Silicon acquisition by bananas (c.V. Grande Naine) is increased in presence of the arbuscular mycorrhizal fungus <Emphasis Type="Italic">Rhizophagus irregularis</Emphasis> MUCL 41833

Aim

This work aimed to investigate the role of arbuscular mycorrhizal fungi (AMF) in the uptake and accumulation of silicon (Si) in banana plants. Si is recognized as a significant element that helps plants resist stresses.

Methods

A pot experiment compared the growth, Si and P accumulation of banana plants pre-colonized or not by an AMF and exposed or not to Si added to the growth substrate.

Results

A marked increase in Si was noticed in pseudostem, leaves and roots of pre-colonized banana plants, in presence as well as in absence of Si added to the growth substrate. Without Si addition, this accumulation was 60 % and 45 % higher in pseudostem and leaves, respectively, while it was 47 % and 41 % in presence of Si added to the substrate. In roots, this increase was 23 % and 52 % in presence and absence of Si added to the substrate, respectively. Phosphorus content in shoots and roots was likewise significantly increased in presence of AMF or Si.

Conclusion

Our findings revealed that pre-colonized banana plants accumulated more Si in shoot and roots than non-mycorrhizal plants and may thus represent a potential novel avenue to explore banana resistance to pests and diseases.

     

<Emphasis Type="Italic">Rhizophagus irregularis</Emphasis> MUCL 41833 can colonize and improve P uptake of <Emphasis Type="Italic">Plantago lanceolata</Emphasis> after exposure to ionizing gamma radiation in root organ culture

Long-lived radionuclides such as ⁹⁰Sr and ¹³⁷Cs can be naturally or accidentally deposited in the upper soil layers where they emit β/γ radiation. Previous studies have shown that arbuscular mycorrhizal fungi (AMF) can accumulate and transfer radionuclides from soil to plant, but there have been no studies on the direct impact of ionizing radiation on AMF. In this study, root organ cultures of the AMF Rhizophagus irregularis MUCL 41833 were exposed to 15.37, 30.35, and 113.03 Gy gamma radiation from a ¹³⁷Cs source. Exposed spores were subsequently inoculated to Plantago lanceolata seedlings in pots, and root colonization and P uptake evaluated. P. lanceolata seedlings inoculated with non-irradiated AMF spores or with spores irradiated with up to 30.35 Gy gamma radiation had similar levels of root colonization. Spores irradiated with 113.03 Gy gamma radiation failed to colonize P. lanceolata roots. P content of plants inoculated with non-irradiated spores or of plants inoculated with spores irradiated with up to 30.35 Gy gamma radiation was higher than in non-mycorrhizal plants or plants inoculated with spores irradiated with 113.03 Gy gamma radiation. These results demonstrate that spores of R. irregularis MUCL 41833 are tolerant to chronic ionizing radiation at high doses.     

Mycorrhizal dependency of banana (Musa acuminata,AAA group) cultivar

Seven banana cultivars (Musa acuminata, AAA group) were inoculated with two species of vesicular arbuscular mycorrhizal (VAM) fungi (Glomus mosseae and Glomus macrocarpum) in a greenhouse experiment. Inoculated plants had generally greater shoot dry weight and shoot phosphorus concentrations compared to the noninoculated plants. A great variation in dependency on mycorrhizal colonization was observed among the banana cultivars. Cv. Williams showed the highest relative mycorrhizal dependency (RMD) and cv. Poyo the lowest. For all the cultivars studied, inoculation with G. macrocarpum resulted in the highest RMD values. Both root dry weight and root hair length or density of the noninoculated plants were inverserly correlated with the RMD values of cultivars.     

Purification and characterization of natural and recombinant human plasminogen activator inhibitor-1 (PAI-1)

Human plasminogen activator inhibitor-1 (PAI-1) was purified from the conditioned medium of endotoxin-stimulated umbilical vein endothelial cell cultures by combinations of zinc-chelate-Sepharose chromatography, gel filtration on Sephacryl S-300 and immunoadsorption on an insolubilized murine monoclonal antibody (MA-7D4). The final product was obtained with a recovery of approximately 20% from conditioned medium containing about 3 micrograms/ml PAI-1. The yield of PAI-1 was 15-100 micrograms/umbilical cord, depending on the culture and harvest conditions. SDS gel electrophoresis revealed a main band with Mr = 46,000 both under reducing and non-reducing conditions. On gel filtration on Sephacryl S-300, however, the material was separated in two fractions, one eluting at the void volume, which contains active PAI-1, and one with Mr = 46,000 containing inactive material that could be reactivated with 12 M urea. SDS gel electrophoresis of the isolated high-Mr fraction revealed several bands including a main 46,000-Mr component, which reacted with anti-(PAI-1) antibodies on immunoblotting and neutralized tissue-type plasminogen activator (t-PA). The active high-Mr fraction and the reactivated low-Mr fraction of PAI-1 inhibited t-PA very rapidly with an apparent second-order rate constant of (1.5-4) x 10(7) M-1 s-1. The cDNA of endothelial cell PAI-1 was cloned and expressed in Chinese hamster ovary cells. The translation product, purified from conditioned medium of transfected cells, also revealed a high-Mr and a low-Mr fraction on gel filtration, which were indistinguishable from the natural proteins by physicochemical, immunochemical and functional analysis. On reduced SDS gel electrophoresis, the high-Mr fraction was separated into the Mr-46,000 low-Mr PAI-1 and two other components with Mr 65,000 and one barely entering the gel. When reactivated low-Mr PAI-1 was added to plasma, PAI activity and PAI-1 antigen eluted with an apparent Mr greater than or equal to 300,000 on gel filtration, indicating that active PAI-1 complexes with one or more binding proteins in plasma.     

Effect of land use on pollution status and risk of fish endocrine disruption in small farmland ponds

To study whether the intensity of agricultural activities affects pesticides loads in pond environment, a large number of Belgian farmland ponds were surveyed in spring 2004. Temporal distribution of pollutants was also investigated over restricted survey ponds sampled three times round year 2007. Sedentary pond Prussian carp juveniles were also captured to determine their brain aromatase activity (AA) and plasma vitellogenin (VTG) levels. Heavy metal distribution was also examined in various pond matrices. Amongst the pesticides analysed, only herbicides were detected. Contamination of pond water by atrazine was frequently observed during spring 2004, while isoproturon and glyphosate were detected round year 2007. Levels of herbicides were inversely related to the distance of ponds to crop field, and values peaked in April or October. Absence of endocrine disruptors in pond water was confirmed by lack of modulation in VTG and AA in male fish. Heavy metals were present in all the pond matrices, but overall contamination levels were low. The results demonstrated that Belgian ponds were mainly contaminated by herbicides and that pond sedentary fish were not affected by endocrine disruptors. They also demonstrated a marked effect of land-use intensity on herbicide pollution which can be mitigated by an adjustment of the buffer zones.     

Evaluation of Restoration Measures in a Shallow Lake through a Comparison of Present Day Zooplankton Communities with Historical Samples

Many shallow lakes have lost a large part of their ecological value during the past decades. Human-induced factors such as eutrophication and inappropriate fish stock management are generally the main causes for this loss. To restore such degraded habitats, several measures are taken, typically involving a reduction of nutrient loading and interventions in the aquatic food web functioning (biomanipulation). In this study, we report on the joint effects of a series of restoration measures in a shallow lake (Lake Kraenepoel, Belgium, 22 ha) and evaluate these effects via three different criteria. The first criterion is that the target condition, being a clearwater phase with submerged macrophytes, was successfully achieved and persisted for a period of at least 5 years after restoration. Second, we detected a substantial change in community structure of cladoceran zooplankton and an associated increase in species richness and conservation value following restoration measures. Finally, we observed that the general structure of the present day cladoceran zooplankton community resembles well that of the preeutrophication period (1929–1931). Current species richness, however, tends to be lower than in the reference period, and some rare species are still lacking. It is conceivable that, when submerged macrophytes develop further, a subset of specialist species may reappear. Overall, the use of historical habitat-specific samples offers a major opportunity for evaluating restoration success in great detail. Community structures may directly be compared, the gain or loss of specific species can accurately be documented, and more insights in the observed patterns be obtained.     

Generation of a stable activated thrombin activable fibrinolysis inhibitor variant

Activated thrombin activable fibrinolysis inhibitor (TAFIa), generated upon activation of TAFI, exerts an antifibrinolytic effect. TAFIa is a thermolabile enzyme, inactivated through a conformational change. The objective of the current study was to generate a stable variant of human TAFIa. Using a site-directed as well as a random mutagenesis approach to generate a library of TAFI mutants, we identified two mutations that increase TAFIa stability, i.e. a Ser305 to Cys and a Thr329 to Ile mutation, respectively. Combining these mutations in TAFI-Ala147-Ile325, the most stable isoform of TAFIa (half-life of 9.4 +/- 0.4 min), revealed a TAFIa half-life of 70 +/- 3.1 min (i.e. an 11-fold increase versus 6.3 +/- 0.3 min for TAFIa-Ala147-Thr325, the most frequently occurring isoform of TAFI in humans) at 37 degrees C. Moreover, clot lysis (induced by tissue plasminogen activator) experiments in which TAFI-Ala147-Cys305-Ile325-Ile329 was added to TAFI-depleted plasma revealed a 50% clot lysis time of 313 +/- 77 min (i.e. a 3.0-fold increase versus 117 +/- 10 min for TAFI-Ala147-Thr325). The availability of a more stable TAFIa variant will facilitate the search for inhibitors and allow further structural analysis to elucidate the mechanisms of the instability of TAFIa.     

Evidence for a pre-latent form of the serpin plasminogen activator inhibitor-1 with a detached beta-strand 1C

Latency transition of plasminogen activator inhibitor-1 (PAI-1) occurs spontaneously in the absence of proteases and results in stabilization of the molecule through insertion of its reactive center loop (RCL) as a strand in beta-sheet A and detachment of beta-strand 1C (s1C) at the C-terminal hinge of the RCL. This is one of the largest structural rearrangements known for a folded protein domain without a concomitant change in covalent structure. Yet, the sequence of conformational changes during latency transition remains largely unknown. We have now mapped the epitope for the monoclonal antibody H4B3 to the cleft revealed upon s1C detachment and shown that H4B3 inactivates recombinant PAI-1 in a time-dependent manner. With fluorescence spectroscopy, we show that insertion of the RCL is accelerated in the presence of H4B3, demonstrating that the loss of activity is the result of latency transition. Considering that the epitope for H4B3 appears to be occluded by s1C in active PAI-1, this finding suggests the existence of a pre-latent conformation on the path from active to latent PAI-1 characterized by at least partial detachment of s1C. Functional characterization of mutated PAI-1 variants suggests that a salt-bridge between Arg273 and Asp224 may stabilize the pre-latent conformation. The binding of H4B3 and of a peptide targeting the cleft revealed upon s1C detachment was hindered by the glycans attached to Asn267. Conclusively, we have provided evidence for the existence of an equilibrium between active PAI-1 and a pre-latent form, characterized by reversible detachment of s1C and formation of a glycan-shielded cleft in the molecule.     

Hyphosphere interactions between Rhizophagus irregularis and Rahnella aquatilis promote carbon–phosphorus exchange at the peri-arbuscular space in Medicago truncatula

Arbuscular mycorrhizal (AM) fungi form a continuum between roots and soil. One end of this continuum is comprised of the highly intimate plant–fungus interface with intracellular organelles for nutrient exchange, while on the other end the fungus interacts with bacteria to compensate for the AM fungus' inability to take up organic nutrients from soil. How both interfaces communicate in this highly complex tripartite mutualism is widely unknown. Here, the effects of phosphate-solubilizing bacteria (PSB) Rahnella aquatilis dwelling at the surface of the extraradical hyphae of Rhizophagus irregularis was analysed based on the expression of genes involved in C-P exchange at the peri-arbuscular space (PAS) in Medicago truncatula. The interaction between AM fungus and PSB resulted in an increase in uptake and transport of Pi along the extraradical hyphae and its transfer from AM fungus to plant. In return, this was remunerated by a transfer of C from plant to AM fungus, improving the C-P exchange at the PAS. These results demonstrated that a microorganism (i.e., a PSB) developing at the hyphosphere interface can affect the C-P exchange at the PAS between plant and AM fungus, suggesting a fine-tuned communication operated between three organisms via two distantly connected interfaces.