Soil functioning in a mosaic of herbaceous communities of a chalky environment: temporal variations of water availability and N dynamics

Background and aims

Although changes in water and nitrogen (N) supply have been largely used to explain modifications in plant communities, the spatio-temporal variability of those factors has been little studied in chalky environments.

Methods

In this study, we explored for 1?year the temporal variations in soil water content, N inorganic forms and net N-mineralization and nitrification for two horizons in three herbaceous communities (short grasslands, tall grasslands, and encroached grasslands) in the Hénouville Nature Reserve (Upper-Normandy, France). Plant available soil water and permanent wilting points of seven plant species were also characterized.

Results

We found that plant available soil water was lower in short grasslands than in tall grasslands and encroached grasslands. Soil water content was below permanent wilting point during four months in short grasslands and only three months in the other communities. Seasonal patterns for inorganic N content and N-mineralization and nitrification were observed with peaks of NH ₄ ⁺ –N in summer and peaks of N-mineralization in spring.

Conclusions

For the studied year, our data highlight the harsh soil desiccation that vegetation endured during the late spring (active growth period) and summer, and show that water shortage is an ecological factor affecting the N cycling in the three successional herbaceous communities.     

Oxidases responsible for resistance to pyrethroids sensitize Helicoverpa armigera (Hübner) to triazophos in West Africa

Helicoverpa armigera (Hübner) is the major insect pest of cotton in Africa, Turkey, Asia, India, Indonesia and Australia. Populations recently developed resistance to pyrethroids in West Africa via the overproduction of cytochrome P450 (oxidases) increasing pyrethroid metabolism. One way to overcome pyrethroid resistance is to use compounds that show negative cross-resistance to pyrethroids. Triazophos is one of these compounds: it is slightly more toxic against pyrethroid resistant larvae of H. armigera than against susceptible ones. Overproduced oxidases transform the non active triazophos into its active form, triazophos-oxon, since this form was significantly more often found in larvae from pyrethroid resistant strain (23%) than in susceptible strain (15%). This suggests that oxidases, which provide resistance by degradation of pyrethroids in the resistant individuals, also activate triazophos in its toxic oxon form resulting in a negative cross-resistance.     

Metabolic correction of congenital erythropoietic porphyria with iPSCs free of reprogramming factors

Congenital erythropoietic porphyria (CEP) is due to a deficiency in the enzymatic activity of uroporphyrinogen III synthase (UROS); such a deficiency leads to porphyrin accumulation and results in skin lesions and hemolytic anemia. CEP is a candidate for retrolentivirus-mediated gene therapy, but recent reports of insertional leukemogenesis underscore the need for safer methods. The discovery of induced pluripotent stem cells (iPSCs) has opened up new horizons in gene therapy because it might overcome the difficulty of obtaining sufficient amounts of autologous hematopoietic stem cells for transplantation and the risk of genotoxicity. In this study, we isolated keratinocytes from a CEP-affected individual and generated iPSCs with two excisable lentiviral vectors. Gene correction of CEP-derived iPSCs was obtained by lentiviral transduction of a therapeutic vector containing UROS cDNA under the control of an erythroid-specific promoter shielded by insulators. One iPSC clone, free of reprogramming genes, was obtained with a single proviral integration of the therapeutic vector in a genomic safe region. Metabolic correction of erythroblasts derived from iPSC clones was demonstrated by the disappearance of fluorocytes. This study reports the feasibility of porphyria gene therapy with the use of iPSCs.     

A plant Shaker-like K+ channel switches between two distinct gating modes resulting in either inward-rectifying or "leak" current

Among the Shaker-like plant potassium channels, AKT2 is remarkable because it mediates both instantaneous "leak-like" and time-dependent hyperpolarisation-activated currents. This unique gating behaviour has been analysed in Xenopus oocytes and in COS and Chinese hamster ovary cells. Whole-cell and single-channel data show that (i) AKT2 channels display two distinct gating modes, (ii) the gating of a given AKT2 channel can change from one mode to the other and (iii) this conversion is under the control of post-translational factor(s). This behaviour is strongly reminiscent of that of the KCNK2 channel, recently reported to be controlled by its phosphorylation state.     

Utilization of mutants to analyze the interaction between Arabidopsis thaliana and its naturally root-associated Pseudomonas

 A model system based on the Arabidopsis thaliana (L.) Heynh. Ws ecotype and its naturally colonizing Pseudomonas thivervalensis rhizobacteria was defined. Pseudomonas strains colonizing A. thaliana were found to modify the root architecture either in vivo or in vitro. A gnotobiotic system using bacteria labelled with green fluorescent protein revealed that P. thivervalensis exhibited a colonization profile similar to that of other rhizobacterial species. Mutants of A.thaliana affected in root hair development and possible hormone perception were used to analyze the plant genetic determinants of bacterial colonization. A screen for mutants insensitive to P. thivervalensis colonization yielded two mutants found to be auxin resistant. This further supports a proposed role for bacterial auxin in inducing morphological modifications of roots. This work paves the way for studying the interaction between plants and non-pathogenic rhizobacteria in a gnotobiotic system, derived from a natural association, where interactions between both partners can be genetically dissected. Received: 6 January 2000 / Accepted: 20 May 2000     

Assembly of plant Shaker-like K(out) channels requires two distinct sites of the channel alpha-subunit

SKOR and GORK are outward-rectifying plant potassium channels from Arabidopsis thaliana. They belong to the Shaker superfamily of voltage-dependent K(+) channels. Channels of this class are composed of four alpha-subunits and subunit assembly is a prerequisite for channel function. In this study the assembly mechanism of SKOR was investigated using the yeast two-hybrid system and functional assays in Xenopus oocytes and in yeast. We demonstrate that SKOR and GORK physically interact and assemble into heteromeric K(out) channels. Deletion mutants and chimeric proteins generated from SKOR and the K(in) channel alpha-subunit KAT1 revealed that the cytoplasmic C-terminus of SKOR determines channel assembly. Two domains that are crucial for channel assembly were identified: i), a proximal interacting region comprising a putative cyclic nucleotide-binding domain together with 33 amino acids just upstream of this domain, and ii), a distal interacting region showing some resemblance to the K(T) domain of KAT1. Both regions contributed differently to channel assembly. Whereas the proximal interacting region was found to be active on its own, the distal interacting region required an intact proximal interacting region to be active. K(out) alpha-subunits did not assemble with K(in) alpha-subunits because of the absence of interaction between their assembly sites.     

Beticolins, nonpeptidic, polycyclic molecules produced by the phytopathogenic fungus Cercospora beticola, as a new family of ion channel-forming toxins

Beticolins are toxins produced by Cercospora beticola, a phytopathogenic fungus responsible for the leaf spot disease of sugar beet. They form a family of 20 nonpeptidic compounds (named B0 to B19) that share the same polycyclic skeleton but differ by isomeric configuration (ortho- or para-) and by a variable residue R (bridging two carbons in one of the six cycles). It has been previously shown that B0 assembles itself into a multimeric structure and forms ion channels into planar lipid bilayers (C. Goudet, A.-A. Very, M.-L. Milat, M. Ildefonse, J.-B. Thibaud, H. Sentenac, and J.-P. Blein, Plant J. 14:359-364, 1998). In the present work, we investigate pore formation by three ortho-beticolins, B0, B2, and B4, and their related (i.e., same R) para-isomers, B13, B1, and B3, respectively, using planar lipid bilayers. All beticolins were able to form ion channels with multiple conductance states, although the type of cyclization (ortho- or para-) and residue (R) result in variations of channel conductance and ionic permeability, respectively. Channel formation by beticolins is likely to be involved in the biological activity of these toxins.     

Organophosphorus insecticides synergize pyrethroids in the resistant strain of cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) from West Africa

Helicoverpa armigera (Hübner) populations from West Africa recently developed resistance to pyrethroid insecticides through enhanced metabolism by mixed-function oxidases. The combination index method was used to study the synergism of pyrethroids by organophosphorus insecticides. Several mixtures of insecticides currently registered to control cotton pest complex in West Africa were tested, including: cypermethrin/ethion, cypermethrin/profenofos, deltamethrin/ triazophos, deltamethrin/chlorpyriphos, cyfluthrin/chlorpyriphos, and betacyfluthrin/chlorpyriphos. In the resistant strain, the organophosphorus insecticides significantly increased the toxicity of pyrethroids suppressing the resistance effect, either by additive or synergistic effects. Significant synergism was shown for the following mixtures: cypermethrin/ethion, deltamethrin/triazophos, and deltamethrin/chlorpyriphos. The use of synergism from these insecticide mixtures should prove to be an additional tool in the overall resistance management strategy because the pyrethroid resistance in H. armigera from West Africa is not yet stable, decreasing between cotton seasons and increasing with treatments. In absence of selection, the susceptibility of H. armigera to insecticides should be restored.     

H Cotransports in Corn Roots as Related to the Surface pH Shift Induced by Active H Excretion

The surface pH shift induced by active H⁺ excretion in corn (Zea mays L.) roots was estimated using acetic acid influx as a pH probe (H Sentenac, C Grignon 1987 Plant Physiol 84: 1367-1372). At constant bulk pH, buffering the medium strongly reduced the magnitude of the surface pH shift. This was used to study the effect of surface pH shift on H⁺ cotransports. In the absence of buffers, the surface pH shift increased with the bulk pH. Buffers decreased ³²Pi influx and this effect was stronger at pH 7.2 than at pH 5.8, and stronger in the absence than in the presence of an inhibitor of the proton pump (vanadate). Buffers exerted a similar depressive and pH-dependent effect on net NO₃⁻ uptake. They hyperpolarized the cell membrane, and stimulated ⁸⁶Rb⁺ influx, K⁺:H⁺ net exchange, and malate accumulation. These results are consistent with the hypothesis that H⁺ accumulation at the cell surface is effective in driving H⁺ reentry. We concluded that the surface pH shift due to proton pump activity is involved in the energetic coupling of H⁺ cotransports.