In vitro propagation of oak (Quereus robur L.) and linden (Tilia cordata Mill.)

Rapid multiplication of axillary shoots of oak and linden has been achieved on broad-leaved tree medium (BTM) and woody plant medium (WPM) containing low level of cytokinin (BAP 0.2–1.0 mg l^-1). High rooting percentages (80–95%) were obtained on low salt, low sucrose media, containing low level of auxins. Rooted plants were transplanted into pots containing a mixture of peat and perlite. Most of the plants (90–95%) survived the transfer. After the hardening off period the new plants were planted in the field.     

Seasonal variations of gas exchange, photosynthetic pigments, and antioxidants in Turkey oak (Quercus cerris L.) and Hungarian oak (Quercus frainetto Ten.) of different age

Seasonal changes in physiological and biochemical parameters were studied in 35-, 55- and 140-year-old trees of Turkey oak (Quercus cerris L.) and Hungarian oak (Q. frainetto Ten.), growing in natural stands in Eastern Balkan Mountains (Bulgaria). During the seasonal drought period (August), assimilation activity, transpiration rate, stomatal conductance and water potential had a seasonal minimum in all the studied tree ages and species. The foliar concentrations of glutathione, ascorbate, α-tocopherol, as well as photosynthetic pigments in oak leaves were significantly affected by season. With the increasing age of the studied trees, we observed a decrease of the physiological activity and an increase of the antioxidants’ accumulation. Both the species were drought tolerant and anisohydric, where Q. frainetto exhibited higher rates of gas exchange than Q. cerris. Moreover, they differed in the extent of increase in the foliar antioxidants and carotenoids.     

Dendroclimatic regions of pedunculate oak (Quercus robur L.) in Belarus

In this study, we present the boundaries of five dendrochronologically homogeneous regions of pedunculate oak (Quercus robur L.) and the master chronologies for them for the period 1703–2018 based on 27 tree-ring chronologies from different locations around Belarus. The average length of the master chronologies ranges from 70 to 310 years. Our research shows that the radial increment of a pedunculate oak in Belarus is primarily limited by the previous year’s August-September temperature, by the current year’s May temperature and by June precipitation. Oak growth in northern Belarus is mostly limited by air temperature, whereas precipitation plays more significant role in central and southern parts of the country. The most unfavorable years for oak growth in Belarus were 1940 and 1952, both of which were years when a very frosty winter was followed by a droughty summer. The results provide an important achievement in Belarusian dendrochronology, since they represent a significant breakthrough for Eastern Europe, where there is a considerable lack of dendroclimatic researches of broadleaved species.     

栎属植物体细胞胚胎发生研究现状

总结了影响栎属植物体细胞胚胎发生的主要可控因素及体细胞胚的遗传变异,组织学研究现状。目前,已能从成年组织上诱导出体细胞胚,但诱导率较低。重复性体胚发生系统已被认为是一种可资利用的繁殖途径,倍受关注。应用DNA分子标记分析表明：体胚细胞系内存在遗传变异。成熟和较低的萌发率是这一技术广泛应用的瓶颈。     

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.     

Phylogeographical variation of chloroplast DNA in holm oak (Quercus ilex L.)

Variation in the lengths of restriction fragments (RFLPs) of the whole chloroplast DNA molecule was studied in 174 populations of Quercus ilex L. sampled over the entire distribution of this evergreen and mainly Mediterranean oak species. By using five endonucleases, 323 distinct fragments were obtained. From the 29 and 17 cpDNA changes identified as site and length mutations, respectively, 25 distinct chlorotypes were distinguished, mapped and treated cladistically with a parsimony analysis, using as an outgroup Q. alnifolia Poech, a closely related evergreen oak species endemic to Cyprus where Q. ilex does not grow. The predominant role of Q. ilex as maternal parent in hybridization with other species was reflected by the occurrence of a single very specific lineage of related chlorotypes, the most ancestral and recent ones being located in the southeastern and in the northwestern parts of the species’ geographical distribution, respectively. The lineage was constituted of two clusters of chlorotypes observed in the ‘ilex’ morphotyped populations of the Balkan and Italian Peninsulas (including the contiguous French Riviera), respectively. A third cluster was divided into two subclusters identified in the ‘rotundifolia’ morphotyped populations of North Africa, and of Iberia and the adjacent French regions, respectively. Postglacial colonization probably started from three distinct southerly refugia located in each of the three European peninsulas, and a contact area between the Italian and the Iberian migration routes was identified in the Rhône valley (France). Chlorotypes identical or related to those of the Iberian cluster were identified in the populations from Catalonia and the French Languedoc region, which showed intermediate morphotypes, and in the French Atlantic populations which possessed the ‘ilex’ morphotype, suggesting the occurrence of adaptive morphological changes in the northern part of the species’ distribution.     

Variability of latewood-widths and -stable isotope ratios in a sessile oak tree (Quercus petraea (Matt.) Liebl.)

Axial variation of latewood-widths and -stable isotopes (δ¹³C and δ¹⁸O) of a single sessile oak tree (Quercus petraea (Matt.) Liebl.) was analyzed. The influence of sampling height was found to be weak for both, latewood-widths and -stable isotopes in the latewood. The observed trends with stem-height show clearly that possible effects due to cambial aging are overrated by annual effects. In addition, relations between samples from different stem-heights are strong for identical calendar years while they are poor for identical cambial age. This indicates that samples from both, living trees and historical wood can be taken from any height position of a trunk or beam as long they are free from anatomical abnormities. Simple correlation and Gleichläufigkeit were used for validation.     

Genetic transformation of selected mature cork oak (Quercus suber L.) trees

A transformation system for selected mature cork oak (Quercus suber L.) trees using Agrobacterium tumefaciens has been established. Embryos obtained from recurrent proliferating embryogenic masses were inoculated with A. tumefaciens strains EHA105, LBA4404 or AGL1 harbouring the plasmid pBINUbiGUSint [carrying the neomycin phosphotransferase II (nptII) and -glucuronidase (uidA) genes]. The highest transformation efficiency (4%) was obtained when freshly isolated explants were inoculated with A. tumefaciens strain AGL1. Evidence of stable transgene integration was obtained by PCR for the nptII and uidA genes, Southern blotting and expression of the uidA gene. The transgenic embryos were germinated and successfully transferred to soil.Abbreviations BA N⁶-Benzyladenine - GUS -Glucuronidase - MSSH Expression-proliferation medium - NAA -Naphthaleneacetic acid - nptII Neomycin phosphotransferase gene - uidA -Glucuronidase gene     

Structure of herbivore communities in two oak (Quercus spp.) hybrid zones

Summary We examined patterns of density and species diversity for leaf-mining Lepidopterans and gall-forming Hymenopterans in two oak (Quercus spp.) hybrid zones: Quercus depressipes x Q. rugosa and Q. emoryi x Q. coccolobifolia. In both species complexes, hybrid hosts typically supported significantly lower densities and species diversity of parasites than did parental types. This contradicts the findings of Whitham (1989) that suggested that hybrid hosts may act as parasite sinks both in ecological and evolutionary time. We discuss features of hybrid zones that are likely to influence patterns of herbivory.     

Microscale spatial distribution patterns of red oak (Quercus rubra L.) ectomycorrhizae

The microscale spatial distribution patterns of ectomycorrhizal (ECM) morphotypes of red oak (Quercus rubra L.) were analyzed over a 600 × 6 × 3 cm (length x width x depth) soil monolith. For this purpose, the soil monolith was divided into 2 × 2 × 1 cm cuboids. Each cuboid was assigned to an organic sublayer, namely the F- or H-layer. A new classification method was used to combine morphotypes with similar distribution patterns into five different groups. For identification of the ECM fungi, internal transcribed spacer (ITS) regions from nuclear rDNA were sequenced and compared with sequences from the international GenBank. Twenty-eight ECM types were identified in the soil monolith. Using this new classification method, it was found that the majority of morphotypes (16) appeared in a scattered horizontal distribution and that only 5 morphotypes formed clusters. In addition, it was found that many morphotypes had a clear preference for a specific organic sublayer: 14 morphotypes preferred the F-layer, 5 the H-layer, and only 4 morphotypes showed no preference for a given layer. Analysis of the distribution patterns showed that ECM fungi either exclude each other or occur together. The most frequent morphotypes, Cenococcum geophilum and Tomentella spec. 01, were associated with the whole ECM community. In contrast, the frequent morphotype Tomentella terrestris showed the lowest degree of association among the identified morphotypes.     

A quantification of shoot shedding in pedunculate oak (Quercus robur L.)

Almost 37 000 fragments (twigs, peduncles, inflorescences) were shed from a single mature oak tree over one year. Shedding showed strong seasonality with abrupt peaks in June and October, yet only weak correlations to weather parameters. The majority of shed twigs were short shoots, consisting of one or few extension units, which were not older than two years. In many twigs, shedding was preceded by the outgrowth of the terminal bud into either a male inflorescence or a defective extension unit. The large majority of shed twigs belong to a category of summer annual and biannual deciduous short shoots of an exploitative nature, only a minority are explorative long shoots. Implications for the architecture of the tree crown are that twigs of higher branching order are mostly affected by shedding, as they consist of short modules which exhibit a high proportion of flower buds.     

Genetic variation and differentiation in population of Japanese emperor oak (Quercus dentata Thunb.) and Mongolian oak (quercus mongolica fisch. ex ledeb.) in the south of the Russian far east

Allozyme variation of Japanese emperor oak Quercus dentata Thunb. and Mongolian oak Quercus mongolica Fisch. ex Lebed.) was examined in 11 populations of these species from southern Primorye. Analysis of 18 loci showed that in these populations, 66.7% of genes of Mongolian oak are polymorphic, the number of alleles per locus being 2.28. In Japanese emperor oak, these parameters were respectively 59.8 and 2.36%. The observed heterozygosity in both species did not differ from the expected values, constituting 0.198 in Japanese emperor oak and 0.161 in Maongolian oak. The Nei's distance between Quercus mongolica and Q. dentate was 0.053. The Japanese emperor oak and Mongolian oak populations formed separate clusters in an UPGMA dendrogram. Taking into account morphological and ecological isolation of the species, we suggest that the differences in gene frequencies may reflect selection acting in favor of adaptation to different growth conditions and counteracting between-species hybridization.     

Growth and nutation parameters of primary root of pedunculated oak (Quercus robur L.)

The elongation of primary roots of pedunculated oak (Quercus robur L.), which is associated with nutation oscillations, was registered by a time lapse cinematography. The following characteristics were expressed graphically: relative growth rate [mm h-¹], frequency [rev. h-¹] and amplitudes [mm] of nutation oscillations, their direction and location of nutation curvatures. The growth-oscillation mechanism is interpreted as a part of the geocontrol system of the root and was evaluated on the basis of its physiological efficiency. Comparison of nutation periods showed that the control system of pedunculated oak is three times more and two times less efficient than that of pea and maize, respectively. Comparison of the oscillation amplitudes of growing roots gives similar results.     

Inheritance and molecular variations of PCR-SSCP fragments in pedunculate oak (Quercus robur L.)

Single-strand conformaiton polymorphism (SSCP) profiles of six PCR-amplified fragments (250–800 bp) were analyzed in three full-sib families of pedunculate oak (Quercus robur L.) and their parents. Among the six fragments, four were polymorphic and one exhibited complex patterns that were not changed by varying the SSCP conditions. The number of bands for the analyzed fragments varied between two and four among individuals regardless of fragment size. As shown by segregation data, the variation in the number of bands between trees could only be attributed to the allelic composition (homozygotes vs heterozygotes): a genotype that exhibited two bands was presumptively homozygous, wheras a genotype exhibiting three or four bands was heterozygous. Mendelian proportions were observed in all crosses for each polymorphic fragment. In one cross, we could clearly identify a null allele due to a possible mutation at a primer site. Single-base mutations and short insertion-deletions were shown to be the molecular causes of the SSCP polymorphism observed between different alleles. The use of SSCP as a technique to identify co-dominant markers of PCR fragments (up to 800 bp) is recommended for gene diversity studies or for gene mapping.     

Induction,maturation and germination of holm oak (Quercus ilex L.) somatic embryos

Cotyledon explants of Panax ginseng produced somatic embryos directly on solid hormone-free MS medium containing 3% (w/v) sucrose while high concentration of NH₄NO₃ (60 mM) induced embryogenic callus. Ten subcultures of the embryogenic callus on hormone-free MS medium with 40 mM NH₄NO₃ gave hormone-independent proliferation of callus, which exhibited proliferation potential even on MS medium with a standard level of NH₄NO₃ (20 mM). Pulse treatment of callus with exogenous auxin or cytokinin (1.0 mg 1^–1 2,4-D, 1.0 mg 1^–1 kinetin) resulted in the loss of the hormone-independent characteristic and caused the callus to brown. For the suspension culture, embryogenic callus was transferred to MS liquid medium containing 3% (w/v) sucrose in an 500 ml Erlenmyer flask. Embryogenic cell clumps in full-strength MS liquid medium discharged toxic substances, resulting in strong suppression of cell growth. In 1/3-strength MS medium, exudation of toxic material did not occur. Embryogenic cell clumps were mass-grown on a large-scale in a bioreactor (20-1), showing a 7.1 increase of fresh weight in 1/3-strength MS medium with 3% (w/ v) sucrose after 5 weeks of culture. Total ginsenoside content of cultured embryogenic cell clumps was low and 6 times below naturally-cultivated ginseng roots.     

Gas composition and respiration of water oak (Quercus nigra L.) and green ash (Fraxinus pennsylvanica Marsh.) roots after prolonged flooding

Summary We compared the effects of 9.5 months of a continuous flooding treatment with a drained control treatment on one year-old seedlings of green ash (Fraxinus pennsylvanica Marsh.) and water oak (Quercus nigra L.), two tree species common to the bottomland-hardwood forests of eastern North America. The internal root gas composition of the more flood tolerant species, green ash, maintained higher oxygen and lower carbon dioxide concentrations under the flooding treatment than water oak. This apparently resulted in differences in rhizosphere oxidation. The amounts of Fe and Mn and the Fe/Mn ratio of the root coating extracted from trees in reduced soil conditions were much higher for the green ash than the water oak. It is argued that this reflects differences in the ability of these two species to maintain rhizosphere oxidation under prolonged periods of flooding and to prevent the accumulation of reduced potentially phytotoxic compounds. Alcohol dehydrogenase activity increased in the green ash and decreased in the water oak in the flooded treatment. This indicated that the better adapted species was able to rely upon increased anaerobic respiration in order to compensate for the decreased root oxygen supply, but the water oak was unable to maintain previous levels of respiration, probably as the result of sulfide toxicity.     

Small‐scale genetic structure and mating patterns in an extensive sessile oak forest (Quercus petraea (Matt.) Liebl.)

Oaks (Quercus) are major components of temperate forest ecosystems in the Northern Hemisphere where they form intermediate or climax communities. Sessile oak (Quercus petraea) forests represent the climax vegetation in eastern Germany and western Poland. Here, sessile oak forms pure stands or occurs intermixed with Scots Pine (Pinus sylvestris). A large body of research is available on gene flow, reproduction dynamics, and genetic structure in fragmented landscapes and mixed populations. At the same time, our knowledge regarding large, contiguous, and monospecific populations is considerably less well developed. Our study is an attempt to further develop our understanding of the reproduction ecology of sessile oak as an ecologically and economically important forest tree by analyzing mating patterns and genetic structure within adult trees and seedlings originating from one or two reproduction events in an extensive, naturally regenerating sessile oak forest. We detected positive spatial genetic structure up to 30 meters between adult trees and up to 40 meters between seedlings. Seed dispersal distances averaged 8.4 meters. Pollen dispersal distances averaged 22.6 meters. In both cases, the largest proportion of the dispersal occurred over short distances. Dispersal over longer distances was more common for pollen but also appeared regularly for seeds. The reproductive success of individual trees was highly skewed. Only 41 percent of all adult trees produced any offspring while the majority did not participate in reproduction. Among those trees that contributed to the analyzed seedling sample, 80 percent contributed 1–3 gametes. Only 20 percent of all parent trees contributed four or more gametes. However, these relatively few most fertile trees contributed 51 percent of all gametes within the seedling sample. Vitality and growth differed significantly between reproducing and nonreproducing adult trees with reproducing trees being more vital and vigorous than nonreproducing individuals. Our study demonstrates that extensive, apparently homogenous oak forests are far from uniform on the genetic level. On the contrary, they form highly complex mosaics of remarkably small local neighborhoods. This counterbalances the levelling effect of long‐distance dispersal and may increase the species’ adaptive potential. Incorporating these dynamics in the management, conservation, and restoration of oak forests can support the conservation of forest genetic diversity and assist those forests in coping with environmental change.     

Plant regeneration by somatic embryogenesis from cultured immature embryos of oak (Querem robur L.) and linden (Tilia cordata Mill.)

Embryogenic cultures and somatic embryos were obtained from immature zygotic embryos of oak (Quercus robur L.) cultured on a modified MS medium and WPM containing BAP (1 mg·l^–1) and GA₃ (1 mg·l^–1) or BAP and IBA. Germination and conversion of oak somatic embryos into plantlets was achieved on WPM containing a reduced concentration of cytokinin. Linden (Tilia cordata Mill.) somatic embryos developed in embryogenic tissues initiated from immature zygotic embryos cultured on a modified MS medium supplemented with 2,4-D (0.3-2.0 mg·l^–1). Germination of linden somatic embryos and plantlet formation occurred on MS medium containing a low concentration of IBA. Oak and linden plantlets produced from somatic embryos were successfully established in soil. Somatic embryos and plantlets were also regenerated from embryogenic cultures of Quercus petraea and Tilia platyphyllos.Abbreviations BAP 6-benzyIaminopurine - GA₃ gibberellic acid - IBA indole-3-butyric acid - 2,4-D 2,4-dichlorophenoxyacetic acid - MS Murashige and Skoog (1962) - WPM woody plant medium     

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.     

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.