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PubMed2001

Clinal differentiation and putative hybridization in a contact zone of Pinus ponderosa and P. arizonica (Pinaceae).

The widely distributed Pinus subsection Ponderosae is a species complex that has a transition zone among taxa in the southwestern United States. In southern Arizona and New Mexico at least two recognized taxa, Pinus ponderosa var. scopulorum and Pinus arizonica or P. ponderosa var. arizonica, are known to coexist in close proximity. In this study, we report the existence of populations where the taxa are sympatric. One of the key characteristics distinguishing taxa is the number of needles per fascicle; P. ponderosa typically has three, P. arizonica has five. We examined the spatial distribution of needle-number types in a belt transect that covers a transition zone from nearly pure three-needle types at the top of Mount Lemmon to five-needle types downslope, in the Santa Catalina Mountains, Arizona. The spatial distribution is inconsistent with there being both free interbreeding among types and selective neutrality of types. Trees with intermediate types, having combinations of three, four, and five needles and/or mean numbers of needles between 3.0 and 5.0, are spatially concentrated in the middle of the transition zone. The spatial distribution supports the occurrence of hybridization and introgression, and this is consistent with reported crossabilities of the types. The results suggest that selection is acting, either on needle number per se or on other traits of the ecotype with which it may be in linkage disequilibrium, to maintain the observed steep clinal differentiation.

Journal Article↗

PubMedDate not supplied

[Comparative analysis of the structure of SUP2 genes in Pichia pinus and Saccharomyces cerevisiae].

SUP2(SUP35) is an omnipotent suppressor gene, coding for an EF-1 alpha-like protein factor, involved in the control of translational accuracy in yeast Saccharomyces cerevisiae. A SUP2 gene analogue from yeast Pichia pinus was isolated by complementation of temperature-sensitive sup2 mutation of S. cerevisiae. Nucleotide sequence of the SUP2 gene of P. pinus codes for a protein of 82.4 kDa exceeding the SUP2 protein of S. cerevisiae for 6 kDa. The SUP2 gene product of P. pinus is similar to the Sup2 protein of S. cerevisiae by its structure and includes a highly conservative (76%) C-terminal region homologus to EF-1 alpha and a lowly conservative N-terminal region. The relation between the evolutionary conservativity of different regions of the Sup2 protein and their functional significance is discussed.

Amino Acid Sequence↗

PubMed1999

Construction of an AFLP genetic map with nearly complete genome coverage in Pinus taeda.

De novo construction of complete genetic linkage maps requires large mapping populations, large numbers of genetic markers, and efficient algorithms for ordering markers and evaluating order confidence. We constructed a complete genetic map of an individual loblolly pine (Pinus taeda L.) using amplified fragment length polymorphism (AFLP) markers segregating in haploid megagametophytes and PGRI mapping software. We generated 521 polymorphic fragments from 21 AFLP primer pairs. A total of 508 fragments mapped to 12 linkage groups, which is equal to the Pinus haploid chromosome number. Bootstrap locus order matrices and recombination matrices generated by PGRI were used to select 184 framework markers that could be ordered confidently. Order support was also evaluated using log likelihood criteria in MAPMAKER. Optimal marker orders from PGRI and MAPMAKER were identical, but the implied reliability of orders differed greatly. The framework map provides nearly complete coverage of the genome, estimated at approximately 1700 cM in length using a modified estimator. This map should provide a useful framework for merging existing loblolly pine maps and adding multiallelic markers as they become available. Map coverage with dominant markers in both linkage phases will make the map useful for subsequent quantitative trait locus mapping in families derived by self-pollination.

Chromosome Mapping↗

PubMed2002

Chitinase production in pine callus (Pinus sylvestris L.): a defense reaction against endophytes?

In shoot tip-derived tissue cultures of Scots pine (Pinus sylvestris L.), browning and subsequent degeneration of the culture is accompanied by lipid peroxidation and lignification of cells, which are characteristic features of a plant defense reaction. Since chitinases are enzymes acting primarily in plant defense, their expression was studied in pine callus in order to elucidate the defense reaction. Chitinases were present diversely in tissue cultures originating from shoot tips and embryos of P. sylvestris, in contrast to Pinus nigra embryogenic callus, where production of chitinases or browning was not detected. Because endophytic microbes had earlier been detected in buds of Scots pine, their subsequent presence in the tissue cultures was considered a potential cause of the defense reaction. Therefore, the presence of endophytes in the tissue cultures was examined by in situ hybridization. Endophytes were found to colonize heavily in 45% of the tissue cultures of P. sylvestris and to form biofilms, while the P. nigra callus was not found to contain any microbes. The endophytes seemed to propagate uncontrollably once a tissue culture of P. sylvestris was initiated. Regardless of the high level of chitinase production in the callus, the control of the endophytes presumably becomes inadequate during the tissue culture of P. sylvestris.

Biofilms↗

PubMed2002

Effects of trichloroacetic acid on the nitrogen metabolism of Pinus sylvestris--a 13C/15N tracer study.

Trichloroacetic acid (TCA) can be found in various environmental compartments like air, rain and plants all over the world. It is assumed that TCA is an atmospheric degradation product of volatile chloroorganic hydrocarbons. The herbicide effect of TCA in higher concentrations is well known, but not much is known about the phytotoxic effects in environmentally relevant concentrations. It can be shown in this study by using the 13C/15N stable isotope tracer technique that [13C]TCA is taken up by roots of two-year-old seedlings of Pinus sylvestris L. and transported into the needles. At the same time the effect of the substance on nitrogen metabolism can be analyzed by measuring the incorporation of 15NO3- into different nitrogen fractions of the plant. The more [13C]TCA incorporation, the higher the synthesis of 15N labelled amino acids and proteins is. These effects on the nitrogen metabolism are probably based on the activation of stress- and detoxification metabolism. It has to be assumed that there is an influence on N metabolism of Pinus sylvestris caused by the deposition of environmentally relevant TCA concentrations.

Biological Availability↗

PubMed2001

[Genetic effects of acute and chronic ionizing irradiation on Pinus sylvestris L., inhabiting the Chernobyl' meltdown area].

The main results of the 12-year radiation-genetic monitoring of radiobiological, cytogenetic, and genetic parameters in Pinus sylvestris forest plantation from the Chernobyl meltdown area are presented. The acute ionizing irradiation at doses > 1 Gy was shown to induce formation of morphoses and depressed growth; at doses > 2 Gy, the reproductive ability of the trees declined. The radiobiological parameters showed a linear or close to linear dose-dependence relationship. The acute irradiation at a dose of 0.5 Gy induced cytogenetic and genetic effects that were significantly higher than the corresponding control values. The relationship between the cytogenetic effects and the absorbed dose was exponential. The dependence of the mutation frequency at specific loci on the absorbed dose was described by a nonlinear curve. The results of cytogenetic analysis of seedlings obtained from seeds annually collected in zones of slight, moderate, and strong damage of Pinus sylvestris L. are presented.

Chromosome Aberrations↗

PubMed2006

Effects of heavy metal pollution on genetic variation and cytological disturbances in the Pinus sylvestris L. population.

This isoenzymatic and cytogenetic study has shown significant differences in genetic composition between two groups of Pinus sylvestris trees: tolerant and sensitive to heavy metal pollution. Total and mean numbers of alleles and genotypes per locus were higher in the pollution-sensitive group of trees, but heterozygosity (Ho) was lower in this group. Fixation index (F) indicates that trees tolerant for pollution were in the Hardy-Weinberg equilibrium, while the sensitive group had a significant excess of homozygosity. Cytological analyses demonstrated numerous aberrations of chromosomes in meristematic root tissue of seedlings developed from seeds collected from trees in the polluted area. The aberrations included chromosome bridges and stickiness, laggards, retarded and forward chromosomes, and their fragments. The mitotic index was markedly lower in this group of seedlings, as compared to the control. Both isoenzymatic and cytological analyses showed a significant influence of heavy metal ions on the genetic structure of the Pinus sylvestris population.

Chromosome Aberrations↗

PubMed2003

Nearly complete genetic maps of Pinus sylvestris L. (Scots pine) constructed by AFLP marker analysis in a full-sib family.

We have constructed nearly complete linkage maps of Pinus sylvestris (L.) using AFLP markers based on a two-way pseudo-testcross strategy in a full-sib family founded in an advanced breeding program. With 39 primer combinations, a total of 737 markers (320 from the mother and 417 from the father) segregated in a 1:1 ratio, corresponding to DNA polymorphism: heterozygous in one parent and null in the other. In the maternal parent, 188 framework markers were mapped in 12 linkage groups, equivalent to the Pinus haploid chromosome number, with a total coverage of 1,695.5 cM. In the paternal parent, 245 framework markers established a map with 15 linkage groups, spanning a genome length of 1,718.5 cM. The estimated total map length was L(F) = 1,681 cM for the female and L(M) = 1,645 cM for the male using a modified method-of-moment estimator. Combining these values with those estimated from the observed map lengths in both parents, we estimated the genome length in Scots pine to be between 1,600 and 2,100 cM. Our genome coverage was estimated to be more than 98% with a framework marker interval of 20 cM for both parents. Most of the female and male linkage groups were associated through the analysis of the intercross markers.

Chromosome Mapping↗

PubMed2004

Transgenic Pinus radiata from Agrobacterium tumefaciens-mediated transformation of cotyledons.

A method for Agrobacterium tumefaciens-mediated transformation of Pinus radiata cotyledon explants was developed using commercially available open-pollinated seed. Pinus radiata is the most widely planted commercial conifer species in the Southern Hemisphere. Reports on transformation of this species have relied on particle bombardment of embryogenic callus derived from immature embryos. The main drawback to the method is the small number of genotypes that are amenable to transformation and regeneration. Since more than 80% of genotypes of radiata pine can be regenerated using cotyledons from mature seed, cotyledon explants were cocultivated with A. tumefaciens strain AGL1 containing a plasmid coding for the neomycin phosphotransferase II (nptII) gene and the beta-glucuronidase (GUS) gene (uidA). Transformed shoots were selected using either geneticin or kanamycin. Critical factors for successful transformation were survival of the cotyledons after cocultivation and selection parameters. Of the 105 putative transformants that were recovered from selection media, 70% were positive for integration of the nptII gene when analysed by PCR. GUS histochemical assay for uidA expression was unreliable because of reaction inhibition by unidentified compounds in the pine needles. Further, only 4 of the 26 independent transformants characterised by PCR and Southern analysis contained an intact copy of both genes. The remaining 22 transformants appeared to have a truncated or rearranged copy of the T-DNA. It is possible that the truncation/rearrangements are due to the Cauliflower mosaic virus (CaMV) 35S promoter. Analysis of the T-DNA junction sites and sequencing of the introduced DNA will help elucidate the nature of T-DNA insertion so that genetic modification of radiata pine can be targeted effectively.

Agrobacterium tumefaciens↗

PubMed2006

PpRT1: the first complete gypsy-like retrotransposon isolated in Pinus pinaster.

We have isolated and characterized a complete retrotransposon sequence, named PpRT1, from the genome of Pinus pinaster. PpRT1 is 5,966 bp long and is closely related to IFG7 gypsy retrotransposon from Pinus radiata. The long terminal repeats (LTRs) have 333 bp each and show a 5.4% sequence divergence between them. In addition to the characteristic polypurine tract (PPT) and the primer binding site (PBS), PpRT1 carries internal regions with homology to retroviral genes gag and pol. The pol region contains sequence motifs related to the enzymes protease, reverse transcriptase, RNAseH and integrase in the same typical order known for Ty3/gypsy-like retrotransposons. PpRT1 was extended from an EST database sequence indicating that its transcription is occurring in pine tissues. Southern blot analyses indicate however, that PpRT1 is present in a unique or a low number of copies in the P. pinaster genome. The differences in nucleotide sequence found between PpRT1 and IFG7 may explain the strikingly different copy number in the two pine species genome. Based on the homologies observed when comparing LTR region among different gypsy elements we propose that the highly conserved LTR regions may be useful to amplify other retrotransposon sequences of the same or close retrotransposon family.

Amino Acid Sequence↗

PubMed2003

Abundance, diversity, and vitality of mycorrhizae of Scots pine (Pinus sylvestris L.) in lignite recultivation sites.

Scots pine (Pinus sylvestris L.) stands cover large areas in the Lusatian and the Middle German lignite mining districts. Due to adverse chemical substrate conditions, the root systems of the trees are restricted to the ameliorated top-spoil and the organic forest floor layers. To investigate functioning of fine root systems under the prevailing site factors, we studied mycorrhizal colonization rate and frequency as well as mycorrhizal diversity, vitality and growth phases in Scots pine ecosystems along a chronosequence in both mining districts. Mycorrhizal rate was close to 100% in both districts. Mycorrhizal abundance was higher in the organic forest floor layer than the mineral soil layer. In total, 25 morphotypes were recorded. Diversity differed between the districts. The mycorrhizae of Amphinema byssoides, Tuber puberulum, Pinirhiza discolor, Pinirhiza cf. bicolorata and E-type were present in both mining areas. These morphotypes are typical of nutrient-rich soils with high pH values. Compared with the undisturbed sites, vitality of mycorrhizae was very high at the test sites on spoil substrate, correlating with the high growth dynamics of mycorrhizae at recultivation sites. A relatively high carbon flow to the mycorrhizal root systems at these sites seems likely. Thus, mycorrhizal root systems are able to cope with the ameliorated top-spoil and the organic layer. The main reason for the adaptation is the large number of ectomycorrhizal fungal species available in this area where Pinus sylvestris is indigenous.

Biodiversity↗

PubMed2006

Selecting ectomycorrhizal fungi for inoculating plantations in south China: effect of Scleroderma on colonization and growth of exotic Eucalyptus globulus, E. urophylla, Pinus elliottii, and P. radiata.

Plantation forestry with exotic trees in south China needs compatible symbionts to improve the growth of seedlings in nurseries and to enhance establishment and growth in the field. Scleroderma, a potentially suitable symbiont for inoculation, is not being used in containerized nurseries in the region due to poor knowledge of its host range. The ability of 15 collections of Scleroderma, nine from Australia and six from Asia, to colonize and promote growth of four important exotic plantation trees (Eucalyptus globulus Labill., Eucalyptus urophylla ST Blake, Pinus elliottii Engl., and Pinus radiata D. Don) was examined in a nursery potting mix. There was generally low host specificity of Scleroderma between tree genera. At 12 weeks after inoculation, 13 to 14 of the 15 spore collections formed ectomycorrhizas on seedlings of eucalypts or pines. The extent of colonization differed between spore treatments with two or four collections forming abundant mycorrhizas (>50% fine roots colonized) on E. globulus or E. urophylla, respectively, and three or five on P. radiata or P. elliottii, respectively. Three collections from Australia strongly colonized all hosts resulting in 26 to 100% of short roots being colonized. Chinese Scleroderma collections resulted in fewer mycorrhizas on eucalypts than on pines. Inoculation stimulated the growth (shoot height and dry weight) of eucalypt and pine seedlings by up to 105% where Scleroderma mycorrhizas developed. The results suggest that there is a need to source Scleroderma from outside China for inoculating eucalypts in Chinese nurseries whereas Chinese collections of Scleroderma could be used in pine nurseries. Further screening of Australian and Chinese Scleroderma should be performed in Chinese nurseries and in the field before final commercial decisions are made.

Agriculture↗

PubMed2005

The effects of humic substances on Pinus callus are reversed by 2,4-dichlorophenoxyacetic acid.

The reversal of humic matter-induced inhibition of callus growth and metabolism by 2,4-dichlorophenoxyacetic acid (2,4-D) was studied in Pinus laricio. Two forest humic fractions (relative molecular mass (Mr) > 3500), derived from soil under Fagus sylvatica (Fs) and Abies alba (Aa) plantation, were used. Pinus laricio callus was grown for a subculture period (4 weeks) on Basal Murashige and Skoog (MS) medium plus forest humic matters (Fs or Aa), at a concentration of 1 mg C/l, and then was transferred, for an additional four weeks, to a MS medium culture without humic matter, but with different hormones: indole-3-acetic acid (IAA, 2 mg/l) or 2,4-dichlorophenoxyacetic acid (2,4-D, 0.5 mg/l) and/or 6-benzylaminopurine (BAP, 0.25 mg/l). Growth of calluse, glucose, fructose, and sucrose contents, and activities of soluble and bound invertases, glucokinase, phosphoglucose isomerase, aldolase, and pyruvate kinase were monitored. The results show a negative effect of humic fractions on callus growth, due to decreased utilization of glucose and fructose, and decreased activities of glycolytic enzymes. The effects are reversible. Substitution of humic fractions with 2,4-D+BAP or 2,4-D is followed by an increase of glycolytic enzyme activities and, consequently, by the utilization of glucose and fructose that induces a restart of growth. In contrast, the inhibitory effects of humic fractions persist when they are substituted with BAP alone, indicating that only the auxin 2,4-D is capable of reversing the negative effects. A possible competitive action on the auxin-binding site between 2,4-D and the chemical structures in the forest humic fractions is suggested.

2,4-Dichlorophenoxyacetic Acid↗

PubMed2005

Trace element accumulation in the moss Hypnum cupressiforme Hedw. and the trees Quercus ilex L. and Pinus halepensis Mill. in Catalonia.

We studied trace element accumulation in the moss Hypnum cupressiforme and the widely distributed Mediterranean trees Quercus ilex and Pinus halepensis located at increasing distances from the Barcelona Metropolitan Area. Hypnum cupressiforme, Quercus ilex and, to a somewhat lesser extent, Pinus halepensis, have proved to be adequate as possible accumulative monitoring species in relation to trace elements pollution. No significant effects of crown orientation were found. One-year old leaves generally accumulated more trace elements than current-year leaves. All the studied trace elements showed greatest concentrations in the Barcelona Metropolitan Area, with lead, cadmium and arsenic concentrations being especially high. In general, trace element concentrations in biomass were similar or higher than the values reported from other Mediterranean urban areas of Europe. The top soil-layer concentrations were also higher in the Barcelona Metropolitan Area indicating the existence of mechanisms of atmospheric deposition and/or concentration in the soil. The lower values of Pb of airborne origin relative to other elements such as Cd, Cu, Zn and Sb suggest that traffic exhausts are not the only important focus of pollutants in this area. The results of biomass concentrations and of enrichment factor of biomasses respect to bedrock and soils show that atmospheric inputs account for the higher trace element concentrations in the Barcelona Metropolitan Area.

Biomass↗

PubMed2005

Evidence for potential impacts of ozone on Pinus cembra L. at mountain sites in Europe: an overview.

We summarize what is known about the impact of ozone (O(3)) on Pinus cembra in the timberline ecotone of the central European Alps and the Carpathian Mountains. In the central European Alps exposure to ambient and two-fold ambient O(3) throughout one growing season did neither cause any visible injury nor affect the photosynthetic machinery and biochemical parameters in current to 1-year-old needles. By contrast, in the southern French Alps and in the Carpathians 1-year-old needles of Pinus cembra trees showed visual symptoms similar to those observed in O(3) stressed pine stands in southern California. For the southern French Alps the observed symptoms could clearly be attributed O(3) and differences in O(3) uptake seems to be the likely key factor for explaining the observed decline. For the Carpathians however, other reasons such as drought may not be excluded in eliciting the observed symptoms. Thus, the action of O(3) has always to be evaluated in concert with other environmental impacts, determining the tree's sensitivity to stress.

Altitude↗

PubMed2005

Indications of limited altered gene expression in Pinus sylvestris trees from the Chernobyl region.

To evaluate the consequences of irradiation on the vegetation of the Chernobyl region, gene expression was compared in morphologically normal and dwarf needles from the same Pinus sylvestris trees in a region where the absorbed dose was 3-5 Gy. To compare the levels of gene expression, arrays consisting of 373 Pinus taeda cDNAs were hybridized with labeled cDNA derived from normal and dwarf needles of P. sylvestris. Twelve genes were significantly (P<0.01) up- or down-regulated between normal and dwarf needles for all five trees taken together. Five of these, related to stress or development, were up- or down-regulated 1.25-1.7-fold in the dwarf needles. There were no significant differences in (137)Cs content in the normal and dwarf needles, or in elongation growth rate of seedlings raised under controlled conditions from seed derived from trees in the region that had received a radiation dose over the range 2-12 Gy.

Cesium Radioisotopes↗

PubMed2005

Diversity of ectomycorrhizal fungi naturally established on containerised Pinus seedlings in nursery conditions.

The study examined the diversity of ectomycorrhizal fungi, naturally established on roots of containerised Pinus seedlings in a nursery, using PCR-RFLP and sequencing of the nuclear ribosomal internal transcribed spacer. Seventy-two samples, including ectomycorrhizae and fruit bodies, were examined. Molecular typing assigned the fungal symbionts to four ectomycorrhizal Boletales: Rhizopogon rubescens, Suillus bovinus, S. variegatus, and R. luteolus. R. rubescens was abundant (37.5%), while Suillus and R. luteolus species were moderately established (25-26%) and rare (2.8%), respectively. In addition, Rhizopogon species colonised P. nigra ssp. salzmannii seedlings, whereas Suillus species were identified on Pinus nigra ssp. nigra seedlings. The diversity and the ability of these naturally established symbionts under artificial nursery conditions were discussed. The molecular survey investigated here should contribute to successful monitoring of mycorrhizal application under both nursery and plantation conditions.

Basidiomycota↗

PubMed2005

Fatty acids from seeds of Pinus pinea L.: composition and population profiling.

Pinus pinea L. is widely disseminated all over the Mediterranean Basin. Qualitatively, P. pinea fatty acid seed composition is identical and typical of the genus Pinus. This composition is made of unsaturated oil with several unusual polymethylene-interrupted unsaturated fatty acids. Linoleic acid is the major fatty acid followed by oleic, palmitic and stearic acids. Quantitatively, for all Mediterranean populations, total amounts of fatty acids seem to be fairly constant and independent from their origin. When applying principal component analysis, it seems that there is not a distinct geographical variability. Tunisian populations appear to be integral part of the Mediterranean populations without any particular structuring. Taking into account this research and the data reported in the literature, we can confirm that P. pinea expresses no significant variability. This low genetic diversity revealed by fatty acid composition can be explained by anthropogenetic diffusion of genetically homogeneous reproductive material as early as the first explorations.

Fatty Acids↗