivariate regression. Both earlywood and latewood fractions from the fifth annual ring were analyzed for each trait. An interval mapping approach designed for an outbred pedigree was used to estimate the number of QTLs, the magnitude of QTL effects, and their genomic position. Eight unique QTLs influencing cell wall chemistry were detected from multiple peak intensities and/or PLS estimates using the one- and two-QTL models. Significant differences in chemical contents were observed among the populations from North Carolina vs Oklahoma, and results from QTLxenvironment analyses suggest that QTLs interact with environmental location. QTLs should be verified in larger experiments and in different genetic and environmental backgrounds. QTL mapping will help towards eventually identifying genes having a major effect on chemical wood properties.

Journal Article↗

The Distance to Which Wound Effects Influence the Structure of Secondary Xylem of Decapitated Pinus pinea.

Decapitation of stems of annuals and trees for the study of vascular and fiber differentiation with or without hormonal application is a common procedure. There is controversy about whether wound effects play a role in such experiments, and to what distance from the point of decapitation. To examine this question, the distance from the point of decapitation at which apparent wound effects are obvious developmentally, was studied in decapitated 4-year-old Pinus pinea plants. The wound effects just below the cut included differentiation of many traumatic resin ducts, a parenchyma band instead of tracheids, more tracheid files, and a higher proportion of late wood. The increase in the number of resin ducts was still considerable and statistically significant 10 cm below the point of decapitation compared with the nondecapitated control. These results indicate that in pines, wound effects in the first 5 cm below the decapitation point (a common point for tissue examination) cannot be ignored in experiments on the regulation of vascular differentiation.

Journal Article↗

Programmed cell death of Pinus nucellus in response to pollen tube penetration.

In ovules of Pinus densiflora, pollen tubes elongate and branch into the nucellar tissue in the direction of the female gametophyte. After pollination, nucellar cells located around the pollen grain and tube die off. We showed here that the nuclei of the nucellar cells were stained by TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP-fluorescein nick end labeling). The number of TUNEL-positive cells increased during pollen tube growth. The tips of pollen tube branches protruded into the nucellar cells to form a convex-concave junction. At this junction, the cell membrane of nucellar cells was separated from the cell wall and the protoplast shrank. Small vesicles and amorphous materials were released from the protoplast into the space between the cell membrane and wall. Vacuoles were collapsed, chromatin was condensed, and mitochondria and plastids were deteriorated in the shrunken protoplast. Agarose gel analysis of DNA isolated from the ovules showed a DNA ladder, suggesting that the nuclear DNA had undergone internucleosomal cleavage. These results suggest that nucellar cells undergo programmed cell death in response to pollen tube penetration with some features resembling apoptosis and other features peculiar to nucellar cells.

Journal Article↗

Acid phosphatase from needles of Pinus silvestris L. Purification of two interconvertible enzyme forms and characterization of a low-molecular weight factor associated with the enzyme.

Two main forms of acid phosphatase (orthophosphoric-monoester phosphohydrolase (acid optimum), EC 3.1.3.2) have been isolated from needles of Pinus silvestris L. The isoelectric points determined by isoelectro-focusing are 3.6 and 9.4. The molecular weights of both forms were estimated as 68 000. The two forms have similar kinetic properties. Chromatography of the acidic enzyme form on DEAE-cellulose results in conversion to the basic form. Interconversion in the reverse direction was demonstrated to occur on incubation of the basic enzyme form with a pine-needle homogenate or with a low-molecular weight fraction thereof. These findings suggest that the interconvertibility between the main enzyme forms is a result of an association of the basic form with a low-molecular weight factor carrying multiple negative changes. This factor has been purified and tentatively identified as an oligoribonucleotide or a fragment of RNA. The implications of these findings for the application of acid phosphatase isoenzymes as genetic markers in forest trees are discussed.

Acid Phosphatase↗

Isocitrate lyase of conifers (Pinus pinea).

1. Isocitrate lyase has been purified about 60 times from the conifer Pinus pinea. A first characterization was made. 2. The high instability is an important feature of this enzyme from higher plants, this causes serious problems in the purification and characterization. 3. A substantial agreement with the data from the literature was found for what concerns pH dependence of Vmax and pKm, the effect of bivalent cations and the requirement of Mg2+. 4. Kinetic studies gave evidence for a mechanism ordered uni-bi with glyoxylate being the last product released, kinetic constants were calculated, no evidence for cooperative effects was found. 5. Equilibrium constant by Haldane method calculation agrees with value calculated with isocitrate lyase from the bacterium Pseudomonas indigofera.

Cations, Divalent↗

15-hydroperoxydehydroabietic acid--a contact allergen in colophony from pinus species.

A new hydroperoxide, 15-hydroperoxydehydroabietic acid (15-HPDA), with contact allergenic properties has been detected in rosin obtained from Pinus species. Detection was facilitated using a synthetic preparation of 15-HPDA for reference purposes. The synthesis and the detection (HPLC and GC) of 15-HPDA in rosin are described. The allergenic activity of 15-HPDA was studied in an experimental sensitization test on guinea-pigs.

Abietanes↗

Purification of an acidic coniferin-hydrolysing beta-glucosidase from developing xylem of Pinus banksiana.

Two cytosolic, acidic (pI 3.8) glycoproteins (M(r) 110,000 and 90,000) from lignifying xylem of Pinus banksiana were electrophoretically isolated and confirmed by combined gas chromatography-mass spectrometry to be capable of hydrolysing E-coniferin to trans-coniferyl alcohol. These isoenzymes yielded identical N-terminal sequences having negligible homology with documented glucosidases, suggesting that they constitute a novel form of beta-glucosidase.

Amino Acid Sequence↗

Regional variation of selected polyaromatic and chlorinated hydrocarbons over the South Island of New Zealand, as indicated by their content in Pinus radiata needles.

We analysed needles of Pinus radiata along three transects through the South Island of New Zealand for chlorinated hydrocarbons ( alpha- and gamma-HCH, HCB and PCB) and PAH (fluoranthene, 3,4-benzopyrene, benzo(ghi)perylene and indenopyrene). At rural sites we also collected samples of moss (Wijkia extenuata) and lichens (Usnea sp.) in order to compare the pollutant concentrations within these plants. A principal component analysis confirmed findings from moss and snow studies in Europe that the PAH concentrations vary regionally in the same way. The pattern of the principal component loadings of the chlorinated hydrocarbons reflects their heterogeneous immission and their different chemodynamics. The regional pattern indicates an increase of gamma-HCH from remote areas to those used intensively for agriculture and gardening, followed by a decrease towards the inner city of Christchurch. In contrast, PCB and PAH show their highest levels of contamination in the city. The regional distribution of PAH in Christchurch correlates well with traffic density, and particularly with the pattern of smoke from domestic fires. Our study demonstrates that needles of P. radiata can be better monitors of atmospheric pollution in New Zealand than mosses or lichens.

Journal Article↗

Height growth of different European Scots pine Pinus sylvestris L. Provenances in a heavily polluted and a control environment.

Results are presented of height measurements and degree of needle injury on five-year-old plants of Scots pine (Pinus sylvestris L.) growing near a phosphate fertiliser plant that emits SO(2) and fluorides. The populations of Scots pine represented in this experiment originate from 11 countries and were substantially differentiated in height growth and extent of needle necroses. Those populations which grew most rapidly were found to be the most sensitive to pollutant injury. The least productive provenances from the north of the range (Sweden, USSR) are at the same time characterized by lowest decline in height growth, lowest mortality and least extensive necroses. It is proposed that gene banks be established for the best genotypes likely to be eliminated in the heavily polluted conditions of Poland today.

Journal Article↗

Effect of simulated acid rain on quantity of epiphytic microfungi on Scots pine (Pinus sylvestris L.) needles.

The effects of simulated acid rain on the phyllosphere microflora of pine (Pinus sylvestris L.) were studied experimentally in northern Finland during the summer of 1988. Trees were irrigated with artificial acid rain of pH4 and pH3 (H(2)SO(4) and HNO(3), weight ratio of S:N=2.86:1). Untreated trees and trees irrigated with spring water (pH6) were used as controls. Two sampling heights (0.5m and 2m) were used. The needles were colonized exclusively by epiphytic fungi, mainly Aureobasidium pullulans (de Bary) Arnaud. The lower branches had significantly more epiphytic fungi than the upper branches. Compared to the control trees, the numbers of epiphytic fungi were significantly decreased on the needles of trees irrigated with acid rain. Acid rain affected the number of epiphytic fungi equally at both sampling heights. The species composition of the epiphytic fungi was not affected by the acid treatments.

Journal Article↗