Trace elements in soil and coniferous needles.
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Gymnosperms possess two to four phytochrome types which apparently are the result of successive gene duplications in the genomes of their common ancestors. Phytochromes are nuclear-encoded proteins whose genes, contrary to chloroplast, mitochondrion, and rRNA genes, have hitherto rarely been used to examine gymnosperm phylogenies. Since the individual phytochrome gene types implied phylogenies that were not completely congruent to one another, conflicting branching orders were sorted by the number of gene lineages present in a taxon. The Gnetatae (two gene types) branched at the base of all gymnosperms, a position supported by bootstrap sampling (distance and character state trees, maximum likelihood). The Gnetatae were followed by Ginkgo, Cycadatae, and Pinaceae (three gene types) and the remaining conifers (four gene types). Therefore, in phytochrome trees, the most ancient branch of the conifers (Pinatae) seems to be the Pinaceae. The next split appears to have separated Araucariaceae plus Podocarpaceae from the Taxaceae/Taxodiaceae/Cupressaceae group. Structural arrangements in the plastid genomes (Raubeson and Jansen 1992) corroborate the finding that there is no close connection between Pinaceae and Gnetatae as suggested by some publications. The analyses are based on 60 phytochrome genes (579 positions in an alignment of PCR fragments) from 28 species. According to rough divergence time estimates, the last common ancestor of gymnosperms and angiosperms is likely to have existed in the Carboniferous.
A strategy for management of giant sequoia groves is formulated using a conceptual framework for ecosystem management recently developed by Region Five of the USDA Forest Service. The framework includes physical, biological, and social dimensions. Environmental indicators and reference variability for key ecosystem elements are discussed in this paper. The selected ecosystem elements include: 1) attitudes, beliefs, and values; 2) economics and subsistence; 3) stream channel morphology; 4) sediment; 5) water; 6) fire; 7) organic debris; and 8) vegetation mosaic. Recommendations are made for the attributes of environmental indicators that characterize these elements. These elements and associated indicators will define and control management activities for the protection, preservation, and restoration of national forest giant sequoia ecosystems.
The effect of stream geomorphology, maturity, and management of riparian forests on abundance, role, and mobility of wood was evaluated in 20 contrasting reaches in the Agüera stream catchment (northern Iberian Peninsula). During 1 year the volume of woody debris exceeding 1 cm in diameter was measured in all reaches. All large woody debris (phi > 5 cm) pieces were tagged, their positions mapped, and their subsequent changes noted. Volume of woody debris was in general low and ranged from 40 to 22,000 cm3 m-2; the abundance of debris dams ranged from 0 to 5.5 per 100 m of channel. Wood was especially rare and unstable in downstream reaches, or under harvested forests (both natural or plantations). Results stress that woody debris in north Iberian streams has been severely reduced by forestry and log removal. Because of the important influence of woody debris on structure and function of stream systems, this reduction has likely impacted stream communities. Therefore, efforts to restore north Iberian streams should include in-channel and riparian management practices that promote greater abundance and stability of large woody debris whenever possible.
A study was made of cambial activity, the localization of storage starch around the cambium, and the localization and occurrence of microtubules in cambial cells from dormancy to reactivation in locally heated (22-26 degrees C) stems of the evergreen conifer Abies sachalinensis. Heating induced localized reactivation of the cambium in the heated portions of the stem. Erect ray cambial cells resumed cell division 1 d prior to the reactivation of fusiform cambial cells and procumbent ray cambial cells. The re-initiation of the division of fusiform cambial cells occurred first on the phloem side. During the heat treatment, the amount of storage starch decreased in procumbent ray cambial cells and in the phloem parenchyma adjacent to the cambium but increased in fusiform cambial cells. Preprophase bands of microtubules, spindle microtubules and phragmoplast microtubules were observed both in erect ray cambial cells and in procumbent ray cambial cells. By contrast, no evidence of the presence of such preprophase bands of microtubules was detected in fusiform cambial cells. The results suggest that the localized heating of stems of evergreen conifers might provide a useful experimental model system for studies of the dynamics of cambial reactivation in intact 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.
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.
Microsatellite DNA/simple-sequence-repeat (SSR) loci were identified, isolated and characterized in white spruce (Picea glauca) by screening both a non-enriched partial genomic library and a partial genomic library enriched for (AG/TC)n-containing clones. Inheritance and linkage of polymorphic SSR loci were determined in F1 progeny of four controlled crosses. We also assessed the compatibility and usefulness of the P. glauca microsatellite DNA markers in five other Picea species. Twenty-four microsatellites were identified by sequencing 32 clones selected from screens of 5,400 clones from the two libraries. The (AG/TC)n microsatellites were the most abundant in the non-enriched library. Eight microsatellite DNA loci were of the single-copy type, and six of these were polymorphic. A total of 87 alleles were detected at the six polymorphic SSR loci in 32 P. glauca individuals drawn from several populations. The number of alleles found at these six SSR loci ranged from 2 to 22, with an average of 14.5 alleles per locus, and the observed heterozygosity ranged from 0.48 to 0.91, with a mean of 0.66 per locus. Parents of the controlled crosses were polymorphic for five of the six polymorphic SSR loci. Microsatellite DNA variants at each of these five SSR loci followed a single-locus, codominant, Mendelian inheritance pattern. Joint two-locus segregation tests indicated complete linkage between PGL13 and PGL14, and no linkage between any of the remaining SSR loci. Each of the 32 P. glauca individuals examined had unique single or two-locus genotypes. With the exception of non-amplification of PGL12 in P. sitchensis, P. mariana, and P. abies and the monomorphic nature of PGL7 in P. mariana, primer pairs for all six polymorphic SSR loci successfully amplified specific fragments from genomic DNA and resolved polymorphic microsatellites of comparable sizes in P. engelmanni, P. sitchensis, P. mariana, P. rubens, and P. abies. The closely related species P. mariana and P. rubens, and P. glauca and P. sitchensiss could be distinguished by the PGL12 SSR marker. The microsatellite DNA markers developed and reported here could be used for assisting various genetics, breeding, biotechnology, tree forensics, genome mapping, conservation, restoration, and sustainable forest management programs in spruce species.
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.
Thermogenically elevated cone temperatures were measured in two Macrozamia cycad species that differ in their daily heating time. Mathematical models of the cones' thermo-physics were tested for their accuracy in predicting these cone temperatures and for comparison of the energetics of both species and the sexes within species. These models accurately predicted temperatures over approximately 8-h periods with average errors of: 0.46 degrees C for Macrozamia lucida, pollinated by the thrips, Cycadothrips chadwicki, that moves during mid-day concurrent with cone heating; and 0.38 degrees C for Macrozamia machinii, pollinated by the weevil, Tranes sp., that moves after sunset during cone heating. The combination of models and experiments revealed a thermogenic sexual dimorphism in both species. For M. lucida, the estimated female mass specific metabolisms, and their theoretically possible and actual temperature increases due to thermogenic metabolism were only 57, 67, and 76% of males. In addition, female thermogenic metabolisms began and peaked much earlier and lasted significantly longer than males (all differences >1 h), and female metabolic peaks preceded their temperature peaks by 65 vs. 46 min for males. The timing of almost all male cone metabolic peaks was optimized with respect to the diurnal ambient heating cycle so that cone temperatures achieved a maximum temperature gain, whereas most female metabolic peaks occurred much earlier than optimal. In M. machinii, thermogenic sexual dimorphism is much larger since its male peak metabolisms are larger, and its females' peaks are much smaller compared to those of M. lucida. This study provides new information regarding the energetics of cycad cones that is relevant to understanding the interactions of the plant traits with their obligate pollinators' behavior.
Syagrus coronata is an economically important palm tree grown as an ornament, for the oil extracted from its seeds, and the wax from its leaves which has several applications in industry. Silicon biocomposites were analyzed in leaves of S. coronata. Silica bodies were found as extracellular silica masses between the hypodermal-layer cell walls and in granules present in the vacuoles of palisade cells. Scanning electron microscopy of the hypodermal layer of cells showed a collection of spherical bodies embedded in enveloping cavities that outlined the general structure of the bodies. Globular subunits with sharp edges formed the spherical bodies that ranged from 6 to 10 microm in diameter (average, 7.8 microm). X-ray microanalysis detected only silicon and oxygen homogeneously distributed throughout the bodies. Vacuoles of palisade cells contained a large number of granules ranging from 20 nm to 1.2 microm in size (average, 300 nm). Transmission electron microscopy associated with electron spectroscopic imaging and electron energy loss spectroscopy were used to determine the elemental composition of the granules. Vacuolar granules were amorphous and composed of silicon and oxygen, suggesting they consist of amorphous silica biominerals. No nitrogen, indicative of organic matter, was detected in the granules.
Sixty-four silicified fossil woods were collected from the Early Miocene Upper Coal-bearing Formation of Janggi Group in Pohang City, the Korean Peninsula. Out of them 23 specimens were identified as gymnosperms and 27 specimens as dicotyledons. The taxa identified include Picea palaeomaximowiczii Watari, Taxodioxylon cunninghamioides (Watari) Watari, T. sequoianum (Merckl.) Gothan, Fagus hondoensis (Watari) Watari, Cercidiphyllum sp., Camellia japonoxyla Suzuki et Terada, Distylium chiharu-hirayae Suzuki et Terada, Aesculus sp., Wataria miocenica (Watari) Terada et Suzuki and W. parvipora Terada et Suzuki. All of these species are reported for the first time from the Tertiary basins of Korea. Compared with those of the Miocene Formations in Japan, most taxa we found are common between the paleo-floras in Korea and Japan during the Early to Middle Miocene.
We describe details of anatomically preserved fossil glossopterid ovules from the Late Permian of Queensland, Australia, that contain several pollen tubes at various stages of releasing flagellated sperm. Each sperm is approximately 12.7 microm long and 13.9 microm wide, with a conspicuous spiral structure comprised of a series of dots that resemble the position of basal bodies of flagella aligned along the multilayered structure (MLS). This configuration is similar to the helically arranged flagella in the sperm of cycads, Ginkgo, and many pteridophytes. However, the motile gametes of Glossopteris are considerably smaller than those of Ginkgo and cycads, and more similar in size, number of basal bodies, and number of gyres in their helix to pteridophyte forms. Glossopteris thus shares the intermediate stage of motile male gamete formation and apparently that of haustorial pollen tubes with cycads and Ginkgo.
The delta5-unsaturated polymethylene-interrupted fatty acid (delta5-UPIFA) contents and profiles of gymnosperm seeds are useful chemometric data for the taxonomy and phylogeny of that division, and these acids may also have some biomedical or nutritional applications. We recapitulate here all data available on pine (Pinus; the largest genus in the family Pinaceae) seed fatty acid (SFA) compositions, including 28 unpublished compositions. This overview encompasses 76 species, subspecies, and varieties, which is approximately one-half of all extant pines officially recognized at these taxon levels. Qualitatively, the SFA from all pine species analyzed so far are identical. The genus Pinus is coherently united--but this qualitative feature can be extended to the whole family Pinaceae--by the presence of delta5-UPIFA with C18 [taxoleic (5,9-18:2) and pinolenic (5,9,12-18:3) acids] and C20 chains [5,11-20:2, and sciadonic (5,11,14-20:3) acids]. Not a single pine species was found so far with any of these acids missing. Linoleic acid is almost always, except in a few cases, the prominent SFA, in the range 40-60% of total fatty acids. The second habitual SFA is oleic acid, from 12 to 30%. Exceptions, however, occur, particularly in the Cembroides subsection, where oleic acid reaches ca. 45%, a value higher than that of linoleic acid. Alpha-linolenic acid, on the other hand, is a minor constituent of pine SFA, almost always less than 1%, but that would reach 2.7% in one species (P. merkusii). The sum of saturated acids [16:0 (major) and 18:0 (minor) acids principally] is most often less than 10% of total SFA, and anteiso-17:0 acid is present in all species in amounts up to 0.3%. Regarding C18 delta5-UPIFA, taxoleic acid reaches a maximum of 4.5% of total SFA, whereas pinolenic acid varies from 0.1 to 25.3%. The very minor coniferonic (5,9,12,15-18:4) acid is less than 0.2% in all species. The C20 elongation product of pinolenic acid, bishomo-pinolenic (7,11,14-20:3) acid, is a frequent though minor SFA constituent (maximum, 0.7%). When considering C20 delta5-UPIFA, a difference is noted between the subgenera Strobus and Pinus. In the former subgenus, 5,11-20:2 and sciadonic acids are < or =0.3 and < or =1.9%, respectively, whereas in the latter subgenus, they are most often > or =0.3 and > or =2.0%, respectively. The highest values for 5,11-20:2 and sciadonic acids are 0.5% (many species) and 7.0% (P. pinaster). The 5,11,14,17-20:4 (juniperonic) acid is present occasionally in trace amounts. The highest level of total delta5-UPIFA is 30-31% (P. sylvestris), and the lowest level is 0.6% (P. monophylla). Uniting as well as discriminating features that may complement the knowledge about the taxonomy and phylogeny of pines are emphasized.
Dibutyroyl derivatives of monoacylglycerols (DBMAG) from conifer seed oil triacylglycerols (TAG) were prepared by partial deacylation of TAG with ethylmagnesium bromide followed by diesterification with n-butyryl chloride. The resulting mixtures were analyzed by gas-liquid chromatography (GLC) with a 65% phenylmethyl silicon open tubular fused-silica capillary column operated under optimal conditions and separated according to both their fatty acid structures and their regiospecific distribution. Seed oils of 18 species from 5 conifer families (Pinaceae, Taxaceae, Cupressaceae, Cephalotaxaceae, and Podocarpaceae) were analyzed. The chromatograms showed a satisfactory resolution of DBMAG containing palmitic (16:0), stearic (18:0), taxoleic (cis-5,cis-9 18:2), oleic (cis-9 18:1), cis-vaccenic (cis-11 18:1), pinolenic (cis-5,cis-9,cis-12 18:3), linoleic (cis-9,cis-12 18:2), alpha-linolenic (cis-9,cis-12,cis-15 18:3), and an almost baseline resolution of DBMAG containing gondoic (cis-11 20:1), cis-5,cis-11 20:2, sciadonic (cis-5,cis-11,cis-14 20:3), dihomolinoleic (cis-11,cis-14 20:2), juniperonic (cis-5,cis-11,cis-14,cis-17 20:4), and dihomo-alphalinolenic (cis-11,cis-14,cis-17 20:3) acids. We have observed that results for Pinus pinaster and P. koraiensis seed oils obtained with this new simple method compared favorably with literature data established with other usual regiospecific analytical techniques. Delta5-olefinic acids are esterified mainly at the external positions of the glycerol backbone in all cases, in agreement with data obtained by other methodologies allowing validation of the GLC regiospecific method. To date, 45 gymnosperm species (mostly Coniferophytes) from 21 genera belonging to 9 families have been analyzed, all of them showing a definite enrichment of delta5-olefinic acids in the external positions of TAG. These fatty acids (FA), with one exception only, represent between approximately 2 and 8% of FA esterified to the internal positions. For some species, i.e., P. koraiensis and P. pinaster, this asymmetrical distribution was established by at least three analytical procedures and confirmed by stereospecific analysis of their seed TAG.
The fatty acid compositions of the seed lipids from four Ephedra species, E. nevadensis, E. viridis, E. przewalskii, and E. gerardiana (four gymnosperm species belonging to the Cycadophytes), have been established with an emphasis on delta5-unsaturated polymethylene-interrupted fatty acids (delta5-UPIFA). Mass spectrometry of the picolinyl ester derivatives allowed characterization of 5,9- and 5,11-18:2; 5,9,12-18:3; 5,9,12,15-18:4; 5,11-20:2; 5,11,14-20:3; and 5,11,14,17-20:4 acids. Delta5-UPIFA with a delta11-ethylenic bond (mostly C20 acids) were in higher proportions than delta5-UPIFA with a delta9 double bond (exclusively C18 acids) in all species. The total delta5-UPIFA content was 17-31% of the total fatty acids, with 5,11,14-20:3 and 5,11,14,17-20:4 acids being the principal delta5-UPIFA isomers. The relatively high level of cis-vaccenic (11-18:1) acid found in Ephedra spp. seeds, the presence of its delta5-desaturation product, 5,11-18:2 acid (proposed trivial name: ephedrenic acid), and of its elongation product, 13-20:1 acid, were previously shown to occur in a single other species, Ginkgo biloba, among the approximately 170 gymnosperm species analyzed so far. Consequently, Ephedraceae and Coniferophytes (including Ginkgoatae), which have evolved separately since the Devonian period (approximately 300 million yr ago), have kept in common the ability to synthesize C18 and C20 delta5-UPIFA. We postulate the existence of two delta5-desaturases in gymnosperm seeds, one possibly specific for unsaturated acids with a delta9-ethylenic bond, and the other possibly specific for unsaturated acids with a delta11-ethylenic bond. Alternatively, the delta5-desaturases might be specific for the chain length with C18 unsaturated acids on the one hand and C20 unsaturated acids on the other hand. The resulting hypothetical pathways for the biosynthesis of delta5-UPIFA in gymnosperm seeds are only distinguished by the position of 11-18:1 acid. Moreover, 13C nuclear magnetic resonance spectroscopy of the seed oil from two Ephedra species has shown that delta5-UPIFA are essentially excluded from the internal position of triacylglycerols, a characteristic common to all of the Coniferophytes analyzed so far (more than 30 species), with the possibility of an exclusive esterification at the sn-3 position. This structural feature would also date back to the Devonian period, but might have been lost in those rare angiosperm species containing delta5-UPIFA.
Pre-germinated pine seedlings and germinating oat and mung bean seeds were flown on the STS-3 Space Shuttle mission. Overall, the seedlings grew and developed well in space. Some oat and mung bean roots, however, grew upward. Lignin content was slightly lower in flight tissues and protein content was higher.
Young pine seedlings, and mung bean and oat seeds were flown on shuttle flights, STS-3 and STS-51F, in March, 1982 and July/August, 1985, respectively. The plant growth units built to support the two experiments functioned mechanically as anticipated and provided the necessary support data. Pine seedlings exposed to the microgravity environment of the space shuttle for 8 days continued to grow at a rate similar to ground controls. Pine stems in flight seedlings, however, averaged 10 to 12% less lignin than controls. Flight mung beans grew slower than control beans and their stems contained about 25% less lignin than control seedlings. Reduced mung bean growth in microgravity was partly due to slower germination rate. Lignin also was reduced in flight oats as compared to controls. Oats and mung beans exhibited upward growing roots which were not observed in control seedlings. Chlorophyll A/B ratios were lower in flight tissues than controls. The sealed PGCs exhibited large variations in atmospheric gas composition but the changes were similar between flight and ground controls. Ethylene was present in low concentrations in all chambers.