Interactive effects of mycorrhization and elevated carbon dioxide on growth of young pedunculate oak (Quercus robur L.) trees

Pedunculate oak (Quercus robur L.) was germinated and grown at ambient CO₂ level and 650 ppmv CO₂ in the presence and absence of the ectomycorrhizal fungus Laccaria laccata for a total of 6 month under nutrient non-limiting conditions. Mycorrhization and elevated atmospheric CO₂ each supported the growth of the trees. Stem height, stem diameter, and dry matter accumulation of pedunculate oak were increased by mycorrhization. Elevated atmospheric CO₂ enhanced stem height, stem diameter, fresh weight and dry weight, as well as lateral root formation of the trees. In combination, mycorrhization and elevated atmospheric CO₂ had a more than additive, positive effect on tree height and biomass accumulation, and further improved lateral root formation of the trees. From these findings it is suggested that the efficiency of the roots in supporting the growth of the shoot is increased in mycorrhized oak trees at elevated atmospheric CO₂.Abbreviations DW dry weight - FW fresh weight - RWC relative water content     

Transport of organic sulfur and nitrogen in the roots of young mycorrhizal pedunculate oak trees (Quercus robur L.)

Uptake and xylem loading of organic sulfur and nitrogen were analyzed in detached mycorrhizal (Laccaria laccata L.) roots of pedunculate oak (Quercus robur L.) seedlings using radiolabeled reduced glutathione (GSH) and glutamine (Gln) for transport analyses. The experiments showed for the first time that GSH is taken up by plant roots from the nutrient solution and is partially allocated to the shoot. Apparently, GSH produced during mineralization processes in the soil can be used by plant roots as a sulfur source. GSH uptake into the roots showed biphasic kinetics within the concentration range studied (0–500 M) with maximum transport velocities (v _max) and substrate affinities (K _m) that were similar to the kinetics of Gln uptake. GSH uptake kinetics were also in the same range as previously reported for sulfate uptake by mycorrhizal roots of pedunculate oak. It may therefore be assumed that GSH and sulfate uptake can be of comparable significance for sulfur nutrition, provided both sulfur sources are available at similar concentrations at the sites of uptake. Xylem loading of GSH and Gln showed monophasic transport kinetics with v _max significantly lower than observed for the two respective uptake systems and, as indicated by the K _m-values, a substrate affinity between the high and the low affinity uptake systems. The possible nature of the transport systems for GSH and Gln is discussed.     

Radial distributions of Cd in stems of oak trees (Quercus robur L.) re-analyzed after 10 years

Radial distribution patterns of Cd were determined in stems of the same oak trees (Quercus robur L.) in 1983 and again in 1994. On both sampling dates the same distribution patterns were generally observed. Highest concentrations of Cd were found at the sapwood-heart-wood transition. A sharp drop over this boundary towards inner parts of the stems occurred in all trees. In the decade between the two investigations the sapwood-heartwood boundaries had shifted outwards by 9–11 annual rings. The Cd peaks at the boundaries were shifted by approximately the same interval. The results suggest that the described Cd peaks are mobile in a radial direction. The present location of such peaks cannot be used to infer the pollution history of the tree's environment. The Cd accumulation at the sapwood-heartwood boundary is probably affected by physiological processes in the wood. Thus it is concluded that radial distributions of Cd in stems of oak trees are no reliable source of information for retrospective monitoring of past time pollution levels.     

The alpha-tocopherol content of leaves of pedunculate oak (Quercus robur L.)--variation over the growing season and along the vertical light gradient in the canopy

This study was performed in order to investigate whether the actual requirement for defence against photo-oxidative stress is reflected by the alpha-tocopherol (alpha-Toco) content in leaves of pedunculate oak (Quercus robur L.). Antioxidants and pigments were quantified in leaves that were collected on six days between May and September 2000 in a mixed pine/oak forest at canopy positions differing in light environment. Pools of hydrophilic antioxidants and photo-protective xanthophyll cycle pigments (V + A + Z) reflected the anti-oxidative demand, as these pools increased with the average light intensity to which the leaves were acclimated. The photo-protective demand was not the determinant of the alpha-Toco content of oak leaves, as (1) foliage of a young oak, exposed to low light levels in the understorey, contained higher amounts of this lipophilic antioxidant than leaves sampled from semimature oaks at canopy positions with a similar light environment, and (2) a strong increase in the alpha-Toco content over the growing season was detected at each investigated crown position, whereas the V + A + Z pool did not show a concomitant accumulation during leaf ageing. The rate of alpha-Toco accumulation differed distinctly between samples taken at different canopy positions.     

Influence of wind loading on root system development and architecture in oak (Quercus robur L.) seedlings.

The effect of wind loading on seedlings of English oak (Quercus robur L.) was investigated. Instead of using a traditional wind tunnel, an innovative ventilation system was designed. This device was set up in the field and composed of a rotating arm supporting an electrical fan, which emitted an air current similar to that of wind loading. Oaks were sown from seed in a circle around the device. A block of control plants was situated nearby, and was not subjected to artificial wind loading. After 7 months, 16 plants from each treatment were excavated, and root architecture and morphological characteristics measured using a 3D digitiser. The resulting geometrical and topological data were then analysed using AMAPmod software. Results showed that total lateral root number and length in wind stressed plants were over two times greater than that in control trees. However, total lateral root volume did not differ significantly between treatments. In comparing lateral root characters between the two populations, it was found that mean root length, diameter and volume were similar between the two treatments. In trees subjected to wind loading, an accentuated asymmetry of root distribution and mean root length was found between the windward and leeward sides of the root system, with windward roots being significantly more numerous and longer than leeward roots. However, no differences were found when the two sectors perpendicular to the wind direction were compared. Mean tap root length was significantly higher in control samples compared to wind stressed plants, whilst mean diameter was greater in the latter. Wind loading appears to result in increased growth of lateral roots at the expense of the tap root. Development of the lateral root system may therefore ensure better anchorage of young trees subjected to wind loading under certain conditions.     

Inheritance of chloroplast and mitochondrial genomes in pedunculate oak investigated with an efficient PCR method

The restriction patterns of two chloroplast fragments and one mitochondrial DNA fragment, amplified by PCR with universal primers, were studied to determine the mode of inheritance of these organelles in 143 progeny of five intraspecific crosses in pedunculate oak (Quercus robur L.). The results indicate that both genomes are maternally inherited, an observation which agrees with the commonly observed pattern of inheritance in angiosperms. They confirm that both chloroplast DNA and mitochondrial DNA can be used as a source of seed-specific markers for the study of the geographic structure of oaks. This is the first report of organelle inheritance within the Fagaceae, an important and widespread tree family.     

Sapwood estimates of pedunculate oak (Quercus robur L.) in eastern Baltic

Pedunculate oak (Quercus robur L.) is one of the widely used and dendrochronologically investigated species in Europe. Still, it is a problematical dating object if its outermost section is missing partly or totally. Thus, we need sapwood estimation of living trees. As sapwood amount varies geographically, numbers of sapwood rings have been published for different regions in Europe but no such estimation has been done for the Baltic States yet. Therefore, this paper deals with the estimation of pedunculate oak sapwood growing in the eastern Baltic region, i.e. in Finland, Estonia, Latvia, and Lithuania.In total, 668 oak core samples of living trees from 43 stands were investigated. Ring widths were measured and the number of sapwood rings was determined according to two criteria: difference of colour and absence of tyloses in earlywood vessels. The samples were divided into two sets, according to the t_H-values between site chronologies and the major geobotanical sub-provinces. Thus, the nine Finnish and western Estonian sites were attributed to the western region and the 34 eastern Estonian, Latvian and Lithuanian sites to the eastern region.As the result of a statistical analysis, we explain that the number of oak sapwood rings ranges from 4.09 to 20.85 and 6.45 to 18.02 within 95% confidence limits in the western and eastern regions, respectively. For the three Baltic countries and southern Finland in general, we recommend to consider a sapwood estimate of 6.18–18.71 rings. Regarding earlier studies, the general European trend of decreasing sapwood ring number towards the east was confirmed. A geographical pattern of eastward decrease of the median sapwood ring number was noticed in the Baltics as well. The chronology based upon 668 samples of living oak trees from all sites covered the period of 1631–2008.     

Structure and identification of root bark of Quercus robur L.

The root bark structure of Quercus robur L. was analysed at different stages of root development and compared to the structure of stem bark. Root bark thickness varied considerably between different roots. Sclereid quantity decreased with increasing distance from the stem, which means it increased with age. Visible growth increments diminished with increasing distance from the stem. In lateral roots crystal quantity decreased with increasing distance from the stem. In lateral roots secondary phloem fibre length, sieve tube member length, and sieve tube diameter showed no regular trend. There were only a few basic structural differences between root and stem bark. The zone of cell differentiation (cell expansion, lignification) was wider in root bark; sieve tube collapse was delayed. In lateral root bark fewer sclereids were formed. The first-formed periderm often originated from deeper cell layers. Thus, primary elements were lacking after periderm formation. In root bark the phellem cell walls were of equal thickness. Thus, phellem lacked visible growth increments. Root bark phellem cells were slightly larger. The root phelloderm was more distinct. The secondary phloem fibres were slightly shorter than those in stem bark. Sieve tube members of stem and root bark were of similar length and diameter. The qualitative bark anatomical characters of oak root bark are suitable for root identifications. Due to minor structural differences between root and stem bark the characters must be used with care.     

Genetic variability of fragmented stands of pedunculate oak (Quercus robur) in Finland

The genetic structure of 33 natural Quercus robur stands in Finland was studied using 13 allozyme loci to analyze the effects of fragmentation in a wind-pollinated tree species. The present fragmented and discontinuous distribution of oak is a result of both short-term human impact and long-term climatic and geological change, including post-glacial land uplift. In accordance with general expectations, genetic diversity in small populations was lower than that in large populations, and differentiation among small populations was higher than that among large populations. Heterozygote deficiency was more pronounced in large populations, which is proposed to be a Wahlund effect created by either spatial sub-structuring or the existence of synchronized flowering lineages. Also genetic differentiation was higher and diversity lower in Finland than the estimates reported for Central Europe. There were differences in the genetic structure on sites of different geological age. We suggest that on most geologically old sites drift has a prominent effect whereas on younger sites also founder effects may be important.     

Plant regeneration from encapsulated somatic embryos of pedunculate oak (Quercus robur L.)

Summary Cotyledonary Quercus robur L. somatic embryos from two cell lines were encapsulated in 4% (w/v) sodium alginate. An artificial endosperm was provided by the addition of P₂₄ medium plus 3% (w/v) sucrose. Oak somatic embryos and oak synthetic seeds were germinated on P₂₄ medium plus 0.1 μM indole-3-butyric acid and 0.9 μM 6-benzylaminopurine or were dehydrated prior to germination. The highest conversion rates (26%) were obtained with encapsulated somatic embryos as well as artificial endosperm-coated somatic embryos. Encapsulation improved the regeneration into oak plantlets in one of the two cell lines tested. The artificial endosperm had no additional beneficial effect on conversion frequency, but increased germination rate in one cell line tested. Significant higher conversion could be attributed to slow desiccation compared to the non-encapsulated control. Cold storage as a post-maturation treatment had no influence on the germination ability of oak synthetic seeds. Differences in the response of the cell lines with respect to conversion frequencies and timing of germination were observed. Fifty-six well-developed plantlets regenerated 12 wk after germination, and 29 plants were transferred to the greenhouse, where they have been successfully established in substrate.     

Detection of microsatellite instability during somatic embryogenesis of oak (Quercus robur L.)

Five microsatellite loci (QpZAG1/5, QpZAG9, QpZAG36, MSQ4, MSQ13) were used to test for genetic stability of three somatic embryogenic culture lines of Quercus robur L. and plantlets derived therefrom. DNA variation was detected among somatic embryos within all embryogenic lines, whereas no genetic instability was found among the regenerated plants. Two microsatellite loci revealed variation, and a locus-dependent instability was observed. The most polymorphic and useful microsatellite locus for detecting genetic variation was QpZAG9, with 28.5% of the investigated loci being variable.     

Physiological and morphological characteristics during development of pedunculate oak (Quercus robur L.) zygotic embryos

The developmental stages of oak zygotic embryos (ZEs) are characterized here according to morphological and physiological features. Seeds were harvested from June to September in 1-week intervals. Excised embryos were classified into four stages of development by using growth parameters. For physiological characterization, endogenous levels of abscisic acid (ABA), indole-3-acetic acid (IAA), l-proline, starch content and water status were determined. The expression of the oak legumin storage protein gene was tested in immature cotyledonary ZEs before and after ABA treatment. The ABA levels of the embryos showed a significant peak during the intermediate stage of maturation (stage III) and then decreased again at the end of the late maturation phase (stage IV). Concomitant with ABA, the moisture content declined with the maximum embryo size. High IAA levels were found at the beginning of embryo enlargement as exponential growth occurred (stage II) but decreased during further development. Starch accumulated gradually in the course of maturation, whereas significant values were found in stage IV ZEs near shedding. Proline, on fresh weight basis, was high during stages I and II. Osmotic potential increased when, by rapid dry matter accumulation, stage II ZEs reached their maximum size during early intermediate development. Expression of precocious germination was higher on hormone-free medium, in particular, among stage II and stage III ZEs. Variations in phytohormone levels in combination with changes in tissue water status seem to be important factors for oak ZE development.     

Quantitative trait loci controlling water use efficiency and related traits in Quercus robur L.

Genetic variation for intrinsic water use efficiency (W _i) and related traits was estimated in a full-sib family of Quercus robur L. over 3 years. The genetic linkage map available for this F1 family was used to locate quantitative trait loci (QTL) for W _i, as estimated by leaf carbon stable isotope composition (δ ¹³C) or the ratio of net CO₂ assimilation rate (A) to stomatal conductance to water vapour (g _w) and related leaf traits. Gas exchange measurements were used to standardize estimates of A and g _w and to model the sensitivity of g _w to leaf-to-air vapour pressure deficit (sg^VPD). δ ¹³C varied by more than 3‰ among the siblings, which is equivalent to 40% variation of W _i. Most of the studied traits exhibited high clonal mean repeatabilities (>50%; proportion of clonal mean variability in global variance). Repeatabilities for δ ¹³C, leaf mass per area (LMA) and leaf nitrogen content were higher than 70%. For δ ¹³C, ten QTLs were detected, one of which was detected repeatedly for all 3 years and consistently explained more than 20% of measured variance. Four genomic regions were found in which co-localizing traits linked variation in W _i to variations in leaf chlorophyll and nitrogen content, LMA and sg^VPD. A positive correlation using clonal means between δ ¹³C and A/g _w, as well as a co-localisation of QTL detected for both traits, can be seen as validation of the theoretical model linking the genetic architecture of these two traits.     

Stem annual increments as ecobiological indicators in Turkey oak (Quercus cerris L.)

In recent decades, the biostatistical analysis of relationships among tree growth and macroclimatic factors has developed remarkably. In this discipline, the parametrisation of tree growth is generally based on the records of annual ring widths measured on breast height transverse sections of tree stems (ARI). The present research enables the stem volume annual increment (AVI) to be evaluated as a dendroclimatic indicator in comparison with ARI. The research is a part of a broader investigation into the ecobiological characteristics of Turkey oak in the Gargano region (southern Italy). The results indicate that AVI is no better than ARI as a means of highlighting expressive relationships among the examined macroclimatic factors (monthly precipitation and monthly averages of temperature daily maxima and minima) and the Turkey oak annual stem growth. In the examined stands, the most growthinfluencing factors are the autumn and spring rainfalls. Regarding the stem breast height radial increment, the most effective temperature factors are the June averages of daily minima and maxima.     

Nutrient acquisition from different soil depths by pedunculate oak

Eight oak trees (Quercus robur L.) received ³²P at a soil depth of 50 cm and ³³P at a soil depth of 15 cm at the end of June 2002 through plastic tubes inserted into the mineral soil. The phosphorus uptake from different soil depths was estimated by analysing the concentration of ³²P and ³³P in the foliage of oak growing in a mixed stand in southern Sweden. ³²P and ³³P were recovered in the leaves/needles after 21 and 39 days. The recovery of labelled P in oak was higher from 15 cm soil depth than from 50 cm, however, more than 4% of the total amount of labelled P was taken up from 50 cm. This indicates that oak can utilize deep soil layers for nutrient uptake. A study on the uptake of Cs (as an analogue to K) and ¹⁵N into the leaves was performed on the same trees and detectable amounts of ¹⁵N and Cs were recovered in leaves and buds. This indicates that ¹⁵N and Cs can be used to study nutrient uptake of mature trees from the mineral soil.     

Effect of cleaning and thinning on height growth and girth increment in holm oak coppices (Quercus ilex L.)

This article studies the influence of cleaning or thinning intensity and of coppice age at the time of silvicultural treatment, on the growth of shoots remaining on the stool, in Quercus ilex coppices.Two experimental designs were used to answer these questions. The first studied the effect of two cleaning intensities on 4, 8, 15, 20, and 25 year old coppices. The second studied the effect of 4 thinning intensities (26%, 42%, 58%, or 78% removal of the initial basal area) on 43 and 57 year old coppices. In both cases, control plots undergoing no silvicultural treatment were maintained for each age class.The results, 4 and 5 years after silvicultural treatment, concerned girth increment and height growth of coppice shoots.Cleaning or thinning consistently had a positive influence on girth increment. In young coppices, less than 25 years old, mean annual girth increment was approximately 6 mm in the controls and double that under heavy cleaning. In older coppices, girth increment was lower in controls (3 mm per year) but the effect of thinning was very positive and, under very heavy thinning, growth was more than 11 mm per year.Cleaning and thinning often modify height growth. This effect was found to be either positive or negative and no general trend could be defined which was valid for all the age classes.Silvicultural treatments also had a positive effect on the appearance and growth of epicormic shoots on the holm oak trunks.The results were also analysed at the stool level. The number or percentage of shoots removed had a positive effect on the individual growth of selected shoots.     

Organic matter distribution and fluxes within a holm oak (Quercus ilex L.) stand in the Etna volcano

Located at 1100 m above sea level, on the western site of the Etna volcano, the ecophysiology of the Mount Minardo holm oak coppice has been investigated for more then twenty years. In this stand, now 31 years old, the above ground biomass amounts to 15000 g m^-2 of organic material, including leaves and perennial woody material. During these 31 years, the mean annual production has been around 775 g m^-2. The yearly mean litterfall amounts to 310 g m^-2, including 200 g m^-2 of leaves, mostly two years old. The soil surface is covered by a litter layer amounting to 3150 g m^-2. Each year, following Jenny's decomposition rate and field measurements, 290 g m^-2 of the litter turns into CO₂, or becomes incorporated in the soil organic matter.     

Differential activity of catalase and superoxide dismutase in seedlings and in vitro micropropagated oak (Quercus robur L.)

 Catalase (CAT) and superoxide dismutase (SOD) activity profiles were examined by native polyacrylamide gel electrophoresis in different tissues of seedlings and microcuttings of oak (Quercus robur L.) initiated from crown material (NL100A) and from basal epicormic shoots (NL100R), which differ in rooting ability. Two CAT isoforms were differentially active in seedlings and microcuttings; in particular, CAT-2 was activated in the basal callus of rooted microshoots. SOD isoenzymes, Mn-SOD and at least four Cu/Zn-SODs were found to be present, with Mn-SODs particularly active in microcuttings. No differences were found between the electrophoretic profiles of the two lines despite their different ontogenetic origin. The strong activity of CAT-2 in rooted microshoots indicates that this isoform is a protein specifically related to rooting. Received: 21 February 2000 / Revision received: 18 October 2000 / Accepted: 18 October 2000     

An indicator system for identification of sites of high conservation value for saproxylic oak (Quercus spp.) beetles in southern Sweden

The saproxylic beetle fauna on old oaks was sampled in four regions of southern Sweden using two methods: window and pitfall trapping. The aim was to test a way of finding indicator species which can be used to identify sites with high species number or that scored high on a conservation priority species index, based on occurrence of red-listed species. From 92 sites surveyed, in total 164 species of saproxylic beetles were identified. Different sets of indicator species were selected based upon 22 sites from a centrally located region. Predictions of species number and the index for 30 other sites from the same province were made. The correlation between observed and predicted species number and the index increased with increasing number of indicators. When comparing different treatment of species indata, the explanatory power of predictions was strongest for presence/absence data. Indicator sets of species effectively caught with pitfall traps gave overall the best predictions of both species number and the index. Predictions of species number and the index worked well within the same regions but gave varied result for the three other regions which shows that transferability of indicators between regions may be doubtful.