PhD Defense: A. Coince

The defense will be held the 3th october 2013 at 9H30 in Amphitheater 7, Faculté des Sciences, Boulevard des Aiguillettes, Vandoeuvre

 

Diversity and composition of fungi and oomycete communities in beech forest in relation to climatic and edaphic variables: from the stand to the continent 

Forest soils are heterogeneous habitats full of microorganisms. In particular, the filamentous eukaryotes (fungi and oomycetes) exhibit a huge diversity and play essential functions in the dynamic and sustainable growth of the forest ecosystem. However, their diversity and their distribution are poorly known; thus, so are the structuring factors of these microbial communities. The main goals were: (i) use a high-throughput pyrosequencing to study soil microbial communities at a broad geographical scale, and particularly to validate its use for the study of soil forest Pythiaceae communities, (ii) study the diversity and structure of fungal and Pythiaceae communities at several spatial scales, (iii) identify possible climatic and edaphic variables structuring these communities and (iv) estimate the possible response of these microbial communities to climatic variation. A pilot study was undertaken at the stand scale. Then, two additional studies were carried out along environ! mental gradients at the regional and continental scales. The use of the pyrosequencing technique was found appropriate for the fungal communities, but difficulties arose in studying the Pythiaceae community. At the stand scale, results suggested the soil to be a valuable substitute for the roots to access the ectomycorrhizal richness and composition using pyrosequencing. The results along the broad scale gradients suggested that fungal richness may not be affected by climate warming but that the composition would be. Moreover, our work indicated that soil pH is a major factor explaining fungal community composition. The main conclusions are still to be confirmed and deeper knowledge of the response of different fungal phylum, or family, would be required. The detection and thus the diversity estimation of the Pythiaceae in forest soil remains a current challenge.

Diversité et composition des communautés de champignons et d’oomycètes de hêtraies, en relation avec des facteurs climatiques et édaphiques : de la parcelle au continent 

Les sols forestiers sont des habitats hétérogènes, véritables réservoirs de microorganismes. Parmi ces microorganismes, les eucaryotes filamenteux (champignons et Pythiacées) sont très divers et jouent un rôle important dans le fonctionnement et la durabilité des écosystèmes forestiers. Leur diversité et leur répartition spatiale à différentes échelles sont encore peu connues et les facteurs qui sous-tendent cette dispersion sont encore peu étudiés. Aussi, les objectifs étaient (i) d’exploiter le séquençage haut-débit pour des études d’écologie microbienne à large échelle et valider son application aux communautés de Pythiacées en milieu forestier, (ii) de décrire ces communautés microbiennes, en termes de diversité et de structure, à différentes échelles spatiales (locale, régionale et continentale), (iii) de caractériser les variables biotiques et abiotiques structurant ces communautés et (iv) d’évaluer la réponse éventuelle de! s communautés aux variations climatiques.Une première étude pilote à l’échelle de la parcelle a été suivi de deux études à grande échelle spatiale le long de gradients environnementaux. Des gradients d’altitude et un gradient latitudinal, à l’échelle continentale, ont été utilisés comme gradient climatique. L’étude préliminaire a donc validé l’utilisation du pyroséquençage pour les communautés fongiques, et en particulier pour les espèces ectomycorhiziennes, et apporté des éléments pour établir une méthodologie d’échantillonnage couplée à cette technique. L’application de ces outils moléculaires à l’étude des communautés de Pythiacées reste à optimiser. Les résultats obtenus sur les communautés fongiques telluriques suggèrent que dans l’hypothèse d’un réchauffement climatique, la richesse fongique ne serait pas directement affectée mais la composition des communautés le serait. La composition des communautés fo! ngiques est également fortement liée au pH du sol. Ces résultats sont à affiner en étudiant plus en détail divers groupes taxonomiques et écologiques en lien avec des variables climatiques plus précises. Par ailleurs, de nombreuses perspectives sont envisageables pour améliorer la détection des Pythiacées dans les sols forestiers, qui reste un challenge en écologie microbienne.

Mots clés: champignons, mycorhizes, Pythophthora, Fagus sylvatica, gradient, temperature, pH, pyroséquençage

Post-doctoral position

Post-doctoral position on poplar rust fungus effectors at INRA Nancy (France)

A one-year post-doc position is available in the EcoGenomic of Interactions team at INRA Nancy starting in november 2013 on the functional analysis of the poplar-poplar rust fungus pathosystem.  This position is supported POPRUST and the INTEGRARUST projects (french ANR & Labex ARBRE).

Context: Through genomics and transcriptomics, our team has selected a list of promising candidate effectors from the rust fungus pathogen Melampsora larici-populina that we would like to further study to understand their role during host infection. The present project is a follow-up of a former post-doc, thus candidate effectors selection has already been established in the lab.

Duties: In the present project, the successful applicant will focus on the most promising effectors towards their production (recombinant proteins in E. coli, structure and localization, with engaged collaborations for NMR & crystallography) and the search for plant partners both in poplar host plants and heterologous systems.

Qualifications: The applicant should have past experience in molecular biology, host-microbe interaction, biochemistry and microscopy.

Duration:  12 months, starting in November 2013

Place of work:  The EGI Lab is part of the Tree-Microbe Interactions Department (http://mycor.nancy.inra.fr/IAM/) offers a creative and stimulating international environment and is one of six departments making up the cluster of Laboratoires d’Excellence (LabEx) ARBRE (http://mycor.nancy.inra.fr/ARBRE/). Our lab is located in the Lorraine INRA Center (trees and forest ecosystems research center) which is in the middle of the Champenoux forest just 20 kilometers from the city of Nancy in north-eastern France. The successful applicant will be based at the INRA Center in the EGI Lab but will work in close collaboration with biochemists also part of our department at the Université de Lorraine.

How to apply? Please send your CV and a cover letter describing your research experience and research interests as well as your research goals by email to duplessi@nancy.inra.fr. Include reference(s) from former lab(s) and pdf or links to selected publications.

Contact: 

Dr Sébastien Duplessis (INRA scientist)
http://mycor.nancy.inra.fr/GIteam/?page_id=13
e-mail, duplessi@nancy.inra.fr
UMR 1136 Interactions Arbre-Microorganismes
Equipe EcoGénomique des Interactions
Centre INRA Nancy Lorraine
54280 Champenoux
France

Published papers from the team related to the project

• Haquard S, Joly DJ, Lin Y-C, Tisserant E, Feau N, Delaruelle C, Legué V, Kohler A, Tanguay P, Petre B, Frey P, Van De Peer Y, Rouzé P, Martin F, Hamelin RC, Duplessis S (2012) A comprehensive analysis of genes encoding small secreted proteins identifies candidate effectors in Melampsora larici-populina (poplar leaf rust). Molecular Plant-Microbe Interactions25:279-293.

• Duplessis S, Cuomo CA, Lin Y-C, Aerts A, Tisserant E, Veneault-Fourrey C, Joly DL, Hacquard S, et al. (2011) Obligate biotrophy features unraveled by the genomic analysis of rust fungi. Proceedings of the National Academy of Science USA, 108:9166-9171Open Access.

• Duplessis S, Hacquard S, Delaruelle C, Tisserant E, Frey P, Martin F, Kohler A (2011) Melampsora larici-populina transcript profiling during germination and time-course infection of poplar leaves reveals dynamic expression patterns associated with virulence and biotrophy. Molecular Plant-Microbe Interactions24:808-818.

• Duplessis S, Joly DJ, Dodds PN (2012) Rust effectors. Chapter 7 In Effectors in Plant-Microbes Interactions, Martin F and Kamoun S (eds), Wiley-Blackwell, pp-155-193.

• Hacquard S, Petre B, Frey P, Hecker A, Rouhier N, Duplessis S (2011) The poplar-poplar rust interaction: insights from genomics and transcriptomics. Journal of Pathogens Vol. 2011, Article ID 716041, 11 pages (doi:10.4061/2011/716041)

 

Posted in Job

IAM recognized by AgreenSkills mobility programme

Agreenskills-AwardedCampus_CMJNOur department has been recognized by the AgreenSkills mobility programme for the quality of the support we offer postdoctoral research fellows that have been awarded an AgreenSkills Fellowship.

AgreenSkills is a postdoctoral research mobility programme co-funded by the European Commission, in the frame of the COFUND – FP7 People Programme, and coordinated by INRA in cooperation with Agreenium.

Article: Forest Ecology and Management

Effects of the use of biocontrol agent (< i> Phlebiopsis gigantea</i>) on fungal communities on the surface of Picea abies  stumps
E Terhonen, H Sun, M Buée, R Kasanen, L Paulin, FO Asiegbu
Forest Ecology and Management 310, 428-433

Abstract

One of the main objectives of sustainable forestry policies is to phase out, if possible, the use of chemical agents in forestry. The saprotrophic fungus Phlebiopsis gigantea has for several years been used as a biocontrol agent against the pathogen Heterobasidion annosum sensu lato. This pathogen is the major cause of root rot disease in conifers that results in economic losses estimated at 50 million euros to Finnish forestry industry annually. A major problem is that, although the effectiveness of Pgigantea as a bio-control agent has empirically been shown, the long term biological effect of this fungus on conifer trees as well as on other wood microflora has not been empirically proven. We investigated the impact of Pgigantea treatment on stump mycobiota using 454-pyrosequencing approach as this has not done before. Samples from forest sites pre-treated with Pgigantea for 1, 6 and 13 years ago were collected, DNA was isolated and the ITS regions were pyrosequenced. Similarly samples were also collected from untreated stumps within the same forest site over the same period of time. A total of 53,117 fungal sequences were generated by 454-pyrosequencing from 18 wood samples. After data cleaning we had 26,127 sequences representing 49% of the original sequences. Possible impact of the treatment on fungal communities in different study sites was analyzed using Principal Component Analysis (PCA). Also diversity indexes and similarity indexes between controls and treated stumps were calculated. Biocontrol Pgigantea represented 0.43% of all fungal sequences and was found only from stumps after one year post-treatment. The pathogen Hannosum s.l. was not observed in this study. Numbers of different OTUs were always higher in the control stumps. However there was no statistical difference noted between treated and non-treated stumps. Based on our results we can conclude that stump treatment should continue as there is no obvious adverse effect on the other stump mycobiota.

Keywords

  • Stump treatment;
  • Biocontrol;
  • Phlebiopsis gigantea;
  • 454-Pyrosequencing

Seminar: J. Felten

Our weekly seminar will be on wednesday this week instead of the usual fridays. Judith Felten, who used to be a PhD student in our lab and who is now doing her a post-doc in Umea (Sweden) will present her work:

“”Modification of cell wall formation through ethylene signaling in the wood of hybrid aspen”

The seminar will start at 1.30 pm in the main seminar room (administrative building).

Project: “Génomique et Adaptation lors des Invasions Biologiques”

The Project entitled “Génomique et Adaptation lors des Invasions Biologiques” headed by S. De Mita has been selected by the “Région Lorraine”

Summary

Due to anthropic activities and climate change, the rate of emerging diseases threatening natural ecosystems and cultivated crops is increasing. In this context it is essential to understand the parameters limiting and favoring the introduction and propagation of exotic pathogenic species in native ecosystems. We developed an experimental system based on the poplar leaf rust and its causative agent, the fungus Melampsora larici-populina. Based on a combination of population and ecological genomics approaches, we aim to understand the interplay between selective pressures and demographic factors during the invasion of natural poplar populations by M. larici-populina migrant from a known source. To this end, we will take profit of a natural system at the limit of the area of distribution of M. larici-populina where a recurring invasion takes place annually from a known source and along a predictable path.

Effectome meeting

Diapositive1Effectome network is a french network project devoted to study functional effectors of pathogenesis developed by bacteria, fungi, oomycetes, plant pathogens and nematodes. The network proposes to organize meetings of the French laboratories involved in this area.

Effectome 2013 will be held at Lauret the 25, 26 and 27th September. Several IAM members will present their research.  Program Effectome 25-27 September 2013.

Article: JACS

Monothiol Glutaredoxins Can Bind Linear [Fe 3 S 4]+ and [Fe 4 S 4] 2+ Clusters in Addition to [Fe 2 S 2] 2+ Clusters: Spectroscopic Characterization and Functional Implications

J. Am. Chem. Soc., Just Accepted Manuscript
DOI: 10.1021/ja407059n
Abstract:
Saccharomyces cerevisiae mitochondrial glutaredoxin 5 (Grx5) is the archetypical member of a ubiquitous class of monothiol glutaredoxins with a strictly conserved CGFS active-site sequence that has been shown to function in biological [Fe<sub>2</sub>S<sub>2</sub>]<sup>2+</sup> cluster trafficking. In this work, we show that recombinant S. cerevisiae Grx5 purified aerobically after prolonged exposure of the cell-free extract to air or after anaerobic reconstitution in the presence of glutathione, predominantly contains a linear [Fe<sup>3</sub>S<sub>4</sub>]<sup>+</sup> cluster. The excited state electronic properties and ground state electronic and vibrational properties of the linear [Fe<sup>3</sub>S<sub>4</sub>]<sup>+</sup> cluster have been characterized using UV-visible absorption/CD/MCD, EPR, Mössbauer and resonance Raman spectroscopies. The results reveal a rhombic S = 5/2 linear [Fe<sup>3</sub>S<sub>4</sub>]<sup>+</sup> cluster with properties similar to those reported for synthetic linear [Fe<sup>3</sub>S<sub>4</sub>]<sup>+</sup> clusters and the linear [Fe<sup>3</sub>S<sub>4</sub>]<sup>+</sup> clusters in purple aconitase. Moreover, the results indicate that the Fe-S cluster content previously reported for many monothiol Grxs has been misinterpreted exclusively in terms of [Fe<sub>2</sub>S<sub>2</sub>]<sup>2+</sup> clusters, rather than linear [Fe<sup>3</sub>S<sub>4</sub>]<sup>+</sup> clusters or mixtures of linear [Fe<sup>3</sub>S<sub>4</sub>]<sup>+</sup> and [Fe<sub>2</sub>S<sub>2</sub>]<sup>2+</sup> clusters. In the absence of GSH, anaerobic reconstitution of Grx5 yields a dimeric form containing one [Fe<sub>4</sub>S<sub>4</sub>]<sup>2+</sup> cluster that is competent for in vitro activation of apo-aconitase, via intact cluster transfer. The ligation of the linear [Fe<sup>3</sub>S<sub>4</sub>]<sup>+</sup> and [Fe<sub>4</sub>S<sub>4</sub>]<sup>2+</sup> clusters in Grx5 has been assessed by spectroscopic, mutational and analytical studies. Potential roles for monothiol Grx5 in scavenging and recycling linear [Fe<sup>3</sub>S<sub>4</sub>]<sup>+</sup> clusters released during protein unfolding under oxidative stress conditions and in maturation of [Fe<sub>4</sub>S<sub>4</sub>]<sup>2+</sup> cluster-containing proteins are discussed in light of these results.

Seminar: Jianping Xu

Jianping Xu (Department of Biology of McMaster University, Hamilton, Ontario, Canada) will  talk about the ” Population biology of ectomycorrhizal fungi from China.” The Seminar will be held in the meeting room of LGEF building on INRA campus the 13th of september at 1.30 pm.

Article: New Phytologist

Extreme diversification of the mating type–high‐mobility group (MATA‐HMG) gene family in a plant‐associated arbuscular mycorrhizal fungus
R Riley, P Charron, A Idnurm, L Farinelli, Y Dalpé, F Martin, N Corradi
New Phytologist

Summary

  • Arbuscular mycorrhizal fungi (AMF) are important plant symbionts that have long been considered evolutionary anomalies because of their apparent long-term lack of sexuality, but recent explorations of available DNA sequence have challenged this notion by revealing the presence of homologues of fungal mating typehigh-mobility group (MATA-HMG) and core meiotic genes in these organisms.
  • To obtain more insights into the sexual potential of AMF, homologues of MATA-HMGs were sought in the transcriptome of three AMF isolates, and their functional and evolutionary trajectories were studied in genetically divergent strains of Rhizophagus irregularis using conventional and quantitative PCR procedures.
  • Our analyses revealed the presence of at least 76 homologues of MATA-HMGs in R. irregularis isolates. None of these was found to be surrounded by genes generally found near other known fungal mating type loci, but here we report the presence of a 9-kb-long region in the AMF R. irregularis harbouring a total of four tandem-repeated MATA-HMGs; a feature that highlights a potentially elevated intragenomic diversity in this AMF species.
  • The present study provides intriguing insights into the genome evolution of R. irregularis, and represents a stepping stone for understanding the potential of these fungi to undergo cryptic sex.