Laser nephelometry applied in an automated microplate system to study filamentous fungus growth

Joubert, Aymeric; Calmes, Benoît; Berruyer, Romain; Pihet, Marc; Bouchara, Jean‐Philippe; Simoneau, Philippe; Guillemette, Thomas

doi:10.2144/000113399

Cited by 46 publications

(49 citation statements)

References 19 publications

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“…Shannon indices were calculated on the basis of the number and abundance of species for each depth using the vegan package in R. Ecophysiological characterization. Ecophysiological characterization was performed as described by Joubert et al (27). Briefly, suspensions of conidia from selected isolates and dilutions of those isolates were made using PDB to achieve a final concentration of suspended spores of between 1 ϫ 10 5 and 5 ϫ 10 5 per ml.…”

Section: Methodsmentioning

confidence: 99%

“…The presence of genes involved in the biosynthetic pathways of secondary metabolites was investigated. Type I polyketide synthase (PKS I), type III polyketide synthase (PKS III), nonribosomal peptide synthetase (NRPS), PKS-NRPS hybrid, and terpene synthase (TPS) genes were amplified using the following degenerate primer pairs with D-inosine modifications (indicated by i): (i) primers KAF1 (GARKSiCAYGGiACiG GiAC) and KAR1 (CCAYTGiGCiCCRTGiCCiGARAA) (28) were used for amplification of type I PKS genes, (ii) primers XKS1 (TTYGAYGCiB CiTTYTTYRA) and XKS2 (CRTTiGYiCCiCYDAAiCCAAA) (27) were used for amplification of PKS-NRPS hybrid genes, (iii) primers AUG003 (CCGGCACCACCGGNAARCCHAA) and AUG007 (GCTGCATGGCG GTGATGSWRTSNCCBCC) (29) were used for amplification of NRPS genes, (iv) primers TPS1 (GCiTAYGAYACiGCiTGGGT) and TPS2 (RA AiGCATiGCiGTRTCRTC) (30) were used for amplification of TPS genes, and (v) primers CHS1 (GAYTGGGCiVTNCAYCCBGGiGGD) and CHS2 (YTCNAYNKTRAKVCCiGGVCCRAA) were used for amplification of type III PKS genes. Primers targeting type III PKS genes were specifically designed for this study using ClustalW (version 1.83) (26).…”

Section: Methodsmentioning

confidence: 99%

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Species Richness and Adaptation of Marine Fungi from Deep-Subseafloor Sediments

Rédou

Navarri

Meslet‐Cladière

et al. 2015

Appl Environ Microbiol

146

117

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The fungal kingdom is replete with unique adaptive capacities that allow fungi to colonize a wide variety of habitats, ranging from marine habitats to freshwater and terrestrial habitats. The diversity, importance, and ecological roles of marine fungi have recently been highlighted in deep-subsurface sediments using molecular methods. Fungi in the deep-marine subsurface may be specifically adapted to life in the deep biosphere, but this can be demonstrated only using culture-based analyses. In this study, we investigated culturable fungal communities from a record-depth sediment core sampled from the Canterbury Basin (New Zealand) with the aim to reveal endemic or ubiquist adapted isolates playing a significant ecological role(s). About 200 filamentous fungi (68%) and yeasts (32%) were isolated. Fungal isolates were affiliated with the phyla Ascomycota and Basidiomycota, including 21 genera. Screening for genes involved in secondary metabolite synthesis also revealed their bioactive compound synthesis potential. Our results provide evidence that deep-subsurface fungal communities are able to survive, adapt, grow, and interact with other microbial communities and highlight that the deep-sediment habitat is another ecological niche for fungi.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Species Richness and Adaptation of Marine Fungi from Deep-Subseafloor Sediments

Rédou

Navarri

Meslet‐Cladière

et al. 2015

Appl Environ Microbiol

146

117

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“…For routine culture , A. brassicicola was grown and maintained on potato dextrose agar (PDA). The method based on microscale liquid cultivation (from conidial suspensions) and automated nephelometric recording of growth, followed by extraction of relevant variables (lag time and growth rate), was described by [49]. To study the susceptibility of fungal strains to ITCs, allyl-ITC (Al-ITC), benzyl-ITC (Bz-ITC) or phenetyl-ITC (Ph-ITC), all purchased from Aldrich Chemical Co. (Milwaukee, WI), were diluted from stock solutions prepared in methanol at the final desired concentrations.…”

Section: Methodsmentioning

confidence: 99%

Characterization of glutathione transferases involved in the pathogenicity of Alternaria brassicicola

et al. 2015

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BackgroundGlutathione transferases (GSTs) represent an extended family of multifunctional proteins involved in detoxification processes and tolerance to oxidative stress. We thus anticipated that some GSTs could play an essential role in the protection of fungal necrotrophs against plant-derived toxic metabolites and reactive oxygen species that accumulate at the host-pathogen interface during infection.ResultsMining the genome of the necrotrophic Brassica pathogen Alternaria brassicicola for glutathione transferase revealed 23 sequences, 17 of which could be clustered into the main classes previously defined for fungal GSTs and six were ‘orphans’. Five isothiocyanate-inducible GSTs from five different classes were more thoroughly investigated. Analysis of their catalytic properties revealed that two GSTs, belonging to the GSTFuA and GTT1 classes, exhibited GSH transferase activity with isothiocyanates (ITC) and peroxidase activity with cumene hydroperoxide, respectively. Mutant deficient for these two GSTs were however neither more susceptible to ITC nor less aggressive than the wild-type parental strain. By contrast mutants deficient for two other GSTs, belonging to the Ure2pB and GSTO classes, were distinguished by their hyper-susceptibility to ITC and low aggressiveness against Brassica oleracea. In particular AbGSTO1 could participate in cell tolerance to ITC due to its glutathione-dependent thioltransferase activity. The fifth ITC-inducible GST belonged to the MAPEG class and although it was not possible to produce the soluble active form of this protein in a bacterial expression system, the corresponding deficient mutant failed to develop normal symptoms on host plant tissues.ConclusionsAmong the five ITC-inducible GSTs analyzed in this study, three were found essential for full aggressiveness of A. brassicicola on host plant. This, to our knowledge is the first evidence that GSTs might be essential virulence factors for fungal necrotrophs.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-015-0462-0) contains supplementary material, which is available to authorized users.

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“…photometry [6,12]. Alifax automated nephelometric microbiological analyzers carry out a rapid microbiological analysis within just several hours, including rapid screening of urine for contamination [4,7].…”

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confidence: 99%

Coherent Fluctuation Nephelometry: A Rapid Method for Urine Screening for Bacterial Contamination

et al. 2015

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Express-test by the method of coherent fluctuation nephelometry for urine contamination was carried out on two prototype instruments with standard polystyrene photometric cuvettes. We analyzed 209 and 119 urine samples. Due to high sensitivity of the method, up to 50% negative samples were detected within 10 min by initial opacity and 90% negative samples were detected during 3.5 h by registration of the bacterial growth curves.

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Laser nephelometry applied in an automated microplate system to study filamentous fungus growth

Cited by 46 publications

References 19 publications

Species Richness and Adaptation of Marine Fungi from Deep-Subseafloor Sediments

Species Richness and Adaptation of Marine Fungi from Deep-Subseafloor Sediments

Characterization of glutathione transferases involved in the pathogenicity of Alternaria brassicicola

Coherent Fluctuation Nephelometry: A Rapid Method for Urine Screening for Bacterial Contamination

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