Fungistatic Mechanism of Ammonia against Nematode-Trapping Fungus Arthrobotrys oligospora, and Strategy for This Fungus To Survive Ammonia.

Soil fungistasis is a phenomenon in which the germination and growth of fungal propagules is widely inhibited in soils. Although fungistatic compounds are known to play important roles in the formation of soil fungistasis, how such compounds act on soil fungi is little studied. In this study, it was found that ammonia (NH3) induced global protein misfolding marked by increased ubiquitination levels of proteins (ubiquitylome data and Western blot verification). The misfolded proteins should trigger the endoplasmic reticulum (ER) stress, which was indicated by electron microscope image and proteome data. Results from the mutants of BiP and proteasome subunit alpha 7 suggested that ER stress played a mechanistic role in inhibiting conidial germination. Results from proteome data indicated that, to survive ammonia fungistasis, conidia first activated the unfolded protein response (UPR) to decrease ER stress and restore ER protein homeostasis, and the function of UPR in surviving ammonia was confirmed by using mutant strains. Second, ammonia toxicity could be reduced by upregulating carbon metabolism-related proteins, which benefited ammonia fixation. The results that metabolites (especially glutamate) could relieve the ammonia fungistasis confirmed this indirectly. Finally, results from gene knockout mutants also suggested that the fungistatic mechanism of ammonia is common for soil fungistasis. This study increased our knowledge regarding the mechanism of soil fungistasis and provided potential new strategies for manipulating soil fungistasis. IMPORTANCE Soil fungistasis is a phenomenon in which the germination and growth of fungal propagules is widely inhibited in soil. Although fungistatic compounds are known to play important roles in the formation of soil fungistasis, how such compounds act on soil fungi remains little studied. This study revealed an endoplasmic reticulum stress-related fungistatic mechanism with which ammonia acts on Arthrobotrys oligospora and a survival strategy of conidia under ammonia inhibition. Our study provides the first mechanistic explanation of how ammonia impacts fungal spore germination, and the mechanism may be common for soil fungistasis. This study increases our knowledge regarding the mechanism of soil fungistasis in fungal spores and provides potential new strategies for manipulating soil fungistasis.

Proteomic changes in Arthrobotrys oligospora conidia in response to benzaldehyde-induced fungistatic stress.

Soil fungistasis limits the effect of fungal agents designed to control plant-parasitic nematodes. Benzaldehyde is a fungistatic factor produced by soil microorganisms that can suppress conidial germination, but the molecular mechanism of this suppression is unknown. In this study, three conidial proteomes of Arthrobotrys oligospora ATCC24927, a nematode-trapping fungus, were obtained, quantified, and compared. Under benzaldehyde fungistatic stress, conidial protein expression profile changed significantly. Screening with a twofold selection criterion revealed 164 up-regulated and 110 down-regulated proteins. 17 proteins related to protein translation were down-regulated and gene transcription analysis suggested that the repression of proteins translation might be one mechanism by which benzaldehyde inhibites conidial germination. Benzaldehyde also resulted in the down-regulation of respiratory chain proteins and mitochondrial processes, as well as the repression of conidial DNA synthesis. In addition, the conidia up-regulated several proteins that enable it to resist benzaldehyde-induced fungistatis, and this was confirmed by a functional assessment of two knockout mutants. This study reveals putative mechanisms by which benzaldehyde causes fungistasis as well as the proteomic response of conidia to benzaldehyde. SIGNIFICANCE: Soil fungistasis limits the effect of fungal agents designed to control plant-parasitic nematodes. Benzaldehyde is one of fungistatic factors produced by soil microorganisms that can suppress conidial germination. In this study, we found that conidial protein expression profile changed significantly under benzaldehyde fungistatic stress. This research revealed new mechanistic data that describe how benzaldehyde is responsible for fungiststis by inhibiting conidial germination. Moreover, we also found that conidia can resist benzaldehyde by up-regulating proteins such as benzaldehyde dehydrogenase and heat shock proteins. This study also showed that proteomics methods play important roles in addressing soil fungistatic mechanisms.

Quantitative proteomics revealed partial fungistatic mechanism of ammonia against conidial germination of nematode-trapping fungus Arthrobotrys oligospora ATCC24927.

Ammonia is one of the fungistatic factors in soil that can suppress conidial germination, but the molecular mechanism underlying the suppression is unknown. In this study, the proteomes of fungistatic conidia, fresh conidia and germinated conidia of Arthrobotrys oligospora ATCC24927 were determined and quantified. The protein expression profile of fungistatic conidia was significantly different from those in the other two conditions. 281 proteins were down expressed in fungistatic conidia and characterized by GO annotation. Gene transcription analysis and inhibition of puromycin (a protein translation inhibitor) on conidial germination suggested that down expression of 33 protein translation related proteins might well result in repression of protein synthesis and inhibition of conidial germination. In addition, 16 down-expressed proteins were mapped to the Ras/mitogen-activated protein (Ras/MAP) regulatory networks which regulate conidial DNA synthesis. The conidial DNA synthesis was found to be definitely inhibited under by ammonia, and function studies of two Ras/MAP proteins by using knock-out strains provided partial evidence that Ras/MAP pathway regulate the conidial germination. These results suggested that down-expression of Ras/MAP related proteins might result in inhibition of DNA synthesis and finally result in inhibition conidial germination. This study revealed partial fungistatic mechanism of ammonia against conidial germination.

Stress distribution on the mandibular condyle and the distraction area in distraction osteogenesis by finite element analysis.

PURPOSE: Many 3-dimensional (3-D) finite element analysis (FEA) models based on the virtual simulation have been constructed to explore the stress distribution during distraction osteogenesis (DO). The aim of this study is to achieve a better understanding of the biomechanical feature of mandible during DO procedure. MATERIALS AND METHODS: Using CT image data of a goat mandible that already has been elongated for 10 mm, an anatomical 3D model was reconstructed. This model was imported into MSC.Patran for monitoring mechanical parameters, including Von-Mises stress, max-principal stress, and mix-principal stress, on the distraction area and the mandibular condyle. RESULTS: The computed result showed special stress distribution on 9 slopes of the condyle and 3 cross-sections of the distraction area. From 0.8 MP to 17.2 MP, a dramatic increase in Von-Mises stress was discovered on the right side of the condyle compared with the left side. From 22.6 MP to 0.9 MP on the distraction area, Von-Mises stress decreased from the central zone to the peripheral zone gradually. CONCLUSIONS: During the process of mandibular DO, the special stress distribution may be an important factor on the physiopathological change of the mandibular condyle and the generation of new bone in the distraction area. The correlations between the value of the stress and physiopathological changes during DO process should be clarified in future research.