Spatial monitoring of gene activity in extraradical and intraradical developmental stages of arbuscular mycorrhizal fungi by direct fluorescent in situ RT-PCR

Seddas, Pascale; Arnould, Christine; Tollot, Marie; Arias, Cecilia M.; Gianinazzi‐Pearson, Vivienne

doi:10.1016/j.fgb.2008.04.013

Cited by 20 publications

(16 citation statements)

References 63 publications

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“…The expression pattern of GintSOD1 during the fungal life cycle is consistent with earlier analyses of this gene sequence in the Medicago truncatula-G. intraradices association (Seddas et al 2008(Seddas et al , 2009, and resembles the expression profile of the orthologous gene of G. margarita in interactions with M. truncatula and Lotus japonicus (Lanfranco et al 2005). Overall these data indicate that SOD1 expression is strongly induced in the intraradical phase of the fungus, irrespective of the plant species and fungus involved in the association.…”

Section: Discussionsupporting

confidence: 87%

Characterization of a CuZn superoxide dismutase gene in the arbuscular mycorrhizal fungus Glomus intraradices

et al. 2010

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To gain further insights into the mechanisms of redox homeostasis in arbuscular mycorrhizal fungi, we characterized a Glomus intraradices gene (GintSOD1) showing high similarity to previously described genes encoding CuZn superoxide dismutases (SODs). The GintSOD1 gene consists of an open reading frame of 471 bp, predicted to encode a protein of 157 amino acids with an estimated molecular mass of 16.3 kDa. Functional complementation assays in a CuZnSOD-defective yeast mutant showed that GintSOD1 protects the yeast cells from oxygen toxicity and that it, therefore, encodes a protein that scavenges reactive oxygen species (ROS). GintSOD1 transcripts differentially accumulate during the fungal life cycle, reaching the highest expression levels in the intraradical mycelium. GintSOD1 expression is induced by the well known ROS-inducing agents paraquat and copper, and also by fenpropimorph, a sterol biosynthesis inhibitor (SBI) fungicide. These results suggest that GintSOD1 is involved in the detoxification of ROS generated from metabolic processes and by external agents. In particular, our data indicate that the antifungal effects of fenpropimorph might not be only due to the interference with sterol metabolism but also to the perturbation of other biological processes and that ROS production and scavenging systems are involved in the response to SBI fungicides.

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Section: Discussionsupporting

confidence: 87%

Characterization of a CuZn superoxide dismutase gene in the arbuscular mycorrhizal fungus Glomus intraradices

et al. 2010

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“…GintSTE is up-regulated in germinating spores and upon root penetration in G. intraradices Host root exudates are known to enhance fungal transcription during spore germination and especially for genes associated with mitochondrial activity (Tamasloukht et al, 2003), mobilization and transport of glycogen and lipid reserves (Park et al, 2002), protein synthesis or stress responses (Seddas et al, 2008). GintSTE expression levels increased significantly upon spore germination but effects did not differ between water and root exudates, suggesting that the up-regulation process for this gene is not dependent on a diffusable signal from the plant partner.…”

Section: Discussionmentioning

confidence: 99%

An STE12 gene identified in the mycorrhizal fungus Glomus intraradices restores infectivity of a hemibiotrophic plant pathogen

et al. 2008

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Summary Mechanisms of root penetration by arbuscular mycorrhizal (AM) fungi are unknown and investigations are hampered by the lack of transformation systems for these unculturable obligate biotrophs. Early steps of host infection by hemibiotrophic fungal phytopathogens, sharing common features with those of AM fungal colonization, depend on the transcription factor STE12. Using degenerated primers and rapid amplification of cDNA ends, we isolated the full‐length cDNA of an STE12‐like gene, GintSTE, from Glomus intraradices and profiled GintSTE expression by real‐time and in situ RT‐PCR. GintSTE activity and function were investigated by heterologous complementation of a yeast ste12Δ mutant and a Colletotrichum lindemuthianum clste12Δ mutant. Sequence data indicate that GintSTE is similar to STE12 from hemibiotrophic plant pathogens, especially Colletotrichum spp. Introduction of GintSTE into a noninvasive mutant of C. lindemuthianum restored fungal infectivity of plant tissues. GintSTE expression was specifically localized in extraradicular fungal structures and was up‐regulated when G. intraradices penetrated roots of wild‐type Medicago truncatula as compared with an incompatible mutant. Results suggest a possible role for GintSTE in early steps of root penetration by AM fungi, and that pathogenic and symbiotic fungi may share common regulatory mechanisms for invasion of plant tissues.

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“…In situ hybridisation (Kaldorf et al 1998) or in situ RT-PCR (Seddas et al 2008) can be used to detect fungal RNA accumulation in intraradical structures, while antibodies have been used for the immunolocalisation of proteins (Peretto et al 1995). Further functional characterisation of gene products was gained by heterologous expression of the genes in other organisms.…”

Section: Methodologiesmentioning

confidence: 99%

“…Interestingly, the transcripts were never detected in arbuscules, but to a high extent in vesicles of Glomus intraradices colonizing M. truncatula roots (Seddas et al 2008). In pea, however, high amounts of GintSOD RNA were identified in laser-dissected arbuscules and decreased to a basic level at late stages of the symbiosis (Kuznetsova et al 2010).…”

Section: B From Physical Contact To Production Of New Sporesmentioning

confidence: 99%

2 Molecular Approaches to Arbuscular Mycorrhiza Functioning

Franken

Waschke

Requena

2012

Fungal Associations

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Spatial monitoring of gene activity in extraradical and intraradical developmental stages of arbuscular mycorrhizal fungi by direct fluorescent in situ RT-PCR

Cited by 20 publications

References 63 publications

Characterization of a CuZn superoxide dismutase gene in the arbuscular mycorrhizal fungus Glomus intraradices

Characterization of a CuZn superoxide dismutase gene in the arbuscular mycorrhizal fungus Glomus intraradices

An STE12 gene identified in the mycorrhizal fungus Glomus intraradices restores infectivity of a hemibiotrophic plant pathogen

2 Molecular Approaches to Arbuscular Mycorrhiza Functioning

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