Analysis by HPLC – mass spectrometry of the indole compounds released by the ectomycorrhizal fungus <i>Hebeloma hiemale</i> in pure culture

Rouillon, Régis; Bernillon, Jacques; Favre-Bonvin, Jean; Bruchet, G.

doi:10.1139/b86-252

Cited by 24 publications

(10 citation statements)

References 15 publications

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“…This reagent does not react with indole-3-carboxylic acid and indole 3-aldehyde (Rouillon et al, 1986). The specificity of this method was confirmed by HPLC and mass spectrometry of culture filtrates and incubation solutions (see abo\"e).…”

Section: Iaa Quantificationmentioning

confidence: 82%

Mycorrhizal formation by Paxillus involutus strains in relation to their IAA‐synthesizing activity

Rudawska

Kieliszewska-Rokicka

1997

New Phytologist

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SUMMARYIAA production by the ectomycorrhizal fungus Paxillus involutus grown in pure culture was investigated by testing IAA accumulation or IAA-synthesizing activity. All the Paxillus involutus strains studied released indole compounds either when incubated with 0-5 mM tryptophan or in its absence. HPLC analyses of indole compounds al! yielded two main peaks identified by reference to controls as being IAA and indo!e-3-carboxylic acid and a small peak of indole-3-aldehyde. Mass spectrometry unequivocally confirmed IAA production in the culture filtrates and in the incubation solution used to study in vitro IAA-synthesizing activity. The IAA-synthesizing activity was found to vary widely, from 7-6 to 137'4 nmol IAA h"^ g"^ f. wt, between strains. The mycorrhizal activity of these strains with seedlings of Pinus sylvestris cultured under aseptic conditions was tested. Fungal strains that were characterized in pure culture conditions by high IAA-synthesizing activity induced more fine roots and significantly higher numbers of mycorrhizas than strains which showed a low activity of IAA synthesizing enzymes. The increased ectomycorrhiza formation was generally accompanied by an increased growth of seedlings.

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Section: Iaa Quantificationmentioning

confidence: 82%

Mycorrhizal formation by Paxillus involutus strains in relation to their IAA‐synthesizing activity

Rudawska

Kieliszewska-Rokicka

1997

New Phytologist

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“…The monokaryotic strain h1 of the ectomycorrhizal fungus H. cylindrosporum was obtained from single-spore germination (50). It was maintained on agar medium, as described by Rouillon et al (43), that contained modified Melin's medium (33) 4 ⅐ 6H 2 O). The mycelium was harvested after 21 days of incubation at 25°C and washed with distilled water, and the dry biomass was measured.…”

Section: Methodsmentioning

confidence: 99%

Different Patterns of Regulation for the Copper and Cadmium Metallothioneins of the Ectomycorrhizal Fungus Hebeloma cylindrosporum

Ramesh

Podila

Marmeisse

et al. 2009

Appl Environ Microbiol

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Metallothioneins (MTs) are small cysteine-rich peptides involved in metal homeostasis and detoxification. We have characterized two MT genes, HcMT1 and HcMT2, from the ectomycorrhizal fungus Hebeloma cylindrosporum in this study. Expression of HcMT1 and HcMT2 in H. cylindrosporum under metal stress conditions was studied by competitive reverse transcription-PCR analysis. The full-length cDNAs were used to perform functional complementation in mutant strains of Saccharomyces cerevisiae. As revealed by heterologous complementation assays in yeast, HcMT1 and HcMT2 each encode a functional polypeptide capable of conferring increased tolerance against Cd and Cu, respectively. The expression levels of HcMT1 were observed to be at their maximum at 24 h, and they increased as a function of Cu concentration. HcMT2 was also induced by Cu, but the expression levels were lower than those for HcMT1. The mRNA accumulation of HcMT1 was not influenced by Cd, whereas Cd induced the transcription of HcMT2. Zn, Pb, and Ni did not affect the transcription of HcMT1 or of HcMT2. Southern blot analysis revealed that both of these genes are present as a single copy in H. cylindrosporum. While the promoters of both HcMT1 and HcMT2 contained the standard stress response elements implicated in the metal response, the numbers and varieties of potential regulatory elements were different in these promoters. These results show that ectomycorrhizal fungi encode different MTs and that each of them has a particular pattern of expression, suggesting that they play critical specific roles in improving the survival and growth of ectomycorrhizal trees in ecosystems contaminated by heavy metals.

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“…In controlling the elicitor-induced synthesis of defense proteins, hormones, especially auxins that are synthesized by the mycorrhizal fungus (Rouillon et al 1986;Gogala 1991), might be involved. In spruce cells, auxins specifically suppressed the induction of one peroxidase isoform, but did not influence the synthesis of HzO2 (Salzer and Hager 1993) and the extracellular alkalinization (data not shown).…”

Section: Release Of Elicitors From Hyphae Of Hebeloma Crustuliniformementioning

confidence: 99%

Rapid reactions of spruce cells to elicitors released from the ectomycorrhizal fungus Hebeloma crustuliniforme, and inactivation of these elicitors by extracellular spruce cell enzymes

Salzer

Hebe

Reith

et al. 1996

Planta

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Abstract. Elicitors released from hyphae or cell walls of the ectomycorrhizal fungus Hebeloma crustuliniforme (Bull. ex Fries.) Qu~l. induced in suspension-cultured cells of Picea abies (L.) Karst. a set of fast reactions: (i) an immediate efflux of C1-into the medium, followed by a K § efflux; (ii) an influx of Ca 2 § (measured as accumulation of 45Ca2+ in the cells); (iii) a phosphorylation of a 63-kDa protein and dephosphorylation of a 65-kDa protein (detectable by 4 min after elicitor application); (iv) an alkalinization of the medium, and (v) a transient synthesis of H202. The removal of extracellular Ca 2 + by EGTA delayed the elicitor-induced alkalinization. A further reduction of this response could be achieved by TMB-8 an inhibitor of Ca 2 § release from intracellular stores. Moreover, the inhibition of protein kinase activity by staurosporine prevented the extracellular alkalinization completely. However, the effectiveness of the elicitors in inducing the extracellular alkalinization was strongly impaired by constitutively secreted enzymes of spruce cells which cleaved the elicitors to inactive fragments. It is suggested that in ectomycorrhizae the efficacy of elicitors released from fungal cell walls is controlled by apoplastic enzymes of the host; the plant itself is able to reduce the activity of fungal elicitors on their way through the plant cell wall. But those elicitors which finally reach the plasma membrane of host cells induce reactions that are similar to the early defense reactions in plant-pathogen interactions.

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Analysis by HPLC – mass spectrometry of the indole compounds released by the ectomycorrhizal fungus Hebeloma hiemale in pure culture

Cited by 24 publications

References 15 publications

Mycorrhizal formation by Paxillus involutus strains in relation to their IAA‐synthesizing activity

Mycorrhizal formation by Paxillus involutus strains in relation to their IAA‐synthesizing activity

Different Patterns of Regulation for the Copper and Cadmium Metallothioneins of the Ectomycorrhizal Fungus Hebeloma cylindrosporum

Rapid reactions of spruce cells to elicitors released from the ectomycorrhizal fungus Hebeloma crustuliniforme, and inactivation of these elicitors by extracellular spruce cell enzymes

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