Lettuce (Lactuca sativa): a species with a high capacity for cadmium (Cd) accumulation and growth stimulation in the presence of low Cd concentrations

Summary

Cadmium (Cd) is a metal pollutant that accumulates in cultivated soils and has detrimental consequences in terms of food safety. Lettuce (Lactuca sativa) can be characterised as having a high capacity to accumulate Cd in its tissues. An analysis of Cd tolerance and Cd accumulation was carried out using two varieties of lettuce (‘Divina’ and ‘Melina’). A wide range of CdCl₂ concentrations was used (0.0, 0.1, 0.6, 3.0, and 15.0 µM CdCl₂). The lowest concentration (0.1 µM CdCl₂) stimulated growth, while the two highest concentrations resulted in a reduction in biomass. Cadmium concentrations were found to be twice as high in roots as in shoots. ‘Divina’ displayed lower concentrations of Cd than ‘Melina’ in nearly all treatments. A strong negative correlation was observed between Cd concentration and Cd tolerance in the roots and shoots (R² > 0.87) of both ‘Melina’ and ‘Divina’. Lettuce grown in the presence of 15.0 µM CdCl₂ had leaf Cd concentrations that were 100-fold higher than the legal maximum level for vegetable products marketed for human consumption, but showed no symptoms of dehydration, chlorosis, or necrosis. This result represents an important alert for lettuce consumers and growers.     

Activity and metabolism of cyano-oxime derivatives in various strains of Botrytis cinerea

The fungitoxicity of cymoxanil, 4 cyano-oxime analogs (in which the acetylurea group was replaced with various groups: amide, ester, propargyl, and cyanomethyl-amide), and 2 cymoxanil-metabolites was studied against various strains of Botrytis cinerea. The fungicidal effect was measured on germ-tube elongation and mycelial growth. Cymoxanil and the analogs bearing the amide and ester groups showed the best anti-botrytis activity. The strains studied can be classified into 3 groups according to the germ-tube sensitivity to cyano-oximes. These groups fitted well with the 3 phenotypes of sensitivity to cymoxanil previously characterized in our laboratory: CyaS₁ (highly sensitive phenotype); CyaS₂ (moderately sensitive phenotype); and CyaR (tolerant phenotype). The bio-transformations of all the cyano-oximes were monitored in the culture-medium of the different strains using HPLC- and IP-HPTLC-methods. HPLC-studies showed that cymoxanil and the analogs bearing the groups amide and ester were quickly metabolized in the culture-medium of the CyaS₁ strain. Moreover, these studies allowed us to correlate disappearance of these cyano-oximes, to their fungicidal activity towards the CyaS₁ strain. This suggests that cyano-oximes and particularly cymoxanil are probably activated in a fungitoxic compound. IP-HPTLC-studies strengthened the precedent results and allowed us to correlate disappearance of cyano-oximes studied with appearance of two acids metabolites of hydrolysis. These metabolites could be the actual active-principles.     

Bark-Beetle Parasitoids Population Surveys Following Storm Damage in Spruce Stands in the Vosges Region (France)

This paper presents the results of a long-term study designed to follow the population dynamics of hymenopteran parasitoids associated with Ips typographus L. (Coleoptera: Scolytidae) in northeastern France (Alsace and Vosges) in spruce stands devastated by the December 1999 storms. Population densities were estimated by periodic samplings in the pre-emerging insect populations developing under the bark of attacked trees. Data were collected between July 2000 and October 2001. I. typographus population density (per m²) increased respectively by 28%, 105%, and 212% in one year in the three sites surveyed. Six hymenopteran ectoparasitoids were found: Coeloides bostrichorum Giraud and Dendrosoter middendorffi Ratzeburg (Hymenoptera: Braconidae); Rhopalicus tutela (Walker), Roptrocerus xylophagorum (Ratzeburg), Roptrocerus mirus (Walker) and Dinotiscus eupterus (Walker) (Hymenoptera: Pteromalidae). After two years, the parasitoid populations increased in terms of average density but parasitism rates remained at a relatively low level, ranging from 0% to 40%, with 60% of the trees having a parasitism rate below 5%. Data outlined the recurrent coexistence of competing parasitoid species.     

Study of the relation between carbofuran degradation and microbial or physicochemical characteristics of some french soils

In France, corn rootworm (Agriotes sp.) is controlled by applying insecticides such as carbofuran to the soil. In recent years, the failure of carbofuran to control this pest has been observed in the south-west area of France in continuous corn cropping. A soil survey was conducted in order to determine the main factors which could influence carbofuran degradation in various soils. Degradation estimates were based on measurement of the release of [ 14 C]carbon dioxide from [carbonyl- 14 C]carbofuran. The enhanced degradation of carbofuran observed in laboratory conditions corresponded to the growth of micro-organisms able to use carbofuran as sole carbon and nitrogen source. The size of this microbial population and the rates of carbofuran degradation were higher in the samples taken from plots with a history of carbofuran use than in samples from previously untreated plots. Abiological degradation was only observed in alkaline soil conditions. Statistical analyses showed that carbofuran-degrading activity was not related to any particular pedological characteristics.     

Virulent and attenuated lines of Leishmania major: DNA karyotypes and differences in metalloproteinase GP63

The Leishmania metalloproteinase GP63 has been reported to play important roles mainly in resistance of promastigotes to complement-mediated lysis and in interaction with macrophage receptors. On the other hand, its function in insect vectors is still unclear. We compared the structure and dosage of gp63 genes and the activity of GP63 in Leishmania major Yakimoff et Schokhor strains and lines differing in virulence for mice and ability to develop in sand flies. The results demonstrate considerable variability in amount and proteolytical activity of GP63 among L. major strains although genomic changes in the gp63 locus were not found. Attenuated LV561/AV line showed low amount and low enzymatic activity of GP63. Serial passages of attenuated parasites through either Phlebotomus duboscqi Neveu-Lemaire or through mice led to a recovery of GP63 proteolytical activity to the level present in virulent LV561/V line. Overexpression of GP63 was found in two L major strains (L119, Neal) with defective lipophosphoglycan (LPG); both these strains were capable to cause mice infection but unable to survive and multiply in sand flies. Differences were found also in karyotypes and in amount of minichromosomes amplified in some lines of the LV561 strain. The results suggest that parasite virulence is not simply correlated with the activity of GP63; however, this enzyme plays a significant role in association with other surface molecules, especially LPG. Overexpression of GP63 can compensate LPG defect in the vertebrate host but in sand flies both molecules fulfil quite different functions and the defect in LPG is lethal for the parasite. On the other hand, linear minichromosomes of about 200 kb found in some lines of the LV561 strain are associated with development in vitro and in the vector but they are not essential for the infection of the vertebrate host.     

Impact of poplar water status on leaf-beetle (Chrysomela populi) survival and feeding

? The aim of this study is to assess the impact of drought on insect-tree relationships. Survival and feeding performances of leaf-beetles, Chrysomela populi put in cages with leaves of host-plants submitted to different levels of water stress were compared.

? Ninety 1-year-old poplar (Populus × euramericana) cuttings were grown in a greenhouse, and distributed into 3 groups submitted to different water treatments during six weeks. Adult C. populi were then encaged for four days with these cuttings, together with the five terminal leaves of a twig. Beetle survival and consumed leaf areas were then determined.

? Predawn leaf water potential values indicated that the drought treatments induced different levels of plant water stress. Plant height growth and total leaf numbers were reduced by plant stress. Beetle survival and leaf areas eaten were also reduced proportionally to plant water deficit.

? In conclusion, this experiment suggests that drought has a negative impact on C. populi survival and feeding.

     

Interaction of Root-knot Nematodes (RKN) and the Bacterium Agrobacterium Tumefaciens in Roots of Prunus Cerasifera: Evidence of the Protective Effect of the Ma RKN Resistance Genes Against Expression of Crown Gall Symptoms

Agrobacterium tumefaciens (AT) is the causal agent of crown gall, a major problem in the family Rosaceae and particularly for Prunus spp. Crown gall symptoms result from the bacterial infection of the cells damaged mechanically at the collar or by root parasitic nematodes. Myrobalan plum (P. cerasifera) is susceptible to AT and is not a host for the root-knot nematode (RKN), M. hapla. Some clones of this plum carry single Ma resistance genes that control M. arenaria, M. incognita and M. javanica. The four above mentioned RKN and Myrobalan progenies segregating for Ma were used in experiments aimed at obtaining a better knowledge of the interaction between AT and RKN in relation to the RKN resistance genes. Prunus rooted cuttings, naturally infected with the bacterium were repotted, grown and inoculated individually with RKN. In a first experiment, Prunus plants were (i) either inoculated with 10,000 juveniles (J2s) of M. arenaria to provide a short inoculum pressure (SIP) or (ii) inoculated by association with one M. arenaria-galled tomato root system that produced a high and durable inoculum pressure of the same nematode species. Four months after RKN inoculation, plants were rated for nematode and bacterial root galling symptoms. RKN and AT galls were more numerous and more homogenous under DIP than under SIP. Nevertheless, for both inoculum regimes, AT galls were present in the RKN-susceptible clones (= carrying none of the Ma genes) and absent in the RKN-resistant clones. Subsequent experiments, conducted under DIP with M. arenaria, M. incognita, M. javanica and M. hapla, also showed, for the three first species, the presence of AT galls only in RKN-susceptible clones whereas Prunus plants inoculated with M. hapla and nematode-free controls were free of AT galls. Consequently RKN act as a wound agent in the AT infection process of Myrobalan plum only when the plant develops a compatible reaction (i.e. when it lacks the Ma resistance genes). Considering that J2s do penetrate the roots of resistant plants, the absence of crown gall symptoms on this material even under durable inoculum pressure strengthens the hypothesis that this nematode stage has a very weak effect on plant cells during the infection process. This is the first evidence of the protective effect of a RKN resistance gene against the expression of root crown gall consecutive to RKN infection. The protective effect of Ma and presumably of other RKN resistance genes against AT is a strong argument for their introgression into Prunus and other Rosaceae or bacterium-susceptible crops.     

In vitro pre-exposure of Haemonchus contortus L3 to blood eosinophils reduces their establishment potential in sheep

Different authors have reported that eosinophils are capable of immobilising infective larvae of different species of nematodes in vitro. However, classifying larvae as mobile or immobile is so subjective that it does not always mean all apparently immobile larvae are dead or those that are mobile are capable of surviving further immune responses if administered to their natural hosts. The objective of this experimental study was therefore to substantiate the role of eosinophils in the killing of Haemonchus contortus infective larvae by comparing the infectivity in sheep of larvae that had been incubated with eosinophil-enriched cell suspensions with control larvae. Since it was not possible to isolate pure eosinophils from sheep blood, we were compelled to evaluate the effects of other blood cells contaminating our eosinophil-enriched suspensions. Although eosinophils and neutrophils were the only cells found adherent to H. contortus infective larvae in vitro, induced eosinophils in the presence of immune serum were primarily responsible for the drastic reduction in larval motility compared to the minor effects of neutrophils and mononuclear cells. Corresponding reductions in faecal egg count and worm numbers were observed when the incubated larvae were transferred intra-abomasally to sheep. Interestingly, the proportion of larvae that failed to establish was much higher following incubation with induced eosinophils compared with other cells or with immune serum alone. Although this study did not address the in vivo role of eosinophils in sheep, the results strongly indicate that sheep blood eosinophils have a larval killing potential in vitro, and a larval mobility test alone may not fully explain the level of damage inflicted on the larvae.