Characterization of a Tymovirus Isolated from Anagyris foetida as a Strain of Scrophularia Mottle Virus

A virus isolated from Anagyris foetida and infecting species in the Leguminosae and Solanaceae had typical properties of a tymovirus. It sedimented as two components (49 S and 103 S) and induced the formation of double membrane-bounded invaginations in the chloroplasts of infected cells. Large numbers of ‘empty shells’ were found in the nuclei. The coat protein had a molecular weight of c. 20.000 and the RNA consisted of c. 5500 nucleotides. Crystallization of the virus in laminal crystals could be achieved by precipitation with 10 % polyethylenglycol 6000 and 1 % NaCl. Serologically, the virus was closely related to Scrophularia mottle, Ononis yellow mosaic and Plantago mottle viruses. The four viruses which all infect leguminous hosts are separated by serological differentiation indices which are mostly between 1 and 3. It is therefore suggested that they all should be considered as strains of the same virus which for reasons of priority should have the name Scrophularia mottle virus (ScrMV). The proposed Anagyris strain clearly differs from the Scrophularia mottle, Ononis yellow mosaic and Plantago mottle strains of ScrMV in host range, symptomatology, electrophoretic mobility serological properties and some cytopathogenic effects. It is not clear why the Anagyris strain infects A. foetida systemically in nature, but only locally under greenhouse conditions.     

Chemotaxonomy and evolution of Plantago L.

In continuation of our investigations of the genus Plantago L. (Plantaginaceae), sixteen species were investigated with respect to water-soluble glycosides. The iridoids auroside, strictoloside and globularicisin, as well as poliumoside, 3-[(4--D-glucopyranosyloxy)phenyl]propionic acid and 2-[4-(-D-glucopyranosyloxy)phenyl]acetic acid were isolated from Plantago for the first time. The latter compound has not previously been isolated as a natural product. Sorbitol was the main carbohydrate in all the species investigated. The distribution of iridoids correlates well with the morphological classification of Rahn and also with a recently published molecular phylogenetic study of nuclear ribosomal and plastid DNA sequences. A new chemotaxonomic finding is an abundance of iridoid glucosides present in one of the two groups within subgenus Coronopus section Coronopus, while the other group lacks iridoids, supporting a subdivision of this section. Moreover, in subgenus Albicans, 10-benzoylcatalpol is a characteristic constituent of section Gnaphaloides, while the corresponding cinnamoyl ester globularin occurs in section Lanceifolia. In biosynthetic experiments, labelled epideoxyloganic acid and deoxygeniposidic acid were incorporated into aucubin and geniposidic acid in Plantago ovata, consistent with earlier findings, but no incorporation into asperuloside was observed. The evolution of biosynthetic pathways in Plantago is discussed.     

Chemotaxonomic aspects of iridoids occurring inPlantago subg.Psyllium (Plantaginaceae)

Within the genusPlantago, subg.Psyllium is characterized by the accumulation of the iridoids plantarenaloside and bartsioside. The latter compound was identified as constituent ofP. sempervirens for the first time. The structures of iridoids were determined by UV, IR,¹³C-NMR and¹H-NMR. Both compounds are of chemotaxonomic value, as they are typical only of subg.Psyllium withinPlantago. The distribution of plantarenaloside within subg.Psyllium suggests the heterogeneity of this taxon.     

Evolution of flower shape in <Emphasis Type="Italic">Plantago lanceolata</Emphasis>

Plantago lanceolata produces small actinomorphic (radially symmetric), wind-pollinated flowers that have evolved from a zygomorphic, biotically pollinated ancestral state. To understand the developmental mechanisms that might underlie this change in flower shape, and associated change in pollination syndrome, we analyzed the role of CYC-like genes in P. lanceolata. Related zygomorphic species have two CYC-like genes that are expressed asymmetrically in the dorsal region of young floral meristems and in developing flowers, where they affect the rate of development of dorsal petals and stamens. Plantago has a single CYC-like gene (PlCYC) that is not expressed in early floral meristems and there is no apparent asymmetry in the pattern of PlCYC expression during later flower development. Thus, the evolution of actinomorphy in Plantago correlates with loss of dorsal-specific CYC-like gene function. PlCYC is expressed in the inflorescence stem, in pedicels, and relatively late in stamen development, suggesting a novel role for PlCYC in compacting the inflorescence and retarding stamen elongation in this wind pollinated species.     

A phylogenetic study of the Plantaginaceae

In a study based on morphological, embryological and chemical data of the Plantaginaceae, within the subclass Sympetalae or Asteridae, the superorder Lamianae is shown to be monophyletic. However, it was not possible to reconstruct the phylogeny within Lamianae or to find a sister-group for the monophyletic Plantaginaceae – Hydrostachyaceae for the latter is rejected. Three or rarely four genera have previously been recognized within Plantaginaceae, but in both cases Plantago appears as polyphyletic, which is considered unacceptable. Six clades are recognized as subgenera within Plantago: subgen. Plantago (c. 131 species), subgen. Coronopus (c. 11 species), subgen. Littorella (three species), subgen. Psyllium (c. 16 species), subgen. Bougueria (one species), and subgen. Albicans (51 species). Within P. subgen. Plantago, the paraphyletic sect. Plantago (c. 42 species) is found in all parts of the world except South America, New Zealand, Australia, and New Guinea, where sect. Oliganthos and sect. Mesembryniae vicariate. It is not possible to infer the phylogeny within sect. Plantago, for example between the endemic species from distant Pacific Islands; their common ancestor might be a species that once had a very wide distribution. Keys to genera, subgenera, sections and series are given. Only one genus, Plantago L., is recognized. The following proposals are made in the Appendix: P. ser. Oliganthos Rahn, ser. nov.; P. ser. Carpophorae (Rahn) Rahn, stat. nov. (= sect. C.); P. ser. Microcalyx (Pilg.) Rahn, stat. nov. (= sect. M.); P. unibrackteata Rahn, nom. nov. (=P. uniflora Hook.f. non L.); P. subgen. Littorella (P. J. Bergius) Rahn, stat. nov. (=Littorella P. J. Bergius); P. araucana Rahn, nom. nov. (=Littorella australis Griseb. non Plantago australis Lam.); P. americana (Fernald) Rahn, comb. nov.(=Littorella a.); P. subgen. Bougueria (Decne.) Rahn, stat. nov. (=Bougueria Decne.); P. nubicola (Decne.) Rahn, comb. nov. (=Bougueria n. Decne.); P. subgen. Albicans Rahn, subgen. nov. Ten lectotypes are selected.     

Hair anatomy inPlantago subg.Psyllium (Plantaginaceae)

The hairs of seven representatives ofPlantago L. subg.Psyllium were studied. Three types of headless hairs and three types of headed hairs were observed. Clavate hairs and hairs with several-celled stalk and unicellular head are characteristic of subg.Psyllium and provide evidence for the distinctness of this taxon. Based on our studies it can be assumed thatRahn's idea of enlarging subg.Psyllium by six sections from subg.Plantago sensuPilger is incorrect.     

Phenological records as a complement to aerobiological data

Phenological studies in combination with aerobiological studies enable one to observe the relationship between the release of pollen and its presence in the atmosphere. To obtain a suitable comparison between the daily variation of airborne pollen concentrations and flowering, it is necessary for the level of accuracy of both sets of data to be as similar as possible. To analyse the correlation between locally observed flowering data and pollen counts in pollen traps in order to set pollen information forecasts, pollen was sampled using a Burkard volumetric pollen trap working continuously from May 1993. For the phenological study we selected the main pollen sources of the six pollen types most abundant in our area: Cupressaceae, Platanus, Quercus, Plantago, Olea, and Poaceae with a total of 35 species. We selected seven sites to register flowering or pollination, two with semi-natural vegetation, the rest being urban sites. The sites were visited weekly from March to June in 2007, and from January to June in 2008 and 2009. Pollen shedding was checked at each visit, and recorded as the percentage of flowers or microsporangia in that state. There was an association between flowering phenology and airborne pollen records for some of the pollen types (Platanus, Quercus, Olea and Plantago). Nevertheless, for the other types (Cupressaceae and Poaceae) the flowering and airborne pollen peaks did not coincide, with up to 1 week difference in phase. Some arguments are put forward in explanation of this phenomenon. Phenological studies have shown that airborne pollen results from both local and distant sources, although the pollen peaks usually appear when local sources are shedding the greatest amounts of pollen. Resuspension phenomena are probably more important than long-distance transport in explaining the presence of airborne pollen outside the flowering period. This information could be used to improve pollen forecasts.     

Spring staging in Brent Geese Branta bernicla: feeding constraints and the impact of diet on the accumulation of body reserves

Summary The diet composition of Brent Geese Branta bernicla on a salt-marsh was quantified. Puccinellia maritima was the principal food species, while Plantago maritima and Triglochin maritima were less commonly taken. Festuca rubra only acted as a substitute for Puccinellia when production of the latter species dropped. The metabolizable energy of the food plants ranged from 5 to 11 kJ·g^–1. By assessing the ingestion rates of geese feeding on different food species, the net intake rate could be derived. Plantago and Triglochin appeared to be the most profitable plants to eat. The proportion of these species in the diet was restricted by (1) the capacity of the alimentary tract, since high intake rates combined with high water contents of the food plants easily led to overfill; and (2) the limited distribution of these plants, in combination with their rapid depletion by grazing geese. These latter factors led to an unequal allocation among individual geese. Most Plantago and Triglochin was obtained by dominant pairs within the flocks. The high quality of Puccinellia allowed geese to gain mass in spring, but the metabolizable energy of this plant species declined during the staging period, and Plantago and Triglochin increased in importance in supplying the geese with components with which to build their body reserves. The timing of the onset of spring growth of the various food species differed between years, and plant phenology was shown to have a profound effect on the final body reserves of the geese.     

Effects of CO2 and nutrient enrichment on tissue quality of two California annuals

The effects of CO₂ enrichment and soil nutrient status on tissue quality were investigated and related to the potential effect on growth and decomposition. Two California annuals, Avena fatua and Plantago erecta, were grown at ambient and ambient plus 35 Pa atmospheric CO₂ in nutrient unamended and amended serpentine soil. Elevated CO₂ led to significantly increased Avena shoot nitrogen concentrations in the nutrient amended treatment. It also led to decreased lignin concentrations in Avena roots in both nutrient treatments, and in Plantago shoots and roots with nutrient addition. Concentrations of total nonstructural carbohydrate (TNC) and carbon did not change with elevated CO₂ in either species. As a consequence of increased biomass accumulation, increased CO₂ led to larger total pools of TNC, lignin, total carbon, and total nitrogen in Avena with nutrient additions. Doubling CO₂ had no significant effect on Plantago. Given the limited changes in the compounds related to decomposibility and plant growth, effects of increased atmospheric CO₂ mediated through tissue composition on Avena and Plantago are likely to be minor and depend on site fertility. This study suggests that other factors such as litter moisture, whether or not litter is on the ground, and biomass allocation among roots and shoots, are likely to be more important in this California grassland ecosystem. CO₂ could influence those directly as well as indirectly.     

<Emphasis Type="Italic">Plantago pyrophila</Emphasis> (Plantaginaceae), a new species from the cerrados of Eastern Bolivia

A new species Plantago pyrophila Villarroel & J. R. I. Wood is described from the Serranias de Santiago de Chiquitos in the cerrados of eastern Bolivia. It is clearly adapted to survival in the cerrado environment where grassland fires are a frequent event. The new species is remarkable for being geographically isolated from other species of the genus, being the only species of Plantago known from the cerrado biome.     

Pollen wall and tapetum development in Plantago major L. (Plantaginaceae): assisting self-assembly

We have attempted to elucidate the underlying mechanisms of sporoderm development and pattern determination in Plantago major through a detailed ontogenetic study, using transmission electron microscopy (TEM) and scanning electron microscopy (SEM). We aim to compare our observations and interpretation with those on other species. Our study of sporoderm development in Plantago from the early tetrad stage to mature pollen grains has shown that pure physical processes, including self-assembly, which are not under direct genetic control, play an important role and represent evidently one of the instruments of evolution. Our observations fit well with the sequence of self-assembling micellar mesophases and show reiteration of some of them, confirming our self-assembly hypothesis. Some attention was also paid to the possible role of rough and smooth endoplasmic reticulum in the cortical cytoplasm of the developing microspores. The tapetum and Ubisch bodies development are also traced. The importance of detailed ontogenetic studies for understanding the establishment of complex pollen walls in any species and for understanding mechanisms underlying sporoderm development was demonstrated. We also present a simulation, obtained in vitro experiments by self-assembly, mimicking pollen grain of Plantago major. It is clear that, in pollen wall development, biological processes and purely physical factors work in tandem.     

Resource allocation patterns of two California-Sonoran desert ephemerals

Summary The patterns of allocation of structural and nonstructural carbon were followed in the co-occurring desert ephemerals Plantago insularis and Camissonia boothii. Patterns of biomass distribution were determined from material harvested at biweekly intervals as were levels of nonstructural sugar and starch. Seasonal patterns of growth and reproduction differed markedly with Plantago allocating significantly more structural and nonstructural carbon to reproduction early in the season. Plantago completed its life cycle in less than 60 days but Camissonia continued both vegetative and reproductive growth to over 100 days. The longer growing season of Camissonia was possible because more energy was allocated to vegetative tissues and storage presumably as investment toward longer life and higher levels of reproduction.     

Lipid Composition of the Roots of Plantago Species: Response to Alteration of the Level of Mineral Nutrition and Ecological Significance

In a study of the adaptation of Plantago species to their specific environment the lipid composition of the roots of several species: Plantago major L. ssp. major, Plantago major L. ssp. pleiosperma Pilger, Plantago lanceolata L., Plantago media L., Plantago maritima L., Plantago coronopus L. was studied as well as the effect of the nutritional regime. Upon exposure to low-salt conditions Plantago major L. ssp. major L. and Plantago maritima L. maintained the level of free sterols in the roots, despite a depressed level of total sterols, and the root lipids were more saturated than under high-salt conditions. Both factors may reduce nutrient leakage from the roots to the low-salt condition. Upon exposure to low-salt conditions, all Plantago species showed a decreased level of galactolipid (exception: Plantago coronopus) and a decreased level of sitosterol (exception: Plantago maritima); the latter being compensated by an elevated level of cholesterol + tocopherol in Plantago major ssp. pleiosperma, Plantago lanceolata and Plantago media. Plantago coronopus was the only species which under low-salt conditions showed an increased level of free sterols, among which cholesterol was the most important; thus indicating a high degree of regulation of membrane permeability under alternating nutritional conditions. The level of sulfolipid was kept constant in all Plantago species, with the highest level observed in Plantago maritima. The role which various lipids may play in maintenance of membrane integrity under alternating nutritional conditions is discussed.     

2-Deceno-δ-lactone-producing Fungi,Strains of Fusarium solani,Isolated by Using a Medium Containing Decano-δ-lactone as the Sole Carbon Source

A screening procedure for microorganisms which have an ability to produce a desired compound as their secondary metabolite is first proposed. In some cases, the target microorganisms can be expected to grow and be enriched in a medium containing the desired compound, namely one of the secondary metabolites of the microorganisms, as the sole source of carbon, degrading and assimilating the compound to the primary metabolites. This approach was applied to isolate alkano-δ-lactones producing fungi by using a medium containing alkano-δ-lactones as the sole source of carbon. We isolated Fusarium solani and Trichoderma viride that had the ability to biosynthesize 2-deceno-δ-lactone (massoialactone) and 2,4-decadieno-δ-lactone(6-pentyl-δ-pyrone), respectively, in a glucose medium.     

Hair morphology inPlantago sect.Coronopus (Plantaginaceae)

The presence of bottle-like hairs has been described in four hitherto uninvestigated taxa ofPlantago sect.Coronopus sensuDietrich. This hair type is a feature characteristic of sect.Coronopus.     

Polymorphic microsatellite loci in Plantago lanceolata

The genus Plantago is particularly interesting for evolutionary studies because of its wide range of mating systems. We have developed primers for five highly polymorphic microsatellite loci isolated from P. lanceolata. All five loci amplified and were polymorphic in the two populations examined, Lowsteads Beach in the United Kingdom and Duke in the United States. These new markers will allow a comparison of population structure between the outcrossing species P. lanceolata, and the highly selfing species P. major.     

Chloroplast DNA variation within and among five Plantago species

Chloroplast DNA variation was studied within and among five Plantago species. We determined polymorphism by surveying 86% of the chloroplast genome using 9 or 11 restriction enzymes for intra or interspecific variation, respectively. All five species were polymorphic for both site and length mutations. The outcrossing P. lanceolata has six chloroplast haplotypes of which some were found in the Netherlands as well as in the United States. The highly selfing P. major had five haplotypes and Dutch and United States populations were differentiated from each other in cpDNA. Plantago species are highly differentiated from each other. Of 252 restriction-sites, 52% were variable among species. Most of this variation was localized in part of the large single copy region. We derived a molecular phylogeny of the five species from restriction-site variability using PAUP 3.0. P. coronopus and P. maritima form a group as do P. major and P. media. P. lanceolata is more distantly related to the other four species. In a consensus tree both P. major haplotypes and both P. media haplotypes were connected to one node.     

Anti-mitotic and anti-genotoxic effects of Plantago lanceolata aqueous extract on Allium cepa root tip meristem cells

Plantago is the most important genus of Plantaginaceae family and is used in traditional medicine around the world for different purposes. Plantago coronopus L., Plantago major L., Plantago media L. and Plantago lanceolata L. are most commonly used species of Plantago in traditional medicine in Turkey. The main goal of this study was to investigate the eventual anti-mitotic and anti-genotoxic effects of P. lanceolata L. leaf aqueous extracts (15 g/L and 30 g/L) on Allium cepa L. root tip meristem cells which were treated with 0.7% hydrogen peroxide. For this purpose, two different experiments were performed under the same conditions. In the first experiment, Allium cepa onion bulbs were treated with 0.7% H₂O₂ for 1 h. After the H₂O₂ treatment, the onion bulbs were treated with two different concentrations (15 g/L and 30 g/L) of P. lanceolata extracts for 24 h. In the second experiment, A. cepa onion bulbs were treated with two different extract concentrations (15 g/L and 30 g/L) for 24 h and then with 0.7% H₂O₂ for 1 h. The test concentrations were determined according to doses which are recommended in alternative medicinal usage by people. As positive and negative control 0.7% H₂O₂ and tap water was used, respectively. As a result, it was determined that aqueous extracts reduced mitotic index and chromosome aberrations in treatment groups in comparison with controls. These results showed that P. lanceolata aqueous extracts have anti-mitotic and anti-genotoxic effects.     

Palynological differences in the pollen content of Eucalyptus honey from Australia,Portugal and Spain

A melissopalynological study was carried out on 75 samples of Eucalyptus honey from different countries (22 from Australia, 13 from Portugal and 40 from Northwest Spain), in which a total of 145 different pollen types were identified. Of such, only Eucalyptus, Acacia, Brassica, Echium, Ligustrum, Plantago, Rumex, Taraxacum, Trifolium and Vicia pollen are present in the honeys from all three countries.

Among the most abundant pollen types in Australian honeys, those belonging to the following families stand out: Myrtaceae (Angophora and Melaleuca), Fabaceae, Asteraceae, Proteaceae and Euphorbiaceae, along with Raphanus, Echium and Citrus.

In the Portuguese honeys, the presence of Fabaceae, Rosaceae and Cistaceae stands out, with Castanea and Erica as secondary pollen types, while Fabaceae, Asteraceae, Boraginaceae, Cistaceae and Scrophulariaceae are the families with the highest quantity of pollen types in the samples from Spain. In the latter, the most important secondary pollen types are: Castanea, Cytisus, Erica, Lotus, Salix and Rubus.     

Microorganisms and nematodes increase levels of secondary metabolites in roots and root exudates of Plantago lanceolata

Plant secondary metabolites play an important role in constitutive and inducible direct defense of plants against their natural enemies. While induction of defense by aboveground pathogens and herbivores is well-studied, induction by belowground organisms is less explored. Here, we examine whether soil microorganisms and nematodes can induce changes in levels of the secondary metabolites aucubin and catalpol (iridoid glycosides, IG) in roots and root exudates of two full-sib families of Plantago lanceolata originating from lines selected for low and high constitutive levels of IG in leaves. Addition of soil microorganisms enhanced the shoot and root biomass, and the concentration of aucubin in roots of both Plantago lines without affecting IG levels in the rhizosphere. By contrast, nematode addition tended to reduce the root biomass and enhanced the stalk biomass, and increased the levels of aucubin and catalpol in root exudates of both Plantago lines, without affecting root IG concentrations. The Plantago lines did not differ in constitutive levels of aucubin and total IG in roots, while the concentration of catalpol was slightly higher in roots of plants originally selected for low constitutive levels of IG in leaves. Root exudates of “high IG line” plants contained significantly higher levels of aucubin, which might be explained by their higher root biomass. We conclude that soil microorganisms can induce an increase of aucubin concentrations in the roots, whereas nematodes (probably plant feeders) lead to an enhancement of aucubin and catalpol levels in root exudates of P. lanceolata. A potential involvement of secondary metabolites in belowground interactions between plants and soil organisms is discussed.