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Effect of mecoprop drift on some plant species of conservation interest when grown in standardized mixtures in microcosms.

There has been an increasing awareness of potential impacts of herbicide drift on to vegetation in nature reserves and field margin habitats adjacent to treated areas. Previous work using single species bioassays has suggested that the effects of a single drift event are confined close to the sprayer (< 10 m). In the present study eight native dicotyledonous species, with and without a perennial grass (Lolium perenne), were grown in standardized microcosms in order to study (1) the effects of herbicide drift where plants were exposed to competition, and (2) the effects of a second spray application. The microcosms were arranged downwind (0-8 m) of a standard agricultural sprayer applying mecoprop at recommended rates. The effects of the herbicide drift on foliar symptoms of plant damage and end-of-season yield were assessed in each of two years. The main conclusions were that (1) growth of Stachys sylvatica and Lolium perenne (where sown) was enhanced near the sprayer and, (2) six other species (Digitalis purpurea, Galium mollugo, Hypericum hirsutum, Lychnis flos-cuculi, Primula veris and Ranunculus acris) showed some evidence of reduction in either performance (assessed non-destructively after a single exposure) or yield after two exposures. Three species (Lychnis flos-cuculi, Primula veris and Ranunculus acris) showed a reduction in flowering performance. Thus, mecoprop drift affected the aesthetic appearance, possibly the fecundity of some species and the balance between species in these microcosm experiments. The implications of these results for the persistence of attractive plant communities in sensitive areas are discussed.

Journal Article↗

Digestion of Pollen Components by Larvae of the Flower-Specialist Bee Chelostoma florisomne (Hymenoptera: Megachilidae).

Pollen digestion in larvae of the solitary bee Chelostoma florisomne (Megachilidae), a pollen-specialist (oligolectic) species on Ranunculus spp. (Ranunculaceae), was investigated using histological techniques. Serial sections from the anterior-, median-, and posterior-midgut/hindgut regions of the larval alimentary canal were selectively stained and examined for disappearance of lipids, proteins, and carbohydrates from ingested Ranunculus pollen grains. During digestion, pollen contents were observed to gradually extrude through the grain apertures. Most of the digestion of lipids and proteins occurred in the anterior- and median-midgut, whereas carbohydrates disappeared more slowly and mainly in the median- and posterior-midgut. In the hindgut, pollen comprised mainly empty exine shells, which were crushed and often broken, suggesting that some components of the wall are degraded during digestion, causing collapse of the outer, chemically resistant wall layer (exine). Lipids were completely digested, but small quantities of proteins and carbohydrates were often still present in grains in the posterior midgut/hindgut. The pollenkitt appeared to be completely absorbed, with none evident in the posterior midgut or in larval feces. The findings indicate that this oligolectic bee has the digestive equipment necessary to effectively remove and absorb the nutrients present in the pollen of its restricted food source. Comparison with similar studies in adult honey bees suggests that differing digestive abilities might be an underlying factor in pollen specialization in bees.

Journal Article↗

Toxic and feeding deterrent effects of native aquatic macrophytes on exotic grass carp (Ctenopharyngodon idella).

Declines of amphibians have been attributed to many factors including habitat degradation. The introduction of grass carp (Ctenopharyngodon idella) as a biological agent for aquatic plant control in ponds and lakes managed narrowly for human recreation has likely contributed to amphibian declines through massive plant removal and associated habitat simplification and thus degradation. This research examined the interactions among grass carp and three Midwestern aquatic plants (Jussiaea repens, Ranunculus longirostris, and R. flabellaris) that may be of value in rehabilitation of habitats needed by amphibians. The feeding preference study found that C. idella avoided eating both J. repens and R. longirostris. Ranunculus species studied to date contain a vesicant toxin called ranunculin that is released upon mastication. The study that compared the effects of R. flabellaris, J. repens and a control food administered by tube feeding to C. idella found significant lesions only in the mucosal epithelium of the individuals exposed to R.flabellaris. The avoidance by C. idella of J. repens and R. longirostris in the feeding preference study, and the significant toxicity of R. flabellaris demonstrated by the dosing study, indicate these plants warrant further examination as to their potential effectiveness in aquatic amphibian habitat rehabilitation.

Amphibians↗

[Commercial identification and resources survey on Chinese drug radix Ranunculi ternati].

Based on the collection of commercial products field investigation in distribution areas and main production places, and consultation of over 200 specimens the identification of traditional Chinese drug Maozhaocao was carried out. The results show that the present source of commercial "Maozhaocao" derives from 2 species of genus Ranunculus. The main species is Ranunculus ternatus, while R. polii is used only in a few regions. A resources survey of the drug was also conducted.

Conservation of Natural Resources↗

Agricultural effects on yield and nutritive value of grassland under a limited fertilisation rate on sandy soils in Flanders.

From 1999 to 2002, we studied the yield production and quality parameters of grassland installed on sandy soils (1) under normal management (NM), (2) under management with respect to meadow birds (MBM) -first mowing date 15 June or later- and (3) under botanical management (BM) -first mowing date 1 July or later. The experiments were conducted at two locations, Aalter and Geel. The biomass production under MBM and BM decreased by resp. 19% and 46% as compared to the NM. The nitrogen content in the vegetation of the delayed first cut (without fertilisation) of MBM (2.4%) and BM (2.3%) was much lower than under NM (3.3%). Since the first cut makes the major part of the total annual dry matter yield (43% for MBM and 59% for BM), the average nitrogen content of the overall DM yield dropped substantially if the first cut was delayed. The same phenomenon occurred for the digestibility of the organic matter: overall 66% for MBM and 59% for BM as compared to 69% for NM. The digestibility was partially influenced by the botanical composition. MBM resulted in a shift in botanical composition, favouring Poa trivialis L., Ranunculus repens L., and Holcus lanatus L. as compared to NM. BM mainly favoured Holcus lanatus L., and Ranunculus repens L. as compared to NM. Under the management types with delayed first cut, problems with residual mineral soil N never occurred.

Agriculture↗

Seasonality of root fungal colonization in low-alpine herbs.

Arbuscular mycorrhizal (AM) and dark septate endophytic (DSE) fungal colonization of Alchemilla glomerulans, Carex vaginata, Ranunculus acris ssp. pumilus and Trollius europaeus growing in low-alpine meadows in the Finnish subarctic were studied at different times during the growing season. Fungal colonization was correlated to soil soluble phosphorus (P) concentration. The influence of flower bud removal on fungal colonization was investigated in A. glomerulans, C. vaginata and R. acris and the correlation between AM and DSE colonization was studied. The fungal colonization patterns were found to be species-specific. R. acris maintained a relatively high rate of fungal colonization throughout the summer, while the rates of colonization of T. europaeus were lower and decreased towards the end of the season. A. glomerulans had constant arbuscular and vesicular colonization throughout the summer, but hyphal and DSE colonization declined towards the end of the season. C. vaginata did not form arbuscular mycorrhiza, but was colonized by DSE fungi and hyaline septate hyphae throughout the season. The soil soluble P concentration showed some seasonal variation, but was also highly variable between the study sites. Bud removal decreased arbuscular colonization of R. acris, but no unique effects were seen in any other parameters or the other species studied. The root fungal parameters correlated with soil P in some species at some sites, but no consistent trend was found. DSE colonization was positively correlated with root vesicular and hyphal colonization in some cases. The differences in fungal colonization parameters may be related to species-specific phenologies.

Alchemilla↗

Effects of elevated ozone on leaf delta13C and leaf conductance of plant species grown in semi-natural grassland with or without irrigation.

Stable carbon isotope ratios (delta(13)C) and leaf conductance (g(s)) were measured (2002, 2003) in Holcus lanatus L., Plantago lanceolata L. Ranunculus friesianus (Jord.), and Trifolium pratense L. at two levels of ozone (O(3)) with or without irrigation. In non-irrigated control plots, R. friesianus showed the least negative delta(13)C, and the smallest response to the treatments. Irrigation caused more negative delta(13)C, especially in H. lanatus. Irrespective of irrigation, O(3) increased delta(13)C in relationship to a decrease in g(s) in P. lanceolata and T. pratense. The strongest effect of O(3) on delta(13)C occurred in the absence of irrigation, suggesting that under field conditions lack of moisture in the top soil does not always lead to protection from O(3) uptake. It is concluded that in species such as T. pratense plants can maintain stomatal O(3) uptake during dry periods when roots can reach deeper soil layers where water is not limiting.

Agriculture↗

Does competition for phosphate supply explain the invasion pattern of Elodea species?

Two invasive aquatic plants, Elodea canadensis and Elodea nuttallii, occurred in north-eastern France. In this study, we examine the influence of phosphorus availability in soft water streams to explain the invasion pattern of exotic species (E. nuttallii and E. canadensis) compared to native plants (Callitriche platycarpa, Ranunculus peltatus). Total phosphorus was measured in these four aquatic macrophytes. Sediment total phosphorus and water-soluble reactive phosphorus were also analysed each season in 2001. Phosphorus content in the two invasive species and in R. peltatus was higher than in C. platycarpa. Elodea species are adapted to the seasonal phosphorus fluctuations as well as R. peltatus and exhibited high phosphorus storage ability. The high fluctuation availability of resources in space or/and time favoured the spread of the invasive plants and confirms the theory of invasibility of Davis et al. [2000. Fluctuating resources in plant communities: a general theory of invasibility. J. Ecol. 88, 528-534]. The eutrophication process increases the invasibility of E. nuttallii's, while inducing competition between E. nuttallii and native macrophyte species.

Ecosystem↗

Effects of submerged macrophytes on sediment resuspension and internal phosphorus loading in Lake Hiidenvesi (southern Finland).

The effects of submerged macrophytes on sediment resuspension and internal phosphorus loading in the shallow Kirkkojärvi basin of Lake Hiidenvesi were studied by sedimentation traps and sediment and water samples. During the 83 d study period, 793 g DW m(-2) of sediment was resuspended within the stand formed by Ranunculus circinatus, Ceratophyllum demersum and Potamogeton obtusifolius. Outside the stand, 1701g DW m(-2) sediment resuspension was measured during the same period. Water turbidity and concentration of suspended solids (SS) were significantly lower within the plant bed compared with the surrounding water area. Despite the higher concentration of inorganic suspendoids, the concentration of chlorophyll a was higher in the open water than within the submerged plant bed, owing to the enhanced nutrient recycling rate in the absence of submerged plants. With the resuspended sediment, 11.8 mg Pm(-2)d(-1) was brought into the water column within the stand and 24.5 mg Pm(-2)d(-1) outside the stand. Within the macrophyte stand, resuspended particles absorbed phosphorus from the water (indicated by the inverse relationship between SS and soluble reactive phosphorus), which was probably connected to the lowered phosphorus concentration of surface sediment due to uptake by macrophytes.

Absorption↗

To succeed globally, disperse locally: effects of local pollen and seed dispersal on tetraploid establishment.

Newly formed tetraploid plants in sympatry with their diploid progenitors should face significant obstacles to persistence and population establishment because of low-fitness triploids formed by cross-ploidy pollinations. Prior models have found restrictive conditions for a minority tetraploid subpopulation to persist. A stochastic spatial model, parameterized using snow buttercups (Ranunculus adoneus), was used to examine the influence of limited seed and pollen dispersal distances on the success of minority tetraploids and the interaction of these factors with different rates of self-pollination and tetraploid advantage. Higher rates of self-pollination and increased tetraploid advantage increase the probability of tetraploid persistence. Limiting the dispersal of seeds and pollen further increases the positive impact of any given level of self-pollination and tetraploid advantage. Taxa with short-distance seed and pollen dispersal should face much less stringent barriers to sympatric polyploid speciation than taxa with long-distance dispersal patterns. With short-distance seed and pollen dispersal, polyploid speciation should be possible in the absence of ecological differentiation or recurrent polyploid formation through unreduced gametes.

Crosses, Genetic↗

Effects of four generations of density-dependent selection on life history traits and their plasticity in a clonally propagated plant.

Life history evolution of many clonal plants takes place with long periods of exclusively clonal reproduction and under largely varying ramet densities resulting from clonal reproduction. We asked whether life history traits of the clonal herb Ranunculus reptans respond to density-dependent selection, and whether plasticity in these traits is adaptive. After four generations of exclusively clonal propagation of 16 low and 16 high ramet-density lines, we studied life history traits and their plasticities at two test ramet-densities. Plastic responses to higher test-density consisted of a shift from sexual to vegetative reproduction, and reduced flower production, plant size, branching frequency, and lengths of leaves and internodes. Plants of high-density lines tended to have longer leaves, and under high test-density branched less frequently than those of low-density lines. Directions of these selection responses indicate that the observed plastic branching response is adaptive, whereas the plastic leaf length response is not. The reverse branching frequency pattern at low test-density, where plants of high-density lines branched more frequently than those of low-density lines, indicates evolution of plasticity in branching. Moreover, when grown under less stressful low test-density, plants of high-density lines tended to grow larger than the ones of low-density lines. We conclude that ramet density affects clonal life-history evolution and that under exclusively clonal propagation clonal life-history traits and their plasticities evolve differently at different ramet densities.

Adaptation, Biological↗

Experimental life-history evolution: selection on growth form and its plasticity in a clonal plant.

The growth form along the continuum from compact phalanx plants to more loosely packed guerilla plants is an important life-history trait in clonal plants. Prerequisite for its evolution is heritable genetic variation. Starting with 102 genotypes of the stoloniferous herb Ranunculus reptans, we performed one selection experiment on spatial spread per rosette as measure of guerillaness (broad-sense heritability 0.198) and another on plasticity in this trait in response to competition (broad-sense heritability 0.067). After two generations, spatial spread was 36.9% higher in the high line than in the low line (realized heritability +/- SE 0.149 +/- 0.039). Moreover, compared with the low line genotypes of the high line had fewer rosettes, a lower proportion of flowering rosettes, a higher proportion of rooted rosettes, more branches per rosette, longer internodes and longer leaves. In the second experiment, we found no significant direct response to selection for high and low plasticity in spatial spread (realized heritability +/- SE -0.029 +/- 0.063), despite a significant correlated response in plasticity in the length of the first three stolon internodes. Our study indicates a high potential for further evolution of the clonal growth form in R. reptans, but not for its plasticity, and it demonstrates that the clonal growth form does not evolve independently of other clonal life-history characteristics.

Analysis of Variance↗

Genomic compatibility occurs over a wide range of parental genetic similarity in an outcrossing plant.

The theory of inbreeding and outbreeding suggests that there is a hump-shaped relationship between the genetic similarity of sexually reproducing parents and the performance of their offspring. Inbreeding depression occurs when genetic similarity is high, whereas hybrid breakdown is expected when genetic similarity is low. Between these extremes, the effect of genetic similarity on fitness is unclear. We studied the shape of this relationship by crossing 65 target genotypes of the clonal, self-incompatible Ranunculus reptans with partner genotypes spanning a broad scale of genetic similarity, ranging from crosses within populations to between-population crosses and hybridisation with a closely related species. Offspring were raised in outdoor tubs. Results revealed a quadratic relationship between parental genetic distance and offspring performance, with the clonal component of fitness more strongly hump-shaped than the sexual component. Optimal genetic similarity encompassed a broad range of within-population and between-population crosses. This pattern of genomic compatibility has important implications for the evolution of mating systems and mate choice.

Analysis of Variance↗

Experimental life-history evolution: selection on the allocation to sexual reproduction and its plasticity in a clonal plant.

Allocation to sexual reproduction is an important life-history trait in clonal plants. Different selection pressures between competitive and competition-free environments are likely to result in the evolution of specialized genotypes and to maintain genetic variation in reproductive allocation. Moreover, selection may also result in the evolution of plastic allocation strategies. The necessary prerequisite for evolution, heritable genetic variation, can best be studied with selection experiments. Starting from a base population of 102 replicated genotypes of the clonal herb Ranunculus reptans, we imposed selection on the proportion of flowering rosettes in the absence of competition (base population: mean = 0.391, broad-sense heritability = 0.307). We also selected on the plasticity in this trait in response to competition with a naturally coexisting grass in a parallel experiment (base population: 14% lower mean in the presence of competition, broad-sense heritability = 0.072). After two generations of bidirectional selection, the proportion of flowering rosettes was 26% higher in the high line than in the low line (realized heritability +/- SE = 0.205 +/- 0.017). Moreover, genotypes of the high line had 11% fewer carpels per flower, a 22% lower proportion of rooted rosettes, and a 39% smaller average distance between rosettes within a clone. In the second experiment, we found no significant responses to selection for high and low plasticity in the proportion of flowering rosettes (realized heritability +/- SE = -0.002 +/- 0.013). Our study indicates a high heritability and potential for further evolution of the proportion of flowering rosettes in R. reptans, but not for its plasticity, which may have been fixed by past evolution at its current level. Moreover, our results demonstrate strong genetic correlations between allocation to sexual reproduction and other clonal life-history characteristics.

Biological Evolution↗

Molecular phylogenetic study of the Ranunculaceae: utility of the nuclear 26S ribosomal DNA in inferring intrafamilial relationships.

There are only a small number of molecular markers currently proven to be useful for phylogenetic inference within the flowering plants. We demonstrate that the 5' end of the 26S ribosomal DNA (ca. 1100 bp) is of great value for investigating generic to subfamilial relationships. We analyzed DNA sequences from 31 species of the Ranunculaceae and four species of the Berberidaceae to test phylogenetic relationships within the Ranunculaceae. The inferred phylogeny strongly supports the concept that the Thalictrum chromosome group is not monophyletic, but consists of three independent lineages: (1) Hydrastis, (2) Xanthorhiza and Coptis, and (3) Thalictrum, Aquilegia, and Enemion. Based on comparison with conventional taxonomic characters, we propose a hypothesis that the third group also includes the rest of the Thalictrum chromosome taxa that have a base chromosome number of seven. For the Ranunculus chromosome group, our study suggests several relationships that have not been recognized by conventional systematics. The inferred 26S rDNA topology is compared with results from two previously published molecular data sets: DNA sequences from rbcL, atpB, and 18S rDNA genes and restriction fragment length polymorphism data from chloroplast DNA. The three topologies are highly congruent and agree with karyological characters, but not with fruit type, both of which have often been used for the higher classification of the Ra- nunculaceae.

Base Sequence↗

Antimicrobial activity of protoanemonin, a lactone from ranunculaceous plants.

Protoanemonin, a component of Ranunculus bulbosus, was tested as an antifungal agent on selected strains of dermatophytes and yeasts. The minimum inhibitory concentrations ranged from 2.0 to 7.5 X 10(-4) M and the minimum lethal concentrations from 3.8 X 10(-4) M to greater than 1.0 X 10(-3) M. The most sensitive dermatophyte tested was Epidermophyton floccosum, and the most sensitive yeast Rhodotorula glutinis. The effects of different culture media and of light on the sensitivity of Rhodotorula glutinis to protoanemonin were also tested. Structural analogies between protoanemonin and other cytotoxic unsaturated lactones, and the reversal by the amino acid cysteine of the antifungal action suggest a possible mechanism of action.

Arthrodermataceae↗

Large differences in the fraction of active bacteria in plankton, sediments, and biofilm.

Generally, only a small fraction of free-living pelagic bacteria are metabolically active, while particle-associated bacteria usually exhibit a larger proportion of active bacteria. Most previous studies on the active fraction of bacteria focus on planktonic communities, and there are only a few studies on sediment and epiphytic biofilm bacteria. We compared the active fraction of the total number of bacteria in three different habitats of the littoral zone of Lake Erken, Sweden, including the sediments, the epiphytic biofilm on the submerged macrophyte Ranunculus circinatus, and the water column. Active bacteria were detected as those with an active electron transport system, identified by the capacity to reduce the tetrazolium salt CTC (5-cyano-2,3-ditolyltetrazolium chloride) into its fluorescent, water insoluble state. There were large differences between habitats. The active fraction of the total number of bacteria detected by fluorescence microscopy (annual mean +/- SD) in the sediments was 46 +/- 10%, on R. circinatus 37 +/- 18%, and in the water column 4 +/- 4%. The abundance of CTC-reducing cells was correlated with total bacterial abundance, and the fraction of CTC-reducing bacteria generally increased with total bacterial abundance, for all the habitats. Consequently, the difference in the fraction of CTC-reducing bacteria between the habitats could be attributed to different densities of bacteria, with a larger proportion of active bacteria at higher bacterial densities.

Animals↗

An Ophiovirus isolated from lettuce with big-vein symptoms.

Big-vein is a widespread and damaging disease of lettuce, transmitted through soil by the chytrid fungus Olpidium brassicae, and generally supposed to be caused by Lettuce big-vein virus (LBVV; genus Varicosavirus). This virus is reported to have rigid rod-shaped particles, a divided double-stranded RNA genome, and one capsid protein of 48 kD, but has not been isolated or rigorously shown to cause the disease. We provide evidence that a totally different virus, here named Mirafiori lettuce virus (MiLV), is also very frequently associated with lettuce showing big-vein symptoms. MiLV was mechanically transmissible from lettuce to Chenopodium quinoa and to several other herbaceous test plants. The virus was partially purified, and an antiserum prepared, which did not react with LBVV particles in decoration tests. As reported for LBVV, MiLV was labile, soil-transmitted and had a single capsid protein of 48 kD, but the particles morphologically resembled those of ophioviruses, and like these, MiLV had a genome of three RNA segments approximately 8.5, 1.9 and 1.7 kb in size. MiLV preparations reacted strongly in Western blots and in ISEM with antiserum to Tulip mild mottle mosaic virus, an ophiovirus from Japan also apparently Olpidium-transmitted. They reacted weakly but clearly in Western blots with antiserum to Ranunculus white mottle virus, another ophiovirus. When lettuce seedlings were mechanically inoculated with crude or partially purified extracts from MiLV-infected test plants, many became systemically infected with MiLV and some developed big-vein symptoms. Such plants did not react in ELISA using an LBVV antiserum or an antiserum to tobacco stunt virus, and varicosavirus-like particles were never seen in them in the EM after negative staining. We conclude that MiLV is a hitherto undescribed virus assignable to the genus Ophiovirus. The cause or causes of lettuce big-vein disease and the properties of LBVV may need to be re-evaluated in light of our results.

Blotting, Northern↗