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Daylength and temperature during seed production interactively affect adaptive performance of Picea abies progenies.

Adaptive traits in Picea abies (Norway spruce) progenies are influenced by the maternal temperatures during seed production. Here, we have extended these studies by testing the effects of maternal photoperiod and temperature on phenology and frost hardiness on progenies. Using eight phytotron rooms, seeds from three unrelated crosses were made in an environmental 2 x 2 factorial combination of long and short days and high and low temperatures. The progenies were then forced to cease growth rapidly at the end of the first growing season. An interactive memory effect was expressed the second growth season. Progenies from high temperature and short days, and from low temperatures and long days, started growth later in spring, ceased shoot growth later in summer, grew taller and were less frost hardy in the autumn than their full siblings from low temperatures and short days, and from high temperatures and long days. Norway spruce has developed a memory mechanism, regulating adaptive plasticity by photoperiod and temperature, which could counteract harmful effects of a rapidly changing climate.

Adaptation, Biological↗

Ectomycorrhizal community structure varies among Norway spruce (Picea abies) clones.

In northern boreal forests, the diversity of ectomycorrhizal (ECM) species is much greater than that of their host trees. This field study investigated the role of individual trees in shaping the ECM community. We compared ECM communities of eight Norway spruce (Picea abies) clones planted in a clear-cut area in 1994 with a randomized block design. In 2003, the ECM fungi were identified from randomly sampled root tips using denaturing gradient gel electrophoresis (DGGE) and rDNA internal transcribed spacer (ITS) sequence similarity. ECM diversity varied among clone groups, showing twofold growth differences. Moreover, according to detrended correspondence analysis (DCA), ECM community structure varied not only among but also within slow-growing or fast-growing clones. Results suggest that ECM diversity and community structure are related to the growth rate or size of the host. A direct or indirect influence of host genotype was also observed, and we therefore suggest that individual trees are partly responsible for the high diversity and patchy distribution of ECM communities in boreal forests.

Ecosystem↗

Cytological instability in tumors of Picea glauca.

Smear preparations of cells taken from primary explants of normal and adjacent tumor wood of Picea glauca showed completely regular mitotic behavior in the normal cells, with the great majority of cells diploid (22 chromosomes), a few tetraploid, but almost none aneuploid. Tumor tissue was extremely unstable, with numbers ranging from 3 to more than 70, with a high proportion of aneuploids but otherwise normal-appearing mitoses. The relation of this mitotic instability to other data on these tumors is pointed out.

Aneuploidy↗

SOMATIC MITOSES IN CELLS OF PICEA GLUCA CULTIVATED IN VITRO.

The cytology of one strain of tumor tissue from the spruce tree, Picea glauca, grown in vitro for more than a year was examined. The cells of this strain are characterized by a uniform chromosome number of 24, and the strain appears to be quite stable. The implications of the results of this study and previous studies on similar material are discussed.

Cell Division↗

Molecular profiling of rhizosphere microbial communities associated with healthy and diseased black spruce (Picea mariana) seedlings grown in a nursery.

Bacterial and fungal populations associated with the rhizosphere of healthy black spruce (Picea mariana) seedlings and seedlings with symptoms of root rot were characterized by cloned rRNA gene sequence analysis. Triplicate bacterial and fungal rRNA gene libraries were constructed, and 600 clones were analyzed by amplified ribosomal DNA restriction analysis and grouped into operational taxonomical units (OTUs). A total of 84 different bacterial and 31 different fungal OTUs were obtained and sequenced. Phylogenetic analyses indicated that the different OTUs belonged to a wide range of bacterial and fungal taxa. For both groups, pairwise comparisons revealed that there was greater similarity between replicate libraries from each treatment than between libraries from different treatments. Significant differences between pooled triplicate samples from libraries of genes from healthy seedlings and pooled triplicate samples from libraries of genes from diseased seedlings were also obtained for both bacteria and fungi, clearly indicating that the rhizosphere-associated bacterial and fungal communities of healthy and diseased P. mariana seedlings were different. The communities associated with healthy and diseased seedlings also showed distinct ecological parameters as indicated by the calculated diversity, dominance, and evenness indices. Among the main differences observed at the community level, there was a higher proportion of Acidobacteria, Gammaproteobacteria, and Homobasidiomycetes clones associated with healthy seedlings, while the diseased-seedling rhizosphere harbored a higher proportion of Actinobacteria, Sordariomycetes, and environmental clones. The methodological approach described in this study appears promising for targeting potential rhizosphere-competent biological control agents against root rot diseases occurring in conifer nurseries.

Bacteria↗

Application of ISSR, RAPD, and cytological markers to the certification of Picea mariana, P. glauca, and P. engelmannii trees, and their putative hybrids.

Picea glauca (white spruce) and P. engelmannii (Engelmann spruce) are so similar and integrated that it is impossible to distinguish between them and their hybrids using morphological characteristics. Although natural hybrids between P. glauca and P. mariana (black spruce) do not generally occur, even though the 2 species are sympatric in North America, a first-generation hybrid, called the Rosendahl spruce, has been reported in the literature. In this study, several inter-simple sequence repeat (ISSR) markers were developed, as were randomly amplified polymorphic DNA (RAPD) markers, to certify spruce trees and their hybrids. ISSR fingerprinting was more efficient than RAPD assay; it detected 70% polymorphic DNA markers among the spruce species analyzed, whereas RAPD fingerprinting detected only 53%. Species-diagnostic ISSR and RAPD markers differentiating P. glauca from P. engelmannii and P. mariana were cloned and sequenced. Molecular certification of the spruce samples analyzed confirmed that all the seeds from interior spruce populations were true hybrids of P. glauca and P. engelmannii. But the analysis of seeds derived from the putative Rosendahl spruce indicated that this tree is likely a pure P. glauca genotype, rather than a hybrid of P. glauca and P. mariana. These data were confirmed by cytological analyses. Further analysis, using a more sensitive DNA amplification method with designed primers flanking the species-diagnostic ISSR and RAPD markers, revealed that such sequences are not generally species-specific because they are present in other spruce species.

Base Sequence↗

New abietane-type diterpenes from the heartwood of Picea morrisonicola.

New abietane-type diterpenes, 15-acetoxy-7-oxodehydroabietic acid (1), picealactones A (2), B (3), and C (4), together with the known 7-oxodehydroabietic acid (5) were isolated and identified from the heartwood of Picea morrisonicola. The structures of 1-4 were determined on the basis of spectral data explanation. Compounds 2-4 possessed a rare 5-dehydro-18, 6-olide functionality. Compounds 1 and 2 were first isolated from natural source.

Abietanes↗

New constituents from the heartwood of Picea morrisonicola HAYATA.

Three new constituents, including a p-menthane type monoterpene, trans-p-menthane-7,8,9-triol (1), an aromatic, 2,6-dihydroxy-3,4-dimethylbenzoic acid methyl ester (2), and a lignan, (+)-morrisonicolanin (3), were isolated from the low polar layer of heartwood extracts of Picea morrisonicola, and their structures were elucidated on the basis of spectroscopic analysis. Of these compounds identified, 1 was obtained as its diacetylated derivative 1a, and 2 was the chemical entity first isolated from a natural source.

Benzoates↗

Formation of trans-verbenol and verbenone from alpha-pinene catalysed by immobilised Picea abies cells.

Both enantiomers and the racemate of alpha-pinene were transformed by Picea abies cells immobilised on alginate. The main products were cis- and trans-verbenol, the later being further transformed to verbenone. The enantiomeric purity of each product more or less corresponded to that of the substrate. Transformation by free cells was faster than that by the immobilised cells. The ratio of products differed to some extent between the transformation by free and immobilised cells.

Bicyclic Monoterpenes↗

Comparison of terpene composition in Engelmann spruce (Picea engelmannii) using hydrodistillation, SPME and PLE.

Terpenes emitted by conifer trees are generally determined by analysing plant extracts or essential oils, prepared from foliage and cones using steam distillation. The application of these procedures limits experiments to cut plant materials. Recently headspace techniques have been adopted to examine terpene emission by living plants. This paper deals with the application of solid-phase micro-extraction (SPME) for the analysis of terpenes emitted by conifers foliage of Engelmann spruce (Picea engelmannii), including its seedlings. The compositions of SPME extracts obtained for destroyed and non-destroyed old and juvenile spruce needles were compared with the compositions of essential oils and pressurised liquid extraction (PLE) extracts corresponding to the same plant materials. No substantial differences have been found in the qualitative terpene composition estimated by analysing essential oil and PLE and SPME extracts from non-destroyed old and juvenile foliage. The disintegration of spruce needles results in the formation of a significant amount of myrcene in the case of the old conifer foliage and non-terpenoic compounds in the case of juvenile conifer foliage. This phenomenon can be attributed to enzymatic reactions occurring in the destroyed plant cells.

Gas Chromatography-Mass Spectrometry↗

Multilocus patterns of nucleotide diversity, linkage disequilibrium and demographic history of Norway spruce [Picea abies (L.) Karst].

DNA polymorphism at 22 loci was studied in an average of 47 Norway spruce [Picea abies (L.) Karst.] haplotypes sampled in seven populations representative of the natural range. The overall nucleotide variation was limited, being lower than that observed in most plant species so far studied. Linkage disequilibrium was also restricted and did not extend beyond a few hundred base pairs. All populations, with the exception of the Romanian population, could be divided into two main domains, a Baltico-Nordic and an Alpine one. Mean Tajima's D and Fay and Wu's H across loci were both negative, indicating the presence of an excess of both rare and high-frequency-derived variants compared to the expected frequency spectrum in a standard neutral model. Multilocus neutrality tests based on D and H led to the rejection of the standard neutral model and exponential growth in the whole population as well as in the two main domains. On the other hand, in all three cases the data are compatible with a severe bottleneck occurring some hundreds of thousands of years ago. Hence, demographic departures from equilibrium expectations and population structure will have to be accounted for when detecting selection at candidate genes and in association mapping studies, respectively.

Chromosomes, Plant↗

Purine and pyrimidine nucleotide synthesis and degradation during in vitro morphogenesis of white spruce (Picea glauca).

In the last few years, somatic embryogenesis and organogenesis of white spruce (Picea glauca) have been used as model systems to investigate biochemical and physiological events related to morphogenesis. This has been possible because studies conducted in vitro allow for manipulations of the culture conditions, in which changes in morphogenetic events can be easily related to physiological alterations. De-novo synthesis, salvage, and degradation of both purine and pyrimidine nucleotides are operative at all stages of somatic embryo maturation and germination. Fluctuations in the activity of these pathways delineate important morphogenic events. The early phases of embryo development are accompanied by a decreased salvage activity of purine nucleotides, which reflects a reduction of cell proliferation and the initiation of organized growth. Activities of the salvage enzymes are present throughout the maturation period, and also during the imposition of the drying period, which is required for successful embryo germination. The operative salvage pathway in dried embryos is needed for the enlargement of the nucleotide pool necessary to sustain the reactivation of the overall cellular metabolism at germination, before the reactivation of the de-novo pathway, which is a later event. Manipulations of the culture medium which improve the germination frequency of the embryos also result in increased salvage activity. Similar changes in nucleotide synthesis were also observed during the initiation of shoot development from epicotyl explants of white spruce and cotyledons of radiate pine. Results from these studies can be used for improving growth and development in culture.

Morphogenesis↗

[Adaptive significance and possible origin of B-chromosomes in Picea Glauca (moench.) voss = P.canadensis B.S.P].

The karyotype (2n = 24 + 1B) was detected in 8 trees of Picea glauca (Moench.) Voss. = P. canadensis B.S.P., and the karyotype (2n = 24) was detected in 2 trees of spruce from the same stand in the centre of Voronezh. The majority of seeds, collected from these trees, were empty. From 150 selected seeds, which were full, four plantlets were obtained. One of them died, another one had no mitotic figures, and the third and fourth plantlets had karyotypes (2n = 24 + 1B, and 2n = 24, respectively). A possible mechanism of formation of B-chromosomes and their adaptive role are discussed.

Adaptation, Biological↗

[Karyological studies on Picea ajanensis (Lindl. et Gord.) Fisch. ex Carr. from different regions].

Results of karyological study of Picea ajanensis (Lindl. et Gord.) Fisch. ex Carr examined from 13 provenances are presented. In addition to the cytotypes with typical chromosome number (2n = 24), P. ajanensis displays cytotypes with one or two B-chromosomes (2n = 24 + 1 - 2B). Among A-chromosomes, there are 8 pairs of long metacentrics and 4 pairs of shorter meta- or submetacentrics. Among B-chromosomes there are two types of chromosomes: metacentric (B1) and submetacentric (B2) ones. There are many nucleolar chromosomes. Several chromosomes have secondary constrictions. Patterns of B-chromosome distribution within P. ajanensis are have been discussed.

Chromosomes, Plant↗

[Effects of light and temperature on seed germination of Picea asperata and Betula albo-sinensis].

The effects of different light and temperature conditions on seed germination of two keystone species (Picea asperata and Betula albo-sinensis) in subalpine coniferous forests were studied by simulating environmental conditions in climatic chambers. The results indicated that the optimal temperature of seed germination for both species was 25 degrees C, with the germination rate of 88.8 +/- 8.4% and 30.8 +/- 10.4%, respectively. Through multiple comparisons with Tukey's HSD, B. albo-sinensis was more sensitive to temperature than P. asperata. Seeds of P. asperata could germinate when the temperature reached 10 degrees C, but those of B. albo-sinensis could not, which implied that the latter as a pioneer species of subalpine coniferous forest areas needed higher lowest temperature for seed germination. Day and night temperature difference (15 degreesC/10 degrees C, 20 degrees C/15 degrees C and 20 degrees C/10 degrees C) had significant effect on the germination rates of tested seeds, which was the highest under 20 degrees C/15 degrees C. A wide range of day and night temperature difference seemed to be unfavorable to the seed germination of B. albo-sinensis. Light intensity also had a marked effect on the germination rates of test seeds. Under 20% transmittance, the germination rate of P. asperata and B. albo-sinensis was the highest, being 83.0 +/- 6.6% and 29.2 +/- 5.5%, respectively, and the initiation time and regularity of germination were the best as well. Furthermore, B. albo-sinensis was less sensitive to light than P. asperata, which was determined by the life behavior of tree itself.

Betula↗

[Genetic control of isozymes in European spruces (Picea abies (L) Karst) of the Ukrainian Carpathian mountains].

Genetical control of the enzymes GOT, GDH, DIA, MDH, SOD, FDH, ADH, ACP and LAP has been studied in nine natural Carpathian populations of Norway spruce (Picea abies (L.) Karst.) using polyacrylamide gel elecrophoresis and analysis of isozyme variability in 346 trees. Seventy one allel products of 20 gene loci have been clearly established. Segregation analysis of the revealed allele variants confirms their monogenic inheritance.

Alleles↗

Two-dimensional electrophoresis of acidic proteins isolated from ozone-stressed Norway spruce needles (Picea abies L. Karst): separation method and image processing.

The effect of ozone stress (< 10,200 or 400 micrograms m-3) on the protein pattern of Picea abies L. Karst needles was studied by fumigating potted grafts from mature trees. Clonal trees of Norway spruce were fumigated in open-top chambers for one growing season. Proteins, soluble and membrane bound, were extracted from the needles in a four-step procedure: (i) grinding in liquid nitrogen and then in a pH 5.0 buffer with thiourea added to inhibit phenol oxidase and polyvinylpolypyrrolidone (PVP) added to bind phenolic compounds; (ii) precipitation of proteins with acetone at -20 degrees C; (iii) solubilization in a modified lysis buffer; and (iv) dialysis against the modified lysis buffer. Proteins were separated by two-dimensional electrophoresis according to O'Farrell's method. The changes in the silver-stained protein pattern were evaluated by using image processing methods. Electrophoretograms from stressed and nonstressed trees are presented.

Carboxylic Acids↗

Vicilin-like seed storage proteins in the gymnosperm interior spruce (Picea glauca/engelmanii).

A seed storage protein cDNA was characterized from a library of interior spruce (Picea glauca/engelmanii complex) cotyledonary stage somatic embryos. The deduced amino acid sequence predicts a 448 amino acid (50 kDa) polypeptide with 28-38% identity with angiosperm vicilin-like 7S globulins. XXC/G codon usage is low (47%) relative to monocot angiosperms while pairwise comparisons show that spruce, monocot, and dicot vicilins are approximately equal in amino acid divergence. Although small by comparison, the spruce vicilin contains an N terminal hydrophilic region characteristic of angiosperm 'large' vicilins. Genomic Southern blotting predicts that the cDNA is encoded by a gene family.

Amino Acid Sequence↗