The Plastids of the Yew Tree (Taxus baccata L.): Ultrastructure and Immunocytochemical Examination of Chloroplastic Enzymes

The various types of plastids occurring in assimilatory and non-green tissues of Taxus baccata L. were investigated with respect to their ultrastructure and their content of starch, chlorophyll, Rubisco and the plastidic coupling factor CF₁. Chlorophyll was estimated from fluorescence intensity, and the presence of both enzymes was examined by electron microscopy after immunogold labelling. All kinds of plastids, irrespective of their location in the tree, contained stromal membranes. Due to their content of CF₁ and chlorophyll (except those of the roots) these membranes were addressed as thylakoids. Since the plastids also contain Rubisco, they represent chloroplasts. However, a decreasing gradient of all investigated chloroplastic characters from the plastids of the assimilatory tissue of the needles to those of axial tissues and of the roots was obvious. All kinds of plastids accumulated large amounts of starch especially in spring prior to bud burst but were virtually free of starch during the winter months.     

Production of taxanes by Taxus baccata plant cells

In callus cultures of Taxus baccata grown on agar media according to Murashige and Skoog supplemented with different growth hormones 8 taxol analogues were identified.     

Production of taxanes by Taxus baccata plant cells

Summary In callus cultures of Taxus baccata grown on agar media according to Murashige and Skoog supplemented with different growth hormones 8 taxol analogues were identified.     

Artichoke Leaf Morphology and Surface Features in Different Micropropagation Stages

Artichoke (Cynara scolymus L.) leaf size and shape, glandular and covering trichomes, stomatal density, stomata shape, pore area and epicuticular waxes during micropropagation stages were studied by scanning electron microscopy (SEM) and morphometric analysis with the aim to improve the survival rate after transfer to greenhouse conditions. Leaves from in vitro shoots at the proliferation stage showed a spatular shape, ring-shaped stomata, a large number of glandular trichomes and juvenile covering hairs, but failed to show any epicuticular waxes. Leaves from in vitro plants at the root elongation stage showed a lanceolated elliptic shape with a serrated border, elliptical stomata, decreased pore area percentage, stomatal density, and mature covering trichomes. One week after transfer to ex vitro conditions, epicuticular waxes appeared on the leaf surface and stomata and pore area were smaller as compared to in vitro plants. Artichoke acclimatization may be improved by hormonal stimulation of root development, since useful morphological changes on leaves occurred during root elongation.     

Microsporogenesis in Taxus baccata L.: The Development of the archaesporium

An ultrastructural and cytochemical investigation of the developmentof the mierosporangium of Taxus baccata reveals many resemblancesto comparable events in other land plants, but also severaldistinctive features. During early development the archaesporiumcontains groups of degenerate cells which ultimately disappear,presumably by resorption. Scattered elsewhere in the cells ofthe archaesporium are highly ordered bodies, possibly crystal-likeformations of tubulin. The tapetum differentiates from the outerlayer of the archaesporium. Mitoses coincidentally cease tobe detectable within the sporangium and the sporogenous cellsbecome recognizable about three weeks before their nuclei entermeiosis. Although, as in other gymnosperms investigated, theearly development of the archaesporial cells is not synchronized,there is a tendency for synchrony to be acquired by the sporogenouscells and diad formation takes place more or less simultaneouslythroughout the sporangium. Measurements by microfluorometry of RNA levels during developmentof the archaesporium show a progressive rise in the RNA contentof the cells up to the beginning of meiosis. Microsporogenesis, Taxus baccata, archesporium     

Modeling response curves of European yew (Taxus baccata L.) using HOF models along the environmental gradient in north of Iran

One of the important issues in plant autecology is the analysis and understanding of species-environment relationships and response of species to ecological gradients. Understanding how species respond to environmental variables is useful for predicting the environmental and geographic distribution of species. The main objective of this research is to study the response curve of European yew to identify the most important climatic and physiographic gradient in the northern forests of Iran. In the present study the Huisman–Olff–Fresco function (HOF) was used to study the response curve of the European yew in relation to each of the environmental variables individually and extract the optimum values. In order to select the optimal model, the Akaike Information Criterion (AIC) was used. Based on the results of this study, 41% of the models have either monotonically increasing or decreasing behaviors (models 2 and 3), 27% of the models have a unimodal symmetric behavior and 32% of models have a skewed behavior. European yew in 59% of models has a unimodal response curve. Based on the variables used in the study, it can be said that the European yew is a species that prefers oceanic climates, midlands altitudes, and north-facing slopes for establishment in temperate forests of Iran. Together, these findings advance niche theory about the European yew tree and may be used for generating better predictive models of European yew distributions under future climate change.     

Tuber-forming Substances in Jerusalem Artichoke (Helianthus tuberosus L.)

Four tuber-forming substances in Jerusalem artichoke were isolated from the leaves. The structures were established by spectroscopic methods as jasmonic acid (2), methyl β-D-glucopyranosyl tuberonate (3), and two new polyacetylene compounds, methyl β-D-glucopyranosyl helianthenate A (4, C₁₉H₂₄O₈) and B (5, C₁₇H₂₂O₈).     

Yew (Taxus x media Rehd.) cell suspension cultures as a source of taxanes

Natural resources of paclitaxel, an effective anticancer compound, were threatened with extinction soon after the discovery of this valuable substance. Cell suspension cultures derived from different Taxus species have rapidly become an alternative source of paclitaxel and other taxanes. In this paper we provide some insight into cell growth characteristics in cell suspension culture of Taxus x media cv. Hicksii, with emphasis on the effects of jasmonic acid (JA) on taxane production in cell lines with different initial taxane content. Additionally cell growth characteristics of two cell lines was followed during cultivation of cell suspension culture of Taxus x media cv. Hicksii. Packed cell volume (PCV) was shown to be a reliable and efficient alternative for measuring cell growth instead of fresh and dry weight. The initial total taxane content was screened in a number of cell lines, followed by observing the effect of JA on cell mass and total taxane production of selected lines. We showed a great variability in initial taxane content in different cell lines, which decreased during cell suspension maintenance. JA was shown to inhibit cell growth and increase total taxane production (14 to 106 fold).     

Catechin production in cultured cells of Taxus cuspidata and Taxus baccata

The main polyphenols in callus and cell suspension cultures of Taxus cuspidata and T. baccata were (+)-catechin and (−)-epicatechin, while lignans, such as (+)-taxiresinol, (+)-isotaxiresinol, (+)-isolariciresinol and (−)-secoisolariciresinol, were present in trace amounts. T. cuspidata cells contained 1.7% (+)-catechin and 2.4% (−)-epicatechin on dry wt basis but when stimulated with methyl jasmonate produced 3.4% catechin and 5.2% epicatechin. These are the highest levels of these metabolites obtained in plant cell cultures.     

Megasporogenesis and the Subsequent Cell Lineage Within the Ovule of Taxus baccata L.

Megasporogenesis in Taxus led to a T-shaped tetrad, the mitochondriaand plastids being largely confined to the chalazal megaspore,the only spore to show further development. Germination of themegaspore followed very soon after its formation. The femalegametophyte was initially coenocytic but after about one monthit became cellular, new walls arising between the nuclei inpositions marked out by accumulations of cisternae of endoplasmicreticulum. No ‘secondary spindles’ or other formsof microtubular participation in the siting of these walls couldbe detected. The development of the archegonia could not be resolved withcertainty, but it seemed likely that the cell analogous to thecentral cell of the pteridophyte archegonium eventually becamethe egg cell. The cytoplasm of the egg cell was clearly zoned.Prominent in the outer cytoplasm were granular bodies, 2–5µm in diameter, identifiable with the ‘grandes inclusions’described from the egg cells of conifers. The evidence pointedto their being derived from hypertrophied plastids which hadencapsulated part of the cytoplasm, a view strengthened by thecomplete absence of recognizable plastids from the mature cell.Bundles of microtubules radiating from the nuclear envelopewere a conspicuous feature, possibly stabilizing the zonationof the cytoplasm. Megasporogenesis, Taxus baccata, egg cell, ‘grandes inclusions’     

Leaf age as a factor in anatomical and physiological acclimative responses of Taxus baccata L. needles to contrasting irradiance environments

To evaluate the acclimative ability of current-year and previous-year needles of a shade tolerant conifer Taxus baccata L. to contrasting irradiance conditions, seedlings were raised under 27% solar irradiance and at 3 years of age they were transferred to an experimental garden and grown for one season under full irradiance (HL), 18% irradiance (ML) or 5% irradiance (LL). Whereas previous year needles did not change anatomically, current year needles in HL were thicker and had a thicker palisade and spongy mesophyll, and greater leaf mass per area than ML or LL needles. LL needles had greater nitrogen concentration than HL needles irrespective of age but only previous year LL needles also had an increased N per area content, thanks to their lack of reduction in LMA. Adjustment of chlorophyll and carotenoid content occurred in both needle age classes with LL and ML needles having much higher concentrations but, in current year needles, only slightly higher per area content than HL needles. Chlorophyll a/b ratio was not affected by age or irradiance. These modifications had no significant effect on photosynthetic capacities, which did not significantly differ between the age classes in HL or LL treatment and between treatments. On the other hand, high growth irradiance resulted in a greater photochemical yield, photochemical quenching, apparent electron transport rate and inducible non-photochemical quenching in needles formed in the current season. In previous year needles, however, only inducible NPQ was enhanced by high irradiance with other parameters remaining identical among treatments. To test sensitivity to photoinhibition, at the end of the summer plants from the three irradiance levels were transferred to a HL situation and F _v/F _M was determined over the following 18 days. Sensitivity to photoinhibition was negatively related to growth irradiance and previous year needles were less photoinhibited than current year needles. Thus, differences in acclimation ability between needle age classes were most pronounced at the level of anatomy and light reactions of photosynthesis, both of which showed almost no plasticity in previous year needles but were considerably modified by irradiance in current year needles.     

Sex influences the taxanes content in Taxus baccata

Like other species of the genus Taxus, European yew trees contain taxanes, including paclitaxel (T) and its precursor 10-deacetylbaccatin III (10-DAB). Taxanes are one of the most effective anticancer drugs. This study was undertaken to describe the levels and patterns of taxane variation in the Sudetian region (SW Poland). Paclitaxel (T) and 10-deacetylbaccatin III (10-DAB) concentrations were analysed in five populations. Needles and twigs were analysed from 60 individuals (30 males and 30 females) in each population. In addition, morphometric measurements were taken in the populations to obtain light intensity coefficients (specific leaf area, SLA). High variability in the taxane contents at both intra and interpopulational levels was found. Nevertheless, females had a significantly higher taxane content compared to males. Because taxanes are carbon-based secondary metabolites, females have higher rate of gas exchange of females compared to males. This was probably an adaptation to greater reproductive effort incurred by females. In this regard, female individuals seem to be better for selecting elite cultivars with a higher taxane production. The relationship between light intensity and taxane content was not significant. Shading, important for optimizing crop production, should not reduce the concentration of taxanes.     

Pollen limitation and fruit abortion in a declining rare tree,the Eurasian yew (Taxus baccata L.): A reproductive cost of ecological marginality

Populations of rare tree species such as the dioecious and anemophilous yew (Taxus baccata) are especially prone to extinction if they occupy marginal habitats. This is the case of yew populations growing in Mediterranean mountains under dry climate and a severe anthropogenic disturbance regime, which show insufficient regeneration. We examined two of the likely mechanisms driving this population decline. The study was conducted in a central Spanish yew population in 2005 and 2006. On one hand, we tested for the effect of the amount of pollen received by female flowers by means of a pollen addition experiment. On the other hand, we tested for the effect of initial fruit set on abortion and mature fruit production. Finally, we evaluated the effect of the spatial arrangement of male and female yew trees on realized fecundity. The success of fertilization and fruiting in the experimental population was pollen-limited. Female trees aborted ca. 70% of the fruit set in both years and treatments. Fruit set was significantly influenced by the females' neighbourhood, with fecundity decreasing exponentially with distance to the nearest male in both years. Overall, our results indicate that factors associated with ecological marginality have a strong influence on yew performance.     

Microsporogenesis in Taxus baccata L.: The Formation of the Tetrad and Development of the Microspores

Following meiosis II in Taxus microsporangia a small proportionof the tetrads regularly degenerated. Despite frequent inequalityin the frequency of ribosomes between the spores of a tetrad,partial degeneration within a tetrad was never observed. Theinitial wall of the young spores was found to resemble the wallof the mother cell in containing a fibrillar layer, and thetwo walls may possess similar isolating properties. The symmetryof the tetrad was regularly iso-bilateral. The formation ofthe sporoderm began as the spores were released into the loculusby the rapid dissolution of the wall of the mother cell. Osmiophilicdroplets emerged from the spore protoplast and entered the wall.The fibrillar layer ceased to be recognizable and the dropletscoalesced to form an outer layer on which up to six sporopolleninlamellae, probably of tapetal origin, were deposited. The accretionof a single layer of sporopollenin droplets, in no recognizablepattern, gave rise to the outer verrucose part of the exine.Cytochemical tests showed that the tapetum was rich in acidphosphatases from the beginning of meiosis. Towards the endof its degeneration the tapetum intruded into the loculus andcould therefore be regarded as partly invasive. Taxus baccata, microsporogenesis, tetrad symmetry, sporoderm     

山荆子腊叶标本表型性状变异分析

为了揭示山荆子表型性状的变异程度和规律,以150份山荆子腊叶标本为材料,选择20个表型性状,统计基本参数并进行聚类分析.结果表明:叶的平均变异系数(CV=32.67%)大于果的平均变异系数(CV=20.7%),果的性状稳定性较高,叶性状中叶柄长变异系数(CV=38%)最大,叶宽位/叶长变异系数(CV=13.29%)最小;果实性状中果径宽位/果柄长(CV=29.37%)变异最大,变异最小的性状是果长/果径(CV=13.10%);选用平均欧氏距离对各地方居群的20个表型性状进行UPGMA聚类,在距离5阈值处,可以划分为5类,在欧氏距离6阈值处,可划分为3大类.     

苹果属山荆子遗传多样性的RAPD分析

采用RAPD分子标记对东北、华北地区山荆子8个天然居群的137株个体进行了遗传多样性研究,10个引物共得到72个扩增位点,其中多态性位点63个.多态位点百分率为87.50%,有效等位基因数(Ne)为1.602 1,Nei's基因多样性(H)为0.338 6,Shannon信息指数(J)为0.496 1,表明山荆子遗传多样性水平较高.基因流(Nm)为1.735 3,说明山荆子各居群间存在一定的基因交流.居群间基因分化系数(Gst)值为0.223 7,说明虽然山荆子居群的遗传变异主要存在于居群内,但各居群间也存在着较高的遗传分化.