Andrea Y. Mansilla

Trichoderma spp. as elicitors of wheat plant defense responses against Septoria tritici

Abstract Leaf blotch of wheat is a widespread and highly active disease that affects wheat production. In addition to the use of chemicals and proper cultivation methods, microbial antagonists are used to control plant pathogens. Trichoderma spp. stimulate a systemic induced response in plants. Therefore, the efficacy of Trichoderma spp. against wheat leaf blotch was evaluated under greenhouse conditions. The susceptible plants were sprayed with Septoria tritici conidiospores. In order to select an efficient method of pretreatment with Trichoderma spp., leaf spraying and seed coating with 14 isolates were tested in 2003 and 2004. The extent of leaf necrosis area and pycnidial coverage was estimated. Antagonism was assessed by the capacity of each Trichoderma spp. isolate to restrict the progress of leaf blotch, 21 days after inoculation. Of the two methods, seed coating was more efficacious against leaf blotch than leaf spraying. Amongst the 14 isolates tested, the isolate prepared from T. harzianum (Th5) produced the highest level of protection. None of the treatments caused changes in plant stem diameter or dry weight. Trichoderma spp. did not get into leaves while S. tritici was present, even in asymptomatic leaf extracts. In addition, the leaf apoplast antifungal proteolytic activity was measured in plants 7, 15, and 22 days after sowing. This antifungal action decreased in plants only inoculated with S. tritici, but increased in those grown from seeds coated with the T. harzianum (Th5) isolate. This increase conferred resistance to the susceptible wheat cultivar. The endogenous germin-like protease inhibitor coordinated the proteolytic action. These results suggest that T. harzianum stimulates a biochemical systemic induced response against leaf blotch.

Nitric‐oxide‐mediated cell death is triggered by chitosan in Fusarium eumartii spores

BACKGROUND The genus Fusarium comprises a heterogeneous group of fungi important for agriculture. Fusarium solani f. sp. eumartii (F. eumartii), historically considered to be a fungal pathogen of potato, has also been associated with tomato disease. Currently, chitosan and its derivatives have been receiving more attention as environmentally friendly antimicrobial compounds in sustainable practices. The aim of the present work was to characterize downstream events associated with the mode of action of chitosan, including nitrosative reactive species, in order to identify new biomarkers of its cytotoxic action. RESULTS Data indicated that chitosan-mediated nitric oxide (NO) production might lead to conidial death, concomitant with the strong reduction in fungal pathogenicity in tomato plants. Following chitosan applications, a notably dose-dependent reduction in conidial viability was demonstrated in F. eumartii. Thereafter, the infectivity of chitosan-treated spores was tested by a bioassay using tomato seedlings. CONCLUSION All these data highlight NO valuable properties as a quantitative and qualitative biomarker of cytotoxic action of chitosan in conidial cells. In addition, these findings place the chitosan assayed here as a fungicide with a high potential of application in sustainable horticultural practices.

Preparation, Characterization, and In Vitro Testing of Nanoclay Antimicrobial Activities and Elicitor Capacity

Clay-based nanocomposites (nanoclays) are interesting systems to hold a wide type of active substances with a wide field of industrial applications. Bentonite-chitosan nanoclay was obtained via cationic exchange of natural bentonite (Bent) with an aqueous solution of chitosan (CS). Their physicochemical and morphological properties were discussed under the light of Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetric analysis, and scanning electron microscopy. Bent-CS characterization indicated that CS was intercalated in 10% (w/w). This polycationic polymer was oriented mostly in a monolayer arrangement, interacting by electrostatic forces between Bent sheets. The antimicrobial action of Bent-CS nanoclay was assayed onto phytopathogens, the bacterium model Pseudomonas syringe pv. tomato DC3000 ( Psy) and the necrotrophic fungus Fusarium solani f. sp. eumartii ( F. eumartii). In addition to demonstrating cell death on both microorganisms, Bent-CS exerted elicitor property on tomato plantlets. The biological actions of this natural nanomaterial might make it proper to be used in crops.

Chitosan as Source for Pesticide Formulations

Late blight and wilt caused by the oomycete, Phytophthora infestans, and the fungus, Fusarium solani f. sp. eumartii, respectively, are severe diseases in Solanaceae crops worldwide. Although traditional approaches to control plant diseases have mainly relied on toxic chemical compounds, current studies are focused to identify more sustainable options. Finding alternatives, a low molecular weight chitosan (LMWCh) obtained from biomass of Argentine Sea’s crustaceans was assayed. In an attempt to characterize the action of LMWCh alone or in combination with the synthetic fungicide Mancozeb, the antimicrobial properties of LMWCh were assayed. In a side-by-side comparison with the SYTOX Green nucleic acid stain and the nitric oxide–specific probe, diaminofluorescein-FM diacetate (DAF-FM DA), yielded a similar tendency, revealing LMWChmediated cell death. The efficacy of LMWCh, Mancozeb, and the mixture LMWCh– Mancozeb was in turn tested. A synergistic effect in the reduction of F. eumartii spore germination was measured in the presence of subinhibitory dosis of 0.025 mg ml−1 LMWCh and 0.008 mg ml−1 Mancozeb. This mixture was efficient to increase the effectiveness of the single treatments in protecting against biotic stress judged by a drastic reduction of lesion area in P. infestans–inoculated tissues and activation of the potato defense responses.