Article: Frontiers in Plant Science

Effector proteins of rust fungi
B Petre, DL Joly, S Duplessis
Frontiers in Plant Science 22, 23

Abstract

Rust fungi include many species that are devastating crop pathogens. To develop resistant plants, a better understanding of rust virulence factors, or effector proteins, is needed. Thus far, only six rust effector proteins have been described: AvrP123, AvrP4, AvrL567, AvrM, RTP1 and PGTAUSPE-10-1. Although some are well established model proteins used to investigate mechanisms of immune receptor activation (avirulence activities) or entry into plant cells, how they work inside host tissues to promote fungal growth remains unknown. The genome sequences of four rust fungi (two Melampsoraceae and two Pucciniaceae) have been analyzed so far. Genome-wide analyses of these species, as well as transcriptomics performed on a broader range of rust fungi, revealed hundreds of small secreted proteins considered as rust candidate secreted effector proteins (CSEPs). The rust community now needs high-throughput approaches (effectoromics) to accelerate effector discovery/characterization and to better understand how they function in planta. However, this task is challenging due to the non-amenability of rust pathosystems (obligate biotrophs infecting crop plants) to traditional molecular genetic approaches mainly due to difficulties in culturing these species in vitro. The use of heterologous approaches should be promoted in the future.

Article: Microbial Ecology

Pairwise Transcriptomic Analysis of the Interactions Between the Ectomycorrhizal Fungus Laccaria bicolor S238N and Three Beneficial, Neutral and Antagonistic Soil Bacteria
A Deveau, M Barret, AG Diedhiou, J Leveau, W de Boer, F Martin, … Microbial ecology, 1-14

Abstract

Ectomycorrhizal fungi are surrounded by bacterial communities with which they interact physically and metabolically during their life cycle. These bacteria can have positive or negative effects on the formation and the functioning of ectomycorrhizae. However, relatively little is known about the mechanisms by which ectomycorrhizal fungi and associated bacteria interact. To understand how ectomycorrhizal fungi perceive their biotic environment and the mechanisms supporting interactions between ectomycorrhizal fungi and soil bacteria, we analysed the pairwise transcriptomic responses of the ectomycorrhizal fungus Laccaria bicolor (Basidiomycota: Agaricales) when confronted with beneficial, neutral or detrimental soil bacteria. Comparative analyses of the three transcriptomes indicated that the fungus reacted differently to each bacterial strain. Similarly, each bacterial strain produced a specific and distinct response to the presence of the fungus. Despite these differences in responses observed at the gene level, we found common classes of genes linked to cell–cell interaction, stress response and metabolic processes to be involved in the interaction of the four microorganisms.

Article: Eucaryotic cell

Cross Kingdom Functional Conservation of the Core Universally Conserved Threonylcarbamoyladenosine tRNA Synthesis Enzymes
PC Thiaville, B El Yacoubi, L Perrochia, A Hecker, M Prigent, JJ Thiaville, …
Eukaryotic Cell, EC. 00147-14

Abstract

Threonylcarbamoyladenosine (t6A) is a universal modification located in the anticodon stem loop of tRNAs. In yeast, both cytoplasmic and mitochondrial tRNAs are modified. The cytoplasmic t6A synthesis pathway was elucidated and requires Sua5p, Kae1p and four other KEOPS complex proteins. Recent in vitrowork suggested that the mitochondrial t6A machinery of S. cerevisiae is composed of only two proteins, Sua5p and Qri7p, a member of Kae1p/TsaD family. Sua5p catalyzes the first step leading to the threonyl-carbamoyl-AMP intermediate (TC-AMP) while Qri7 transfers the threonyl-carbamoyl moiety from TC-AMP to tRNA to form t6A. Qri7p localizes to the mitochondria, but Sua5p was reported to be cytoplasmic. We show that Sua5p is targeted both to the cytoplasm and to the mitochondria through the use of alternative start sites. Import of Sua5p into the mitochondria is required for this organelle to be functional since the TC-AMP intermediate produced by Sua5p in the cytoplasm is not transported into the mitochondria in sufficient amounts. The conservation of this minimal t6A pathway was characterized in vitro and, for the first time, in vivoby heterologous complementation studies in E. coli. The data revealed a potential for TC-AMP channeling in the t6A pathway as co-expression of Qri7p and Sua5p is required to complement the essentiality of E. coli tsaD. Our results firmly established that Qri7p and Sua5p constitute the mitochondrial pathway for the biosynthesis of t6A and bring additional advancement in our understanding of the reaction mechanism.

Article: Nature communications

Latent homology and convergent regulatory evolution underlies the repeated emergence of yeasts
LG Nagy, RA Ohm, GM Kovács, D Floudas, R Riley, A Gácser, M Sipiczki, …
Nature Communications 5

Abstract

Convergent evolution is common throughout the tree of life, but the molecular mechanisms causing similar phenotypes to appear repeatedly are obscure. Yeasts have arisen in multiple fungal clades, but the genetic causes and consequences of their evolutionary origins are unknown. Here we show that the potential to develop yeast forms arose early in fungal evolution and became dominant independently in multiple clades, most likely via parallel diversification of Zn-cluster transcription factors, a fungal-specific family involved in regulating yeast–filamentous switches. Our results imply that convergent evolution can happen by the repeated deployment of a conserved genetic toolkit for the same function in distinct clades via regulatory evolution. We suggest that this mechanism might be a common source of evolutionary convergence even at large time scales.

Project: FUNTUNE

This project has been selected by the French National Research Agency and involves in particular F. Martin, C. Fourrey, A. Kohler, E. Martino and E. Morin of our department.

SUMMARY

The development of a bio‐economy based on sustainable processes to transform renewable carbon sources as an alternative to fossil carbon chemistry is a major challenge. The lignocellulose contained in plant biomass is the most abundant biopolymer on earth and provides a renewable resource for bio‐energy as well as for platform molecules aimed at new value‐chains in bio‐industry. However, cost‐effective transformation of plant biomass is limited by the recalcitrance of lignocellulose and by its diversity in chemical composition. Plant biomass recalcitrance is mainly due to the crystalline structure of cellulose and to the presence of lignin, a polyphenolic polymer that restricts the accessibility of cellulosic enzymes to polysaccharides and strengthens the cell wall structure. Thermo‐chemical pre‐treatments are currently used to make the polysaccharide fractions amenable to enzymatic hydrolysis. In addition to their cost, these pre‐treatments have negative environmental impacts. One promising alternative to thermo‐chemical treatments is the development of eco‐friendly enzymatic processes able to efficiently harness the recalcitrant lignocellulose. The second challenge is the chemical diversity of biomass feedstock. One response to this challenge is the development of enzymatic cocktails with high efficiency on a range of diverse biomasses.

In this project we propose to explore fungal enzymatic machineries in order to design new fungi‐inspired enzyme cocktails able to mitigate recalcitrance of plant biomass from diverse sources. Plant‐associated fungi have evolved enzymatic toolboxes to adapt to diverse host plants and lignocellulosic substrates. These enzymatic toolboxes are the key factors for finely tuned modification of plant cell walls during fungal growth. While fungal wood decayers use a large range of carbohydrate‐acting enzymes (CAZymes) and oxidoreductases to degrade plant cell walls, symbiotic and biotrophic pathogenic fungi cause limited and targeted damage to plant cell walls leading to non ‐ disruptive cell wall loosening. When switching from biotrophy to destructive necrotrophic growth, hemi‐biotrophic plant pathogens secrete specific sets of enzymes that dramatically alter plant cell walls. Our purpose here is to identify and exploit the different sets of fungal enzymes associated with these different levels of plant cell wall deconstruction.

Using comparative analyses of available genomics and transcriptomics data, we will identify the sets of enzymes that are produced simultaneously by fungi when they alter plant cell wall structure or integrity. Fungal enzymes co‐expressed upon growth on plant tissues will be used to develop optimised enzyme cocktails for the in vitro release of high value molecules from plant biomass. One original aspect of the project lies in the concomitant analysis of enzymes active on cellulose, hemicelluloses and lignin, as recent evidence has shown that synergistic effects arise from the combined action of enzymes on different cell wall components. As genetics tools are available for model plant pathogens and symbiotic fungi, the integration of these species into the study will allow further in planta functional analyses. The activity of the newly designed enzyme cocktails will be tested on model biomasses and on agriculture and forest co‐products at the cell level as well as at the whole biomass level. Ultimately, we will identify high‐value molecules released from each type of plant biomass after enzymatic treatment. Besides providing new enzymatic tools for green chemistry, the project will elucidate how wood decayers, symbiotic and pathogenic fungi modify plant cell walls to successfully establish within host tissues.

Article: Frontiers in Plant Science

On the current status of Phakopsora pachyrhizi genome sequencing

M Loehrer, A Vogel, B Huettel, R Reinhardt, V Benes, S Duplessis, … Frontiers in Plant Science 5, 377

Abstract:

Recent advances in the field of sequencing technologies and bioinformatics allow a more rapid access to genomes of non-model organisms at sinking costs. Accordingly, draft genomes of several economically important cereal rust fungi have been released in the last three years. Aside from the very recent flax rust and poplar rust draft assemblies there are  genomic data available for other dicot-infecting rust fungi. In this article we outline rust fungus sequencing efforts and comment on the current status of Phakopsora pachyrhizi (Asian soybean rust) genome sequencing.

Article: Journal of Biological Chemistry

Structural and spectroscopic insights into BolA-glutaredoxin complexes
T Roret, P Tsan, J Couturier, B Zhang, MK Johnson, N Rouhier, …
Journal of Biological Chemistry, jbc. M114. 572701

Abstract

BolA proteins are defined as stress-responsive transcriptional regulators but they also participate to iron metabolism. Although they can form [2Fe-2S]-containing complexes with monothiol glutaredoxins (Grx), structural details are lacking. Three Arabidopsis thaliana BolA structures were solved. They differ primarily by the size of a loop referred to as the variable [H/C] loop which contains an important cysteine (BolA_C group) or histidine (BolA_H group) residue. From 3D modeling and spectroscopic analyses of A. thaliana GrxS14-BolA1 holo-heterodimer (BolA_H), we provide evidence for the coordination of a Rieske-type [2Fe-2S] cluster. For BolA_C members, the cysteine could replace the histidine as a ligand. NMR interaction experiments using apo-proteins indicate that a completely different heterodimer was formed, involving the nucleic acid binding site of BolA and the C-terminal tail of Grx. The possible biological importance of these complexes is discussed considering the physiological functions previously assigned to BolA and to Grx-BolA or Grx-Grx complexes.

Article: Advances in Botanical Research

Genomics of arbuscular mycorrhizal fungi: out of the shadows. M Malbreil, E Tisserant, F Martin, C Roux, FM Martin. Advances in Botanical Research 70, 259-290

Abstract

Arbuscular mycorrhizal (AM) symbiosis is the most widespread mutualistic association. It concerns 80% of land plants and involves fungi belonging to the phylum Glomeromycota. Benefits to the host plants due to this symbiosis range from nutrient supply to protection against pathogens. AM fungi are important components of the soil microbiome and are of great interest for managing sustainable agriculture, provided that their life cycle is better understood. Recently, major advances in the genomics of the model AM fungus Rhizophagus irregularis DAOM197198 have been published, offering new tools to investigate the biology of this symbiosis. In this chapter, we provide an overview of the efforts that were necessary to reach these results, from the discovery of these fungi and the description of their mutualistic incidence to their in vitro cultivation and on to genomics. The genome of DAOM197198 is estimated at ca. 150 Mb. It is haploid and less polymorphic than expected. Although it is an obligate biotrophic fungus, very little gene loss was observed. We put the Rhizophagus gene repertoire in perspective with previous investigations performed on the physiology of AM fungi: germination, early signalling with host plants, plant invasion, metabolism (phosphorous, carbon and nitrogen) and sexuality. Clearly, the publication of the genome of R. irregularis DAOM197198 is a turning point in the study of AM fungi, and large areas of their biology that still remain to be elucidated will now become accessible for investigation.

Article: Mycorrhiza

Forty years of inoculating seedlings with truffle fungi: past and future perspectives

C Murat, Mycorrhiza July 2014

Abstract

The first commercialization of seedlings inoculated with truffle fungi occurred in 1973. Over the last 40 years, considerable progress has been made relative to quality control for inoculated seedlings. A recently published paper by Andrés-Alpuente and colleagues (Mycorrhiza 24:29–37, 2014) reviewed and tested the different methods of mycorrhization assessment currently used in Europe. The aim of this paper is to augment their findings by adding information to the discussion about the most important steps in seedling quality control. Additionally, the history of seedlings inoculated with truffles, procedures currently used in France for seedling control quality, and a reflection on future research aimed at increasing truffle production will be presented.