Induced systemic resistance (ISR) is a process elicited by telluric microbes, referred to as plant growth-promoting rhizobacteria (PGPR), that protect the host plant against pathogen attacks. ISR has been defined from studies using Pseudomonas strains as the biocontrol agent. Here, we show for the first time that a photosynthetic Bradyrhizobium sp. strain, ORS278, also exhibits the ability to promote ISR in Arabidopsis thaliana, indicating that the ISR effect may be a widespread ability. To investigate the molecular bases of this response, we performed a transcriptome analysis designed to reveal the changes in gene expression induced by the PGPR, the pathogen alone, or by both. The results confirm the priming pattern of ISR described previously, meaning that a set of genes, of which the majority was predicted to be influenced by jasmonic acid or ethylene, was induced upon pathogen attack when plants were previously colonized by PGPR. The analysis and interpretation of transcriptome data revealed that 12-oxo-phytodienoic acid, an intermediate of the jasmonic acid biosynthesis pathway, is likely to be an actor in the signaling cascade involved in ISR. In addition, we show that the PGPR counterbalanced the pathogen-induced changes in expression of a series of genes.

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The establishment of arbuscular mycorrhizal associations causes major changes in plant roots and affects significantly the host in term of plant nutrition and resistance against biotic and abiotic stresses. As a consequence, major changes in root transcriptome, especially in plant genes related to biotic stresses, are expected. Potato microarray analysis, followed by real-time quantitative PCR, was performed to detect the wide transcriptome changes induced during the pre-, early and late stages of potato root colonization by Glomus sp. MUCL 41833. The microarray analysis revealed 526 up-regulated and 132 down-regulated genes during the pre-stage, 272 up-regulated and 109 down-regulated genes during the early stage and 734 up-regulated and 122 down-regulated genes during the late stage of root colonization. The most important class of regulated genes was associated to plant stress and in particular to the WRKY transcription factors genes during the pre-stage of root colonization. The expression profiling clearly demonstrated a wide transcriptional change during the pre-, early and late stages of root colonization. It further suggested that the WRKY transcription factor genes are involved in the mechanisms controlling the arbuscular mycorrhizal establishment by the regulation of plant defence genes.

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Biological control of Rhizoctonia solani with Trichoderma harzianum has been demonstrated in several studies. However, none have reported the dynamics of expression of defence response genes. Here we investigated the expression of these genes in potato roots challenged by R. solani in the presence/ absence of T. harzianum Rifai MUCL 29707. Analysis of gene expression revealed an induction of PR1 at 168 h post-inoculation (hpi) and PAL at 96 hpi in the plants inoculated with T. harzianum Rifai MUCL 29707, an induction of PR1, PR2 and PAL at 48 hpi in the plants inoculated with R. solani and an induction of Lox at 24 hpi and PR1, PR2, PAL and GST1 at 72 hpi in the plants inoculated with both organisms. These results suggest that in the presence of T. harzianum Rifai MUCL 29707, the expression of Lox and GST1 genes are primed in potato plantlets infected with R. solani at an early stage of infection.

Macrophomina phaseolina is a soil-borne fungal pathogen with a wide host range that causes charcoal rot in soybean [Glycine max (L.) Merr.]. Control of the disease is a challenge, due to the absence of genetic resistance and effective chemical control. Alternative or complementary measures are needed, such as the use of biological control agents, in an integrated approach. Several studies have demonstrated the role of arbuscular mycorrhizal fungi (AMF) in enhancing plant resistance or tolerance to biotic stresses, decreasing the symptoms and pressure caused by various pests and diseases, including M. phaseolina in soybean. However, the specific contribution of AMF in the regulation of the plant response to M. phaseolina remains unclear. Therefore, the objective of the present study was to investigate, under strict in-vitro culture conditions, the global transcriptional changes in roots of premycorrhized soybean plantlets challenged by M. phaseolina (+AMF+Mp) as compared with nonmycorrhizal soybean plantlets (-AMF+Mp). MapMan software was used to distinguish transcriptional changes, with special emphasis on those related to plant defense responses. Soybean genes identified as strongly upregulated during infection by the pathogen included pathogenesis-related proteins, disease-resistance proteins, transcription factors, and secondary metabolism-related genes, as well as those encoding for signaling hormones. Remarkably, the +AMF+Mp treatment displayed a lower number of upregulated genes as compared with the -AMF+Mp treatment. AMF seemed to counteract or balance costs upon M. phaseolina infection, which could be associated to a negative impact on biomass and seed production. These detailed insights in soybean-AMF interaction help us to understand the complex underlying mechanisms involved in AMF-mediated biocontrol and support the importance of preserving and stimulating the existing plant-AMF associates, via adequate agricultural practices, to optimize their agro-ecological potential.

Root colonization by arbuscular mycorrhizal (AM) fungi is a dynamic process involving major changes in plant gene expression. Here, the expression of a phosphate transporter gene (PT3) and several defense genes, already known to be involved in the various stages of AM establishment, were monitored in the mycelium donor plant (MDP) in vitro culture system associating potato plantlets with an AM fungus. This system allows fast and homogenous mycorrhization of seedlings at their early stage of development by growing the plantlets in active mycelial networks, but has never been validated for gene expression analysis. Here, QRT-PCR analyses were conducted in parallel to pre- (1 day), early (2 and 3 days), and late (6, 9, and 15 days) stages of root colonization. We observed the induction of a plant gene marker of AM root colonization (PT3) at the late stage and the induction of MAPK and PAL genes at the early and late stages of root colonization. We also demonstrated the induction of PR1 and PR2 genes at pre- and late stages and of GST1 and Lox genes at a late stage of root colonization. These results validated the MDP in vitro culture system as an optimal tool to study gene expression analysis during the AM fungi establishment. This system further opened the door to investigate gene networks associated with the plants-AM fungi symbiosis.

In this work, we described an in vitro system adequate for investigating the pathosystem soybean/arbuscular mycorrhizal fungi (AMF)/Fusarium virguliforme. Pre-mycorrhized plantlets with Rhizophagus irregularis were infected by F. virguliforme either locally via a plug of gel supporting mycelium (Method 1) or via a macroconidia suspension applied to the medium surface (Method 2). Root colonization by the AMF and infection by the pathogen were similar to the usual observations in pot experiments. Within a period of 18 days, more than 20% of the roots were colonized by the AMF and infection by the pathogen was observed in all the plants. In presence of AMF, a decrease in symptoms and in the level of root tissue infection was noticed. With Method 1, smaller necrotic lesions were observed in the pre-mycorrhized plantlets. In Method 2, pathogen infection was slower but more homogenous. These results demonstrated the suitability of the in vitro cultivation system to study the pathosystem soybean/AMF/F. virguliforme. We propose this in vitro cultivation system for studying the mechanisms involved in the biocontrol conferred by AMF against F. virguliforme in soybean.