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Vitality fertilization of Scots pine stands growing along a gradient of heavy metal pollution: short-term effects on microbial biomass and respiration rate of the humus layer.

In 1992 forest vitality fertilization experiments were established on a heavy metal deposition gradient with four treatments in three replications at distances of 0.5, 4 and 8 km from a Cu-Ni smelter in order to estimate their impact on the disturbed forest ecosystem. The increase in Cu concentration in the humus (F/H) layer of the Calluna site type Scots pine (Pinus sylvestris) stands from ca. 300 to 8000 mg kg(-1) d.m. (dry matter) along the 8 km long transect towards the smelter resulted in declining soil microbial biomass and soil respiration activity. Three independent measurements of microbial biomass: C(mic)-FE (fumigation extraction), C(mic)-SIR (substrate induced respiration), and ATP have been used together with an indicator of fungal biomass (ergosterol) and microbial activity (soil respiration). Within this Cu pollution range, all the measured microbial biomass levels declined to 10%-28% of the control plot values and activity assessed by respiration was lowered to 16%. Liming has increased the C(mic)-SIR and respiration rate. Treatments with test fertilizer, made from grounded apatite, did not result in different microbial biomass and respiration rate values compared to the respective controls along the whole gradient. Nitrogen + lime treatments resulted in similar changes to lime alone. No changes, as compared to the respective control, could be detected with nitrogen fertilization at the less polluted end of the gradient.

Journal Article↗

PROFILE: Impending Recovery of Kirtland's Warbler: Case Study in the Effectiveness of the Endangered Species Act

/ The Endangered Species Act (ESA) has received a large amount of criticism in recent years by conservative landowners and others who believe that it has infringed on property rights. It also has been criticized by those who think it has been costly and ineffective in reaching its goal of preventing extinction and recovering species. Recent evidence, however, shows that the ESA has stabilized or increased the populations of over a third of the listed species. In addition, its chief administrator, the US Fish and Wildlife Service, has been increasingly flexible in implementing the ESA. After reviewing the administrative machinery of the ESA, this paper provides a case study of one endangered species, the Kirtland's warbler (Dendroica kirtlandii). This particular recovery program actually began before passage of the federal ESA, when biologists alerted the Michigan Department of Natural Resources of the perilously low population of this bird, which only breeds under jack pine (Pinus banksiana) trees in Michigan. By the time an ESA Recovery Team was formed for this bird in 1975 (the first such team created under the ESA), a legacy of consensus and interagency cooperation was well established. This has led to successful efforts at habitat management and control of its nest parasite, the brown-headed cowbird (Molothrus ater). While the Kirtland's warbler is not yet recovered, its population is near an all-time high, and its recovery is possible within the next decade. When (and if) this happens, it will be clearly attributable to this successful model of federalism for natural resources management.KEY WORDS: Endangered species; Kirtland's warbler; Brown-headed cowbird; Jack pine; Endangered Species Act

Journal Article↗

Models to Assess the Risk of Snow and Wind Damage in Pine, Spruce, and Birch Forests in Sweden.

/ Each year damage to forests caused by snow and wind causes high economic losses. In Sweden, approximately 4 million m3 are damaged annually by snow and wind, roughly corresponding to a value of US$150 million, and in Europe, the damage amounts to hundreds of millions of US dollars each year. To help to reduce these losses, tools for risk assessment within forest management have been developed. Predictions were developed of the risk of damage from snow and wind to Scots pine (Pinus sylvestris L.), Norway spruce [Picea abies (L.) Karst.] and Birch (Betula spp. L.) plots using tree, stand, and site characteristics. The data were obtained from 6756 permanent sample plots within the Swedish National Forest Inventory, which were inventoried twice at five-year intervals between 1983 and 1992. Input data for model development used measurements from the first inventory of tree characteristics for the largest sample tree, stand, and site data, and records of snow and wind damage from the second inventory. Models were developed for three different regions for pine- and spruce-dominated sites, while models for the whole country were developed for birch sites. In general the estimated proportion of damaged plots was highly overestimated (31.7%-56.2%), compared with the observed proportion of 3.4%-11.9%. The models for Norway spruce comprising tree, stand, and site data show the best predictability of damaged plots, with 60.6%-67.6% of plots correctly classified. It is concluded that the models developed can be used to detect sites with a high probability of damage from snow and wind, and thus be used as tools to reduce future damage and costs in practical forestry.KEY WORDS: Birch; Norway spruce; Risk assessment; Risk management; Scots pine; Silviculturehttp://link.springer-ny.com/link/service/journals/00267/bibs/24n2p209.html

Journal Article↗

The chlB gene encoding a subunit of light-independent protochlorophyllide reductase is edited in chloroplasts of conifers.

ChlB is one of three chloroplast genes shown so far to be required for light-independent chlorophyll synthesis. It occurs in some algae, lower plants, and gymnosperms, but not in angiosperms. We have demonstrated, for the first time in conifer chloroplasts, the presence of two internal C to U editing sites in this transcript. In the chlB transcript of Pinus sylvestris, the editing of the second position in a CCG codon leads to an amino-acid substitution from proline to leucine. Editing of a nearby CGG codon, resulting in an arginine to tryptophan substitution, has also been observed. The nucleotide sequence of this region has been compared with other species of gymnosperms. Out of seven species analysed, editing at both sites has only been detected in spruce, while in Larix only the editing which results in the Arg to Trp substitution was found. In other cases, both leucine and tryptophan are encoded by cpDNA, suggesting that conservation of these amino acids, through encoding by DNA or by editing of the RNA, is critical for the protein function. Transcripts are partially edited at the CGG codon and the relative abundance of cDNA molecules with the edited C is species-specific. The possible involvement of RNA editing in the regulation of gene expression in different organs of pine seedlings is discussed.

Amino Acid Sequence↗

Antifungal activity of stilbenes in in vitro bioassays and in transgenic Populus expressing a gene encoding pinosylvin synthase.

The effect of two stilbene compounds, pinosylvin and resveratrol, on the growth of several fungi was evaluated in plate tests. Wood decay tests were carried out with birch and aspen samples impregnated with the two stilbenes. In plate experiments, resveratrol had an enhancing effect on growth at concentrations where pinosylvin was already enough to prevent the growth of most fungi studied. Pinosylvin impregnated at 0.2% (w/w) concentration significantly reduced the decay caused by all fungi except Phellinus tremulae. In contrast, a resveratrol content of 0.8%, did not protect the wood from decay. A pinosylvin-synthase-encoding gene from Pinus sylvestris was transferred into aspen ( Populus tremula) and two hybrid aspen clones ( Populus tremulax tremuloides) by Agrobacterium tumefaciens-mediated transformation. Transgenic plants accumulated pinosylvin synthase-specific mRNA and showed stilbene synthase enzyme activity in vitro. Transgenic aspen line H4 showed increased resistance to Phellinus tremulae, while two hybrid aspen transformants decayed faster than the control trees. However, we were unable to detect the accumulation of stilbenes in the transgenic plantlets.

Acyltransferases↗

Encoding of host and non-host plant odours by receptor neurones in the eucalyptus woodborer, Phoracantha semipunctata (Coleoptera: Cerambycidae).

In the eucalyptus woodborer, Phoracantha semipunctata, responses from single receptor neurones to host ( Eucalyptus globulus) and non-host ( Pinus pinaster, Olea europeae) plant odours were examined, using gas chromatography linked with electrophysiological recordings. Thirty-two compounds elicited responses that appeared as increased firing rates correlating with the elution of the active components. A subset of neurones classified as type A ( n=17) responded to one compound, a second subset B ( n=15) responded to two compounds and a third subset C ( n=9) responded to three or more compounds. For each neurone of type B or C, the active chemicals were structurally related. Neurones responding to monoterpenes, sesquiterpenes and non-terpenoids, respectively, showed non-overlapping response spectra, whereas neurones within one chemical group showed some overlap. The results suggest that most neurones, which receive information about plant odours, are narrowly rather than broadly tuned, each specialised for the detection of one or two related compounds. Although most neurones responded to compounds released by host and non-host species, a few responded specifically to compounds of only one species. This suggests a chemical "fingerprint" mechanism for discrimination between odour blends of the different plants, where only a few compounds may be specific for a particular species or individual.

Animals↗

Identification of plant odours activating receptor neurones in the weevil Pissodes notatus F. (Coleoptera, Curculionidae).

Plants release complex mixtures of volatiles important in the interaction with insects and other organisms. In the search for compounds that contribute to the perception of odour quality in the weevil Pissodes notatus, single olfactory receptor neurones on the antennae were screened for sensitivity to naturally produced plant volatiles by the use of gas chromatography linked to single cell recordings. We here present 60 olfactory neurones responding to 25 of the numerous compounds released by host and non-host plants. All the neurones show high selectivity and are classified into 12 distinct types. The two most abundant types respond to alpha-pinene, beta-pinene, and 3-carene ( n=17), and to isopinocamphone and pinocamphone ( n=17), respectively. Other neurone types respond to limonene ( n=9), beta-phellandrene ( n=3), and fenchone ( n=4). Responses to beta-caryophyllene ( n=1) and to ethanol ( n=4) are also shown. Except for two pairs, the neurone types do not show overlap of the molecular receptive range. The active compounds are present in the host, Pinus pinaster, as well as in non-hosts, supporting the idea that plant odour quality is mediated by the ratio of the compounds rather than specific odorants.

Animals↗

Identification of processes governing long-term accumulation of 137Cs by forest trees following the Chernobyl accident.

The regularities of 137Cs distribution in trees (Pinus sylvestris and Betula pendula) growing in different types of forest ecosystems were investigated. High levels of heterogeneity of 137Cs activity concentrations in different parts of the trees, resulting from their varied metabolism have been shown. The data obtained demonstrate a non-uniform character of 137Cs distribution along the trunks, which can be explained by radionuclide fixation by the xylem vessel walls and by geometry changes along the tree trunk. It has been found that the radial distribution of 137Cs in the tree trunk is dependent on the availability of 137Cs in soil, which governs the transfer of this radionuclide via xylem sap and on the properties of the xylem. The accumulation of 137Cs by trees was influenced by the vertical distribution and availability of 137Cs in the soil as well as by the root biomass distribution in different soil horizons. A bioavailability factor, which takes into account the vertical distribution of radiocesium in soil, bioavailability of this radionuclide and distribution of root biomass in different soil horizons is proposed for comparative analyses of 137Cs transfer from soil to trees in different types of forest ecosystems.

Cesium Radioisotopes↗

Regulation of amino acid uptake in conifers by exogenous and endogenous nitrogen.

Although an accumulating amount of research clearly indicates that plants are capable of taking up exogenous amino acids, the actual importance of such organic N sources for plant N nutrition is under debate. In this study, we show that amino acid uptake by Scots pine (Pinus sylvestris L.) is significantly decreased by elevated internal NH(4)(+) levels, while it increases following exposure to exogenous amino acids. Furthermore, amino acid uptake is larger in N-deficient plants than in plants grown with a large access of N. The regulatory pattern of amino acid uptake shows important similarities to the regulation of NO(3)(-) and NH(4)(+) transport as well as to the regulation of yeast amino acid transporters. In addition, our data suggest that uptake may be regulated by factors not originating from N metabolism. The up-regulation of uptake in response to N deficiency suggests that amino acid uptake may be a significant contributor to the N economy of P. sylvestris.

Adaptation, Physiological↗

Repeated freeze-thaw cycles induce embolism in drought stressed conifers (Norway spruce, stone pine).

Freezing and thawing lead to xylem embolism when gas bubbles caused by ice formation expand during the thaw process. However, previous experimental studies indicated that conifers are resistant to freezing-induced embolism, unless xylem pressure becomes very negative during the freezing. In this study, we show that conifers experienced freezing-induced embolism when exposed to repeated freeze-thaw cycles and simultaneously to drought. Simulating conditions at the alpine timberline (128 days with freeze-thaw events and thawing rates of up to 9.5 K h(-1) in the xylem of exposed twigs during winter), young trees of Norway spruce [Picea abies (L.) Karst.] and stone pine (Pinus cembra L.) were exposed to 50 and 100 freeze-thaw cycles. This treatment caused a significant increase in embolism rates in drought-stressed samples. Upon 100 freeze-thaw cycles, vulnerability thresholds (50% loss of conductivity) were shifted 1.8 MPa (Norway spruce) and 0.8 MPa (stone pine) towards less negative water potentials. The results demonstrate that freeze-thaw cycles are a possible reason for winter-embolism in conifers observed in several field studies. Freezing-induced embolism may contribute to the altitudinal limits of conifers.

Adaptation, Physiological↗

A small family of novel CuZn-superoxide dismutases with high isoelectric points in hybrid aspen.

Several CuZn-superoxide dismutases (SODs; EC 1.15.1.1) were cloned from hybrid aspen (Populus tremula L. x tremuloides Michx.). Two of the cloned genes encode representatives of a novel type of CuZn-SOD and we named it HipI-SOD because of its high isoelectric point (> or =9). The SODs were cloned by screening a cDNA library with a probe based on a Scots pine (Pinus sylvestris L.) CuZn-SOD that is predominantly located extracellularly. The expression pattern of HipI-SOD was examined using a Northern blot technique and compared with the expression patterns of cytosolic and chloroplastic SODs. Distinct expression patterns were observed for the three types of CuZn-SOD, with HipI-SODs showing strong expression in apical tissues. Southern blots as well as protein analysis suggest that these novel HipI-SODs belong to a small gene family, one member of which might be monomeric.

Amino Acid Sequence↗

Synthesis and oxidative insolubilization of cell-wall proteins during osmotic stress.

The cell walls in the new white roots of jack pine (Pinus banksiana Lamb.) were observed to constrict around the shrinking protoplast of osmotically stressed roots, and pressure was maintained via an apparent adjustment of cell-wall size and elasticity. These elastic alterations of the cell wall permitted the root cells to maintain full turgor despite the loss of most of the water in the tissue. The constriction of the root cell wall around the dehydrating protoplasts to maintain turgor may reflect changes in cell wall structure. We found that these shrinking root cells synthesize and secrete into the intercellular fluid a set of proteins. These proteins become tightly associated (i.e. guanidine HCl- and sodium dodecyl sulfate-insoluble) with the cell wall but can be released from the matrix, after briefly boiling in 0.1% sodium dodecyl sulfate, by the combination of guanidine HCl, CaCl2 and dithiothreitol. However, these cell-wall proteins became insoluble with time. The proteins could subsequently be destructively extracted from the wall with acid NaClO2 treatments. After these proteins were incorporated into the cell walls, the roots adopted a new, smaller maximal tissue volume and elastic coefficients returned to normal levels.

Amino Acid Sequence↗

Myosin, microtubules, and microfilaments: co-operation between cytoskeletal components during cambial cell division and secondary vascular differentiation in trees.

The immunolocalisation of unconventional myosin VIII ('myosin') in the cells of the secondary vascular tissues of angiosperm (Populus tremula L. x P. tremuloides Michx. and Aesculus hippocastanum L.) and gymnosperm (Pinus pinea L.) trees is described for the first time and related to other cytoskeletal elements, as well as to callose. Both myosin and callose are located at the cell plate in dividing cambial cells, whereas actin microfilaments are found alongside the cell plate; actin and tubulin are both associated with the phragmoplast. Myosin and callose also localise to the plasmodesmata-rich pit fields in the walls of living cells, which are particularly abundant within the common walls between ray cells and between ray cells and axial parenchyma cells in the phloem and xylem. In those xylem ray cells that contact developing vessel elements and tracheids, myosin, tubulin, actin and callose are localised at the periphery of developing contact and cross-field pits; the respective antibodies also highlight the bordered pits between vessels and between tracheids. The aperture of the bordered pits, whose diameter diminishes as the over-arching border of these pits develops, also houses myosin, actin and tubulin. Myosin, actin and callose are also found together around the sieve pores of sieve elements and sieve cells. We suggest that an acto-myosin contractile system (a 'plant muscle') is present at the cell plate, the sieve pores, the plasmodesmata within the walls of long-lived parenchyma cells, and at the apertures of bordered pits during their development.

Actin Cytoskeleton↗

Analysis of gymnosperm two-AP2-domain-containing genes.

AINTEGUMENTA (ANT) and APETALA2 (AP2) are transcription factors that are involved in several developmental processes in Arabidopsis thaliana. They are similar in structure, containing two AP2 domains, and have partially redundant functions in reproductive organ development. Expression and functional analyses of ANT and AP2 homologs have been performed previously in almost all angiosperms. In this study, one ANT homolog and two AP2 homologs were isolated from the gymnosperm Pinus thunbergii and were named PtANTL1, PtAP2L1, and PtAP2L2. PtANTL1 is the first reported gymnosperm ANT homologous gene. Based on a gene tree constructed with sequences of all A. thaliana two-AP2-domain-containing genes, it is likely that PtANTL1 and ANT, and likewise PtAP2L1 and AP2, are orthologs. The expression patterns of PtANTL1/PtAP2L1/PtAP2L2 were examined with Southern hybridization of the quantitative RT-PCR products and in situ hybridization. PtANTL1 and PtAP2L1 had almost identical expression patterns in the analyzed organs, and PtANTL1/PtAP2L1/PtAP2L2 were continually expressed in the developing female cone. Our analysis suggests that gymnosperms have orthologs to both ANT and AP2, and that the most recent common ancestor of extant seed plants has two type AP2 subfamily genes, ANT-like and AP2-like, involved in the development of female reproductive organs.

Amino Acid Sequence↗

The family of DOF transcription factors: from green unicellular algae to vascular plants.

This article deals with the origin and evolution of the DOF transcription factor family through a phylogenetic analysis of those DOF sequences identified from a variety of representative organisms from different taxonomic groups: the green unicellular alga Chlamydomonas reinhardtii, the moss Physcomitrella patens, the fern Selaginella moellendorffii, the gymnosperm Pinus taeda, the dicotyledoneous Arabidopsis thaliana and the monocotyledoneous angiosperms Oryza sativa and Hordeum vulgare. In barley, we have identified 26 different DOF genes by sequence analyses of clones isolated from the screening of genomic libraries and of ESTs, whereas a single DOF gene was identified by bioinformatics searches in the Chlamydomonas genome. The phylogenetic analysis groups all these genes into six major clusters of orthologs originated from a primary basal grade. Our results suggest that duplications of an ancestral DOF, probably formed in the photosynthetic eukaryotic ancestor, followed by subsequent neo-, sub-functionalization and pseudogenization processes would have triggered the expansion of the DOF family. Loss, acquisition and shuffling of conserved motifs among the new DOFs likely underlie the mechanism of formation of the distinct subfamilies.

Algal Proteins↗

Retroelements contribute to the excess low-copy-number DNA in pine.

Excess DNA in the single-copy component is rarely recognized as a contributor to the C-value paradox yet the single-copy component of the pine genome is reported to comprise over 3000 Mb of DNA, in large excess over the estimated 100 Mb required for gene expression. Two hypotheses regarding the factors that might contribute to the excess low-copy-number DNA were tested. The first hypothesis proposes that the excess low-copy kinetic component is actually overestimated by reassociation data analysis. To test this, a previously published C0t curve for Pinus strobus was reanalyzed using a new estimate of genome size based on laser flow cytometry. Part of the excess low-copy-number DNA in the pine genome could be attributed to the choice of parameters used in the analysis of the reassociation data. The second hypothesis holds that diverged retrotransposons contribute to the excess low-copy DNA. Sequences randomly sampled from single-copy and low-repetitive kinetic components of the P. taeda genome were characterized. Twelve of 46 fragments cloned from these fractions were found to show sequence similarity to retroelements: hence diverged retroelements contribute to the excess low-repetitive kinetic component in the pine genome. Similarity search was shown to be a conservative method for identifying retroelements, and thus the number of retroelements in the low-copy component was actually underestimated. Most of the retroelements in this fraction were nonfunctional. divergent from known retroelement families and previously reported only for flowering plants. Divergent retrotransposons are thus a major factor contributing to the expansion of the low-repetitive DNA component in higher plants.

Amino Acid Sequence↗

Families of clustered microsatellites in a conifer genome.

Microsatellite clustering may account for genetic maps which do not coalesce into the expected number of linkage groups. Microsatellite organization within the large genome of Pinus taeda (1C = 20,000 Mb) was determined by (1) testing whether repeat motifs were sequestered within the low-copy DNA kinetic component and (2) testing for repeat motif clusters within DNA fragments regardless of copy number. Within the low-copy kinetic component, either (AC)n or (AG)n repeat units were present in 32% of sequences. No repeat motifs were found in the total genome control. Clustered repeat motifs were frequent; the (ATG)n triplet repeat motif was located upstream from a CG-rich trinucleotide microsatellite in 26 out of 44 microsatellite sequences. Fourteen of the clustered (ATG)n sequences could be assembled into four microsatellite sequence families based on similarities in the flanking regions. Consistent with the DNA turnover model, family members shared similar flanking regions but differed in repeat motif composition and length.

Base Sequence↗

Stem respiratory potential in six softwood and four hardwood tree species in the central cascades of Oregon.

Mature and old growth trees of varying sapwood thickness were compared with regard to stem respiration. An increment core-based, laboratory method under controlled temperature was used to measure tissue-level respiration (termed respiratory potential) of ten different tree species. Bark (dead outer and live inner combined), sapwood, and heartwood thickness measurements were used to predict sapwood volume from stem diameter (including bark) for four of the ten species. These predictions of sapwood volume were used to scale respiratory potential to the main-bole level (excluding all branches). On the core level, species that maintained narrow sapwood (8-16% of bole radius) such as Pseudotusga menziesii, Taxus brevifolia, and Thuja plicata, had sapwood respiratory potentials in the lower bole that were 50% higher (P<0.05) than species with wide sapwood (>16% of bole radius), such as Abies amabilis, Pinus monticola, and Tsuga heterophylla. This pattern was not observed for inner bark respiratory potential, or for sapwood respiratory potential within the crown. On the main-bole level, respiratory potential per unit volume was inversely correlated to the live bole volumetric fraction (inner bark plus sapwood divided by whole bole volume) (Adj. R(2)=0.6). Specifically, tree species with 18-20% of the main bole alive potentially respired 1.3-3 times more per unit live bole volume than species with over 40%, suggesting that the live bole was less metabolically active in tree species that maintained large volumes of sapwood.

Cell Respiration↗