The plant host Brassica napus induces in the pathogen Verticillium longisporum the expression of functional catalase peroxidase which is required for the late phase of disease.

The devastating soilborne fungal pathogen Verticillium longisporum is host specific to members of the family Brassicaceae, including oilseed rape (Brassica napus) as the economically most important crop. The fungus infects through the roots and causes stunting and early senescence of susceptible host plants and a marked decrease in crop yield. We show here that V. longisporum reacts to the presence of B. napus xylem sap with the production of six distinct upregulated and eight downregulated proteins visualized by two-dimensional gel electrophoresis. Identification of 10 proteins by mass spectrometry revealed that all upregulated proteins are involved in oxidative stress response. The V. longisporum catalase peroxidase (VlCPEA) was the most upregulated protein and is encoded by two isogenes, VlcpeA-1 and VlcpeA-2. Both genes are 98% identical, corroborating the diploid or "amphihaploid" status of the fungus. Knock downs of both VlcpeA genes reduced protein expression by 80% and resulted in sensitivity against reactive oxygen species. Whereas saprophytic growth and the initial phase of the plant infection were phenotypically unaffected, the mutants were not able to perform the late phases of disease. We propose that the catalase peroxidase plays a role in protecting the fungus from the oxidative stress generated by the host plant at an advanced phase of the disease.

Beech carbon productivity as driver of ectomycorrhizal abundance and diversity.

We tested the hypothesis that carbon productivity of beech (Fagus sylvatica) controls ectomycorrhizal colonization, diversity and community structures. Carbon productivity was limited by long-term shading or by girdling. The trees were grown in compost soil to avoid nutrient deficiencies. Despite severe limitation in photosynthesis and biomass production by shading, the concentrations of carbohydrates in roots were unaffected by the light level. Shade-acclimated plants were only 10% and sun-acclimated plants were 74% colonized by ectomycorrhiza. EM diversity was higher on roots with high than at roots with low mycorrhizal colonization. Evenness was unaffected by any treatment. Low mycorrhizal colonization had no negative effects on plant mineral nutrition. In girdled plants mycorrhizal colonization and diversity were retained although (14)C-leaf feeding showed almost complete disruption of carbon transport from leaves to roots. Carbohydrate storage pools in roots decreased upon girdling. Our results show that plant carbon productivity was the reason for and not the result of high ectomycorrhizal diversity. We suggest that ectomycorrhiza can be supplied by two carbon routes: recent photosynthate and stored carbohydrates. Storage pools may be important for ectomycorrhizal survival when photoassimilates were unavailable, probably feeding preferentially less carbon demanding EM species as shifts in community composition were found.

Defence reactions in the apoplastic proteome of oilseed rape (Brassica napus var. napus) attenuate Verticillium longisporum growth but not disease symptoms.

BACKGROUND: Verticillium longisporum is one of the most important pathogens of Brassicaceae that remains strictly in the xylem during most stages of its development. It has been suggested that disease symptoms are associated with clogging of xylem vessels. The aim of our study was to investigate extracellular defence reactions induced by V. longisporum in the xylem sap and leaf apoplast of Brassica napus var. napus in relation to the development of disease symptoms, photosynthesis and nutrient status. RESULTS: V. longisporum (strain VL43) did not overcome the hypocotyl barrier until 3 weeks after infection although the plants showed massive stunting of the stem and mild leaf chlorosis. During this initial infection phase photosynthetic carbon assimilation, transpiration rate and nutrient elements in leaves were not affected in VL43-infected compared to non-infected plants. Proteome analysis of the leaf apoplast revealed 170 spots after 2-D-protein separation, of which 12 were significantly enhanced in response to VL43-infection. LS-MS/MS analysis and data base searches revealed matches of VL43-responsive proteins to an endochitinase, a peroxidase, a PR-4 protein and a beta-1,3-glucanase. In xylem sap three up-regulated proteins were found of which two were identified as PR-4 and beta-1,3-glucanase. Xylem sap of infected plants inhibited the growth of V. longisporum. CONCLUSION: V. longisporum infection did not result in drought stress or nutrient limitations. Stunting and mild chlorosis were, therefore, not consequences of insufficient water and nutrient supply due to VL43-caused xylem obstruction. A distinct array of extracellular PR-proteins was activated that might have limited Verticillium spreading above the hypocotyl. In silico analysis suggested that ethylene was involved in up-regulating VL43-responsive proteins.