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Isolation and characterization of a cDNA from Quercus robur differentially expressed in juvenile-like and mature shoots.

A full-length cDNA clone named QRCPE (Quercus robur crown preferentially expressed) that is differentially expressed during in vitro culture of mature and juvenile-like shoots of Quercus robur L. was identified by differential display. The deduced amino acid sequence showed that the encoded protein is small, contains a predicted N-terminal hydrophobic signal peptide that targets the protein to the cell wall, and is rich in glycine and histidine residues. Accumulation of QRCPE mRNA was higher in oak microshoots derived from crown branch shoot cultures than in oak microshoots derived from basal shoot cultures at the end of the multiplication and rooting period. Among organs, the highest accumulation of QRCPE transcripts was detected in roots, followed by stems and leaves, with preferential accumulation in specific organs of ontogenetically older shoots. Although QRCPE mRNA was abundant in oak zygotic and somatic embryos, almost no QRCPE mRNA accumulation was detected in nodular callus cells, suggesting a possible role of this gene in embryonic development. In proliferating shoot cultures of two chestnut (Castanea sativa Mill.) clones, the QRCPE homolog was preferentially expressed in crown-derived shoots. On the other hand, expression analysis of QRCPE in juvenile and mature material from soil-grown oak plants indicated that this gene is expressed from the embryonic to mature phases, but is progressively down-regulated during plant maturation. In vitro culture conditions induced changes in QRCPE transcript abundance in both basal and crown shoots in a phase-dependent manner. We conclude that QRCPE expression in oak is correlated with the ontogenetic stage of shoots, and thus this gene may be useful as a potential molecular marker for maturation-related characteristics.

Aesculus↗

Evidence for shared ancestral polymorphism rather than recurrent gene flow at microsatellite loci differentiating two hybridizing oaks (Quercus spp.).

Quercus petraea and Quercus robur are two closely related oak species, considered to hybridize. Genetic markers, however, indicate that despite sharing most alleles, the two species remain separate genetic units. Analysis of 20 microsatellite loci in multiple populations from both species suggested a genome-wide differentiation. Thus, the allele sharing between both species could be explained either by low rates of gene flow or shared ancestral variation. We performed further analyses of population differentiation in a biogeographical setting and an admixture analysis in mixed oak stands to distinguish between both hypotheses. Based on our results we propose that the low genetic differentiation among these species results from shared ancestry rather than high rates of gene flow.

Analysis of Variance↗

Diversity and specificity of ectomycorrhizal fungi retrieved from an old-growth Mediterranean forest dominated by Quercus ilex.

We analysed the ectomycorrhizal (ECM) fungal diversity in a Mediterranean old-growth Quercus ilex forest stand from Corsica (France), where Arbutus unedo was the only other ECM host. On a 6400 m2 stand, we investigated whether oak age and host species shaped below-ground ECM diversity. Ectomycorrhizas were collected under Q. ilex individuals of various ages (1 yr seedlings; 3-10 yr saplings; old trees) and A. unedo. They were typed by ITS-RFLP analysis and identified by match to RFLP patterns of fruitbodies, or by sequencing. A diversity of 140 taxa was found among 558 ectomycorrhizas, with many rare taxa. Cenococcum geophilum dominated (35% of ECMs), as well as Russulaceae, Cortinariaceae and Thelephoraceae. Fungal species richness was comparable above and below ground, but the two levels exhibited < 20% overlap in fungal species composition. Quercus ilex age did not strongly shape ECM diversity. The two ECM hosts, A. unedo and Q. ilex, tended to share few ECM species (< 15% of the ECM diversity). Implications for oak forest dynamics are discussed.

Mediterranean Region↗

[Genetic differentiation of pedunculate oak Quercus robur L. in the European part of Russia based on RAPD markers].

Using randomly amplified polymorphic DNA markers (RAPD), genetic variation and differentiation in four populations of pedunculate oak Quercus robur L. were examined. The populations occupy a large part of the Quercus robur range in the European Russia (Voronezh and Novgorod oblasts; Republics of Mordovia and Bashkortostan). With each of six random primers (A02, A09, A17, B01, B08, B11), 96 DNA samples were analyzed by PCR. In all, 48 putative polymorphic RAPD loci were detected. We failed to reveal population-specific DNA fragments for any primer although the frequencies of 14 fragments were significantly different among populations. The oak populations studied exhibited high variability: 73-90% of genes were polymorphic and the effective allele number was about 1.4. The total genetic variation varied from 0.202 (Vor) to 0.245 (Nov), which corresponded to the estimates for populations of this species from Central and Western Europe. The populations examined showed low among-population differentiation (GST = 0.098); gene flow Nem was 4.61. The proportion of among-population variation of the RAPD loci studied accounted for 7% of the total variability; more than 93% of the total variability was explained by individual and within--population variation.

Genetic Variation↗

[Niche characteristics of plant populations in Quercus aliena var. acuteserrata stands in Qinling Mountains].

Based on the data collected from 31 plots and using Levins, Hurlbert and Pianka formulas, this paper calculated and analyzed the niche breadths and overlaps of 24 tree and 29 shrub populations in Quercus aliena var. acuteserrata stands in Mt. Qinling, Shaanxi. The results showed that the Levins and Hurlbert's niche breadths of Quercus aliena var. acuteserrata , Pinus armandi, Toxicodendron vernicifluum, Populus davidiana and Pinus tabulaeformis in arbor layer were 3.695, 1.695, 1.325, 0.840, 0.702 and 0.036, 0.299, 0.568, 1.721, 2.701 respectively, and those of Corrylus heterophylla, Lespedeza formosa and Smilax vaginata in shrub layer were 1.833, 1.466, 0.984 and 0.111, 0.300, 0.594, respectively. In general, the niche overlaps between the species with bigger niche breadth and other species were higher, and those of the species having similar environmental requirements were also higher.

Biodiversity↗

[Effects of soil water regimes on the growth of Quercus mongolica seedlings in Changbai Mountains].

This paper studied the response of the seedlings of Quercus mongolica, one of the dominant tree species in Changbai Mountains, to the artificially controlled three soil water gradients, including their morphology, biomass and photosynthetic characteristics. The results indicated that various water regimes significantly affected the biomass and its distribution pattern in the leaves, branches and roots, as well as the leaf gas exchange. Under soil water stress, the crown structure changed, and the tree height, groundline diameter, single leaf area, and aboveground and belowground biomass were inhibited. As soil water content decreased, the ratio of belowground and aboveground biomass dry weight significantly increased. Water stress had a negative effect on net photosynthetic rate, CO2 use efficiency and carbon use efficiency. The responses of stomatal conductance, transpiration rate and water use efficiency to water stress were complicated. Only at low soil water content, the stomatal conductance and transpiration rate significantly decreased, while water use efficiency increased. It was demonstrated that Quercus mongolica was the tree species with variable resistance to drought, and the resistance could be improved by long-term soil water stress.

Biomass↗

[Genetic diversity of microsatellites (SSRs) of natural populations of Quercus variabilis].

Genetic diversity of five Quercus variabilis natural populations in four provinces of China was studied with microsatellite (SSR) markers. A relatively high level of genetic diversity was detected in Q. variabilis species with 16 polymorphic microsatellite loci. Average number of alleles (A) and effective number of alleles (N(e)) were 8.4375 and 5.9512 respectively. The mean expected heterozygosity (H(e)) was 0.8059 and Nei diversity index (h) was 0.8041. Higher diversity was found with the populations from the central range of the species in contrast to those from peripheral areas and human activities might decrease the genetic diversity of populations. The majority of genetic variation occurred within populations, which could be concluded from the low coefficient of genetic differentiation (F(st)=0.0455). In addition, significant correlation was found between geographical distance and genetic distance. All these results present a basis to the conservation and utilization of genetic diversity of Quercus variabilis.

Alleles↗

[Physiological-ecological effects of Populus davidiana--Quercus liaotungensis mixed forest in Ziwuling forest area].

This paper studied the soil physical- properties under Populus davidiana, Quercus liaotungensis, and Populus davidiana--Quercus liaotungensis mixed forest in the Ziwuling forest area of Loess Plateau, and the leaf photosysthetic characteristics of these three types of forests. The results showed that soil moisture content in 0 - 300 cm layer was the highest under P. davidiana forest, and obviously increased below 200 cm in depth under P. davidiana--Q. liaotungensis mixed forest, which was 10.5% - 19.76% higher than that under Q. liaotungensis forest. In 0 - 60 cm layer, P. davidiana forest showed the highest soil bulk density and the lowest soil porosity, while P. davidiana--Q. liaotungensis mixed forest presented the lowest soil bulk density and the highest soil porosity, and both of these indices surpassed their corresponding values under pure forests, which indicated that the mixed forest could make effective use of water in deep soil, and obviously improved soil physical and chemical properties. P. davidiana and Q. liaotungensis had a higher content of leaf chlorophyll than P. davidiana--Q. liaotungensis mixed forest, and Q. liaotungensis presented the highest leaf chlorophyll content. Q. liaotungensis had the highest photosynthetic rate and stomatal conductance, followed by P. davidiana, and by P. davidiana--Q. liaotungensis mixed forest. The water use efficiency of the forests ranked in the decreasing order of Q. liaotungensis in pure forest, Q. liaotungensis in mixed forest, P. davidiana in mixed forest, and P. davidiana in pure forest. Q. liaotungensis in mixed forest presented the highest F(v)/F(m) and F(v)/F(o), and did not remarkably differ from those in pure forest, but in the mixed forest, the F(v)/F(m) and F(v)/F(o) of P. davidiana were markedly lower than those of P. davidiana in pure forest. Both the q(p) and NPQ of P. davidiana and Q. liaotungensis in pure forests were higher than those in mixed forest, respectively. In Ziwuling forest area, Q. liaotungensis in Q. liaotungensis--dominant climax community tended to be more stable, and grew better than Q. liaotungensis in P. davidiana--Q. liaotungensis mixed forest, and P. davidiana would be gradually replaced. Compared with P. davidiana in pure forest, P. davidiana in its mixed forest was at a disadvantage in its growth.

China↗

Why nuclear ribosomal DNA spacers (ITS) tell different stories in Quercus.

The molecular systematics of Quercus (Fagaceae) was recently assessed by two teams using independently generated ITS sequences. Although the results disagreed in several remarkable features, the phylogenetic trees for either hypothesis were highly supported by bootstrap resampling. We have reanalyzed the ITS sequences used by both teams (eight taxa) to reveal the underlying patterns of this divergence. Within species, conspicuous length and G + C% divergence were evident in most sequence comparisons. In addition, a high rate of substitutions and deletions involving highly conserved motifs in both ITS spacers were present in a set of sequences. This was coupled with a less thermodynamic stability in the RNA structure, lacking conserved hairpins that are putatively involved in the processing of the RNA transcripts. Compelling evidence suggests that the divergent ITS alleles reported by one team are pseudogenes, i.e., nonfunctional paralogous loci. The hypothesis that the contrasting phylogenetic histories drawn from Quercus using ITS data are not strictly related to technical differences between laboratories, but that they have rather been generated from the analysis of paralogous sequences, best reconciles the available data. The risk of incorporating ITS paralogues in plant evolutionary studies which can distortion the phylogenetic signal should caution molecular systematists. Without a detailed inspection of some basic features of the sequence, including the integrity of the conserved motifs and the thermodynamic stability of the secondary structures of the RNA transcripts, errors in evolutionary inferences could be easily overlooked.

Base Composition↗

Alteration of foliar flavonoid chemistry induced by enhanced UV-B radiation in field-grown Pinus ponderosa, Quercus rubra and Pseudotsuga menziesii.

Chromatographic analyses of foliage from several tree species illustrate the species-specific effects of UV-B radiation on both quantity and composition of foliar flavonoids. Pinus ponderosa, Quercus rubra and Pseudotsuga menziesii were field-grown under modulated ambient (1x) and enhanced (2x) biologically effective UV-B radiation. Foliage was harvested seasonally over a 3-year period, extracted, purified and the flavonoid fraction applied to a mu Bondapak/C(18) column HPLC system sampling at 254 nm. Total flavonoid concentrations in Quercus rubra foliage were more than twice (leaf area basis) that of the other species; Pseudotsuga menziesii foliage had intermediate levels and P. ponderosa had the lowest concentrations of total flavonoids. No statistically significant UV-B radiation-induced effects were found in total foliar flavonoid concentrations for any species; however, concentrations of specific compounds within each species exhibited significant treatment effects. Higher (but statistically insignificant) levels of flavonoids were induced by UV-B irradiation in 1- and 2-year-old P. ponderosa foliage. Total flavonoid concentrations in 2-year-old needles increased by 50% (1x ambient UV-B radiation) or 70% (2x ambient UV-B radiation) from that of 1-year-old tissue. Foliar flavonoids of Q. rubra under enhanced UV-B radiation tended to shift from early-eluting compounds to less polar flavonoids eluting later. There were no clear patterns of UV-B radiation effects on 1-year-old P. menziesii foliage. However, 2-year-old tissue had slightly higher foliar flavonoids under the 2x UV-B radiation treatment compared to ambient levels. Results suggest that enhanced UV-B radiation will alter foliar flavonoid composition and concentrations in forest tree species, which could impact tissue protection, and ultimately, competition, herbivory or litter decomposition.

Flavins↗

SSR markers for Quercus suber tree identification and embryo analysis.

Three Quercus simple sequence repeat (SSR) markers were amplified by polymerase chain reaction (PCR) from nuclear DNA extracts of trees and in vitro-induced haploid embryos from anther cultures of Quercus suber L. These markers were sufficiently polymorphic to identify 10 of 12 trees located in two Spanish natural areas. The same loci have been analyzed in anther-derived haploid embryos showing the parental tree allele segregation. All the alleles were present in the haploid progeny. The presence of diverse alleles in embryos derived from the same anther demonstrated that they were induced on multiple microspores or pollen grains and they were not clonally propagated. Also, diploid cultures and mixtures of haploid-diploid tissues were obtained. The origin of such cultures, either somatic or gametic, was elucidated by SSR markers. All the embryos showed only one allele, corroborating a haploid origin. Allelic composition of the haploid progeny permitted parental identification among all analyzed trees.

Alleles↗

Leaf water potential and stomatal conductance in Quercus pyrenaica Willd. forests: vertical gradients and response to environmental factors.

Two permanent sampling sites were selected at the two extremes of a rainfall gradient in natural forests of Quercus pyrenaica Willd. located in the Sierra de Gata (Salamanca Province, Spain). During 1991 and 1992, diurnal courses of transpiration rate (E), stomatal conductance (g(s)) and leaf water potential (Psi(l)) were studied at different levels in the tree canopy. Leaf water potential fell from a high value in the early morning to a minimum at midday and then increased again in the afternoon. Values of Psi(l) in the early morning and at midday were lower at Fuenteguinaldo (dry site) than at Navasfrías (wet site) and were related to soil water availability. Stomatal conductance increased during the morning and maximum g(s) occurred about 3 h before Psi(l) was at its lowest value. The highest values of g(s) were found at Navasfrías during July. The lowest Psi(l) values were found in the upper parts of the canopy and differences among canopy levels were greatest when soil water availability was low. In August, minimum values of Psi(l) differed among canopy levels by 1.0 MPa at Navasfrías and by 0.8 MPa at Fuenteguinaldo. The maximum difference in g(s) among canopy levels was about 150 mmol m(-2) s(-1) at Navasfrías and about 300 mmol m(-2) s(-1) at Fuenteguinaldo. The results indicate a nonconservative pattern of water use in Quercus pyrenaica.

Journal Article↗

Requirements for in vitro rooting of Quercus robur and Q. rubra shoots derived from mature trees.

Stabilized shoot cultures initiated from crown material of six adult Quercus robur L. trees and from basal epicormic shoots of a Quercus rubra L. tree showed good in vitro rooting capacity. An initial five-day dark period generally improved the rooting response but was detrimental to plantlet quality. There were clonal differences in rooting capacity. The concentration and exposure time of the indolebutyric acid (IBA) treatment were critical for root induction. In both species, best rooting efficiency was achieved by culture in medium containing 25 mg l(-1) IBA for 24 h and subsequent transfer to an auxin-free medium containing 1% activated charcoal. For all clones tested, the charcoal benefited both shoot quality and root system development, the latter being enhanced by the formation of many lateral roots. Total root system area and length, measured with a digital image analyzer, were significantly greater in medium containing charcoal than in medium lacking charcoal. Because darkening the basal part of the shoots with aluminum foil during the rooting phase only caused a small increase in rooting, we conclude that the large effect of charcoal on rooting was the result of adsorption of inhibitory compounds from the medium or the explant or both, rather than of basal darkening. Other factors affecting the rooting response of Q. robur were: (a) the position on the tree of the material from which cultures were initiated (the topophysical effect); and (b) shoot quality. Recycling the same horizontally placed explant on multiplication medium allowed three successive crops of shoots to be obtained, and rootability was typically maintained from crop to crop.

Journal Article↗

Adaptations and responses to drought in Quercus species of North America.

Most North American oaks (Quercus spp.) are adapted to drought-prone sites by an ability either to avoid, or to tolerate, water stress, or both. Generally, they have deep-penetrating root systems, enabling them to maintain relatively high predawn water potentials during drought. Oaks have thick leaves and some have relatively small stomata, both characteristics that favor high water use efficiency. However, some species, from warm regions, have large stomatal pores. The rapid evaporative cooling made possible by large stomata, may be an adaptation to high temperature. Some southeastern species display leaf curling during drought, and Q. douglasii a native of California is drought deciduous. Oaks have a ring-porous xylem anatomy, allowing rapid sap movement in large diameter, early-wood vessels when soil water is plentiful, and slower, but sustained, water movement in narrower, late-wood vessels, which are more resistant to cavitation, during drought. Oaks frequently maintain a higher rate of photosynthesis at low leaf water potentials and high vapor pressure deficits than co-occurring species of other genera. An exception is Quercus rubra, which is generally restricted to relatively mesic sites. During drought, many oak species, especially those native to arid regions, undergo changes in tissue osmotic potential. However, it remains to be shown whether such changes are phenological or drought induced. Reported values for bulk modulus of elasticity vary widely among species and studies, and have been observed to both increase and decrease during drought in a way that is unrelated to region or to changes in predawn water potential or osmotic potential. Diurnal leaf water potential during drought is probably a poor indicator of differences among oak species in gas exchange rate, because of interspecific variation in desiccation avoidance and tolerance.

Journal Article↗

Functional expression of the gene cu, encoding the phytotoxic hydrophobin cerato-ulmin, enables Ophiostoma quercus, a nonpathogen on elm, to cause symptoms of Dutch elm disease.

We studied the involvement of the phytotoxic hydrophobin cerato-ulmin (CU) in pathogenesis and virulence of Dutch elm disease (DED) by expressing its encoding gene (cu) in Ophiostoma quercus, a nonpathogenic species on elm closely related to the DED pathogens O. ulmi and O. novo-ulmi. The production of the toxin was quantitatively determined in culture filtrates and in mycelial extracts of the transformants. Production of CU in vitro was associated with the ability to cause typical DED symptoms, consisting of foliar yellow and wilting and vascular tissue discoloration on a moderately resistant elm genotype. The presence of CU was monitored by enzyme-linked immunosorbent assay in symptomatic leaves of plants inoculated with O. quercus transformants expressing CU and found to be associated with wilt symptoms. In general, the virulence of the cu-expressing transformants, as measured in terms of vascular discoloration and percentage of defoliation, was lower than that of the mildly pathogenic isolate E2 of O. ulmi. However, one transformant (C39) displayed a virulence level intermediate between that of E2 and 182, a highly virulent isolate of O. novo-ulmi. Our results indicate that CU production influences virulence in nonaggressive strains of Ophiostoma fungi.

Ascomycota↗

Evidence of the Photosynthetic Origin of Monoterpenes Emitted by Quercus ilex L. Leaves by 13C Labeling.

The carbon of the four main monoterpenes emitted by Quercus ilex L. leaves was completely labeled with 13C after a 20-min feeding with 99% 13CO2. This labeling time course is comparable with the labeling time course of isoprene, the terpenoid emitted by other Quercus species and synthesized in leaf chloroplasts. It is also comparable with that of phosphoglyceric acid. Our experiment therefore provides evidence that monoterpenes emitted by Q. ilex are formed from photosynthesis intermediates and may share the same synthetic pathway with isoprene. By analyzing the rate and the distribution of labeling in the different fragments, we looked for evidence of differential carbon labeling in the [alpha]-pinene emitted. However, the labeling pattern was quite uniform in the different fragments, suggesting that the carbon skeleton of the emitted monoterpenes comes from a unique carbon source.

Journal Article↗

Superoxide anion scavenge effect of Quercus glauca Thunb. in whole blood of patients with ankylosing spondylitis.

Nine phenolic compounds, catechin (1), epicatechin (2), gallocatechin (3), epigallocatechin (4), procyanidin B-4 (5), catechin-3-O-rhamnoside (6), rutin (7), querglanin (8) and isoquerglanin (9) were isolated from oak leaves (Quercus glauca Thunb. Fagaceae), and the latter two (8, 9) were identified as new compounds. Several Quercus species have been used in folk medicine as an astringent for hemorrhoids and for treatment of inflammation, jaundice, and tumor. In this study, these compounds were tested for scavenging effects of the superoxide anion in the whole blood of patients with ankylosing spondylitis by means of an ultra-sensitive chemoluminescence (CL) analyzer and lucigenin amplification. The results showed that at a concentration of 2.3 x 10(-5) M, isoquerglanin (9) displayed the strongest inhibition activity (73.55%), followed by querglanin (8) (68.81%) and then gallocatechin (3) and epigallocatechin (4) (66.97 and 60.17% inhibition, respectively). In addition, the blood chemoluminescence (CL) level of patients with ankylosing spondylitis was inhibited by superoxide dismutase (SOD) but not by catalase, suggesting that superoxide anion is the major component of reactive oxygen species (ROS) involved in this assay system.

Acridines↗

Laboratory evaluation of molluscicidal activity of extracts from Cotula cinerea (L) and Quercus lusitania var. infectoria galls (Oliv.).

In this work, we have studied the molluscicidal activity of different extracts obtained from Cotula cinerea and Quercus lusitania var. infectoria galls. The hydroalcoholic extract of Cotula cinerea, acetonic extract and gallotanin of Quercus infectoria galls have presented high activity against Bulinus truncatus. The hydroalcoholic extract of Cotula cinerea was fractionated by chromatography on silica gel column. We have isolated two very active fractions at concentrations respectively of 52.5 and 27.5 ppm.

Evaluation Studies as Topic↗