Gérard Marigo

Success in the demographic expansion of Fraxinus excelsior L.

This review examines the ecological traits and ecophysiological characteristics of the common ash (Fraxinus excelsior L.) that help to explain the current spread of ash communities in Europe. As a consequence of anthropogenic disturbance, the common ash has experienced new environmental conditions and the frequency of the species has fluctuated over space and time. Owing to its ubiquity, over the last few decades the common ash has been greatly favoured in areas abandoned by agriculture and also along impounded rivers. F. excelsior is a mesophilic species that usually thrives on alluvial soils and can also survive the strong water deficit on hillslopes. Drought tolerance in ash trees is related to stomatal regulation, a decrease in osmotic potential and also an increase in the elastic modulus ε. Malic acid plays a central role in drought tolerance, first as one of the major solutes involved in osmotic adjustment, and second as an effector for stomatal regulation through a drought-induced increase in its xylem concentration.

Osmotic adjustment in Fraxinus excelsior L . : malate and mannitol accumulation in leaves under drought conditions

Abstract In leaves of Fraxinus excelsior L., malate and mannitol were characterized by 13C NMR spectroscopy and enzymatic specific assays as the major constituents of a soluble carbon fraction involved in an osmotic adjustment. During a summer drought where predawn leaf water potential of adult trees growing in a mesoxerophilic stand fell to – 4 MPa in August, malate and mannitol leaf contents increased by a factor of 1.8 and 2.2 respectively, compared to control trees growing on a flood plain. This drought stress led to concentrations as high as 280 mM and 600 mM for mannitol and malate, respectively. The effects of gradually developing water deficit were also studied in a semi-controlled environment in 3-year-old seedlings. When predawn leaf water potential reached -6 MPa, leaves displayed a low turgor pressure but stomatal conductance was still measurable. Malate and mannitol were also the main osmoticum involved. After rewatering, gas exchange capacities were largely restored. Altogether, these results show that the strong water-stress tolerance of Fraxinus excelsior is in part related to an accumulation of malate and mannitol.

Larvicidal Effect of a Cell-Wall Fraction Isolated from Alder Decaying Leaves

Decaying alder leaves in water from Alpine Aedes breeding sites, particularly their toxicity to larval Culicidae, were investigated and characterized with comparative toxicological and chemical methods. Bioassays that used third-instar Aedes aegypti as a reference species indicated that the larvicidal effect of crude leaf litter varied with decaying age of the litter, while no toxicity was detected from leaching water of the mosquito breeding sites. Ten-month-old leaf litter was the most toxic. Comparison of the different soluble and insoluble fractions obtained after sequential extraction of decomposed litter allowed us to localize the toxicity factor to an insoluble cell-wall fraction. The toxicity seems to be linked to phenolic activity. It is higher than that found for tannic acid solutions used as a reference to mimic the larvicidal effects of the molecules naturally occurring in decaying litter. The pattern of establishing the larvicidal effect of alder leaf litter in water of Alpine Aedes breeding sites is discussed.

Role of vegetable tannins in habitat selection among mosquito communities from the Alpine hydrosystems.

The role of vegetable tannins in habitat selection among mosquito communities in Alpine hydrosystems was investigated through ecotoxicological comparison of 19 arthropod species characteristic of 12 breeding sites known for their abiotic environmental factors and their different riparian vegetation. The toxicity of tannins was experimentally compared among species representative of both the dipteran fauna and the crustacean fauna associated with the mosquito breeding sites. Bioassays using tannic acid solutions at concentrations from 0.1 to 11 mM separated the dipteran taxa into five groups of differential sensitivity and the crustacean taxa into four groups. The different levels of sensitivity among taxa were correlated with the various amounts of total phenolics and tannins found in the most prominent plant types associated with the different breeding sites. This suggested that tannins and, more generally, phenolic compounds may be involved in plant-arthropod interactions in Alpine hydrosystems.

Involvement of Ligninlike Compounds in Toxicity of Dietary Alder Leaf Litter Against Mosquito Larvae

The toxicological characteristics of dietary decomposed alder leaf litter against mosquito larvae were further investigated through enzymatic and chemical purification of a phenoliclike cell-wall fraction isolated from crude litter. The toxicity of the subfractions obtained was controlled by standard bioassays on third instars of Aedes aegypti chosen as a reference target species. Enzymatic hydrolyses of the cell-wall fraction were performed with caylase, pectolyase, esterase, and β-glycosidase, in order to release, respectively, cellulose material and phenolic compounds bound to lignins. These treatments did not affect the larvicidal activity and the phenolic activity of the cell-wall fraction. Chemical alkaline and acid hydrolyses were carried out to break ester and glycosidic bonds of the cell-wall fraction. Comparison of HPLC profiles of the hydrolysates from both toxic and nontoxic fractions did not reveal differences between the phenolic acids released. Aluminum chloride, known for its phenolic complexing activity, counteracted the larvicidal activity of the cell-wall fraction. Altogether, these results suggest the involvement of ligninlike compounds in the toxicity of dietary alder leaf litter against larval mosquitoes. The toxicity of this fraction, which was very sensitive to drastic and smooth oxidations, seemed to be associated with a strong oxidative potential. These results are discussed in relation to a possible mode of action of lignins in the plant–mosquito interactions.

Phenology, growth and ecophysiological characteristics of Fallopia sachalinensis

Fallopia (Polygonum) sachalinensis (Polygonaceae) is native to the Far East from where it was introduced into Europe and Northern America. It has only recently appeared on the large alluvial plains of alpine rivers, while it has formed monospecific stands in many habitats. In this study, we investi- gated some of the biological characteristics of this species in order to predict its invasiveness under the specific environmen- tal conditions of alpine alluvia. When compared to other plants which tend to monopolize space, F. sachalinensis appears to have a life cycle characterized by an early seasonal develop- ment. It exhibits a high growth rate - ca. 4 to 5 cm/day, and its productivity is associated with intrinsic characteristics of the species: abundant leaf cover, the appearance of branches when the leaves reach an optimal size on the main stem, and favourable leaf orientation to capture high light intensity. Another morphological characteristic is the presence of a large medullary air cell containing a liquid rich in mineral salts which can be used by the plant when the demand for evaporation increases. Clonal spread associated with an ex- traordinarily high rate of proliferation of below-ground or- gans, and mechanisms for adaptation to adverse conditions are biological traits rendering F. sachalinensis an invader in alluvial plains along alpine rivers. There is a risk that such species experience a demographic explosion in the geographi- cal region under study.

Comparative Capability to Detoxify Vegetable Allelochemicals by Larval Mosquitoes

In order to confirm the phytotoxicological basis for the ecological specialization of larval culicine fauna among different subalpine mosquito breeding sites, we compared the capability of six different Aedes larval taxa or populations of different ecological origin to detoxify dietary leaf litter originating from the environmental vegetation. Detoxification experiments were performed through in vitro digestion of a toxic leaf litter fraction using larval extracts as the enzymatic sources. Comparison of toxicological and detoxifying properties among the different larval samples indicates an association between their tolerance to leaf litter toxicants and their detoxification capability, which vary according to ecological origin. The fact that the detoxifying factor within the larval extracts appears to be a protein-like compound with a molecular weight bigger than 30 kDa suggests the possible involvement of detoxifying enzymes in larval tolerance to leaf litter toxicants. This is congruent with previous biochemical data that suggests the involvement of cytochrome P450 monooxygenase and esterase activities in the detoxification process.

Hot Extraction And Characterization of a Ligninlike Fraction Involved in Larvicidal Effects of Decomposed Leaf Litter against Mosquito

Hot water-extraction was performed on decomposed leaf litter in order to solubilize the toxic fraction involved in the dietary interaction against mosquito larvae in subalpine breeding sites. The toxic fraction was partially extracted by water with an optimum temperature of 60°C and recovered in an insoluble form. Phytochemical characterization was achieved through differential enzymatic hydrolyses, using the laccase mediator delignifying system, and aluminum chloride chelation monitored by standard bioassays; comparative spectrophotometric analyses in ultraviolet light after solubilization in acetyl bromide; and comparative reversed-phase high-performance liquid chromatography of the phenolic aldehydes after alkaline nitrobenzene oxidation. The results suggested the involvement of ligninlike compounds in the toxicity of the isolated fraction. Toxicity of this fraction appeared far stronger than that of the crude leaf litter. The involvement of this ligninlike fraction in the dietary toxicity of leaf litter against larval mosquito was then investigated.