The recent release of several basidiomycete genome sequences allows an improvement of the classification of fungal glutathione S-transferases (GSTs). GSTs are well-known detoxification enzymes which can catalyze the conjugation of glutathione to non-polar compounds that contain an electrophilic carbon, nitrogen, or sulfur atom. Following this mechanism, they are able to metabolize drugs, pesticides, and many other xenobiotics and peroxides. A genomic and phylogenetic analysis of GST classes in various sequenced fungi--zygomycetes, ascomycetes, and basidiomycetes--revealed some particularities in GST distribution, in comparison with previous analyses with ascomycetes only. By focusing essentially on the wood-degrading basidiomycete Phanerochaete chrysosporium, this analysis highlighted a new fungal GST class named GTE, which is related to bacterial etherases, and two new subclasses of the omega class GSTs. Moreover, our phylogenetic analysis suggests a relationship between the saprophytic behavior of some fungi and the number and distribution of some GST isoforms within specific classes.
The white rot fungus Phanerochaete chrysosporium, a saprophytic basidiomycete, possesses a large number of cytosolic glutathione transferases, eight of them showing similarity to the Omega class. PcGSTO1 (subclass I, the bacterial homologs of which were recently proposed, based on their enzymatic function, to constitute a new class of glutathione transferase named S-glutathionyl-(chloro)hydroquinone reductases) and PcGSTO3 (subclass II related to mammalian homologs) have been investigated in this study. Biochemical investigations demonstrate that both enzymes are able to catalyze deglutathionylation reactions thanks to the presence of a catalytic cysteinyl residue. This reaction leads to the formation of a disulfide bridge between the conserved cysteine and the removed glutathione from their substrate. The substrate specificity of each isoform differs. In particular PcGSTO1, in contrast to PcGSTO3, was found to catalyze deglutathionylation of S-glutathionyl-p-hydroquinone substrates. The three-dimensional structure of PcGSTO1 presented here confirms the hypothesis that it belongs not only to a new biological class but also to a new structural class that we propose to name GST xi. Indeed, it shows specific features, the most striking ones being a new dimerization mode and a catalytic site that is buried due to the presence of long loops and that contains the catalytic cysteine.
The glutathione-S-transferase (GST) proteins represent an extended family involved in detoxification processes. They are divided into various classes with high diversity in various organisms. The Ure2p class is especially expanded in saprophytic fungi compared to other fungi. This class is subdivided into two subclasses named Ure2pA and Ure2pB, which have rapidly diversified among fungal phyla. We have focused our analysis on Basidiomycetes and used Phanerochaete chrysosporium as a model to correlate the sequence diversity with the functional diversity of these glutathione transferases. The results show that among the nine isoforms found in P. chrysosporium, two belonging to Ure2pA subclass are exclusively expressed at the transcriptional level in presence of polycyclic aromatic compounds. Moreover, we have highlighted differential catalytic activities and substrate specificities between Ure2pA and Ure2pB isoforms. This diversity of sequence and function suggests that fungal Ure2p sequences have evolved rapidly in response to environmental constraints.
Bakanae disease is a major rice disease caused by Fusarium fujikuroi. Infected plants show slender and hyper elongated internodes due to the over-production of gibberellic acid. Application of Trichoderma spp. as biocontrol agent is gaining attention due to high capability in hyperparasitize the soil borne pathogen. The studies aimed to screen and evaluate the bio-efficacy of Trichoderma spp. with antagonistic activities against F. fujikuroi and plant growth-promoting properties. All the 65 Trichoderma isolates were isolated from healthy rice rhizosphere soil. Thirty eight out of 65 Trichoderma isolates exhibited more than 45 Percentage of Inhibition Radial Growth (PIRG) against F. fujikuroi in dual culture plate testing. All selected Trichoderma isolates were further in vitro screened for antagonistic testing: volatile compounds production and hydrogen cyanide production and plant growth-promotion properties: IAA production and phosphate solubilization. Twelve Trichoderma isolates were selected for further evaluation on antagonistic activity against F. fujikuroi, germination rate, plumule and radical lengths and vigor index. Finally, seven of the most potential Trichoderma isolates were selected for greenhouse evaluation. The bakanae disease incidence and disease severity in rice plant treated with respective selected Trichoderma isolates were significant reduced as compared with untreated plant. However, there was no significant increase in plant height between Trichoderma inoculated and uninoculated plants. Moreover, rice plant treated with Trichoderma T61 showed significantly increase in total plant dry biomass as compared to untreated plants. The selected Trichoderma isolates have potential to be developed as biological control agent against F. fujikuroi and also as an alternative for bakanae management.
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