Improved RP‐HPLC and anion‐exchange chromatography methods for the determination of amino acids and carbohydrates in soil solutions

In spite of their low concentrations in soil solutions, low–molecular weight organic substances (LMWOS) such as amino acids, sugars, and uronic acids play a major role in the cycles of C and N in soil. With respect to their low concentrations and to possible matrix interferences, their analysis in soil leachates is a challenging task. We established two HPLC (high‐performance liquid chromatography) methods for the parallel determination of amino acids and carbohydrates in soil leachates. The pre‐column derivatization of amino acids with an o‐phthaldialdehyde (OPA) mercaptoethanol solution yields quantitation limits between 0.03 and 0.44 µmol L^–1 and S_D values of <8.3% (n = 9). High‐performance anion‐exchange chromatography (HPAEC) on a Dionex CarboPac PA 20 column with a NaOH acetate gradient combined with pulsed amperometric detection (PAD) was used for the determination of carbohydrates. The calibration curves obtained for 11 carbohydrates showed excellent linearity over the concentration range from 0.02 to 50.0 mg L^–1. Recovery studies revealed good results for all analytes (89%–108%). Interferences from Hg(II) salts and chloroform used for stabilization of the leachates did not occur with both chromatographic methods. The optimized method was successfully used for quantitative determinations of amino acids and carbohydrates in soil leachates.     

Learning performances of honeybees (Apis mellifera L) are differentially affected by imidacloprid according to the season

To establish the sublethal concentrations domain, acute and chronic oral tests were conducted on caged honeybee workers (Apis mellifera L) using imidacloprid and a metabolite, 5-OH-imidacloprid, under laboratory conditions. The latter showed a 48-h oral LD50 value (153 ng per bee) five times higher than that of imidacloprid (30 ng per bee). Chronic feeding tests indicated that the lowest observed effect concentrations (LOEC) of imidacloprid and of 5-OH-imidacloprid on mortality of winter bees were 24 and 120 microg kg(-1) respectively. Behavioural effects of imidacloprid and 5-OH-imidacloprid were studied using the olfactory conditioning of proboscis extension response at two periods of the year. Winter bees surviving chronic treatment with imidacloprid and 5-OH-imidacloprid had reduced learning performances. The LOEC of imidacloprid was lower in summer bees (12 microg kg(-1)) than in winter bees (48 microg kg(-1)), which points to a greater sensitivity of honeybees behaviour in summer bees, compared to winter bees.     

Pyrethroid insecticides derived from 2,2-dimethyl-3-(3-phenoxybenzyloxy)propanal

Fourteen oxime ethers and eleven other unsaturated compounds derived from 2,2-dimethyl-3-(3-phenoxybenzyloxy)propanal were synthesised and their insecticidal activity against Musca domestica, Locusta migratoria, Dysdercus cingulatus, Aedes aegypti, Plutella xylostella, Drosophila melanogaster, Tetranychus urticae and Spodoptera littoralis tested. Structural variations including chain length, branching and unsaturation of the oxime ether O-alkyl group and substituent exchange at the carbon-carbon double bond of the unsaturated compounds were made in order to examine the structure-activity relationship. All biological activities were compared with the activity of the ether pyrethroid MTI-500.     

Patterns of SSR variation in bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) seeds under ex situ genebank storage and accelerated ageing

Maintaining seed viability and germplasm integrity is a challenging task in conservation of plant genetic resources, as seeds under storage will lose viability and genetic changes will occur. Attempt was made to analyze the patterns of genetic changes in wheat germplasm under ex situ genebank storage and accelerated ageing treatments. A set of 16 naturally aged wheat accessions under ex situ genebank storage since 1994 were sampled. Four recently regenerated wheat accessions were selected, four random seed samples were chosen from each accession, and three of them were exposed to three different accelerated ageing treatments. These 32 seed samples in two germplasm sets displayed a range of germination rates from 4 to 98 %. Thirty-seven microsatellite markers representing 21 wheat chromosomes were applied to screen 12 seeds of each sample and 449 SSR alleles were scored. Large SSR variation was found in each germplasm set. There was 73.1 % of the total SSR variation present among the naturally aged samples and 78.2 % present among the accelerated ageing samples. Several analyses for genetic association consistently revealed no clear genetic separations among samples of high or low germination rates in both germplasm sets. Samples under different accelerated ageing treatments did not show much genetic differentiations from the original sample of each accession. Mantel tests revealed non-significant associations between SSR variability and sample germination rates for both germplasm sets. These findings are useful for understanding seed deterioration under different ageing conditions and suggest that genome-wide SSR variability may not provide sensitive markers for the monitoring of wheat seed viability.     

Induction of resistance against downy mildew pathogen in pearl millet by a synthetic jasmonate analogon

The synthetic 1-oxo-indanoyl-l-isoleucine methyl ester (Ind-Ile-Me) represents a highly active mimic of octadecanoic phytohormones, which are involved in plant defenses against pathogens and pests. Seed treatments and foliar spray with Ind-Ile-Me were tested for induced resistance against downy mildew disease caused by the phytopathogenic oomycete Sclerospora graminicola in pearl millet (Pennisetum glaucum) under greenhouse and field conditions. Under greenhouse conditions, a 50% protection level was achieved after seed treatment. Seed treatment in combination with foliar spray resulted in 60% protection. The induction of resistance was correlated with the enhanced activities of defense-related proteins such as phenylalanine-ammonia-lyase, peroxidase, and enhanced level of hydroxyproline-rich glycoproteins. Under field conditions, a maximum protection of 62% was recorded upon seed treatment along with foliar spray. Hence, it infers that Ind-Ile-Me can be used as a valuable protection compound at least in downy mildew disease management.     

Facultative and obligate diapause phenotypes in populations of the European spruce bark beetle Ips typographus

The bark beetle Ips typographus is the most destructive insect pest in Norway spruce-dominated forests. Its potential to establish multiple generations per year (multivoltinism) is one major trait that makes this beetle a severe pest. Ips typographus enters diapause to adjust its life cycle to seasonally changing environments. Diapause is characterized by developmental and reproductive arrest; it prolongs generation time and thus affects voltinism. In I. typographus a facultative, photoperiod-regulated diapause in the adult stage has been described. In addition, the presence of an obligate, photoperiod-independent, diapause has been hypothesized. The diapause phenotype has important implications for I. typographus voltinism, as populations with obligate diapausing individuals would be univoltine. To test for the presence of different I. typographus diapause phenotypes, we exposed Central and Northern European individuals to a set of photoperiodic treatments. We used two ovarian traits (egg number and vitellarium size) that are associated with gonad development, to infer reproductive arrest and thus diapause. We found a distinct effect of photoperiod on ovarian development, with variable responses in Central and Northern European beetles. We observed obligate diapausing (independent of photoperiod) individuals in Northern Europe, and both facultative (photoperiod-regulated) as well as obligate diapausing individuals in Central Europe. Our results show within-species variation for diapause induction, an adaptation to match life cycles with seasonally fluctuating environmental conditions. As the diapause phenotype affects the potential number of generations per season, our data are the basis for assessing the risk of outbreaks of this destructive bark beetle.

     

An alternative method to estimate zero flow temperature differences for Granier's thermal dissipation technique

Calibration of the Granier thermal dissipation technique for measuring stem sap flow in trees requires determination of the temperature difference (DeltaT) between a heated and an unheated probe when sap flow is zero (DeltaT(max)). Classically, DeltaT(max) has been estimated from the maximum predawn DeltaT, assuming that sap flow is negligible at nighttime. However, because sap flow may continue during the night, the maximum predawn DeltaT value may underestimate the true DeltaT(max). No alternative method has yet been proposed to estimate DeltaT(max) when sap flow is non-zero at night. A sensitivity analysis is presented showing that errors in DeltaT(max) may amplify through sap flux density computations in Granier's approach, such that small amounts of undetected nighttime sap flow may lead to large diurnal sap flux density errors, hence the need for a correct estimate of DeltaT(max). By rearranging Granier's original formula, an optimization method to compute DeltaT(max) from simultaneous measurements of diurnal DeltaT and micrometeorological variables, without assuming that sap flow is negligible at night, is presented. Some illustrative examples are shown for sap flow measurements carried out on individuals of Erica arborea L., which has needle-like leaves, and Myrica faya Ait., a broadleaf species. We show that, although DeltaT(max) values obtained by the proposed method may be similar in some instances to the DeltaT(max) predicted at night, in general the values differ. The procedure presented has the potential of being applied not only to Granier's method, but to other heat-based sap flow systems that require a zero flow calibration, such as the Cermák et al. (1973) heat balance method and the T-max heat pulse system of Green et al. (2003).     

Drivers of bryophyte diversity allow implications for forest management with a focus on climate change

Understanding the major drivers of diversity in forests is crucial for ecologists, conservationists, and particularly forest managers. In most forested habitats, bryophytes are diverse and important primary producers. Here we report the first study of the relative importance of regional macroclimate as compared to local variables on abundance, species richness, and community composition of bryophytes growing in soil and in dead wood in a mountain temperate forest. Using PCA axes, we built predictor sets for macroclimate, microclimate, soil attributes, and dead wood availability. The explanatory power of each set was tested using variance partitioning. Abundance and species richness of soil bryophytes was best explained by microclimate, whereas the community composition did not distinctively differ between the environmental sets. In contrast, dead wood bryophyte abundance, species richness, and community composition was clearly driven by macroclimate. Among the single axes, the component represented best by soil moisture was the main driver for soil bryophyte abundance. In contrast, dead wood bryophyte abundance was mostly affected by components correlated with minimum global radiation and minimum temperatures as well as by the component represented by dead wood. The component represented by canopy openness was superior in explaining the community composition of both soil and dead wood bryophytes. We conclude that (1) dead wood amounts should be increased in closed stands to act as a buffer during climate changes, and (2) open canopy, which provides important habitats for soil-inhabiting bryophytes, should be provided by management with slow reforestation after natural or logging openings.