A senescence-delaying pre-culture medium for transcriptomics of Podospora anserina

Benocci, Tiziano; Vries, Ronald P. de; Daly, Paul

doi:10.1016/j.mimet.2018.01.010

Cited by 5 publications

(4 citation statements)

References 17 publications

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“…To simplify the detection of early markers of ligninolytic activity of P. anserina, we used cultivation conditions that would allow us to evaluate changes in both lignin structure and in the fungal secretome [21]. To this end, we incubated the fungus on pre-isolated lignin from wheat straw that was combined with the hemicellulose fraction of the straw, i.e., glucuronoarabinoxylan (GAX), instead of growing the fungus on unfractionated wheat straw.…”

Section: Cultivation Setupmentioning

confidence: 99%

“…Media and growth conditions for P. anserina were based on previously described procedures [21,60]. Waterinsoluble wheat glucuronoarabinoxylan (GAX, Megazyme, Wicklow, Ireland, 100 mg) and GAX (100 mg) combined with wheat straw lignin isolate (100 mg) were dispersed in 5 mL M2 minimal medium in 40-mL flasks and autoclaved prior to inoculation.…”

Section: Growth Of Podospora Anserina On Wheat Straw Ligninmentioning

confidence: 99%

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Evidence for ligninolytic activity of the ascomycete fungus Podospora anserina

Erven

Kleijn

Patyshakuliyeva

et al. 2020

Biotechnol Biofuels

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Background: The ascomycete fungus Podospora anserina has been appreciated for its targeted carbohydrate-active enzymatic arsenal. As a late colonizer of herbivorous dung, the fungus acts specifically on the more recalcitrant fraction of lignocellulose and this lignin-rich biotope might have resulted in the evolution of ligninolytic activities. However, the lignin-degrading abilities of the fungus have not been demonstrated by chemical analyses at the molecular level and are, thus far, solely based on genome and secretome predictions. To evaluate whether P. anserina might provide a novel source of lignin-active enzymes to tap into for potential biotechnological applications, we comprehensively mapped wheat straw lignin during fungal growth and characterized the fungal secretome.Results: Quantitative 13 C lignin internal standard py-GC-MS analysis showed substantial lignin removal during the 7 days of fungal growth (24% w/w), though carbohydrates were preferably targeted (58% w/w removal). Structural characterization of residual lignin by using py-GC-MS and HSQC NMR analyses demonstrated that C α -oxidized substructures significantly increased through fungal action, while intact β-O-4′ aryl ether linkages, p-coumarate and ferulate moieties decreased, albeit to lesser extents than observed for the action of basidiomycetes. Proteomic analysis indicated that the presence of lignin induced considerable changes in the secretome of P. anserina. This was particularly reflected in a strong reduction of cellulases and galactomannanases, while H 2 O 2 -producing enzymes clearly increased. The latter enzymes, together with laccases, were likely involved in the observed ligninolysis. Conclusions:For the first time, we provide unambiguous evidence for the ligninolytic activity of the ascomycete fungus P. anserina and expand the view on its enzymatic repertoire beyond carbohydrate degradation. Our results can be of significance for the development of biological lignin conversion technologies by contributing to the quest for novel lignin-active enzymes and organisms.

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Section: Cultivation Setupmentioning

confidence: 99%

Section: Growth Of Podospora Anserina On Wheat Straw Ligninmentioning

confidence: 99%

Evidence for ligninolytic activity of the ascomycete fungus Podospora anserina

Erven

Kleijn

Patyshakuliyeva

et al. 2020

Biotechnol Biofuels

Self Cite

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“…This tentatively suggests that they may act through the activation of specific pathway(s), rather than having general roles in modulating the redox status of the substrate. This is substantiated by the fact that lignin modulates transcription in P. anserina [74]. Interestingly, improved growth and fertility triggered by lignin may be a general phenomenon in fungi, since it was previously demonstrated that lignin increases growth and fruiting body production in the basidiomycete shitake [75] and that extractives stimulate the production of perithecia in Chaetomium indicum and Chaetomium globosum [76], two fungi closely related to P. anserina.…”

Section: Discussionmentioning

confidence: 93%

Lignin Degradation and Its Use in Signaling Development by the Coprophilous Ascomycete Podospora anserina

Dicko¹,

Ferrari

Tangthirasunun

et al. 2020

JoF

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The filamentous fungus Podospora anserina is a good model to study the breakdown of lignocellulose, owing to its ease of culture and genetical analysis. Here, we show that the fungus is able to use a wide range of lignocellulosic materials as food sources. Using color assays, spectroscopy and pyrolysis–gas chromatography mass spectrometry, we confirm that this ascomycete is able to degrade lignin, primarily by hydrolyzing β–O-4 linkages, which facilitates its nutrient uptake. We show that the limited weight loss that is promoted when attacking Miscanthus giganteus is due to a developmental blockage rather than an inefficiency of its enzymes. Finally, we show that lignin, and, more generally, phenolics, including degradation products of lignin, greatly stimulate the growth and fertility of the fungus in liquid cultures. Analyses of the CATΔΔΔΔΔ mutant lacking all its catalases, pro-oxidants and antioxidants indicate that improved growth and fertility of the fungus is likely caused by augmented reactive oxygen species levels triggered by the presence of phenolics.

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“…This indicated that LB broth medium could be a suitable medium for Iranian Lizard Leishmania culture and recombinant protein production compared with the RPMI1640 medium, brain heart infusion medium and M199 medium [ 3 ]. Another study found that the good growth without senescence of Podospora anserina was observed only on LB medium, which prolonged the health of the mycelium, while liquid cultures without senescence were observed only in LB broth medium [ 4 ]. Taken together, in life science experiments, LB broth medium is the preferred bacterial growth solution for almost all microbes, including bacteria, fungi, and parasites.…”

Section: Introductionmentioning

confidence: 99%

The Metabolomics Changes in Luria–Bertani Broth Medium under Different Sterilization Methods and Their Effects on Bacillus Growth

Wang,

Guo,

Chen

et al. 2023

Metabolites

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Luria–Bertani broth (LB) culture medium is a commonly used bacterial culture medium in the laboratory. The nutrient composition, concentration, and culture conditions of LB medium can influence the growth of microbial strains. The purpose of this article is to demonstrate the impact of LB liquid culture medium on microbial growth under different sterilization conditions. In this study, LB medium with four different treatments was used, as follows: A, LB medium without treatments; B, LB medium with filtration; C, LB medium with autoclaving; and D, LB medium with autoclaving and cultured for 12 h. Subsequently, the protein levels and antioxidant capacity of the medium with different treatments were measured, and the effects of the different LB medium treatments on the growth of microorganisms and metabolites were determined via 16s rRNA gene sequencing and metabolomics analysis, respectively. Firmicutes and Lactobacillus were the dominant microorganisms, which were enriched in fermentation and chemoheterotrophy. The protein levels and antioxidant capacity of the LB medium with different treatments were different, and with the increasing concentration of medium, the protein levels were gradually increased, while the antioxidant capacity was decreased firstly and then increased. The growth trend of Bacillus subtilis, Bacillus paralicheniformis, Micrococcus luteus, and Alternaria alternata in the medium with different treatments was similar. Additionally, 220 and 114 differential metabolites were found between B and C medium, and between C and D medium, which were significantly enriched in the “Hedgehog signaling pathway”, “biosynthesis of plant secondary metabolites”, “ABC transporters”, “arginine and proline metabolism”, and “linoleic acid metabolism”. LB medium may be a good energy source for Lactobacillus growth with unsterilized medium, and LB medium filtered with a 0.22 μm filter membrane may be used for bacterial culture better than culture medium after high-pressure sterilization. LB medium still has the ability for antioxidation and to keep bacteria growth whether or not autoclaved, indicating that there are some substances that can resist a high temperature and pressure and still maintain their functions.

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A senescence-delaying pre-culture medium for transcriptomics of Podospora anserina

Cited by 5 publications

References 17 publications

Evidence for ligninolytic activity of the ascomycete fungus Podospora anserina

Evidence for ligninolytic activity of the ascomycete fungus Podospora anserina

Lignin Degradation and Its Use in Signaling Development by the Coprophilous Ascomycete Podospora anserina

The Metabolomics Changes in Luria–Bertani Broth Medium under Different Sterilization Methods and Their Effects on Bacillus Growth

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