Phytopathogen infection alters primary metabolism status and plant development. The alternative oxidase (AOX) has been hypothesized to increase under pathogen attack preventing reductions, thus optimizing photosynthesis and growth. In this study, two genotypes of Medicago truncatula, one relatively resistant (Jemalong A17) and one susceptible (TN1.11), were infected with Fusarium oxysporum and Rhizoctonia solani. The in vivo foliar respiratory activities of the cytochrome oxidase pathway (COP) and the alternative oxidase pathway (AOP) were measured using the oxygen isotope fractionation. Gas exchange and photosynthesis‐related parameters were measured and calculated together with antioxidant enzymes activities and organic acids contents. Our results show that the in vivo activity of AOX (valt) plays a role under fungal infection. When infected with R. solani, the increase of valt in A17 was concomitant to an increase in net assimilation, in mesophyll conductance, to an improvement in the maximum velocity of Rubisco carboxylation and to unchanged malate content. However, under F. oxysporum infection, the induced valt was accompanied by an enhancement in the antioxidant enzymes, superoxide dismutase (SOD; EC1.15.1.1), catalase (CAT; EC1.11.1.6) and guaiacol peroxidase (GPX; EC1.11.1.7), activities and to an unchanged tricarboxylic acid cycle intermediates. These results provide new insight into the role of the in vivo activity of AOX in coordinating primary metabolism interactions that, partly, modulate the relative resistance of M. truncatula to diseases caused by soil‐borne pathogenic fungi.

Fusarium and Rhizoctonia genera are important pathogens of many field crops worldwide. They are constantly evolving and expanding their host range. Selecting resistant cultivars is an effective strategy to break their infection cycles. To this end, we screened a collection of Medicago truncatula accessions against Fusarium oxysporum, Fusarium solani, and Rhizoctonia solani strains isolated from different plant species. Despite the small collection, a biodiversity in the disease response of M. truncatula accessions ranging from resistant phenotypes to highly susceptible ones was observed. A17 showed relative resistance to all fungal strains with the lowest disease incidence and ratings while TN1.11 was among the susceptible accessions. As an initiation of the characterization of resistance mechanisms, the antioxidant enzymes’ activities, at the early stages of infections, were compared between these contrasting accessions. Our results showed an increment of the antioxidant activities within A17 plants in leaves and roots. We also analyzed the responses of a population of recombinant inbred lines derived from the crossing of A17 and TN1.11 to the infection with the same fungal strains. The broad-sense heritability of measured traits ranged from 0.87 to 0.95, from 0.72 to 0.96, and from 0.14 to 0.85 under control, F. oxysporum, and R. solani conditions, respectively. This high estimated heritability underlines the importance of further molecular analysis of the observed resistance to identify selection markers that could be incorporated into a breeding program and thus improving soil-borne pathogens resistance in crops.

A faba bean rhizospheric Pseudomonas aeruginosa isolate RZ9 was used for studying its antifungal activity and protecting effects of faba bean and common bean against the root pathogen Fusarium culmorum strain MZB47. The dual culture tests showed that RZ9 inhibits MZB47 in vitro growth by 56%. When mixing RZ9 cell suspension with MZB47 macroconidia at equal proportion, the macroconidia viability was reduced with 70%. Pathogenicity tests conducted in sterile conditions showed that MZB47 caused an intense root rotting in faba bean 'Aquadulce' plantlets and a slight level in common bean 'Coco blanc'. This was associated to significant decreases in plant growth only in 'Aquadulce', reducing shoot dry weight (DW) by 82% and root DW by 70%. In soil samples, MZB47 caused severe root rotting and induced significant decreases in shoot DW (up to 51%) and root DW (up to 60%) for both beans. It was associated to a decrease in nodule number by 73% and 52% for faba bean and common bean, respectively. Biocontrol assays revealed that the inoculation of RZ9 to MZB47-treated plantlets enhanced shoot DWs (25% and 110%) and root DWs (29% and 67%), in faba bean and common bean, respectively. Moreover, root rotting levels decreased and nodule number increased in treated compared to untreated plantlets. Collected data highlighted the disease severity of F. culmorum and demonstrated the potential of using RZ9 in controlling Fusaria root diseases in beans. Thereby, the current study represents the first report on the biocontrol effectiveness of P. aeruginosa against F. culmorum in beans.Additional keywords: biocontrol; root rot; Vicia faba; Phaseolus vulgaris.

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Iron (Fe) deficiency is one of the major environmental stresses affecting plant production in the world. The selection of tolerant genotypes is considered an effective remediation strategy for this stress. The present study was carried out in order to investigate the biodiversity within Medicago truncatula plants in response to Fe deficiency, to identify tolerant genotypes and to assess the main tolerance mechanisms. To do this, a screening test was performed on 20 M. truncatula genotypes cultivated in minimal medium. Biometric and physiological markers were analyzed, including plant biomass, chlorophyll and root architecture. Results showed a biodiversity among the 20 genotypes. Interestingly, Fe deficiency tolerance was highest in TN8.20 and A17 genotypes. However, the lowest tolerance behavior was observed in TN1.11 and TN6.18. In order to investigate the main tolerance mechanisms, an experiment was conducted in the hydroponic system on already selected genotypes. Assessment of Fe deficiency tolerance was performed mainly on plant growth parameters, Fe (III)‐chelate‐reductase activity, rhizosphere acidification and antioxidant system defense. The relative better tolerance of A17 and TN8.20 to Fe deficiency was positively correlated with their capacity to maintain higher Fe‐acquisition efficiency in roots via rhizosphere acidification and the stimulation of Fe (III)‐chelate‐reductase activity. Moreover, tolerant genotypes showed the lowest decreases in chlorophyll content and photosynthetic activity (CO2 assimilation) compared to the sensitive ones. The efficiency of antioxidant capacity of the tolerant genotypes was revealed in stimulation of catalase (CAT) and peroxidase (POX) activities as well as accumulation of polyphenols, leading to the maintenance of cell integrity under Fe deficiency.

Plants encounter a myriad of microorganisms at the root-soil interface that can invade with detrimental or beneficial outcomes. Promoting a symbiotic relationship may interfere with the restriction responses to pathogens. In this study, we established a tripartite Medicago truncatula- Sinorhizobium meliloti-Fusarium oxysporum interaction to study the effect of the interplay between symbiosis and defense on the symbiosis establishment and efficiency in a resistant (A17) and susceptible (TN1.11) genotype to F. oxysporum infection. Our results showed that Sm induced the expression of allene oxide cyclase 1 and 2 (MtAoc1) and (MtAoc2) at 6 and 24 hours post infection (hpi) in A17 and at 24 hpi in TN1.11. This up-regulation of MtAoc1 and MtAoc2 was also observed when co-inoculating TN1.11 genotype with Fo and Sm. Moreover, in this later treatment the expression level of the gene of the common signaling pathway doesn’t make infection 3 (DIM3) was reduced compared to Sm-inoculated plants. This reduction of DIM3 transcripts was concomitant to a decrease in nodule number (NN), nodule fresh weight, and nitrogen-fixing rate (NFR), and to an accumulation of spermine in nodules. This defectiveness in symbiosis parameters, in the co-inoculation treatment, was also observed in A17 genotype, along with an accumulation of phytohormones in leaves and polyamines in nodules. Our results suggest that in the tripartite interaction, the high expression of MtAoc and the decrease of DMI3 transcripts along with the accumulation of phytohormones and polyamines in leaves overlapped with symbiosis establishment and functioning and thus, reduced NN and NFR.

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